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Original Abstract Book
AFRICAN-AMERICAN STUDIES
A PROPHETESS WITHOUT HONOR: WILLIE MAE FORD SMITH AND THE GOSPEL-BLUES MUSIC
REVOLUTION 1930-1939
Jamar O. Rahming (Dr. George Junne, Jr.), Department of Africana Studies, University of Northern Colorado,
Greeley, Colorado 80639
The 1930 Baptist Gospel Music Convention marked a pivotal event in the evolution of what is known today as the
Gospel-Blues music movement. Blues musician Thomas A. Dorsey had a religious awakening and decided to return
to his roots: the gospel sound. However, Dorsey’s homecoming did not involve giving up the blues. Dorsey
synchronized the embellishments and rhythms of the blues with conventional protestant hymnals and early Negro
spirituals, creating Gospel-Blues music. Though Dorsey was the creator of this exuberant sound, he did not popularize
it nor was he its sole founder. Archival evidence collected in 5 states and oral history interviews with 27 associates of
the Gospel-Blues movement affirms that St. Louis gospel heroin Willie Mae Ford Smith played a founding role in the
movement by popularizing it and carrying it to the Black churches. Previous researchers point at Dorsey as the
leading founder and Mahalia Jackson as the source of its dissemination. Consequently, Smith has received only
residual acknowledgement as being a founder in this movement. Smith’s profound contribution as a founder of
Gospel-Blues music has gone unnoticed. Smith, unlike many early gospel singers, chose not to take part in the hit
record circuit. She believed that the primary purpose of the gospel singer was to proselytize the Christian faith. The
late 1930’s marked the advent of commercialism and the hit record industry for gospel singers. Many gospel singers
found producing records more lucrative than working the conventions and church revivals. Smith’s contempt for
commercialism and endearment to evangelical work encumbered her from receiving the recognition that her influence
merited. Through the analysis of oral historical evidence, this work will demystify contributions of such a seminal, yet
highly unrecognized founder in gospel music.
NEGOTIATING TRIPLE-CONSCIOUSNESS: FINDING SELF IN COMMUNITY
Jenna S. Hanson (Dr. Veena Deo), Department of English, Hamline University, Saint Paul, MN 55104
Warring identities, being American, black, and a woman, plague the characters in bell hooks’ Bone Black: Memories
of Girlhood, Toni Morrison’s Beloved, and Gloria Naylor’s The Women of Brewster Place. Each author presents a
definition of an “ideal” community in which these women would be able to reconcile their disjointed
triple-consciousness—the extension to black women of W.E.B. Dubois’ notion of double-consciousness. Naylor’s
ideal community is one in which women join together for personal healing despite their differing economic class and
sexual identities. For Morrison, racial unity is integral to move forward and heal past wounds caused by slavery and
institutionalized racism. hooks focuses on the transformation of American society into an integrated space in which
race can be discussed and cross-cultural connections can be made. These three ideas of community can be seen as
a set of concentric circles; hooks’ community cannot be achieved without first Naylor’s and then Morrison’s. Once the
ideal community is realized space can be created to host open communication on race and gender, one in which
equal access to opportunity is given.
1
AFRICAN-AMERICAN STUDIES
The Unfinished Revolution: Bettie Mae Fikes and the Ongoing Civil Rights Movement
E. Shane Steward, Research Sponsor-Elizabeth McRae, History, Western Carolina University, Cullowhee, NC, 28723
The year is 1965, and a young woman stands up in front of a church and begins to sing. She is not just singing in a
Sunday morning service, but is singing in what is becoming a quick tradition of African Americans in the South, a
mass meeting. She throws back her head and begins to sing “This Little Light of Mine.” She sings with only the soul of
a girl whom has had to endure a lifetime of repression and segregation. For this young girl, Bettie Mae Fikes, a time
has come for a change, and little by little the life of repression that she has known for most of her life, looks better. In
2005, some 40 years later, Fikes stands in the reception room of one of Selma, Alabama’s greatest symbols of the old
South, The St. James Hotel. She is now part owner of this landmark, and in her mind things are better, yet she is
aware that equality—economic, social, and political has yet to be achieved. She is still fighting the fight to improve
race relations and poverty in her hometown of Selma. To her the fight is ongoing and will only be won, when
everyone, including Selma’s black and white leaders, confronts the historical legacies of poverty and racism and its
effects on economic development. Using oral histories, personal interviews, discography, archival collections, and
newspapers, this paper argues that by looking at the life of Fikes, we learn that the civil rights movement did not end
in the 1960s. Her life also demonstrates the legacies of grassroots activism in framing the lives of those who
participated in the struggle for freedom. Fike’s journey illuminates the victories of the 1960s, the persistent struggle
for social justice, and the continuation of the civil rights movement. Her history challenges us to rewrite the narrative
of the movement, acknowledging the unfinished nature of this revolution.
2
AMERICAN STUDIES
Dysfunctional Father-Son Relationships in American Political Fiction
JP Gritton, Michael Reynolds, English, Hamline University 1536 Hewitt Ave/St. Paul, MN/55104
There are few themes as recurrent in American fiction as that of the dysfunctional father-son relationship. And there
are few ideas more generally accepted in American political fiction than the pervasive sense that the state is not "us."
The father-son relationship in American political fiction embodies conceptions about the relationship between the
self/citizen and the "fatherland"/state. A precursory study has revealed a few consistent themes. For starters, fathers
are generally incompetent, neglectful, ridiculed, or simply omitted in American political fiction. To name just a few,
Pappy Stark in All The King`s Men, or Governor John McKay in The Candidate. The father-son relationship is also
defined by what might be called a fantasy of immortality; consistently, father-figures in American political fiction
attempt to be simultaneously fathers and sons (to beget themselves): Phillip Jessup in Sinclair Lewis’ It Can`t Happen
Here, or Willie Stark in All The King`s Men, who is both raising a son and a self incarnate. Lastly, these relationships
are defined by the theme of filial sacrifice. In one way or another and for one reason or another, fathers consistently
sacrifice their sons in American political fictions: Judge Irwin sacrifices a functional relationship with his son for his
reputation in All The King`s Men, while Major Carteret almost loses his son to the frenzied mob he has created in The
Marrow of Tradition. Authors of American political fiction convey a distance between the self and the state through
dysfunctional father-son relationships, affirming the old maxim that politicians are a "bunch of bastards."
THE AMERICAN EXPANSIONARY IMPULSE AND GOLD IN THE BLACK HILLS
Eric Matul (Dr. Gretchen Buggeln), Christ College, Valparaiso University, Valparaiso, Indiana 46383
The main focus of this project involves the discovery of gold in the Black Hills of present-day South Dakota, the
subsequent gold rush to the area, and the conflict that arose with the Sioux that owned the land. George Armstrong
Custer`s expedition explored the region in 1874. The supposed purpose was the establishment of a military fort in the
Hills; however, it is clear that there was an interest in the wealth that could be extracted from the area as well. The
paper includes a description of the expedition through personal accounts by newspaper correspondents, Lt. Col.
Custer, Private Theodore Ewert, and Lt. James Calhoun. These participants show the tendency to see land as a
commodity that was being wasted by the Indians who were not reaping all of its economic potential. Newspaper
accounts illustrate the hype created, which would guarantee a rush to the Hills. In addition to the actual expedition,
the paper explores the economic and political atmosphere that contributed to the gold rush. Economically, the Panic
of 1873 left Americans clamoring for relief, which a gold rush would help supply. Politically, President Grant’s Peace
Policy with the Native Americans was losing ground, because the Indians represented an obstacle to expansion and
the health of the nation. Americans, therefore, wanted the Hills opened for public use. The Laramie Treaty of 1868,
however, cited the Black Hills as an area in which non-Indians were prohibited. The paper discusses the attempts at
negotiating with the Sioux, a daunting task due to divisions between those living on reservations and those following
Sitting Bull. No agreements on the sale of the Hills could be reached and President Grant removed the military
presence from the area, which allowed miners to enter illegally without any major barriers. Indians that were not on
reservations were ordered in the winter of 1875-1876 to return to the agencies no later than January 31. By not doing
so, they were considered hostile, making a war unavoidable. Many of the Indians, however, had joined the hostile
bands due to corrupt agencies, controversy over selling the Hills, and measures to reduce their hunting territories.
These numbers proved deadly to Custer at the Battle of the Little Bighorn in 1876. The purpose of the paper is not to
place blame on Custer for the gold rush; rather, it is to show the many factors contributing to these historical events.
In particular, the American expansionary impulse and the desire to fulfill the principles embodied in “manifest destiny”
caused the rush and Custer’s death.
3
AMERICAN STUDIES
BUILDING CHARACTER AND EIGHT-SIDED HOMES: THE OCTAGON CRAZE IN CONTEXT
Netha A. Cloeter (Dr. Gretchen Buggeln), Christ College (The Honor’s College), Valparaiso University, Valparaiso,
Indiana 46383
Domestic architecture plans and their constructed counterparts reflect ideal and actual extensions of our individual
and social selves. Using this principle as the conceptual framework for inquiries, this presentation examines the
decade-long octagon house “craze” that emerged on the mid-nineteenth century American architectural landscape.
Three sources are considered: the cultural context, the initial plan as prescribed by renowned phrenologist Orson S.
Fowler, and a surviving example of the octagonal form applied to domestic architecture. The octagon house plan
thrived in an era of idea books and architectural experimentation, and represents a unique manifestation of nineteenth
century ideals. A close reading and analysis of Orson Fowler’s 1848 octagon design manifesto, The Octagon House:
A Home for All, reveals both the strange and revolutionary ideas he advanced in connection with the octagon form.
References to the “science” of phrenology and an implied connection between character and domestic architecture
add a dimension of cultural significance to the text. The principles espoused in Fowler’s manifesto are compared to
an actual application of Fowler’s plan: a surviving octagon house in Valparaiso, Indiana, built in 1856. New
information regarding the construction and history of this house is presented, along with its noticeable deviations from
Fowler’s text. Though Fowler’s own home was the only structure that fully conformed to his proposed model, the
basic pattern and principles reverberated throughout the built environment of nineteenth century America and provide
insight into the domestic ideals and concerns of this era. This illustrated presentation unravels the cultural
significance, surrounding socio-historical forces, and continued preservation of this architectural enigma.
Yellow Men Can`t Jump: Defining Community Through Japanese American Basketball Leagues
Dana L. Heatherton (Dr. La`Tonya Rease-Miles, Dr. Valerie Matsumoto, Dr. Henry Yu), Department of English,
University of California Los Angeles, Los Angeles, California 90095-1541
Japanese American Basketball Leagues have been a cultural institution within the Japanese American population of
Los Angeles since the 1930’s and currently boast 10,000 participants within the county. Originating after the
community’s release from WWII Internment Camps, these leagues attempted to compensate for the geographic
fragmentation of the community. Originally the leagues restricted eligibility to only full-blooded Japanese Americans;
however, as the Japanese American population became increasingly mixed with other races and the leagues gained
popularity outside their community, the rules subsequently needed to change. Currently the leagues have assumed a
more multi-culturist image but their culturally specific mission has maintained the same.
Through a textual examination of the league`s enrollment policies, relevant newspaper articles, personal
interviews, as well as a visual assessment of documentaries, I will demonstrate how racial exclusivity, while not
explicitly stated within the leagues policies, still exists. By identifying the rhetorical strategies used to cleanse the
racialized vocabulary from its policies, I will demonstrate how the leagues craft a façade of inclusivity to ostensibly
accommodate the changing racial diversity of its community, but ironically, also use these strategies to disguise the
perpetuation of a mono-racial agenda. By highlighting the leagues’ tension to preserve a cultural mission that doesn’t
align with the community it currently caters to, these policies will help serve as a lens to understand the larger struggle
for Japanese Americans to find a cohesive cultural identity.
4
ARCHITECTURE
ST. MICHAEL`S CHURCH: EXPLORATIONS OF TRADITION AND MODERNITY
Maria C. Wiering (Dr. Victoria Young), Department of Art History, University of St. Thomas, St. Paul, Minnesota,
55105.
Completed in 2005, St. Michael’s Church in St. Michael, Minnesota is the newest church in the Archdiocese of
Minneapolis and St. Paul. Unusual to the Western world, the church’s design is influenced mainly by Eastern Catholic
rite architecture, such as the Hagia Sophia in present-day Istanbul, Turkey. As such, the building is a visual testament
to the universality of the Catholic Church. A universal design approach is also favored by the Church through utilizing
documents such as Built of Living Stones (2000) and the General Instruction of the Roman Missal (2003), among
others. However, many architects are in disagreement over the documents’ correct architectural and artistic
interpretations. Traditionalists believe a church’s style should reflect the Church’s rich history while modernists argue
that design should represent present or future culture. Because St. Michael’s Church incorporates both traditional and
modern elements, it is destined to become a player in the contemporary debate on sacred space and its implications
for the Catholic faithful. Through examining church documents, interviews with the makers of St. Michael’s Church,
and documenting the building itself, I explored the integration of not only tradition and modernity in the church’s
design and present debate, but also the cross-cultural architectural and artistic notions present in this twenty-first
century religious space.
THE EPIPHANY SYSTEM
Michael Taylor (Ms. Anne Mooney), College of Architect, The University of Utah, Salt Lake City, Utah 84112
The Epiphany System is an interdisciplinary collaboration between computer science and architecture. Our goal is to
break free from the traditional definitions of how a building is designed. We will provide the Epiphany System with a
loosely defined architectural form and through its application will achieve numerous distinctive final forms. To
accomplish our goal we defined a virtual camera that allows us to render a video that represents what a person
viewing the architectural form would see as they move around the form. The dimensions of the virtual form will then
be directly related to the camera/person. Every time the camera/person moves to a new point along a virtual path the
physical dimensions of the form will also change. Hence by defining the relationships between the camera and
architectural form coupled with an infinite number of paths the camera/person could take leads to an infinite number
of unique forms free from any preconceived notions about shape and dimensions of the final form.
The form will be designed entirely with the Maya software. The C++ coding language is used to create custom
Maya nodes, allowing us to link the width, length, and height of the individual walls to the camera. In addition Maya
“bones” have allowed us to take the right angles of the base form and create smooth flowing curves across the entire
form. Plans to place openings at predetermined locations has been replaced with the goal of creating openings based
on where the camera is pointed at certain points along the path. At first every expression will be constantly and
dramatically manipulating the form. Expressions are programed to stop one by one. Each time an expression is
stopped the location of camera represents a point of epiphany by finalizing one dimension of the form in such a way
that the relation between the camera/person and the form conform to perfect golden ratios creating an aesthetic final
design unlike any other building.
5
ARCHITECTURE
INDUSTRIAL HEMP AS A NEW SUSTAINABLE FIBER (History and Analysis of Characteristics)
Christy Osborn (Elizabeth Coles), Dept. of Construction Technology - Interior Design Program. Indiana University Purdue University Indianapolis, Indianapolis, Indiana 46202
Textile fibers that are used for commercial fabrics are typically made by using some type of petroleum based
chemical. Recognizing the diminishing supply of fossil fuels, the hunt to discover a renewable natural fiber that could
replace synthetic fibers has become a research issue in many countries. Industrial or low-THC hemp is a natural fiber
that could solve this problem.
Hemp is nature’s longest, strongest, most durable fiber. Per acre, hemp out produces cotton by 200%, but uses no
pesticides or fertilizers and it is biodegradable. However, cotton is the leading user of pesticides, accounting for 60%
of pesticides used each year.
The purpose of this research was to discover commercial uses and applications for industrial hemp in fiber form. The
following items have been identified as necessary initial components of a research project on Industrial Hemp and
these components were investigated in summer, 2005 as the intital aspects of the industrial hemp project.
•
•
•
•
•
•
•
History and Background
Plant Identification
Growing and Cultivation procedures and techniques
Environmental advantages and disadvantages
Product development and uses
Laws and Regulations – past and future
Economic impact
The focus of the later research concentrated on the ability of using Industrial Hemp as a textile within commercial
interior design projects. Testing of fabrics made from industrial hemp was done to determine their suitability and
safeness. The testing procedures included, but were not limited to, fire retardancy, color stability and durability,
abrasion resistance, sun fading resistance and water and moisture resistance. Comparisons of above durability and
safety characteristics included hemp fiber fabrics with those of other cellulosics (cotton and linen) and petroleum
based fabics such as nylon and polyester based textiles.
MAXIMIZING THE ENERGY EFFICIENCY OF A HOUSE
Charlie Stroup, Rex Adelberger, Physics, Guilford College 5800 West Friendly Ave, Greensboro, NC 27410
Energy costs for the average new American home run over $1,200 a year, with $400 of that being wasted through
poor energy efficiency. I will present a computer model of home efficiency that considers parameters such as the
percentage of the surface area occupied by windows, the thermal properties of the insulating materials used, the
amount of heat transfer through air draft, geographical variations in climate, and effects of methods of energy
conservation on the part of the owners. This computer model calculates an energy efficiency statistic and illustrates
which parameters have the strongest impact on this value. With this information, I can recommend a means of
improving the energy efficiency of a house.
6
ART/ART HISTORY
CAPTURING HISTORY ON CLOTH: AFRICAN AMERICAN QUILTS AS CULTURAL AND HISTORICAL
RECORDS
Molly Braswell (Ginger Spivey), Department of Art History, The University of North Carolina at Asheville, Asheville,
North Carolina 28804
Both functional and visually pleasing, quilts also have directly and indirectly served to communicate the victories,
tragedies, and traditions of the African American community. The stories projected on the quilts, either intentionally
or unintentionally, reveal the artists’ histories. This project seeks to prove that African Americans’ quilts are a valid,
though unconventional, form of historical records. Extensive written historical records do not exist as most African
American slaves were forbidden to read or write. While some cultures depend solely on oral tradition and others rely
on written accounts to reveal facts, African Americans have crafted quilts which successfully tell their history. Though
the quilting methods have generally remained the same, the personal and cultural stories portrayed--through design
patterns, color choices, and encoded symbols--have evolved significantly over time, along with the African American
culture. The study of African Americans’ quilts created during the Antebellum South, after the Civil War, and through
the Civil Rights Movement reveals the documentary nature of this craft. Also, contemporary artists, such as Carolyn
Mazloomi and Faith Ringgold, show the recent changes in this practice. Contemporary artists continue to give quilts
meaning as historical records by addressing African Americans’ social and political concerns through the intentional
use of traditional African colors and techniques. Ironically, African Americans have employed a stable, time honored
tradition to project the changes in a culture. By highlighting the importance and potential of quilts, this project shows
how a community has used available means to encode messages, to provide social exchange, to honor heritage, to
proclaim spirituality, and thus to document their history.
Propaganda as Art: The Political Murals of the Bogside Artists
Abby Sener (Dr. Karen Rosell), Art and Art History, Juniata College, 1700 Moore Street, Huntingdon, PA 16652
There has been a longstanding tradition of murals in Northern Ireland. Openly political, the murals have presented
the sentiments of Unionists since the early 20th century. More recently, the Republicans have begun to create their
own murals using a greater variety of themes than their Unionist counterparts. These murals are a part of a larger
question: can propaganda be considered art? University of Ulster sociologist Bill Rolston argues that the individuals
who create these murals are not artists: "The Republican painters of Northern Ireland are not and will never be
established artists." By implication, he seemingly discounts the murals themselves as art; he considers them to be
mere propaganda. However, I argue that the murals are art, regardless of their overt political nature. In an attempt to
explore this issue, I will focus on a group of men known as the Bogside Artists, whose works are located in Derry,
Northern Ireland. Four of their murals will be presentetd in the context of Jacques Maquet`s three principles of
composition: geometry, organic form, and dynamism. I will also analyze the murals from a fourth dimension, that of
universality. The works of these artists will then be compared to political artworks by Goya, Delacroix, Rivera, and
Daumier. Through this analysis, I hope to provide a compelling argument that the works of the Bogside Artists
should, first and foremost, be considered art rather than propaganda. Finally, they deserve as much respect in the
world of art as works by historically renowned artists.
7
ART/ART HISTORY
CINDY SHERMAN AND SUZANNE VALADON: BOTH SIDES OF THE CANVAS
Holly B. Brown (Dr. Karen J. Rosell), Art History, Juniata College, 1700 Moore Street, Huntingdon, PA 16652
In the world of art, contemporary artist Cindy Sherman and 19th century painter Suzanne Valadon are unique, in that
both women were influenced by their experiences as artists and models. Through their artworks, lives, and aesthetic
theories I intend to explore how each employed different methods to express a critique of the traditions of the art
world.
First, I will examine Cindy Sherman`s Untitled Film Stills in comparison to some of her later works, especially her
History Portraits. In the Film Stills Sherman depicts herself in varying guises with seriousness and untheatrical poses
meant to capture a moment in the life of her characters, who nevertheless are theatrical and false by nature. The fact
that she attempts to portray actresses of her childhood more seriously than she treats respected portraits of "real"
people, reveals the distinction of the Film Stills.
In contrast to Sherman, Suzanne Valadon hated the theatricality of modeling for some of the most famous 19th
century artists. Her work can be categorized by the paintings that show femininity in a modern, positive light, as well
as paintings that depict models in the obvious act of posing and being uncomfortable. Her paintings of women,
especially the female nude, relate most to Sherman`s Film Stills in appearance and in theory.
Both artists attempt to change the anti-feminist traditions they experienced as children. But where Valadon
endeavors to do this through a new, independent art form depicting women and their myriad of emotions, Sherman
enters the media she is trying to subvert only to destroy it from the inside. Ultimately, however, both practices can be
considered effective and innovative for their respective time periods.
INCAN USAGE OF WARI AND TIWANAKU ART TO PROMOTE A NATIONAL IDENTITY
Krista L. Garcia (Dr. Sara Orel), Division of Fine Arts, Truman State University, Kirksville, Missouri 63501
In less than a century, the Inca created the largest empire in the pre-Columbian Americas, complete with an
impressive infrastructure allowing their empire to efficiently function administratively and economically. The imperial
state controlled much of the art production, in particular the most valued textiles, pottery, and the architectural
expansion of the empire as seen in roads, walls, and other stonemasonry. The Inca conquered millions and worked to
formulate an imperial style in the art created. This cultural transmission went only in one direction; the artistic styles
of those the Incans controlled were rarely adopted. However, the Incan state did employ symbolism and styles from
the Wari and Tiwanaku empires, both of which had collapsed more than four centuries prior to the Incan ascent in the
Andean region. To avoid the art from subjugated peoples in the Incan empire, yet utilize elements from cultures
which existed centuries prior demonstrates significant ideological and political motivations in the Incan ruling elite.
This paper will examine the relationship between artistic transmission between the Wari, Tiwanaku, and Incan
Empires, emphasizing the Incan desire to create ties to earlier political powers as part of the formation of a national
identity.
8
ART/ART HISTORY
THE ART OF SYPHILIS IN SIXTEENTH CENTURY ITALIAN PRINTMAKING
Margarita S. Berg (Professor Michael Gaudio), Department of Art History, University of Minnesota, Minneapolis,
Minnesota 55455
This work examines the theme of the “Danger of Women” in Sixteenth Century Italian printmaking, focusing on the
association of venereal diseases with the male concern about the threats that women bestow upon them. The project
investigated documents and folklore sources concerning the scientific knowledge and popular belief of the society
about the disease and its representations in printmaking. Diseases, such as syphilis, were considered highly topical
during the Renaissance, an era of significant medical research. Already in the sixteenth century syphilis became the
topic of distinguished literary and medical treaties, as well as a suitable subject for artistic representations. Poems
and pamphlets on the disease were often illustrated with prints, providing the public with a visual testimony of the new
disease. In Italy, syphilis gained particular importance at the courts of several ruling families, including that of Duke
Este of Ferrara, whose numerous relatives were infected with the disease. This project focuses on prints intended
specifically for connoisseurs, i.e. educated members of the elite and early print collectors. Works, such as Giulio
Campagnola’s "Venus Reclining" and Marcantonio Raimondi’s "Dream", in part served to warn the male viewers of
the threats of sexuality and the deceptive nature of disguised female polluters, represented by the generic nude
women in the engravings. Since males were often unaware of the woman’s hidden condition, the disease became
associated with disguise. In these prints, images of passive and highly erotic female bodies presented before the
active male beholder carry across a sense of hidden danger, which is reflected in the gloomy landscapes around
them. These images do not speak of the possession of beautiful women, but rather warn about the threats of such
desire and the danger of the triumph of the feminine over the masculine. Through particular iconographic symbols
and the overall character of compositions, these prints, produced around the first decade of the sixteenth-century,
demonstrate social concerns about the threat of sexually transmitted diseases, in particular, syphilis.
Helping or Exploiting: The Fine Line Documentary Photographers Walk
Rachel Souza, (Dr. Rebecca Bailey), School of the Arts, Meredith College, 3800 Hillsborough St., Raleigh, North
Carolina 27604, Dr. Rebecca Bailey, Art Department Meredith College, 3800 Hillsborough St., Raleigh, NC, 27607
One week after Hurricane Katrina, I traveled to Biloxi, MS, to capture the stories of survivors and document the
destruction. Biloxi was littered with memories and remnants of peoples lives; the sense of invading the sacred was
overwhelming. Brokenness stretched as far as I could see; teddy bears, wedding pictures and clothes in the mud; I
couldn`t handle any more. Yet, something inside me screamed to press on, as if sacrificing my emotions and
continuing on would do justice to the lives that were destroyed.
This experience of taking something intensely personal from someone through my camera led me to
self-evaluation and exploration of my intentions. Documentary photographers have to ask where the line is between
exploiting and helping the individuals in our images. What purpose is served by photographing situations of human
suffering? Do the images benefit the people in them? Are we responsible for the way the images are displayed? The
media plays an important role in deciding where the line between helping and exploiting lies. It has become
increasingly difficult for society to react to disturbing images. We are bombarded by dramatic, emotional, disturbing
images on a daily basis. This repetition of images has caused us to become desensitized to the situations they
portray. I will briefly explore the media`s role in coverage of Hurricane Katrina, juxtaposing those images with my
own.
This paper will explore these issues through images of Biloxi compared to historical images by Matthew Brady
from the Civil War. I will also reference contemporary work of international photographers, including Jane Atwood`s
land mine and prison series, as well as images by Chris Hondros from Sierra Leone, the West Bank and Iraq.
9
ART/ART HISTORY
GLASSIFICATION: REEXAMINING WASTE
Hayden Wilson (Rick Maas), Department of Art and Environmental Studies, University of North Carolina at Asheville,
North Carolina, 28804
Glaze chemicals contain various heavy metals proven to be toxic to humans. Current disposal techniques for
unwanted glaze materials require a high hazardous waste handling fee. This project will determine whether the
process of glassification can be utilized effectively to encapsulate and immobilize toxic heavy metals present in waste
glaze in order to reclassify them as non-hazardous. To verify the effectiveness of this procedure a system for refining
the glaze materials was created. The first step was the construction of wooden tables on which wet glaze could be
spread and dried. A ball mill was then fabricated to pulverize the dried chunks of glaze. Once the glaze was in
powdered form, it was mixed with different amounts of glass powder (batch) and solid glass pieces (frit). A blast
furnace was then built to melt the glaze/glass mixture in a crucible. Once melted, the mixture was poured out and
placed in water to be fractured into frit. This frit was then tested using TCLP (Toxicity Characterization Leaching
Procedure) to determine the efficacy of the glass in immobilizing the toxic materials. Finally the results were entered
into an experimental design formula in order to deduce the most effective glaze to glass ratios.
DESIGNERS, ARTISTS, ICONS AND EXHIBITS: FASHION AND ART IN NEW YORK CITY, 1996-2001
Emily M. Orr (Professor Mary Ann Calo) Department of Art and Art History, Colgate University, Hamilton NY 13346
At the end of the twentieth century in New York, fashion became a headliner on the art scene when the Guggenheim
mounted the Armani retrospective, the Ace Gallery exhibited the creations of Issey Miyake, and the Costume Institute
at the Metropolitan Museum of Art displayed the attire of pop-culture icons of Rock Style, as well as the classic
garments of Jackie Kennedy, whose estate was also auctioned off by Sotheby’s with tremendous success. These
institutions have challenged and stretched the definition and implications of fashion’s place in our culture and society
in various ways. Issey Miyake’s designs represent his purist, truly creative attitude towards the construction of
clothing as an art form. The Costume Institute historicizes fashion by examining how cultural icons have promoted
and made themselves famous through their stylish images. The Guggenheim and Sotheby’s attract thousands with
celebrity names.
Advertisements, magazines, photography, models and even New Yorkers themselves make fashion into a desirous
commodity and object of display on a daily basis. But during this period, museums, galleries, and auction houses
allowed their visitors to indulge in fashion as an artistic act. This paper will examine how the popular culture of this
era has created an image-conscious public that would be drawn towards exhibits of the style idols and the designers
who outfit them. As wearers and constant exhibitors of fashion themselves, visitors literally saw their spending and
superficiality validated and glorified when clothing appeared behind glass and in exhibition and auction house
catalogs of elite organizations. However, this exaltation of fashion did not come without consequence. I will
investigate how and why the spirit of the urban community of this period elevated clothing and costume to museum
and auction-worthy status, evoking both critical acclaim and immense criticism.
10
ART/ART HISTORY
MONARCHS, MAVENS, AND MERCHANTS: SPHERES OF COLLECTING IN SIXTEENTH-CENTURY
EUROPE
Megan C. Boyer (Dr. Nadja Aksamija), Department of Art and Art History, Colgate University, Hamilton, New York
13346
The art of collecting in sixteenth-century Europe transcended geographic and social boundaries. Eclectic collections,
which consisted of paintings, prints, books, natural wonders, and automata, sprung up across Europe, representing
the various desires and motivations of their patrons. The impulse to order, classify, and display had taken a hold of
Europe during the Renaissance, yet the motivations that directed the creation of collections and the physical contents
of collections differed between social spheres. Royal collections were stimulated by the allure of prestige; these
collections were all-encompassing, containing items such as delicately incised gemstones, paintings by Renaissance
masters, and exotica from conquered lands. Princely collectors, including Philip II of Madrid and Rudolf II of Prague,
gave visual form to their wealth and power through the amassing of collections. Scientists, like Ulisse Aldrovandi of
Bologna, collected thousands of natural specimens in their pursuit of knowledge. Merchants, such as the Fugger
family of Augsburg, were driven by economic gain and represented the “middlemen” in the exchange from natural
sources to royal Kunstkammern. A closer examination of the objects contained within Renaissance collections reveals
that some of these marvels are curiously devoid of the qualities of logic and restraint that are seen as characteristic of
Renaissance art. There is nothing rational about the sixteenth-century obsession with “unicorn horns”, and nothing
subdued about a nautilus shell embellished with gold and gemstones. How are these curiosities to be situated within
the framework of the Renaissance art? This paper will engage the different sociohistorical contexts which produced
royal, scientific, and merchant collections and will attempt to position natural and artificial oddities within the broader
structure of Renaissance visual culture.
The Hellenic Tarot
Martha Watson (Ginger Sheridan), Jacksonville University, 2800 University Blvd. North, Jacksonville, Florida 32211,
Ginger Sheridan, Computer Art Department, Jacksonville University, 2800 University BLVD, Jacksonville FL 32211
The use of Tarot cards has been a spiritual guidance device practived since antiquity. The Tarot methodology is a
part of a belief that spiritual energy exists and can be tapped into through the use of a symbolically organized system
consisting of 74 cards. I am investigating a correlation of the Tarot deck system and Hellenic mythology. My goal is
to create a Hellenic Tarot card system, themed with Greek Gods, by researching the history and design of Tarot card
decks(specifically the Rider-Waite system), how the decks relate to the Kabbalah(the Jewish Tree of Life) and the
meaning of symbols used in their design. In designing my reference deck, I will reseach Greek religion and chose
from the most prominent of their copious gods and goddessses as a source of symbolism and imagery. The deities
will correspond with the meanings of the cards as representative figures. Http://www.darkless.net/tarot/
The product of this thesis research will result in original Tarot Card designs of the Major Arcana. I will create the
imagery as pencil illustrations, scan them into a computer and manipulate color with software. The final imagery will
be printed on cardstock.
ALMOST THERE...
Georgia Johnson, Caroline Madden, Computer Art Department, Jacksonville University, 2800 University Blvd. North,
Jacksonville, FL 32211
Body Language is a powerful vehicle for communication much of which operates at a subconscious level. Influenced
by hearing impairment and inspired by contemporary animation and the silent movie era, this animated short entitled
“Almost there…” illustrates how human nature overcomes obstacles. The storyline deals with a fisherman’s response
when besieged by a comedy of errors on his return to shore. The genre is slapstick comedy reminiscent of silent films;
however the use of color will render the work contemporary.
Research of Chris Bailey’s contemporary animation work as well as silent films, especially those of Charlie Chaplin
and Buster Keaton serve as primary resources for this project. The slapstick of the Three Stooges is also considered.
Facial expressions and body language used in response to stimuli are explored and compared. The most visually
communicative of these are used in the animation “Almost there…” to provide a body language that communicated
clearly without sound.
11
ART/ART HISTORY
CENTERS AND PERIPHERIES: MEXICAN ECCLESIASTICAL ARCHITECTURE OF THE EIGHTEENTH
CENTURY
Clare C. Weber (Dr. Nadja Aksamija), Department of Art and Art History, Colgate University, 13 Oak Dr, Hamilton,
NY, 13346
Scholars writing on the eighteenth-century colonial architecture of Mexico have struggled with classifying it within
traditional Baroque parameters. Though this architecture has been discussed as “Baroque,” it has often been
considered peripheral because of its location outside of Western Europe. My research, focusing on the churches in
Mexico City, Puebla de los Angeles, and smaller towns in the Puebla region, addresses whether it is even appropriate
to call these churches “baroque,” and what would be a more suitable term to describe them. Previously used terms
do not properly express the style of these buildings because of the negative connotations attached to them, such as
peripheral, mestizo (a racially charged term), or acculturated form (a concept which places all creative power in the
hands of the conqueror, the Spanish). Instead, the term “hybrid” and the notion of transculturation are better models
for describing what these buildings exemplify: the melding of two cultures to produce another unique form, as
opposed to an imitation of the buildings of European centers.
This paper focuses on Mexico City, Puebla de los Angeles, and the region of Puebla because of their sizes and
relations to each other. Mexico City was, and still is, a large metropolitan city, whereas Puebla de los Angeles is
smaller, close to Mexico City, but still filled with impressive sacred structures. In addition, the surrounding areas of
both cities have innovative churches of their own. Because of the relative sizes and locations of these cities, the
center-periphery model proposed by scholars such as Enrico Castelnuovo, Ljubo Karaman, and Immanuel Wallerstein
can be applied. However, another goal of my research is to evaluate whether this model is even useful for
understanding the architecture of colonial Mexico, without demeaning it in the process. Some of the buildings
examined are the Sacristy (Sagrario Metropolitano) in Mexico City; San Francisco Acatepec in Puebla de los Angeles;
Our Lady of Ocotlan Sanctuary in Tlaxcala; and Santa Maria Tonantzintla in Tonantzintla, which together offer a
geographic and stylistic array. Although scholars have engaged in theoretical and contextual investigations of hybrid
theory and eighteenth-century Mexican architecture, focused studies on the ecclesiastical buildings in these areas are
lacking, mostly because there is still great dissension in the field about how to best classify them.
SALLY MANN AND LEWIS CARROLL: ALIKE IN INTENTIONS
Sarah C. Sankovich (Ginger Sheridan, Cheryl Sowder, Dana Chapman, Janet Haavisto) Department of Visual Arts,
Jacksonville University, 2800 University Blvd. N, Jacksonville, Florida 32211
Compared to the history of other art forms, the history of photography is relatively short since it spans only about one
hundred and fifty years. Despite this brief history, certain patterns have emerged. By studying the photographs of the
Victorian photographer Lewis Carroll alongside the photographs of contemporary photographer Sally Mann, the
similarity of subject matter is obvious; both photographed children. What may be less apparent is that while Carroll
and Mann lived in different centuries, each artist often portrayed children in ways that elicited social concern. Based
on Carroll’s photographic and social involvement with young girls, members of society have branded him a pedophile.
The work of Sally Mann is often viewed as having underlying sexual content, and it is thought to be pornographic due
to the nudity and poses of her young subjects. With more in-depth research of both primary and secondary sources,
this paper will argue that, despite the controversy, both artists were motivated by pure and similar intentions; each
endeavored to capture the true nature and innocence of childhood through the medium of photography.
12
ART/ART HISTORY
Saving Place: an Installation
Nina Stolz, Julie Weitz, Art Department, University of South Florida, 4202 E. Fowler Ave., Tampa, FL 33620
Research for my thesis focuses on understanding the concept of installation, looking in depth at various
installation artists, and narrowing down these artists to a select few that are particularly pertinent to my subject matter
(thus far I have found Ann Hamilton, Kara Walker, and Allan Kaprow the most influential). The research also includes
physical investigation of materials, space, visual interpretation of ideas, and most importantly, my family history in
order to construct my own installation as the culmination of my research.
I have come to the point in my research where I am now simply working on improving specific attributes of my
installation and putting together the written presentation. The general idea of the installation involves the recreation of
the action of hanging-to-dry sheets and pillowcases; these are the most intimate and telling of personal possessions.
On each of these garments are depictions of my own experiences using collage, drawing, and sewing. The installation
also includes hanging leaves to simulate their falling and leaves on the floor to represent the passage of time.
Participants will be guided through the installation by a maze of the specifically placed garments, leaves and sounds
of family dinner conversations. They will hopefully be reminded of their own experiences and enticed by the ideas
presented about memory and the remembering process.
Images of the work in progress can be viewed here: http://nstolz.blog.usf.edu/gallery/Saving-Place%3A-an-Installation
EXPRESSIONS OF CULTURE: VISUAL FORMS OF THE KUBA
Karen J. Willis (Dr. Mary F. Francey), Curatorial Services - Utah Museum of Fine Arts, The University of Utah, Marcia
& John Price Museum Building 410 Campus Center Drive, Salt Lake City, Utah 84112.0350
This project involves the research, development, and installation of an exhibition of African cultural material drawn
from the Utah Museum of Fine Arts collections. Often museum visitors are unaware of, or underestimate the
complexity of the African nations, yet the diversity of this continent is evident in the social, economic, aesthetic, and
religious aspects of everyday African life. The project developed from a philosophical and ethnographic framework,
connecting the physical, aesthetic, and spiritual elements that are critical to the production of cultural material.
Objects produced by a people are a way of contesting and negotiating relationships, for they help affirm and
reproduce those connections. As cultures change, visual forms may remain stylistically constant, although the
meanings attached to them may vary. By examining cultural material as the representation of a society, complexities
are revealed – the artistic and social constructs become apparent in the functional and sacred objects made and
utilized by a culture.
Kuba culture is rich with ritual and symbolism; this heritage is striking in the language it presents – a visual language
of varied forms and geometric patterns. A variety of works represent a range of designs reflecting indigenous beliefs.
Examples illustrate that disparate objects often serve to convey, support and perpetuate different parts of this culture’
s belief system. In displaying common objects alongside those of the sacred and sublime, the correlations and
ubiquity of this visual language can readily be observed. Visual forms also reinforce other types communication, as
exemplified by the story of Woot, a figure from the Kuba origin myths. Woot, the first ancestor, is symbolized by a
geometric pattern used to embellish carvings and textiles; representations also occur in oral histories and the
performances of dance-dramas. The various references maintain this character as a central figure of the culture. This
exhibition illustrates how the visual forms of the Kuba people present strong statements that carry traditional ideas;
consistencies and changes found in these works are explored – demonstrating that both styles and meanings are
dynamic.
13
ART/ART HISTORY
ABORIGINAL CHILD ARTISTS OF THE CARROLUP NATIVE SETTLEMENT: THE COMPLICATED
LEGACY OF THE “NOBLE SAVAGE”
Molly L. Cyphers (Dr. Nadja Aksamija), Department of Art and Art History, Colgate University, Hamilton, NY 13346
An incredible collection of one hundred and thirteen drawings, created by Aboriginal children of Western Australia
during the late 1940s and early 1950s, was recently rediscovered at the Picker Art Gallery at Colgate University.
These works were produced at the Carrolup Native Settlement, which was founded in 1915 by the Australian
government to encourage the process of Aboriginal assimilation into white society. The settlement was directed and
staffed by various Europeans, whose decisions regarding the education of the Aboriginal children were centered
around exploiting the assumed innate creativity of the “noble savage.” The resulting “Carrolup drawings” were widely
exhibited, inspiring an outpouring of literature that featured racially-charged language.
This paper contends that the immense popularity of the artists was a product of a white Western classification of the
Carrolup children as the infamous “other” on two levels: not only were they dark-skinned, of a non-Western culture
that was considered “primitive,” but they were also children, and thus even further removed from the traditional notion
of the artist. While it is generally believed that the various exhibitions of the 1950s did much to debunk the
assumptions held by society at large concerning the work of child artists and the notion of “primitive art,” this paper
will investigate how the display of these drawings, and the accompanying literature generated as a result of such
exhibitions, did little more than to create a role that the Aboriginal child artists were expected to play. This paper will
aim to unpack the problematic legacy of writings on the subject of the Aboriginal child artists through the examination
of crucial and thus far overlooked discrepancies that exist between the actual events surrounding the lives of the
Carrolup children and ways in which they were understood by Western society.
DEFINING THE ROLE OF ARTISTS’ BOOKS IN A NEW CENTURY
Kristie Etson and Paula Johns (Ms. Susan Hogue), Honors College and Department of Art, The University of South
Carolina, Columbia, South Carolina, 29208
Before the advent of Gutenberg’s printing press in the mid-15th century, books were delightful hand printed pages of
colored and decorated text. Due to the nature of the printing press, books became less artistic, yet still contained the
same information. In the recent artists’ book movement, one can see a return to blending beauty, text, and content.
The purpose of this research was to determine the characteristics of the modern artists’ book and to compare this with
pre-Gutenberg manuscripts. Although the Livre D’Artiste movement seemed to emulate the values of artists’ book,
and inspired the modern artists’ book, it is not defined as a true artists’ book. This project is a result of undergraduate
research begun in SCCC 384K- Art of the Book, a class that the Honors College of the University of South Carolina
offered. In this class, it was discovered that an artists` book is not a linear traditional story formatted by rules; they are
books bound only by the author’s imagination. Unlike a linear book, an artists` book involves blending form, function,
and content, visually and literally. They resume the artistic development of books where the pre-Gutenberg
manuscripts left off. Artists’ books were researched through studying artists` books, reading about artists` books, in
class presentations by established book artists, by experimenting with book construction and content techniques,
learning to operate a nineteenth century printing press, art discussions, and finally the creation of artists’ books.
Through our research, we found that artists` book are a collaboration of a visual and written story that requires a
perfect blend of form and function.
14
ART/ART HISTORY
WILLIAM POPE.L, CAPITALIST MASCULINITY, AND THE BLACK MALE BODY
Allison Phillips (Professor Cris Hassold), Department of Humanities, New College of Florida, Sarasota, Florida 34243
This paper will explore a series of public performances (Crawls and Schlong Journey) by contemporary artist William
Pope.L. Pope.L’s Crawl pieces, in which the artist (frequently dressed in a good suit) crawls along the ground, both
explore the proliferation of homelessness within urban areas and the concomitant cultural blindness surrounding these
prostrate bodies. At the same time, Pope.L’s prostrate body presents a striking counterpoint to the skyscraper
dominated urban landscapes in which he performs, highlighting the ways that Western culture privileges the phallic
and vertical. This combination of Freudian gender analysis and Marxist inflected commentary on capitalist structures
is located within the space of Pope.L’s own black male body. Pope.L’s uses his body in order to work with and
explore the peculiar position of the black male body: oscillating between presence (masculinity) and absence
(blackness). In this paper I will examine these Crawl pieces (and the related performance, Schlong Journey) using
Pope.L’s own theory about masculinity and its troubled relationship to black male bodies. I will show that the work of
William Pope.L attempts not only to bring to light socio-economic structures within the public urban realm in which he
performs, but also rethink the black male body outside of the restrictive bounds of masculinity.
THE ICONOGRAPHY OF THE SARCOPHAGUS OF JUNIUS BASSUS
Lauren Sapikowski (Dr. Kathleen Schowalter). Department of Art, Washington and Lee University, Lexington, Virginia
24450.
The Sarcophagus of Junius Bassus is a prominent example of early funerary Christian art, completed in 395 CE. It
was made specifically for Junius, the son of a consul who followed his father’s footsteps to become prefect of Rome.
Given the very Christian nature of the sarcophagus, it is curious that Junius was not baptized until he was on his
deathbed. This is probably because he was in public office; most public officials were pagan at the time and so were
most Romans. Most of the sarcophagus has damaged, but one of the more interesting sides remains intact. This side
contains scenes from the Old Testament as well as the life of Christ. Despite the fact that this side of the sarcophagus
has been highly studied, art historians have yet to clear some remaining questions about the arrangement of the
scenes on this panel. I will argue that the Sarcophagus images are arranged in a pattern where like images were
placed in opposition to one another. For example, the sacrifice of Isaac in the upper left panel relates directly to the
sacrifice of Daniel in the lower right. Connecting these images creates a cross (X). The arrangement of the scenes in
such a cross was no mistake; it has symbolic meaning for Christians as it represents the Chi in the Chi-Rho. The
Chi-Rho is the symbol of Christ which is significant, particularly at the time of Junius’ life. In fact, Constantine dreamt
he would win a major battle if he rode under the XP. The battle was indeed won, which inspired his conversion and
the legalization of Christianity. Having a Chi, and potentially Rho, as the image pattern for the sarcophagus is
noteworthy. For the newly baptized Junius, it was important that his show of faith last for an eternity. The longevity of
his devotion to Christianity would not only be important to him, but make an incredible statement for the religion itself.
POLITICAL GRAFFITI ART IN LATIN AMERICA AND REEVALUATING THE ART HISTORICAL
STRUCTURE
Rose T Candela (Virginia Spivey), Department of Art-Art History, The University of North Carolina at Asheville,
Asheville, North Carolina 28804 USA
Latin America has been marked by revolutions throughout the 20th century, such as the Mexican Revolution of 1910
and the Cuban Revolution of 1953. Art within these cultures frequently became a platform for communicating socialist
and Communist ideology rooted in the desire to overthrow capitalist elitist driven societies. Studies exist of well known
individuals such as Mexican artist Diego Rivera, who promoted a Communist perspective in his commissioned
murals; but what about lesser known collectives who also used art to spread their political message? One example is
the RPB or the Ramona Parra Brigade, working in Chile 1970s. Named after a young Chilean woman shot while
protesting, this graffiti group was dedicated to the ideology of Communism as a release from oppressive military
dictatorships. Why haven’t groups like the RPB been recognized and admitted within art history and museums, while
other political artists with similar ideological beliefs such as Diego Rivera or artists are found throughout the canon of
art history? The difference may be found in the impermanent nature of the material utilized in graffiti art. The
structure of art history prefers the identifiable and tangible object to analyze and posses; and thus the Chilean group
does not conform to the structure of art history. Where, then, does the RPB belong? This research challenges and
reevaluates the system of art history, while acknowledging the validity of grassroot artists, activists, and organizations
to be accepted into the history of art.
15
ART/ART HISTORY
LUMINIST LANDSCAPES: THE OVERLOOKED AMERICAN ART
Annah R. Lee (Virginia Spivey), Department of Art, The University of North Carolina at Asheville, Asheville, North
Carolina 28804
In the late nineteenth century, an American art movement, now called Luminism, had the potential to give credibility
to painting in the United States. Though uniquely American, Luminism was unable to break the dominating European
artistic influence. It would take another 50 years for American art to be freed of European constraints when Abstract
Expressionism gained acceptance as the first American painting style of international importance. This research
project focuses on the question, why have the Luminists failed to achieve the status appropriate to their
accomplishment? The failure of the Luminists may have been caused by their use of small canvases, their calm, still,
dreamlike landscapes, or their reliance on light, not brushstroke, to create emotion in their intimate spiritual scenes.
Furthermore, Luminist paintings did not appeal to an American public that was accustomed to the more European
tradition of landscape painting, characterized by monumental scale and dramatic depictions of nature, popularized by
the Hudson River School of the mid-nineteenth century. Some recent scholarship has given Luminist artists such as
John Frederick Kensett, Martin Johnson Head, and Fitz Hugh Lane more recognition than they experienced in their
lifetime. However, the tendency to group together all American landscape painters of this period has undermined the
important innovations of the Luminist artists. This project will examine the possible reasons for Luminism’s lack of
acceptance through a visual analysis of Luminist images and comparisons to the more popular landscape paintings of
the time. An historiographic analysis of the time period and a critical analysis of recent scholarship on nineteenth
century American art will seek to further understand why Luminism is the overlooked American art.
THE PHOTOMONTAGES OF HANNAH HOECH: IDEOLOGICAL CONSTRUCTS IN VISUAL
REPRESENTATIONS OF GENDER.
Sarah Mills, (Virginia Spivey), Department of Art History, University of North Carolina at Asheville, Asheville, NC
28804
Between 1918 and 1938 the German artist Hannah Hoech created photomontages representing gender ambiguously.
Through a formal analysis of her work and a closer look at the historical background of Germany in the early 20th
century, it can be seen how Hoech’s visual language provides a profound perception of society. Her image
translations probe deeper into the interrelationships of culture and gender. Hoech created photomontage by cutting
out photographs from popular media sources and arranging them on a paper based material. By dissembling mass
media images Hoech manipulates the very power structures of society that she wishes to question. The
photomontage technique also conveys the technological and consumer revolutions of the historical period that were
generating the changed perceptions of gender roles. Hoech’s iconographic content functions as an insight to the
symbols that became embedded within the society through the constructs of external institutions, in which the female
artist works. Coming from ideologies, arising from a certain socio-economic context, these visual forms produce
stereotypes of the traditional and modern woman, which Hoech captures ironically and cynically in her photomontage.
Hoech’s visual context depicts conflicting myths of this “New Woman” in a way that is intrinsic to the practice of art. A
semiotic analysis will be used to reveal how her visual code naturalizes a broader ideological message of her time.
16
ART/ART HISTORY
CRANACH’S MADONNA: FROM NAZI PRIZE TO JEWEL IN NORTH CAROLINA MUSEUM OF ART
Mary L. Moody (Virginia Spivey), Department of Art, The University of North Carolina, Asheville, NC 28804
Adolph Hilter’s ambitious dream of building a museum of the world’s greatest art never came to fruition, although
before he fell from power his Nazi henchmen had confiscated over 600,000 works of art. These various objects were
eventually sold to unsuspecting collectors around the world. Today, numerous art museums are faced with difficult
questions surrounding works with clouded provenance. This research asks how do museums unearth the lost history
of their collections when the provenance is ambiguous, and should the artwork be returned when relatives of prewar
owners present bona fide claims? The unique case study that is explored reveals the inspiring procedure that the
North Carolina Museum of Art (NCMA) undertook when it received a claim that one of its prized paintings, Lucas
Cranach’s Madonna and Child in a Landscape (1518) was a victim of Nazi looting. This paper will present the results
of the investigation regarding the history of this Northern Renaissance masterpiece. Examination of the NCMA’s
archives revealed the process that the museum underwent to determine proof of the prior ownership. In tracing the
history of this work, further investigation of the documents found the record of the confiscation of this painting from
the collection of a Jewish collector, its subsequent placement in the home of a top Nazi official in Vienna and its final
destination in the collection of the North Carolina Museum of Art. This presentation will conclude with discussion of
the exceptional resolution that the museum developed, and the researcher’s position that this provides a potential
prototype to be used by art museums around the world.
17
ATMOSPHERIC/ENVIRONMENTAL STUDIES
FRAMING THE STORM: USING ROBERT M. ENTMAN`S CASCADING NETWORK ACTIVATION
MODEL TO EXPLAIN MESSAGE FRAMES USED DURING HURRICANE KATRINA
Sayward C. MacInnis (Dr. Marilyn Sarow), Department of Mass Communication, Winthrop University, Rock Hill, South
Carolina 29733.
Using Robert M. Entman`s Cascading Network Activation Model, this paper analyzes the message framing of
politicians and the media during the Hurricane Katrina crisis. Entman`s model contains five steps: from top to
bottom, strongest to weakest, administration, power elites, the media, news frames, and the public. Each element
affects the other. In fact, lower steps, such as the media or public, can create the frame by pushing their desired
frame upward through the cascade. But a waterfall does not flow upward without great effort; likewise, the influence
of the public on framing is most often rare and weak.
Using content analysis of two newspapers, The New York Times and The Charlotte Observer, and press releases
from President Bush, FEMA, Mississippi Governor Hayley Barbour, Mississippi Senator Trent Lott, Louisiana
Governor Kathleen Blanco, Louisiana Senators Mary Landrieu and David Vitter, and New Orleans Mayor C. Ray
Nagin, a two-week period was studied to determine how the media`s and administration`s frames differed and why.
This paper also analyzes how the media determined Katrina`s frame rather than politicians; though, according to
Entman`s model, administration should have had the greater power. Public opinion will be used to support the data.
THE PSYCHOLOGICAL IMPACT OF HURRICANE KATRINA
Sang-Eun Park (Ms. Oona Cha and Dr. Norbert Schwarz), Department of Psychology, The University or Michigan at
Ann Arbor, Michigan 48109
The purpose of this project was to find out how natural disasters like hurricane Katrina impact on political psychology.
By comparing the president speeches delivered since one week before the event of Katrina and the speeches after
hurricane Katrina, the researcher wanted to see if there were differences between regular speeches and the president
speeches during the period of crisis in the following categories: if the president speeches after Katrina were
emotionally more appealing than other president speeches and if the speeches in the crisis period tend to emphasize
on the nation’s unity by using the words that contain the concept of `we` more often than the words for the concept of
individualism, such as “I.” The president speeches needed for the project were collected from the official White House
website, for president, and all speeches were strictly concerned about the effects of Hurricane Katrina or about the
issues on the disaster recovery. In terms of the methodology, a main device of the project was a software called,
“Linguistic Inquiry and Word Count (LIWC),” which is "a textual analysis application designed by Professor James
Pennebaker and colleagues to delve into the way people write and what their writing reveals about their states of
mind." (LIWC website). Twenty president speeches, ten speeches from each period, were examined by LIWC and the
words fallen into the categories of emotion as well as the unity of nation were counted for an analysis.
Analysis of the data by LIWC showed that the speeches during the crisis time contain more words that express
emotional psychological process than the speeches in peaceful time. Besides, the researcher found that the president
speeches after Katrina tend to prefer the words that indicate the concept of groupism than the concept of
individualism.
18
ATMOSPHERIC/ENVIRONMENTAL STUDIES
THE EFFECT OF AN EXTREME FLOODING EVENT ON LONG-TERM WATER QUALITY IN THE
SOUTHERN APPALACHIANS
Brian S. Evans (Steven C. Patch), Department of Environmental Studies, University of North Carolina at Asheville, NC
28804
In September 2004, the remnants of three hurricanes, Frances, Ivan and Jeanne, brought high levels of rainfall and
intense flooding to Western North Carolina. The goal of this study was to assess the effects of the hurricanes on the
water quality of three subwatersheds of the French Broad River, and determine if there have been any long-term
effects. This study also determines how land-use, slope, and rainfall influenced flood severity and impact. Stream
samples collected between October 2003 and August 2004 were analyzed and compared with samples collected
between October 2004 and August 2005. Water samples were tested for pH, alkalinity, turbidity, total suspended
solids, heavy metals, and nutrient content. Post-hurricane benthic monitoring was done at sites with pre-hurricane
data using the standard NC Division of Water Quality (DWQ) “EPT” collection method, where only the most
pollutant-intolerant groups are collected, mayflies, stoneflies and caddis flies. These data were compared with past
data obtained from DWQ. Areas of stream bank erosion were located by aerial photography, personal interviews with
local officials associated with the monitoring of each subwatershed, and GIS-based modeling of stream channel
morphological characteristics using the HEC-GeoRAS model. USGS Stream flow data, along with precipitation data
supplied by the National Weather Service, was used to estimate the flood intensity for each subwatershed. Land
Cover data were separated into three categories: agricultural, urban and forested. Mixed linear models were used to
assess differences in water quality pre- and post-flooding and the effects of land-cover on the degree of variation.
Using overlay analysis in ArcGIS, the results of each of the water quality tests were compared with land-cover
classification in order to estimate the effects of land cover on response to the hurricanes.
TROPICAL CYCLONE HYPERACTIVITY IN THE EASTERN AND CENTRAL CARIBBEAN SEA DURING
THE 2005 ATLANTIC HURRICANE SEASON
Amy Harless (Christopher C. Hennon) Department of Atmospheric Sciences, The University of North Carolina at
Asheville, Asheville, NC 28804
The 2005 hurricane season produced an unprecedented number of hurricanes in the Atlantic Ocean. With
twenty-three named storms, the pre-determined list of storm names was exhausted; and for the first time since
record-keeping, an auxiliary list was implemented. In addition, a second record was broken when Hurricane Wilma’s
minimum pressure hit 882 millibars, exceeding the record low of Hurricane Mitch (1998). One aspect of the overall
hyperactivity was the elevated number of tropical cyclone formations in the eastern and central Caribbean Sea. This
research focuses on the tropical cyclone activity in this area and examines the reasons contributing to this increased
frequency. The North Atlantic covers a vast amount of area, and different regions of this area have distinctive
atmospheric conditions that make the creation and development of hurricanes more favorable. Results will be
presented that examine these specific conditions. The uniqueness of this season, which made this region more
conducive to hurricane formation and intensification, will be compared to the past climatology. The driving forces
investigated are mid-latitude baroclinic factors, sea surface temperatures, depth of the region’s oceanic mixed layer,
and vertical wind shear. The Chi-Square statistical test will be applied in order to give an indication as to whether or
not there is a difference in the number of hurricanes in and crossing through the area of study compared to past
seasons. This research may lead to more accurate forecasting for the origination and tracking of a season’s
hurricanes in the eastern and Central Caribbean Sea. This may enable forecasters to more skillfully project how
individual regions will be affected by tropical systems, thus increasing warning lead times and better protecting
society against the destruction of hurricanes.
19
BIOCHEMISTRY
Oxidative Stress in Batten Disease
Crosby A Sommers (1), Jared W Benedict (1,2), and David A Pearce (1,2)., 1 Center for Aging and Developmental
Biology, 2 Department of Biochemistry and Biophysics, University of Rochester School of Medicine and Dentistry, 601
Elmwood Avenue, Rochester, NY 1464
Batten disease is a childhood neurodegenerative disorder caused by a mutation in CLN3. Oxidative stress has been
implicated in the pathology of many neurodegenerative disorders. One possible disease mechanism in Batten
disease is oxidatively induced damage to proteins and other cellular structures along with decreased antioxidant
defense systems. Specific regions of the central nervous system of Cln3-knockout (Cln3-/-) mice showed increased
protein oxidation measured by protein carbonylation. In addition, glutathione levels were significantly lowered in
diseased brain tissue. The brain regions affected are part of the motor loop, which is specifically targeted in Batten
disease, suggesting oxidative stress is a contributing pathological mechanism.
Effect of Antiprogesterone Org 31710 on Ovarian Cancer Cell Growth
Eddie A Dickes (Alicia A Goyeneche and Carlos M Telleria), Division of Basic Biomedical Sciences, University of
South Dakota School of Medicine, Vermillion, SD 57069
Ovarian cancer is the deadliest of the gynecologic cancers and is the fifth leading cause of cancer death among
women in the United States. Today, 50% of women diagnosed with ovarian cancer die from it within five years. Poor
prognosis is due to late diagnosis and lack of efficient chemotherapy treatments. Ovarian cancer patients are
primarily treated with a combination of platinum agents, such as cisplatin, and microtubule disrupters, such as
paclitaxel. The problem with current treatments is that patients quickly develop resistance to these drugs, creating a
need to study more efficient ways to treat the disease. Our laboratory studies have demonstrated that mifepristone
(RU486), which binds with high affinity to progesterone receptors and glucocorticoid receptors, inhibits growth of
various ovarian cancer cell lines without triggering cell death. Org31710 is a new synthetic steroid with mostly
progesterone antagonistic activity. It binds with high affinity to progesterone receptors and with very low affinity to
glucocorticoid receptors. This is significant because the potential usage of Org31710 as a cytostatic in humans would
have lesser side effects than mifepristone, such as management of glucose metabolism and stress and immune
responses, which are mediated through glucocorticoid receptors. The purpose of this study is to study the efficacy of
Org 31710 as a cell growth inhibitor in the ovarian cancer cell line 2008. Our results indicated that Org 31710 is an
effective antiproliferative agent on the 2008 ovarian cancer cell line. A time course as well as flow cytometry
experiments indicated that inhibition of cell growth is present after 24 h of exposure to the drug, and the effect is
maintained thereafter. As a result of treatment, 2008 cells appear to be arrested in the G1 phase of the cell cycle.
Inhibition of growth in 2008 by Org 31710 is associated with increased expression of the cyclin-dependent kinase
inhibitors p21 and p27. Cells that detached as a consequence of Org 31710 treatment, presented fragmented DNA,
whereas cells that remained attached after treatment, did not show DNA fragmentation. Collectively our data suggest
that Org 31710 can be an effective agent for ovarian cancer therapeutics.
20
BIOCHEMISTRY
THE EFFECT OF ESTROGEN, DAIDZEN, AND GENISTEIN ON PC-12 CELL VIABILITY FOLLOWING
HYPOXIC/ISCHEMIC EXPOSURE
Desiree Bryant(Dr. Deborah Rayfield)Natural Science Department, Bowie State University, Bowie, Maryland 20715
The overall incidence of cardiovascular disease is significantly lower in premenopausal women when compared to
age-matched men. Thus, cardiovascular disease is largely a disease of the postmenopausal women. Considering that
women will spend more than a third of their lives in the post menopausal years, preventative measures are of
paramount importance. The Women’s Health Initiative spearheaded a large randomized study to look at primary
prevention of cardiovascular disease in menopausal women. The study was prematurely ended when interim analysis
demonstrated an unacceptable risk profile related to an increase in the incidence of breast cancer, with no
cardiovascular protection. Data from this study also suggest an increase in blood clotss, strokes, and coronary
disease. In light of these findings, more women are searching are searching for alternative treatments for menopause
relates vasomotor symptoms and prevention of postmenopausal cardiovascular disease. A common denominator
among these disorders is hypoxia. Hypoxia plays a major role in cardiomyocyte disfunction and impaired neurological
function. The intent of the study is to measure the effect of geistein, 17-beta estradiol, and daidzen on changes in
PC-12 cell viability following exposure to hypoxia and ischemia. PC-12 cells were chosen for use in this study due to
their enhanced oxygen sensing capabilities.
The specific aims of this study are to (1) measure the effect of genistein, 17-beta estradiol, and daidzen on
PC-12 cells exposed to time intervals of hypoxic and ischemic exposures (2) determine the role of tyrosine inhibition
on observed responses, and to (3) measure the ability of genistein, daidzen, and 17-beta estradiol to prevent
thapsigargin induced apoptosis in PC-12 cells. Cell death will be monitored by measuring the release of lactate
dehydrogenase enzymatic activity into the culture supertanant. Differentiated and undifferentiated PC-12 cells will be
maintained in a suspension culture until sued for assay. Ischemic conditions will be simulated by exposing cells to an
aglycemic and hypoxic conditions. Results will be expressed as mean +/- S.E. Data will be analyzed using a one way
analysis of variance with Bonferroni corrections for multiple comparisons or by using the Student’s test P values <
0.05 will be considered statistically significant.
Role of the Central Complex on Transitional Behaviors of Blaberus Discoidalis
Yidi Xu, Angela Ridgel, Roy Ritzmann, Department of Biology, Case Western Reserve University, 10900 Euclid Ave.,
Cleveland, OH 44106-7080
Insects are able to navigate through a wide variety of complex terrain by climbing and turning to overcome and avoid
barriers; these are known as transitional behaviors. They do this through tactile and visual sensing of their
environment using head sensors, the antennae and the eyes respectively. Sensory input is sent to the brain,
processed, and used to direct appropriate actions in the legs. The specific area of the brain that controls transitional
behaviors is not yet fully understood, and therefore of particular interest.
Previous findings have suggested that the central complex (CC), a cluster of interconnected neuropils located on
the midline of the protocerebrum (comparable to the cerebellum), is important in turning behaviors. Contact with a
barrier can result in an imbalance of sensory information coming from the left and right antennae. We hypothesize
that the CC processes this information and sends descending instructions to the thoracic ganglia, a collection of
sensory and motor neurons in the leg segments, which then directs the necessary leg movements to move around the
barrier. The goal of this study is to disrupt small regions of the CC to alter descending signals and affect transitional
behaviors.
Discrete electrolytic lesions were created in the CC of cockroaches of the species Blaberus discoidalis to damage
specific brain regions, and the subsequent behaviors of the cockroaches were observed and compared to their
performance in the normal state. Abnormal behaviors included those that affected walking, turning, and climbing.
Some cockroaches displayed no abnormal behavior. Although all lesions were targeted to the CC, each lesion hit a
different area of the brain, with some missing and some hitting within the CC. It was found that lesions in the CC
caused significant abnormalities in turning and climbing behaviors, while those outside of the CC caused abnormal
walking. Motor deficits varied in type and degree of severity depending on the exact lesion location. These results
not only further demonstrate the role of the CC in locomotion, but also show that different regions within the CC have
distinct effects on transitional behaviors.
21
BIOCHEMISTRY
THE ROLE OF THROMBOMODULIN IN PROSTATE CANCER PROGRESSION
Jonathan Hathaway and Tyler Walker (Dr. Laura Glasscock and Dr. Chris Teigland), Department of Biology, Winthrop
University, Rock Hill, SC 29733 and Carolinas Medical Center, Charlotte, NC 28232.
An association between blood coagulation and metastatic dissemination has long been recognized. Thrombomodulin
(TM) is a receptor found on endothelial cells that is regulates blood coagulation. TM binds to the serine protease,
thrombin (IIa), and the TM-IIa complex activates the serine protease, protein C (pC), into the anticoagulant serine
protease, activated Protein C (APC). It has also been ascertained that number of TM receptors on prostate cancer
cells (CaP) are elevated compared to normal prostate epithelial cells. It has also been shown that TM regulates the
proliferation of CaP cells in vitro. Since TM is elevated in patients with CaP, regulates CaP cell proliferation, and is
involved with pC, APC, and IIa in coagulation, we chose to study what effect these TM associated serine proteases
have on the proliferation of PC-3, LNCaP and DU-145 CaP cell lines. We determined that as IIa concentrations
increased (0-400 nm), the rate of proliferation of DU-145 cells decreased up to 43%. However, this negative trend
was not seen with PC3 CaP cell lines until 300 nm IIa. Unlike with IIa, as pC concentrations increased (0-100 nm), the
proliferation of both cell lines decreased. In PC3 cells, proliferation decreased up to 35% and in DU-145 cells up to
21%. Similarly, as APC concentrations increased (0-100 nm), the proliferation rates also decreased in PC3 cells up to
27% and in Du-145 cells up to 41%. We concluded that TM, pC, APC, and to some degree, IIa, negatively regulate
proliferation of PC3, DU-145, and LNCaP CaP cell lines.
PRODUCTION OF RECOMBINANT MACPF COMPLEMENT C8&#945; IN SF9 INSECT CELLS FOR
PROTEIN PRODUCTION AND PURIFICATION
Zainab Ghadiyali(Dr Chasta Parker), Dept of Chemistry,Geology and Physics,Winthrop University, Rock Hill, SC
29733
Composed of over 35 proteins, enzymes, and regulatory molecules, the complement system works in conjunction with
the immune system to destroy and remove foreign substances (i.e. bacteria, virally infected particles, fungi, and
parasites) from the body. The objective of this project is to produce a recombinant form of the putative
membrane-binding region (MACPF) of the 64 kDa C8&#945; subunit of the eighth component of human complement
(C8) for structural analysis. C8 is an oligomeric protein made up of three non-identical subunits, C8&#61537; (64
kDa), C8&#61538; (64 kDa), and C8&#61543; (22 kDa). The C8&#61537; and C8&#61543; subunits are covalently
bonded through a disulfide bond while C8&#61538; is non-covalently associated with C8&#61537;. C8&#61537; and
C8&#61538; are homologous to one another and together with C6, C7, and C9 comprise the MAC protein family. The
MACPF is considered to be self-folding, is ~ 43 kDa in size, and contains two disulfide bonds and no carbohydrates.
The MACPF region has been cloned into the vector pIEx-4 and the sequence verified. The cloning experiments, the
production and initial purification steps will be presented.
This work was supported by the SC-INBRE NIH grant and Winthrop University.
EXPRESSION OF ASCORBATE PEROXIDASE FROM SWITCHGRASS
Jody A. Elson (Frank Kovacs), Deparment of Chemistry, The University of Nebraska at Kearney, Kearney, Nebraska
68849
The goal of this present work was to fully characterize the gene sequence of a possible ascorbate peroxidase and
then clone this gene into a plasmid containing a 6x-histidine tag (pET28a). Panicum virgatum, more commonly
known as switchgrass, was chosen by the United States Department of Energy as a target species for development as
a biofuel crop. As a result there has recently been considerable interest in better characterizing the biochemistry of
this plant. Ascorbate peroxidase (APX) is a heme-containing enzyme that plays a vital role in hydrogen peroxide
detoxification. We obtained a gene from a complementary DNA library prepared from the stems and leaves of
switchgrass. The plasmid with the gene was purified, quantified and sent for sequencing. The sequence data showed
remarkable similarity to the sequence of an APX gene from another organism called Pennisetum glaucum, more
commonly referred to as millet. There is a difference of three amino acids in the code from the P. glaucum protein
and our protein. However, the differing amino acids have very similar properties. Using the sequence data, we
designed PCR primers to introduce the NdeI and EcoRI cut sites to the beginning and end of the gene respectively.
Polymerase Chain Reaction (PCR) was used to amplify the gene. The PCR product was then inserted into a TOPO®
plasmid. The TOPO® plasmid was transformed into chemically competent TOP10 Escherichia coli cells. Finally,
the gene was cut from the TOPO® plasmid using restriction enzymes and inserted into a pET-28a plasmid. In further
studies, we will express this protein in E. coli (BL21 (DE3)) and use the histidine tag for affinity purification.
22
BIOCHEMISTRY
Determining the Structure of STIV-A81: a Protien from a Novel Crenarcheal Virus
Nathanael G. Lintner1,3, Eric T. Larson1,2, Mark Young1,4 and C. Martin Lawrence1,2 (C. Martin Lawrence1,2)
1Thermal Biology Institute, 2Dept. of Chemistry and Biochemistry, 3Dept. of Cell Biology and Neuroscience, 4Dept. of
Plant Sciences and Plant Patho
Sulfolobus Turreted Isocahedral Virus (STIV) is a virus that infects Sulfolobus Solfataricus of the Domain Archaea
and the Kingdom Crenarcheota. STIV was isolated from a hot spring in the Midway Geyser Basin in Yellowstone
National Park. The hot spring ranged in pH from 2.9 to 3.9 and had a temperature range of 72°C to 92°C. STIV has
a circular 18 kilobase genome with 36 open reading frames (ORF). A BLAST search gives little information on the
function of the open reading frames. Therefore, we are attempting to determine the structures of STIV proteins using
X-ray crystallography. Structural similarities between members of the STIV proteome and those of known function
will facilitate annotation of the STIV proteome. Current efforts are focused on ORF A-81. This gene has been cloned
into the P-14 gateway plasmid with a C-terminal 6x His tag and transformed into E. coli (BL-21). The protein has
been expressed and purified using heat denaturation and Ni-NTA affinity chromatography. This protein was further
purified by size exclusion chromatography using a superdex S-200 column with buffer exchange into 5 mM Tris, pH
8.0, 25 mM NaCl.. The protein was concentrated to 11 mg/mL and used for sitting drop vapor diffusion crystallization
trials. Crystals of A-81 were found with a well solution consisting of 1.1 M trisodium citrate, 0.1 M HEPES pH 7.75,
final pH 8.0. The crystals show anisotropic diffraction patterns that extend to 3.0 Å in to two dimensions, but only to
6.0 Å in the third, indicating that the crystals are disordered in one dimension. Optimization of crystallization
conditions, including the use of additive screens, is ongoing. Further progress towards the structure determination of
STIV A-81 will be presented.
The Role of Methylated Groups in the Binding of High Mobility Group A
Kelly E. Pace, (Dr. Takita F. Sumter), Chemistry Department, Winthrop University, 520 Cherry Road SIMS 101, Rock
Hill, SC 29733
The high mobility group A, HMGA, proteins are a family of proteins
involved in transcriptional regulation, retroviral integration, and
cancer initiation. The family members are all over expressed in cancers and have established oncogenic properties.
The precise mechanisms involved in HMGA-mediated transformation are unclear. All HMGA proteins posses three
characteristic AT hook motifs and an acidic carboxy-terminal tail. Previous studies suggest that in transcription, these
AT hook motifs act cooperatively to facilitate the binding of the HMGA proteins to their DNA targets while the acidic
amino acid residues located within the C-terminal tail portends binding specificity. More recently, it has been noted
that arginine residues located within the AT hook DNA binding domains of HMGA proteins possess covalently linked
methyl groups in proteins isolated from cancerous specimens. It is
possible that this arginine methylation plays is involved in binding of these proteins to oncogenic DNA targets. To
study the role of arginine methylation in DNA binding, we overexpressed HMGA1a in prokaryotic and eukaryotic
expression systems. Prokaryotic expression allows rapid and high level production of HMGA lacking the
posttranslational methyl residues. We also overexpressed our protein in Sf9 insect cells to produce proteins bearing
the methylated arginine residues. Since the methylated groups are formed through post translation modification, to
make protein without them cells incapable of post translational modification will be used; bacterial cells meet this
criteria. The
yields and methods of protein isolation will be discussed and future studies will entail the development of a
fluorescence assay to quantitate the binding of HMGA proteins to its DNA targets. It is likely that these and other
studies will lead to specific and more effective cancer therapies.
23
BIOCHEMISTRY
Determination of The Protein Content of Isolated Trichomes from Arabidopsis thaliana
Michael Waldt (Dr. Ross Jilk), Department of Chemistry, University of Wisconsin - River Falls, Centennial Science Hall
410 South Third Street, River Falls WI 54022
We report here on our efforts to determine the protein composition of trichomes from Arabidopsis thaliana.
Determining the protein makeup of trichomes will provide new information on cell fate determination in leaf cells as
well as provide a greater understanding of cell differentiation in general. Previous genetic research has discovered
three transcription factors that together form an initiation complex involved in trichome development. However, it has
yet to be determined what proteins are expressed in response to this complex. Trichomes are removed from the leaf
through a combination of chemical and mechanical means, using 50 mM EGTA and vortexing in the presence of
small glass beads. Once the trichomes have been removed from the leaves, the soluble proteins are extracted. This
allows for direct analysis of the protein content of trichomes. Through this, we hope to gain better knowledge of the
proteins involved in the development of trichome differentiation. Primary protein characterization will be
accomplished through the use of SDS-PAGE and two dimensional gel electrophoresis.
IDENTIFICATION OF THE MOTIF THAT IS REQUIRED FOR THE SUPERCOILING ACTIVITY OF DNA
GYRASE
Valerie M. Kramlinger (Dr. Hiroshi Hiasa), Department of Pharmacology, University of Minnesota Medical School-Twin
Cities, Minneapolis, Minnesota 55455
DNA gyrase is the only topoisomerase that can introduce negative supercoils into DNA. It is thought that the binding
of conventional type II topoisomerases, including topoisomerase IV, to DNA takes place at the catalytic domain
across the DNA gate, whereas DNA gyrase binds to DNA not only at the amino-terminal catalytic domain but also at
the carboxyl-terminal domain (CTD) of the GyrA subunit. The binding of the GyrA CTD to DNA allows gyrase to wrap
DNA about itself and catalyze the supercoiling reaction. Recent structural studies, however, have revealed striking
similarities between the GyrA CTD and the ParC CTD, as well as the ability of the ParC CTD to bind and bend DNA.
Thus, the molecular basis of gyrase-mediated wrapping of DNA needs to be reexamined. Here, we have conducted a
mutational analysis to reveal the functional role of the ‘GyrA-box’, a 7-amino acid-long motif unique to the GyrA CTD,
in determining the DNA binding mode of gyrase. Either a deletion of the entire GyrA-box or a substitution of the
GyrA-box with 7 Ala residues abolishes the ability of gyrase to wrap DNA about itself and catalyze the supercoiling
reaction. However, these mutations do not affect the relaxation and decatenation activities of gyrase. Thus, it is the
presence of a GyrA-box that makes gyrase a unique topoisomerase, which is capable of wrapping DNA and
catalyzing the supercoiling reaction.
The Institute of Technology, University of Minnesota, Minneapolis, Minnesota 55455
24
BIOCHEMISTRY
C-11 BODIPY, A FLUORESCENT PROBE FOR DETECTING FREE RADICALS, PHOTOPRODUCES
REACTIVE OXYGEN SPECIES AND IS TOXIC TO CELLS
Pulsar Li, Piotr Bilski, and Colin F. Chignell, Laboratory of Pharmacology and Chemistry, NIEHS, RTP, NC 27709
When oxidized by free radicals in membranes, the highly fluorescent lipophilic probe C11-BODIPY changes its
fluorescence from red to green, which is caused by the oxidation of the unsaturated side chain. We have found that
C11-BODIPY itself produces singlet oxygen (1O2) and free radicals upon irradiation with visible light. Upon prolonged
irradiation, the C11-BODIPY red fluorophore was slowly photobleached in organic solutions and in aqueous micellar
solutions, which was usually associated with a rise of green fluorescence. Both the photobleaching rate and the
production of green fluorescence increased considerably in the presence of sodium azide because C11-BODIPY is
able to photooxidize the azide anion to the azide radical. The azide radical was trapped using DMPO, which produced
a strong spectrum of the DMPO/&#9679;N3 radical adduct. Even in the absence of azide, C11-BODIPY oxidized
vitamin E and ascorbyl palmitate to their corresponding radicals, which could involve 1O2. The quantum yields of
1O2 production by BODIPY range from 0.04 to ca. 0.1, depending on the solvent and oxygen concentration.
C11-BODIPY quenched 1O2 physically with the rate constant kq=6.9x10^6 M-1s-1 in acetonitrile. While 1O2 does not
oxidize C11-BODIPY directly, 1O2 is able to oxidize unsaturated fatty acids in membrane, thus starting a radical
oxidation that would contribute to the C11-BODIPY response. In addition, both LDH and MTS cell toxicity assays
suggest that C11-BODIPY presents moderate phototoxicity towards cell membranes in both light and dark
environments. Our findings show that C11-BODIPY is a moderate photosensitizer that may be prone to produce
false-positive results when used to detect free radicals in membranes.
---end of abstract--*note, due to exponents and postscript formatting, this abstract is best viewed using a Word document format. please
let me know to whom I should send the abstract in word format. thank you.
EXPRESSION AND NMR STRUCTURE DETERMINATION OF RECOMBINANT HUMAN TROPOMYOSIN
ISOFORM 5 AND ITS MUTANTS
Oscar R. Rebolledo(1), Carlos J. Vera(2), Xuemei Huang(1), Sandy Go(1), Stanley J. Opella(1), L. Amy Sung(2),
Department of Chemistry and Biochemistry(1), Department of Bioengineering(2), University of California at San Diego,
La Jolla, CA
An erythrocyte’s membrane has only a thin protein network underneath its lipid bilayer. The actin-spectrin network has
a quasi-hexagonal topology, with six long spectrin molecules radiating from a ~37 nm central short actin protofilament
at the junctional complex. The length of the protofilament may be partially attributed to a “molecular ruler,” a complex
of globular erythrocyte tropomodulin (E-Tmod) and rod-like tropomyosin (TM) isoforms 5/5b. The topology and
properties of the network provides the mechanical stability for the lipid bilayer and the remarkable elasticity of
erythrocytes. A model of hTM5 is presented in which the six actin-binding domains (6dAc) are defined. Inserting or
deleting certain domains through overlap extension PCR yields hTM5 of varying lengths. Here we report the
expression and characterization of full-length hTM5 and its mutants with 3dAc and 4dAc. The pET-3d/hTM5 and
mutant plasmids were each transformed into BL21(DE3)pLysS competent cells and induced to express in M9 minimal
media in the presence of (15NH4)2SO4 using IPTG . The recombinant hTM5 proteins were purified by heat
denaturation, ammonium sulfate precipitation, and membrane dialysis. TM exists as two-chain, &#945;-helical
coiled-coils. Preliminary structural analysis was done through circular dichroism (CD) and 1H-15N HSQC NMR
experiments. CD results showed that hTM5 and the mutant hTM5-4dAc contain an average value of 58% and 61%,
respectively, of &#945;-helical content. NMR showed overlapping signals in the full length and mutant’s spectra. This
investigation will determine the secondary structure of hTM5 and its mutants, and the constructs will be used to
generate and test in vitro and in vivo their impact on erythrocyte`s network topology.
25
BIOCHEMISTRY
The Isolation, Purification, and Characterization of an Antifreeze Protein From the Sea Smelt
(Hypomesus japonicus)
Ayanah Duhaney and Mohit Sirohi (Loyd D. Bastin), Department of Chemistry. Widener University, One University
Place, Chester, Pennsylvania 19013
Many organisms are able to survive at nearly subzero temperatures. However, at these temperatures, bodily fluids
are normally susceptible to freezing. Therefore, many organisms have adapted to these lower temperatures through
the synthesis of antifreeze proteins. Antifreeze proteins are proteins capable of binding to ice and making further ice
crystal growth energetically unfavorable thus depressing the freezing point non-colligatively and allowing the
organism to endure harsher conditions. The Sea Smelt is capable of surviving frigid temperatures. Here, we report
the isolation, purification, and thermodynamic studies of an antifreeze protein from the muscle of sea smelt. Through
the study of the structure and function of antifreeze proteins, we hope to determine the mechanism by which proteins
inhibit ice crystal growth in the sea smelt.
Progress towards crystal-structure of stabilized HIV gp120 core protein in the CD4-bound
conformation as a basis for vaccine development and further
Martin Lawrence, Martin Teintze, Bjoern Traenkle , Martin Lawrence, Chemistry/Biochemistry,, Montana State
University Bozeman, Leon Johnson Hall 728
The objective of this project is to stabilize the HIV coat protein gp120 in a conformation specific to when it is bound to
the cellular receptor CD4 and to determine its 3-dimensional x-ray structure. This will allow further insight into the
interaction between binding sites of receptor and coat protein.
HIV gp120 is known to fold into 2 major conformations dependent on whether it is bound to the receptor or not. The
unbound conformation, in which the recognition site is occluded by other parts of the protein, predominates over the
bound one. The inability of the immune system to create broadly reactive antibodies is therefore not only due to the
low fidelity of the viral reverse transcriptase causing many mutations in gp120, but also to the inaccessibility of the
binding site. In order to maintain receptor binding ability some residues need to be conserved and are common
among all HI-viruses.
The gp120 core protein used for this studies is created by removing variable parts of the gp120 gene in order to
minimize the number of flexible loops, since these can prevent crystallization. Analyzing earlier structures of the
related SIV gp120 reveals positions most suitable for site directed mutagenesis to introduced cysteine residues to
form stabilizing disulfide bonds. A eukaryotic expression system is used for protein expression. Several types of
chromatographic methods are used for purification. Crystallization conditions are screened using automated
procedures and best conditions will be further optimized to obtain diffraction quality crystals. Diffraction data will be
collected and the structure will be solved using molecular replacement.
Protein stabilized in the bound conformation could be used to create universal antibodies recognizing the binding site
of gp120. Results will be useful in order to understand how an effective vaccine will have to look like.
26
BIOCHEMISTRY
ISOLATION AND CHARACTERIZATION OF ALL-TRANS-RETINOIC ACID ISOMERS PRODUCED BY
ULTRAVIOLET LIGHT EXPOSURE
Andres Chang (Dr. Melissa Kelley), Department of Chemistry, University of Central Arkansas, Conway, AR, 72034
The purpose of this project was to isolate and characterize isomers produced when all-trans retinoic acid (t-RA) is
exposed to ultraviolet light (UV) radiation. All-trans-retinoic acid and its isomer 9-cis-retinoic acid (9-cis-RA) are
biologically active metabolites of vitamin A (retinol) and play an important role in many critical life processes including
vision, reproduction, cellular development, and epithelial cell differentiation by mediating gene expression through
nuclear receptors that bind t-RA and 9-cis-RA. Retinoids are light-sensitive compounds that isomerize when exposed
to light. In this project, t-RA was dissolved in solvents of different sizes and polarities (methanol, ethanol, hexane,
acetone, and DMSO) and exposed to ultraviolet light (350nm). Samples were collected at 5 minutes intervals for 20
minutes. The samples were analyzed using UV/Vis spectroscopy to detect initial changes occurring after exposure to
UV radiation. Reverse-phase high performance liquid chromatography (HPLC) was then used to detect and isolate
different isomers present in the samples. Finally, nuclear magnetic resonance (NMR) spectroscopy was used to
determine the molecular structure of the isomers isolated by HPLC. UV/Vis and HPLC data indicate that t-RA rapidly
isomerizes in the presence of UV light. Isomers were formed when t-RA was exposed to UV light in methanol,
ethanol, and hexane. However, some isomers produced initially experienced further reaction, leading to a decrease in
concentration of the initial isomers at 10, 15, and 20 minute time periods. Those reactions appear to occur faster in
small polar solvents, with non-polar bulkier solvents having a slower rate. While the initial isomers were detected after
exposing the hexane samples to UV light for 20 minutes, those same isomers were not detected in the methanol or
ethanol samples that were exposed over the same period. Our data suggests that solvent polarity and size plays a
significant role in all-trans-retinoic acid isomer production.
Evaluation of Streptomyces H7667 in Glycogen Synthase Kinase 3beta assay
1Adam P. Foss, 1Wei Xiang, 2Fo Sek Hin, 2Ho Coy Choke (1Leng Chee Chang), 1Department of Chemistry and
Biochemistry, College of Science and Engineering, University of Minnesota, Duluth, Duluth, Minnesota 55812,
2School of Science and Technology, Universit
www.d.umn.edu/~lcchang/chem4342/ppt/ click on abs-mod_3[1].pdf
Studies on signal transduction pathways have indicated various promising molecular targets for therapeutic
intervention in cancer therapy, including glycogen synthase kinase-3 beta (GSK-3&#946;). This kinase plays a role in
tumorigeneses, apoptosis, cell proliferation, and various other cellular functions. In addition, deregulation of
GSK-3&#946; has resulted in cancer and other diseases such as diabetes. Natural products play a vital role in drug
discovery having lead to several new GSK-3&#946; inhibitors. In this study, mammalian GSK-3&#946; cloned into
yeast is used as a primary screen to search for GSK-3beta inhibitors. Natural product extracts from the Natural
Products Branch, Developmental Therapeutic Program of the National Cancer Institute are being evaluated using this
bioassay. Species of Streptomyces, fungi, and other microbes are being evaluated. The crude organic extract of
Streptomyces H7667, obtained through a materials transfer agreement with Professor Ho Coy Choke from the
University Malaysia Sabah, has been selected for further study, since preliminary screening of the organic extract at a
concentration of 80 ug/20 &#956;L shows a clear zone of inhibition (12-15 mm) in the GSK-3&#946; assay. Several
active fractions have been obtained from S. H7667 by bioassay-guided fractionation through column chromatography
and other techniques. Isolation and biological evaluation of these active fractions will be presented.
27
BIOCHEMISTRY
IDENTIFICATION AND CHARACTERIZATION OF BIO-WARFARE AGENTS IN COMPLEX MATRICES
Sarah C. Zimmermann, Faithhan Choi, (Gail Gaspirich, Steven Lev), Department of Physics, Astronomy, and
Geosciences, Towson University, Towson, MD., 21252
Currently, detection of biological agents is fraught with problems of too many false positives and false negatives. A
major reason for these problems is the variable background of biological and non-biological particles that exist in the
atmosphere. As part of an effort to develop a direct method for detecting a biological-agent in the presence of a
complex natural matrix (i.e. atmospheric dust), the chemical composition of counted batches of Bacillus subtilis in
suspension was determined using a direct injection ICP-MS method. Previous work at Towson University using the
simulant B.subtilis, has determined that this bacterial species has a diagnostic inorganic fingerprint that is
reproducible and easily detected using direct injection ICP-MS. Laboratory cultured, counted and fingerprinted
batches of B. subtilis were mixed with a natural proxy for atmospheric dust at different proportions (e.g.1 B.
subtilis:1background, 2 B. subtilis:1 background, 3 B. subtilis:1 background, 5B. subtilis:1 background) in order to
determine if the direct injection ICP-MS method can detect the presence of a bio-agent in the presence of dust. The
results from these experiments suggest that as few as 2.55x1010 cells of B.subtilis can be detected in the presence of
1 gram of background dust. This type of sensitivity suggests that monitoring the inorganic composition of
atmospheric dust using direct injection ICP-MS may be a viable method of point detection for biological-agents.
EFFECTS OF STRUCTURAL VARIATION ON SUBSTRATE ENANTIOSELECTIVITY IN MANDELAMIDE
HYDROLASE
David J. Savage and Drew P. Kelly (Dr. Betty H. Stewart) Chemistry Department, Austin College, Sherman, TX
75090
Mandelamide hydrolase (MAH) is a recently discovered enzyme of the mandelate pathway for energy production in
the soil bacterium Pseudomonas putida. MAH catalyzes the hydrolysis of both enantiomers of mandelamide to
mandelate and an ammonium ion at an optimum pH of 7.8. MAH has a molecular weight of 53, 814 Daltons and is
comprised of 507 amino acids, of which four are cysteines. This study focused on the expression, isolation, and
characterization of MAH mutants in which cysteines at positions 124 and 175 were converted to alanines. Both the
wild and mutant type enzymes were genetically engineered to express a hexahistidine tag that simplified the
purification process while not affecting enzymatic activity. The activity of the enzymes was measured
spectrophotometrically using a coupled kinetic assay. Under our experimental conditions, the wild type and mutant
enzymes demonstrated significant enantioselectiviy for (R)-mandelamide. Also, the MAH C124A and C175A mutants
exhibited decreases in catalytic efficiency of 5% and over 50%, respectively, when compared to the wild type
enzyme. These findings suggest that the cysteine at position 124 is not critical to overall enzymatic activity, whereas
the cysteine at position 175 appears to be important for function.
The role of AT1 and AT2 receptors in AII stimulation of cardiac fibroblasts
Myo-Pale’ Aye (Dr. Thomas Peeler), Biology Department, Susquehanna University, Selinsgrove, PA 17870
Angiotensin II (AII) has various physiological functions including the regulation of blood pressure and body fluid
homeostasis. AII also specifically stimulates cell division in cardiac fibroblasts through its specific subtype receptors.
AII stimulation is mediated by two cell surface receptors, AT1 and AT2. The present research investigates whether
AT1 receptors or AT2 receptors are responsible for the AII stimulated increase in focal adhesion abundance in cardiac
fibroblasts. Cardiac fibroblasts were isolated from neonatal rats using enzymatic dispersion and cultured in vitro.
Cardiac fibroblasts were serum-starved for 48 hours prior to treatment with AII. The serum-starved cardiac fibroblasts
were exposed to 10-8 M AII for 24 hours in the presence of either the AT1 receptor antagonist, losartan, or the AT2
receptor antagonist, PD 123319. After treatment with AII, cells were fixed on the cover slips and incubated with
antiphosphotyrosine antibody to visualize the abundance of focal adhesions. An increase in focal adhesion
abundance was observed in the cells treated with AII in the presence of the AT2 inhibitor, PD 123319 and in the
absence of both inhibitors. However, no change in the abundance of focal adhesions was observed in cardiac
fibroblasts treated with AII in the presence of the AT1 inhibitor, losartan. Results from this experiment demonstrate
that the AT1 receptors are essential for the AII stimulated increase in focal adhesion abundance in cardiac fibroblasts.
28
BIOCHEMISTRY
ß-GLUCAN CAUSES LUNG MACROPHAGES AND EPITHELIAL CELLS TO RELEASE
PRO-INFLAMMATORY MEDIATORS
Suzanne M. Crumley, Zvezdana Vuk-Pavlovic, and Andrew H. Limper, Thoracic Diseases Research Unit, Mayo Clinic,
Rochester, Minnesota 55905
Lung inflammation is a serious threat to persons infected with life-threatening diseases, such as AIDS or other
illnesses involving an immunocompromised state. The fungal parasite Pneumocystis aggravates lung inflammation
through the ß-glucan component of its cell wall. After the fungus has invaded the lungs, the body defends itself by
the subsequent release of cytokines such as TNF-alpha, IL-8, and others. In these investigations it was found that
both epithelial cells and macrophages respond to ß-glucan in a concentration-dependent manner in their release of
pro-inflammatory mediators. However, in excess, TNF-alpha often causes more damage to the lungs than
Pneumocystis itself. Therefore, it is important to understand exactly how macrophages are stimulated to release
TNF-alpha. The mechanism of dectin-1 is one of the many receptors present on alveolar macrophages that
recognizes ß-glucan and stimulates macrophages to secrete TNF-alpha. ß-glucan interaction with dectin-1 receptors
outside the cell is yet unknown. Crosslinking of dectin-1 receptors could be involved in this activation. In this
investigation, it was first reconfirmed that the ß-glucan does stimulate TNF-alpha release through the dectin-1
receptor. By showing that anti-dectin-1 antibody blocked ß-glucan binding to dectin-1 and inhibited TNF-alpha
release. Lung collections SP-A and SP-D also play a role in the pulmonary inflammation. In an infected or damaged
lung, the surfactant proteins SP-A and SP-D affect the release of pro-inflammatory mediators, yet, this role is not
completely understood. Human airway epithelial cells (HAEo-) were found to enhance IL-8 release when
pre-incubated with SP-A and SP-D. This means that the epithelial cells and macrophages may interact with collectins
through the same mechanism. These observations give insight into how cellular conditions can affect inflammatory
response.
Luther College, Decorah, Iowa 52101
Pheromone Binding Proteins in Elephant Urine
Roman Z. Beyga (Dr. Randall A. Kopper), Department of Chemistry, Hendrix College, Conway, Arkansas 72032
Pheromones are important in the propagation of the African elephant. They help male elephants determine the stage
of the menstrual cycle of the female elephant, as well as showing which elephants are in musth. Aside from temporal
secretions, elephants have pheromones in their urine. With their trunks, Male elephants take up some of the urine
they find and bring it to their vomeronasal organ. When a male elephant encounters respective pheromones, he starts
the mating process. Pheromones by themselves, however, are volatile compounds, and do not stay in solution for
extended periods of time. This reasoning has prompted researchers to hypothesize that there are pheromone binding
proteins in urine. We will look for potential pheromone binding proteins in elephant urine by gel electrophoresis and
optimize its degradation by protease digestion. Any pheromones released will be detected and characterized by
GC/MS. Elephant trunk mucus will be investigated for potential protease activity to explain the release of
pheromones. Results will show whether or not there is a pheromone binding protein in African elephant urine.
29
BIOCHEMISTRY
ROLE OF THE LARGE AND SMALL SUBUNITS OF CHLAMYDOMONAS REINHARDTII PERIPLASMIC
CARBONIC ANHYDRASE ON THE CATALYTIC PROPERTIES OF THE ENZYME
Evgenia Nikolova (Dr. H. David Husic), Department of Chemistry, Lafayette College, Easton, Pennsylvania 18042
The significance of the subunit constitution and quaternary structure in Chlamydomonas reinhardtii periplasmic
carbonic anhydrase (CAH1) on the catalytic properties of the enzyme was studied. CAH1 is an oligomeric zinc
metalloenzyme consisting of two disulfide-bound large (36 kDa) subunits, each of which connects to a small (4 kDa)
subunit by a disulfide bond. The dithiothreitol (DTT)-induced subunit dissociation of the protein and its effect on
catalysis was examined under denaturing and non-denaturing conditions. Gel filtration chromatography of CAH1
treated with 5 M guanidine hydrochloride (GuHCl) and 10 mM DTT resulted in almost complete subunit dissociation
and 91% inactivation of catalytic activity. The fluorescence enhancement of dansylamide at 470 nm due to the
coordination of this aromatic sulfonamide to the active site zinc ion was used to monitor protein renaturation under
various conditions. Following CAH1 denaturation with 5 M GuHCl, a 64 % recovery of the fluorescence characteristic
of the dansylamide-CAH1 complex was observed upon dilution to reduce the concentration of GuHCl, while no
fluorescence recovery was detected when 10 mM DTT was included during denaturation. The effect of 10 mM DTT
treatment on CAH1 subunit association, as judged by SDS-PAGE, and inactivation of catalytic activity, showed
similar trends with up to 61% decrease after 100 min. However, DTT alone caused only a 20 % reduction of
fluorescence enhancement, indicating a smaller effect of subunit dissociation on dansylamide binding to the active
site, than on catalytic activity. These results indicate that disulfide bonds and subunit association are important for
catalysis, but may not be required to allow for the formation of the dansylamide-CAH1 complex, indicating some
retention of active site structure after treatment. Supported by NSF grant 041876.
Characterization of ARRD-I gene involved in late-set progressive retinal degernation in mice
Andrius Giedraitis (1), Md Nawajes Mandal (1), Bo Chang (2), John Heckenlively (1), Radha Ayyagari (1), , (1)
Department of Opthalmology and Visual Sciences, University of Michigan, Ann Arbor, MI; (2) The Jackson
Laboratories, Bar Harbor, ME
The purpose of this study is to characterize the antibodies raised against ARRD-I protein and determine the
localization of this protein in the retinal tissue of mouse and human.
We identified a mouse model with age-related progressive retinal degeneration (ARRD) segregating in an autosomal
recessive manner. Clinical evaluation of affected mice showed sub-normal rod and cone electroretinograms (ERGs)
at the age of eight months. By 22 months, ERG response was not detectable. The chromosomal region responsible
for this trait was identified by genetic mapping and the candidate gene, ARRD-I, was identified by mutation analysis.
Characterization of this gene revealed that it has multiple transcripts composed of four to fourteen exons.
Interestingly, a nonsense or stop codon mutation in exon eight was found in all mice that carried this disease, but no
mutations were observed in the control mice. By RT-PCR, we detected a significant level of expression of this gene in
the mouse retina, and by in situ hybridization, the transcripts were found to localize to the outer and inner nuclear
layer of the retina. We have raised antibodies against this protein and plan to investigate the specificity and sensitivity
of these antibodies. We will subsequently determine the localization of this protein in the retinal tissue of mouse and
human by immunohistochemical analysis using the ARRD-I antibodies. In performing this study, we hope to garner a
better understanding of the role of the protein in retinal tissue and the mechanisms involved in age-related retinal
degeneration, one of the leading clauses of blindness. Further comprehension of the underlying mechanisms can
allow for future response in combat of this disease.
30
BIOLOGY
STUDENT DIRECTED WEST NILE VIRUS PROGRAM GENERATES MOSQUITO DISCOVERIES AND
COMMUNITY GOODWILL
Steven P. Moberly (Dr. Claude Baker), Biology Department, Indiana University Southeast, 4201 Grantline Rd, New
Albany, IN 47150
West Nile virus (WNV), first reported in New York in 1999, began spreading across the nation reaching Indiana in
2001. Indiana University Southeast initiated a student directed WNV research and monitoring program in 2002. The
purpose of this study was to monitor the spread of WNV in three southern Indiana counties (Floyd, Clark and
Harrison). Since its inception, over 200 students, with the author as project director, have been involved with the
program; conducting studies on mosquito biodiversity and WNV cycling. More than 50,000 mosquitoes were
identified in 2004 and 2005. Most specimens were tested for WNV at the Indiana University Southeast laboratory or
at the state reference lab in Indianapolis, Indiana. I found the first Asian Rock Pool mosquitoes, Ochlerotatus
japonicus, in Indiana in 2004. While working to uncover the distribution of the newly introduced mosquito, the author
discovered a close correlation between positive WNV mosquito pools and drought conditions (drought amplification).
This research project has generated community goodwill, provided many students with research opportunities, while
concurrently protecting public health through mosquito control efforts and networking with local, state and federal
agencies such as health departments and the CDC. Our research has been funded by Eli Lilly, Indiana University and
indirectly through funding from the individual health departments, the Indiana State Department of Health and the
Centers for Disease Control.
http://www.indiana.edu/~interns/moberly/
http://www.homepages.indiana.edu/082004/text/mosquitoes.shtml
GENETIC LINEAGE TRACKING OF MRSA THROUGH SCCMEC TYPING
Authors: DaNette Bushma, Megan Maciel, & Justin Rhees, Research Sponsor: Weber State University
Undergraduate Research Committee and LDS Hospital., Insitution: Weber State University, 3905 University Circle,
Ogden UT 84408
New strains of Methicillin Resistant Staphylococcus aureus (MRSA) are emerging with increased virulence and
resistance to antibiotics, causing great concern in the medical community. Two genes have been shown to play a
significant role in the virulence of this organism: the Staphylococcal Cassette Chromosome mec (SCCmec) gene and
the Panton-Valentine Leukocidin (PVL) gene. The SCCmec gene codes for one of five cassette types which can be
used for genetic tracking of certain MRSA strains. This study will determine the SCCmec cassette type of MRSA
isolates, in an attempt to track their geographical origins, whether within the community or hospital acquired within our
population. Research performed on 200 frozen MRSA isolates from Intermountain Health Care’s Latter-Day Saint
Hospital (Salt Lake City, Utah) will determine each isolates cassette type found on the SCCmec gene. The isolates
will be extracted using the Qiagen method, and tested by real time polymerase chain reaction (PCR) using the Roche
light-cycler. The accumulated data on the PVL gene and antimicrobial resistance obtained last year by Weber State
students will be complied to form a research database. An infection control team could then use this database as a
tool to track the genetic lineage of MRSA and minimize serious outbreaks.
31
BIOLOGY
MICROSATELLITE MUTATION RATES IN DNA MISMATCH REPAIR-PROFICIENT AND -DEFICIENT S.
CEREVISIAE
Alexandriana T. Spivey&#916;, Jayne Boyer&#911;, Kevin Lehner&#911;, Rosann Farber&#911;ß,
&#916;Department of Biology, Shaw University, Raleigh, NC; &#911;Department of Pathology and Laboratory
Medicine, University of North Carolina-CH, Chapel Hill, NC; ß
Approximately 90% of hereditary non-polyposis colorectal cancer (HNPCC) cases are caused by mutations in genes
that encode enzymes involved in DNA mismatch repair (MMR). Inactivation of these enzymes may lead to a high
frequency of mutations in other genes responsible for cell proliferation and growth regulation. MMR is a cellular
process that identifies and corrects frameshift errors in mismatched bases and microsatellites that occur during DNA
replication. In eukaryotes microsatellites are one class of repetitive DNA sequences and are defined by their repeat
unit size of 1-6 bp; in the human genome, microsatellites are made up of 6-30 repeat units. Since all eukaryotes have
a similar MMR system, we examined the effects of MMR on microsatellite mutation rates in the yeast Saccharomyces
cerevisiae. Previously in our lab the stability of a 16.5 repeat polyGT tract was measured using a reporter gene
URA3-GT, in which the microsatellite was inserted in-frame into the yeast URA3 gene. Examination of several strains
containing the reporter gene at chromosomes III, V, and XII showed that the efficiency of DNA mismatch repair varies
in different regions of the genome, perhaps reflecting some aspect of chromosome structure. To further evaluate the
efficiency of MMR at different chromosomal sites, we used yeast strain CEN5::URA3-GT, which has the reporter gene
URA3 in-frame near the centromere of chromosome V. We constructed an isogenic MMR-deficient strain
(CEN5::URA3-GT Dmsh2) and determined the microsatellite mutation rates of this strain versus MMR-proficient strain
(CEN5::URA3-GT) by fluctuation analysis. We found a 100-fold increase in mutation rates in the MMR-deficient
strain at the polyGT tract as it compares to MMR proficient strain. Therefore, the absence of MMR near the
centromere at chromosome V in Saccharomyces cerevisiae results in a significant increase in microsatellite mutation
rates.
INVESTIGATION OF A PATHWAY IMPLICATED IN ALZHEIMER’S DISEASE: IDENTIFICATION AND
ANALYSIS OF G12 MUTANT PROTEINS IMPAIRED IN BINDING TO PP2A
Robert Tate (Ted Meigs), Department of Biology, The University of North Carolina at Asheville, Asheville, NC 28804
Trimeric G proteins are critical intracellular proteins that serve as transmitters of external chemical information to the
interior of cells, and are involved in an enormous variety of cellular events and changes (or responses). The focus of
this research is a trimeric G protein subunit termed Ga12 that has been implicated in the development of cancer and
the progression of existing cancer toward malignancy and metastatic invasion. The Meigs’ laboratory at
UNC-Asheville has formed a collaboration with a research group at Harvard Medical School headed by Dr. Bradley
Denker, whose group has made the recent discovery that Ga12 interacts strongly with a protein termed PP2A (protein
phosphatase-2A). Phosphatases, such as PP2A, are important proteins in cells that remove phosphate groups from
other proteins and molecules. PP2A had previously been demonstrated to chemically modify another protein, termed
Tau that appears in the neurofibrillary tangles that are a hallmark in the brain tissue of Alzheimer’s disease patients. It
is thought that Ga12 may stimulate PP2A’s ability to dephosphorylate Tau. Thus, the Ga12-PP2A interaction has
emerged as a significant finding in the scientific effort to understand the molecular mechanism of Alzheimer’s
disease. The focus of this project, which began in the summer 2005, has continued into this academic year, has been
to identify specific subregions of Ga12 that are involved in its molecular interaction with PP2A. This researcher has
produced, purified, and immobilized a hybrid version of PP2A. This researcher has screened this immobilized
version of PP2A for biochemical interaction with a panel of Ga12 mutants. In preliminary results, several Ga12
mutants and their corresponding binding regions have emerged as possible non-interactors with PP2A . This lab is
now attempting to confirm these results and quantitatively analyze the ability of these Ga12 mutants to interact with
PP2A. This information should be important to the ongoing collaboration between UNCA and Harvard Medical
School, and will hopefully provide molecular insights toward an understanding of the critical pathways of Alzheimer’s
disease
32
BIOLOGY
The Search for Protein Interactors with CDCA8
Blase G. Hennessy (Dr. William Taylor), Department of Biological Sciences, University of Toledo, Toledo, OH 43606
Cancer is a disease of cells that have lost the ability to normally regulate rates of cell division, and is the second
leading cause of death in the United States. Many of the genes important in regulating the cell cycle are unknown,
and an acquired knowledge of these genes could lead to important insights about cancer. One gene, cdca8, which has
proven to be involved in the cell cycle, was identified through an Affymetrix gene chip analysis of genes regulated by
p53, a known tumor-suppressing gene. Using a FLAG-tagged version of CDCA8, our lab found that during mitosis of
human cancer cells, CDCA8 becomes localized to the cleavage furrow. Experiments that I conducted showed that
when translation of CDCA8 was inhibited by using small interfering RNA, cells were unable to progress properly
through mitosis, leading to multinucleation. Other groups have confirmed these results. The next important step in
determining exactly how CDCA8 works is to find which proteins, it interacts with. Currently I am using a yeast two
hybrid assay to find potential protein interactors with CDCA8. Using CDCA8 in a pDEST 32 vector cotransfected with
a cDNA library into competent MaV203 yeast cells, I have screened thousands of potential protein interactors. Thus
far, only several extremely weak potential interactors have been isolated, but with further library screenings, I hope to
identify strong, relevant proteins that interact with CDCA8. Continued work will focus on testing the biological role of
these potential interactors.
Enhanced Expression of the CDCA8 Protein During Mitosis
Andrew C. Stiff (Dr. William Taylor), The University of Toledo Office of Research,Department of Biological Sciences,
The University of Toledo, 2801 W. Bacroft Toledo, OH 43607
Cancer is the second leading cause of death in the United States, killing approximately 500,000 people every year.
One of the characteristics of cancer is uncontrolled cell proliferation and a way to understand this is by examining the
cell cycle. The process by which cells divide their nuclear and cytoplasmic contents is called mitosis, composed of the
steps interphase, prophase, metaphase, anaphase, telophase, and cytokinesis. CDCA8 is a recently identified protein
required for chromosome segregation and cytokinesis. We identified two spliced variants transcribed for the cdca8
gene. We tested whether these variants encoded a protein of similar size by transient transfection of epitope tagged
versions. We observed that both variants led to the production of a protein with the same molecular weight.
Interestingly, after cells were treated with nocodazole, a drug that arrests cells in mitosis, higher levels of CDCA8
accumulated. This observation suggests that the CDCA8 protein is stabilized during mitosis. This stability may be
related to the potential phosphorylation of candidate sites, serine165 and threonine 106, that were recently identified.
In several experiments we observed that mutation of these sites showed increased levels of the CDCA8 protein
during mitosis. However, this result was not evident in each transfection suggesting that this may be a subtle effect.
Future experiments are aimed at measuring the stability of the CDCA8 protein during mitosis and the determining the
mechanism of regulation.
33
BIOLOGY
UNDERSTANDING BASAL METABOLIC RATE AND CONDUCTANCE IN ORD’S KANAGROO RAT,
DIPODOMYS ORDII
Nikki A. Smith and Marilyn R. Banta, School of Biological Sciences, University of Northern Colorado, Greeley,
Colorado 80639
Kangaroo rats (family Heteromyidae) are adapted to survive in desert areas including Arizona, California, and
Nevada. Previous researchers have found that most species of kangaroo rats have the physiological means to cope
with the desert’s high temperatures, lack of water, and limited food resources. Ord’s kangaroo rats, Dipodomys ordii,
are widely dispersed, not only in desert environments but also in colder regions, such as northern Colorado. Little is
known about the physiology of individual animals living in cold climates. Therefore, the purpose of this research was
to compare the physiology of a population of D. ordii inhabiting northern Colorado with desert populations using data
published in two previous studies. In the current study, D. ordii were live-trapped, tagged, and brought to the lab to
measure basal metabolic rate (BMR) and conductance. Basal metabolic rate is the amount of oxygen an animal
consumes under comfortable conditions, and conductance is the rate of heat flow from an animal to its surroundings
(inverse of insulation). These quantitative data were collected once monthly for nine months using an oxygen
analyzer and the computer program DATACAN. Analysis of covariance (ANCOVA) and a Kruskal-Wallis test were
used to determine whether there were changes in BMR and conductance across the 9 months of the study. We found
that as the local climate changes, D. ordii respond by raising BMR during spring months but not during winter or
summer months but they do not alter conductance. In general, animals in this population seem to have lower BMR
and lower conductance than animals in desert populations. Our findings suggest that populations of Ord’s kangaroo
rats are adapted to their local climate and can alter their physiology seasonally.
The Localization of Sodium and Potassium Channles Ganglion Cells in Mouse Retina
Natalia Figacz (Dr. Karen Myhr), Wayne State Undergraduate Research, Biological Sciences, 5047 Gullen Mall Detroit
MI, 48202
Neurons send information long distances through the body. Information is sent using electrical signals called action
potentials. Ion channels are vital for this neuronal function. Action potentials depend on types of sodium and
potassium channels that are located throughout the neuron. Sodium’s positive charge is responsible for the
depolarization of the negative interior of the cell, while potassium maintains the resting potential of the cell. Without
ion channels, information would not be sent from one neuron to another. Yet little is known about the specific location
of the ion channels in retinal ganglion neuronal cells. The mouse retina is an excellent model system, since it is
easily accessible. The ganglion cells of the retina, whose axons form the optic nerve, use action potentials to send
information long distances. Thus, the ganglion cells are an excellent neuron in which to study the ion channels that
generate action potentials. Specific questions we are addressing include where the ion channels are in the neurons,
and in which subtypes of ganglion cells they are found. Ion channels in the axons help send information long
distances, whereas channels in the dendrites and cell bodies help integrate signals from other neurons. Subtypes of
ganglion cells perform unique functions. To determine ion channel distribution, we used immunohistochemistry, in
which antibodies are used to label the proteins that make up the ion channels. The labeled pieces of retina are then
viewed with fluorescence microscopy, which allows localization of the labeled ion channels. Our results indicate that
ion channel location is characteristic for a specific ion channel. Sodium channels are localized to axons and cell
bodies; and different types of potassium channels are found in different subtypes of neurons. These results enhance
our understanding of the generation of action potentials.
34
BIOLOGY
THE EFFECTS OF ANTIBIOTIC INTRODUCTION ON THE FEEDING EFFICIENCY OF THE AMERICAN
DOG TICK, DERMACE
Stephanie Pratt, Jenny Sandler, and Kristi Teal (Dr. Laura Fielden), Division of Science, Truman State University,
Kirksville, Missouri 63501.
This project was designed to study whether antibiotic use in domesticated host animals affected the intestinal
microbial communities of tick species. We hypothesized that the antibiotic use in host species would be a detriment
to the tick – since many arthropods rely on microbial commensals for digestive processes. The aim of this study was
to determine the effect of two different types of antibiotics on feeding efficiency. Instead of feeding ticks directly on
hosts subject to antibiotic treatment, this study employed an artificial feeding method to introduce the antibiotic into
the tick. Unfed adult females of Dermacentor variabilis were artificially fed for twelve hours using capillary tubes
containing equine serum albumin and the antibiotics Chlortetracycline and Sulfadiazine. After antibiotic introduction,
ticks were placed on a rabbit host and allowed to complete feeding (approx 7 days). All engorged ticks were removed
from the host and monitored for egg laying performance. The preliminary data has shown that the antibiotics do not
have a negative impact on the feeding efficiency (i.e., mass gain during blood feeding). Mass of engorged ticks
pre-fed with Sulfadiazine (530.34±16.62 mg) was not significantly different than the control group (528.05±25.54 mg).
However, ticks fed with Chlortetracycline (604.86±9.04 mg) were significantly heavier (t= 1.86, d.f. = 36, P< 0.05)
than the control group indicating the Chlortetracycline treatment had a beneficial effect on the feeding efficiency. The
differences shown between the two antibiotic groups may result from the biochemical properties of the antibiotic.
Sulfadiazine destroys bacteria that synthesize folic acid whereas Chlortetracycline inhibits protein synthesis within the
microbe. Future studies will aim towards isolating bacterial fauna from tick guts to determine whether the antibiotic is
effective against bacteria endogenous to the tick intestine or those introduced via the host blood.
POLLEN ONTOGENY IN EPHEDRA AMERICANA (GNETALES)
Allison S. Doores (Dr. Jeffrey M. Osborn), Division of Science, Truman State University, Kirksville, Missouri 63501
Ephedra, or Mormon’s Tea, along with Welwitschia and Gnetum, comprise the gymnospermous order Gnetales.
Historically, the Gnetales has been regarded as the sister group to angiosperms, or flowering plants, though recent
molecular studies have allied the Gnetales with conifers, such as pines, spruces, and junipers. Several investigations
have focused on mature pollen of Ephedra; however, little is know about pollen development in the genus. The
objective of this research was to comprehensively study pollen ontogeny in Ephedra americana. Using combined
light, scanning electron, and transmission electron microscopy, all major developmental stages have been
documented, including the microspore mother cell, tetrad, free microspore, and mature stages. Developmental
events including the deposition of callose, primexine, tectum, infratectum, foot layer, and endexine lamellae will be
discussed. Significant pollen wall deposition occurs during the tetrad stage, although additional deposition and
compression of endexine lamellae occur during the free microspore stage. At the end of the tetrad stage, the
infractectal layer is composed of small granules, the foot layer is relatively thin, and the endexine is composed of
robust lamellae. Mature grains are elliptic to ellipsoidal in shape and have 5-17 longitudinally oriented plicae. These
plicae have distinct ridges and furrows. Pollen grains are inaperturate, although the exine is considerably thinner
within the furrows, which have a straight or undulated morphology. These developmental characters will be discussed
regarding systematic and phylogenetic interpretations of Ephedra and Gnetales.
35
BIOLOGY
CHARACTERIZATION OF DROSOPHILA MELANOGASTER TRANSGENIC FOR HUMAN OTK18
Anjeza Pashaj (Darby J. Carlson and Dr. Kimberly A. Carlson), Department of Biology, University of Nebraska at
Kearney, Kearney, Nebraska 68849
OTK18 is a human gene that has been characterized as transcriptional suppressor containing 13 zinc finger motifs
and both Krüppel-associated boxes (KRAB) A and B. This transcriptional regulator is assumed to play multiple roles
within human development, specifically in immune and neuronal processes. As has been done for many human
genes, further characterization of OTK18 is being carried out in a simpler model system. The main thrust of the
proposed research was to create, balance, and express human OTK18 in transgenic Drosophila melanogaster lines.
In this study, a stably balanced line (Line 14, 3rd chromosome) was created and characterized. Utilizing the
GAL4-UAS system, Line 14 containing the UAS sequence was crossed to a “driver” line (hsp70-1822 or hsp70-1799)
containing the GAL4 sequence. OTK18 expression was turned on by heat-shocking the mated females, eggs, larva,
and pupa at 37&#730;C for 1 hour. The resulting progeny expressed OTK18 RNA and appeared to display a unique
phenotype. A secondary objective was to clone, express, and purify OTK18 protein for use in the characterization of
the transgenic lines. In the future, we hope to define a mechanism for OTK18 gene regulation that can be related to
human developmental processes.
Synthesis of a Series of Bicyclic Diquinones: Potential Cancer and Leukemia Drugs
Jesse Van Heukelom (Dr. Grigoriy Sereda), Department of Chemistry, University of South Dakota, Vermillion, South
Dakota 57069
Bicyclic quinones and diquinones are attracting attention from the world of chemistry as building blocks for organic
synthesis, i.e. mimicking intramolecular photoelectron transfer. They have shown a wide spectrum of biological
activities: anti-inflammatory, anti-malaria activity, and cytotoxicity against resistant leukemia L1210 cell lines. Upon
the addition of various functional groups on the third benzene ring, the project is expecting to control the
pharmacokinetics properties and to chemically link them to porphins rings for entry into the human body. The
purpose of my research is to synthesize a series of new bicyclic diquinones and evaluate the role of the non-quinone
benzene ring in the cytotoxicity of bicyclic diquinones, to explore the relationship between the cytotoxicity of
triptodiquinones toward leukemia and ovarian cancer cells, and to optimize photodynamic and biological properties of
the system by varying the diquinone fragment and its linkage with the porphyrin moiety. In addition, the project will
examine whether lone benzene rings play a role in the cytotoxicity, and what implications they may have. Each
quinone’s cytotoxicity will also be assessed in relationship to their red-ox potentials to find the quantitative
structure-activity relationships of each compound. Upon synthesis, the quinones will have their cytotoxicities
analyzed against ovarian cancer cells at the University of South Dakota’s School of Medicine and the L1210 leukemia
cells at South Dakota State University. Currently, my colleagues and I have synthesized and analyzed five new
bicyclic quinones in the series of the desired eight new quinones with promising results of cyotoxicity toward the
particular cancer lines during primary testing.
36
BIOLOGY
PLASTRON RESPIRATION IN TICKS (ACARI: IXODIDAE)
Bach Q. Ha and Susan M. Villarreal (Dr. Laura Fielden and Dr. Phil Ryan), Division of Math and Computer Science,
Division of Science, Truman State University, Kirksville, Missouri 63501.
Ticks are blood-feeding arthropods that are well known for their survivability. Although ticks are terrestrial organisms,
they can survive extended periods of submergence under water as for example after heavy rainfall or flooding. A
plastron is a physical gill consisting of a thin layer of air trapped by hydrophobic hairs or other cuticular projections.
Hence a plastron is an alternate respiration system that can absorb oxygen from water. The complex spiracular plates
of ticks have been postulated to serve as plastrons but until now, this has not been verified. In this study, we confirm
the existence of plastron respiration in the dog tick Dermacentor variabilis. Adult dog ticks can survive submergence
in water for over two weeks. Wetting the spiracular plate with alcohol, thereby debilitating any potential plastron
function lowered survival to less than three days. Biomathematical studies currently in progress are modeling the
efficiency of the spiracular plate as a plastron. We have developed a mathematical model to predict survivability of
submerged ticks under water. This model requires determination of a suite of physical and biological parameters
including volume of the air film contained within the spiracular plate, the plastron air/water interface area and the
oxygen consumption and biomass of the submerged tick. It is hoped that this model can be successfully used in the
future to predict underwater survivability of other species of ticks which show both interspecific and intergeneric
morphological variation in spiracular plate structure. This study provides the first example of plastron respiration in the
Ixodidae.
PREDICTING OCCURRENCE OF MISSOURI BLADDERPOD (LESQUERELLA FILIFORMIS):
MODELING OVER SPACE AND TIME
Elizabeth R. Bobzien and William B. Leeds (Dr. Michael Kelrick and Dr. Hyun-Joo Kim), Divisions of Science and
Mathematics and Computer Science, Truman State University, Kirksville, Missouri 63501
Missouri bladderpod (Lesquerella filiformis) is a rare and federally threatened plant species found in southwest
Missouri and northern Arkansas. Understanding how Missouri bladderpod abundance changes through time, and in
relation to its heterogeneous glade habitat, is crucial for successful conservation of the species. Abundance of
Missouri bladderpod and percent cover of several of its habitat’s attributes were recorded within an orthogonal grid of
cells on Bloody Hill Glade at Wilson’s Creek National Battlefield in Springfield, MO in 1997, 1998, 2003 and 2005.
Coherently integrating both the spatial and temporal information within this data set is challenging, as there are few
well developed analytic methods to accomplish this. In this study, we analyzed this spatially explicit ecological data
using various statistical approaches. Logistic regression, ANOVA, repeated measures analysis, and log-linear
modeling were used in temporal analyses of the abundance of Missouri bladderpod, while the spatial component of
the data set was probed within a Geographical Information System software environment. Our results showed 2005 to
be a significantly higher abundance year than previous years with several 5x5-m cells supporting more than 500
plants; areas of change were visualized using GIS map algebra. Hop clover, an exotic species, tended to occupy
microhabitat types that also supported Missouri bladderpod, indicating the possibility of competition between the two.
However, in 2005, Missouri bladderpod was observed in many cells formerly occupied solely by hop clover, and also
increased its abundance in cells that it had occupied previously. This finding is of particular interest in conserving
Missouri bladderpod; although it is often assumed that exotics would threaten the well-being of native species, this
may not be the case in this instance. The techniques and approaches we used can be applied to many different
problems in habitat modeling and conservation biology.
37
BIOLOGY
CELLULAR FRACTIONATION OF WILD TYPE AND PCM MUTANT ESCHERICHIA COLI STRAINS
Madiha Salim (Dr. Jonathan Visick), Department of Biology, North Central College, Naperville, Illinois 60540.
Escherichia coli can survive various environmental conditions with the help of repair enzymes. We are investigating
damage to cellular proteins resulting from spontaneous formation of isoaspartyl (isoAsp) residues from aspartate or
asparagine. This damage can be repaired by the L-isoaspartyl protein carboxyl methyltransferase (PCM), and the
absence of PCM results in survival defects when E. coli is starved (unable to make new proteins) and subjected to
protein-denaturing stresses.
Surprisingly, pcm mutants do not accumulate more isoAsp over 10 days in stationary phase than wild-type. PCM
appears to be a cytoplasmic enzyme, so we hypothesized that it may limit the amount of IsoAsp accumulation in the
cytoplasm of wild type cells, but that this difference may be masked by isoAsp accumulation in the periplasm, outer
and/or inner membrane of the cell. We would also like to determine how active PCM is and how much of its substrate
and an inhibitory product are present at various points during starvation and recovery.
We fractionated E.coli cells and used marker enzyme assays to determine the purity of each cellular compartment.
We have obtained periplasmic and cytoplasmic fractions with less than 10% contamination by other fractions, and are
developing methods for effectively testing membrane fractions. We are continuing to fractionate cells grown under
various conditions and will analyze our samples chemically using the HPLC to determine PCM activity, SAM, SAH
and isoAsp content in each fraction.
DEVELOPMENT OF A PREDICTION EQUATION FOR TOTAL BODY MASS ACROSS THREE SIZES OF
EQUIDS (Equus caballus)
Kathryn M. Smith, Tammy R. Bragg and Alana N. Walker (Dr. R. Charles Apter and Dr. Dean DeCock), Department
of Agricultural Science, Department of Math and Computer Science, Truman State University, Kirksville, Missouri,
63501
As an extension of an ongoing study in allometry, this project concerns the development of a prediction equation for
computing total body mass of the domestic horse. Previously, nine body dimensions, including body mass, were
determined for 43 draft horses, 82 miniature horses, and 43 stock type horses. Each type of horse measured was
chosen to represent a different body classification that is recognized by the horse industry. The sample pool did not
exclude animals based on gender, because the previous allometric study showed that, for the study’s purpose, there
was no significant difference between male and female horses. The developed equation utilizes the easily measured
body dimensions of heartgirth and body length. Inspiration for this formula came from viewing the horse torso as a
cylinder, with radius being derived from heartgirth, and body length representing the height of the cylinder. Linear
regression analysis of torso volume and total body mass yielded a strong linear relationship (Fobs = 28,912, p < .001)
with an R2 value of 0.994. Development of an accurate prediction equation for total body mass utilizing
easily-measured body dimensions would serve the horse industry well since nutritional requirements and dosages of
various medications and anthelmintics are typically expressed on a per-unit-body mass basis.
Control of photosynthesis in Sphagnum, why the canopy matters.
Samantha L. Glover (Dr. Steven Rice), Biology Department, Union College, Schenectady, NY 12308
Each leaf in Sphagnum (the genus of peatmosses) is photosynthetic; as shoots grow vertically and each leaf gets
deeper in the canopy, it is subject to lower light levels. However, even in this environment, interior leaves contribute
to whole-plant photosynthetic carbon gain. Sphagnum canopies differ in the density and physiology of canopy
elements (i.e., branches) and this variation should have an effect on the overall physiological performance. In this
study, physiological comparisons from three Sphagnum species (S. fallax, S. magellanicum, and S. fuscum) are used
to understand the underlying mechanisms that cause variation in canopy photosynthesis. I will study light attenuation,
analyze chlorophyll concentrations, measure nitrogen content, chlorophyll fluorescence, and assess photosynthetic
rate on both the canopy as a whole, and at different depths within the canopy. To measure photosynthetic rate, we
will monitor CO2 uptake in a closed chamber system and assess variation in the canopy using a chlorophyll
fluorescence technique. In terms of comparison between species, the species with an overall higher rate of
photosynthesis likely has a higher rate of photosynthesis extending deeper into the canopy. Overall through
comparisons of nitrogen content, light attenuation, chlorophyll analysis, and photosynthetic rates at different positions
within the canopy, we will be able to understand how variation in canopy structure influences photosynthetic
performance in Sphagnum.
38
BIOLOGY
CLASSIFYING TEMPORAL PROPERTIES OF CONE RETINAL PHOTORECEPTORS IN THE LIZARD
ANOLIS SAGREI USING ELECTRORETINOGRAPHY
Marisa S. Zarchy (Dr. Leo Fleishman), Department of Biology, Union College, Schenectady, New York 12308
The purpose of this project was to determine the temporal properties of cone photoreceptors in the lizard retina.
Anoline lizards have four classes of cones in their retina all of which contribute to color vision. Lizards use their
color-vision system for courting behaviors, hunting prey, and communication. Therefore, it is important to understand
the physiological properties of photoreceptors to learn how lizards respond in their natural environment. One
pertinent property is temporal response. Photoreceptors measure light intensity by totaling the number of photons
absorbed over a particular time interval, known as the retinal integration time. The duration of the retinal integration
time determines how rapidly the eye can respond to changes in the visual image, and is therefore a critical feature of
temporal response. Integration time was measured using electroretinography. An electroretinogram (ERG) is
recorded by
placing an electrode on the surface of the cornea of an anesthetized lizard and stimulating the eye with flashes of
light. ERG amplitude is proportional to the intensity of response of the photoreceptors. We stimulated the eye with
very brief flashes of light (initiallly <4 msec) and then gradually increased stimulus flash duration, which increased
the number of photons absorbed and thus increased the amplitude of the ERG. When the flash duration equals or
exceeds the retinal integration time ERG amplitude no longer increases with stimulus duration. However, temporal
response is affected by many factors. We examined the effect of stimulus light intensity and state of adaptation by
using two sources of light (stimulus flash light and adaptive light) to attain various light intensities. By using stimulus
lights of different wavelengths we explored the response of the different types of cone photoreceptors classes. By
considering all of these variables, we will complete a descriptive study characterizing temporal response of each type
of photoreceptor in the lizard visual system.
CURCUMIN MODULATES ANGIOGENIC FACTORS THROUGH INHIBITION OF NF-&#922;B IN
OVARIAN CANCER CELLS
Sarah K. Fogoros, Miheon Choi, and (J. Rebecca Liu), Division of Gynecologic Oncology, Department of Obstetrics
and Gynecology, University of Michigan, Ann Arbor, Michigan 48109
Objectives: Curcumin has potent anti-neoplastic activity in several tumor types, and is thought to exert
anti-inflammatory effects in part through inhibition of NF-&#61547;B. Inhibition of NF-&#61547;B has been linked to
inhibition of the cytokine IL-8, production of PGE2, and vascular endothelial growth factor (VEGF). IL-8, PGE2, and
VEGF are expressed in ovarian cancers, and are associated with increased angiogenesis and poor prognosis. We
sought to determine if curcumin inhibits expression of NF-kB as well as production of these angiogenic factors in
ovarian cancer cells.
Methods: Constitutive activation of endogenous NF-kB was determined in a panel of ovarian cancer cell lines.
Cytotoxic activity of curcumin in ovarian cancer cells was determined by sulforhodamine assay. NF-kB activation was
determined by transfecting cells with a NF-kB-dependent reporter plasmid, and measuring luciferase activity.
Constitutive activation of endogenous COX-2 was determined by immunoblotting. IL-8 PGE2, and VEGF levels were
determined using an enzyme-linked immunosorbent assay (ELISA).
Results: Curcumin treatment decreased cell viability in all ovarian cancer cell lines tested in a dose dependent
manner. NF-&#954;B and IKK were constitutively active in a subset of cell lines, and curcumin down-regulated
NF-&#954;B and IKK activity. In cell lines with high endogenous levels of IL-8 and VEGF, curcumin inhibited IL-8 and
VEGF production.
Conclusions: Most ovarian cancer patients die of chemoresistant recurrent disease. Conventional chemotherapeutic
agents induce apoptosis, and resistance to chemotherapy can be promoted by constitutive activation of NF-kB. We
have shown that curcumin is cytotoxic in ovarian cancer cells, and exerts its effects by inhibiting NF-&#954;B
activation, and decreasing levels of IL-8 and VEGF. The inhibition of these angiogenic factors along with the induction
of apoptosis may have broad clinical benefits to ovarian cancer patients.
39
BIOLOGY
EFFECTS OF SHRUB COMPETITION ON PHOTOSYNTHETIC PHYSIOLOGY OF THREE TEMPERATE
TREE SPECIES
Kathryn V. Smith (Dr. Rebecca A. Montgomery), Department of Forest Resources, University of Minnesota, St. Paul
MN 55108
The pine forest of Minnesota historically had frequent ground fires that opened up the forest understory. These fires
suppressed dominant shrub species like hazel (Corylus cornuta). Currently, hazel has little to no suppression and
trees have poor regeneration rates. Shrubs compete with seedlings for light, water and nutrients. Poor regeneration
rates may result if seedlings are poorly equipped to cope with the high resource competition from hazel and cannot
maintain positive carbon balance to survive and grow to maturity. We quantified the physiological effects of above
and belowground competition from hazel by determining the photosynthetic rates, specific leaf area (cm2 g-1), and
nitrogen in seedlings of Acer rubrum, Quercus rubra and Betula papyifera under four experimental treatments: (1)
removal of shrubs (2) tieback of shrubs (3) removal of shrubs and addition of shade and (4) control. We found no
difference among species in SLA, but statistically significant differences among treatments (F6,48= 6.34, P=0.001);
Individuals in the control plots had the highest SLA (674 ± 41 S.E.) and those in shrub removal plots had lowest (428
±41 S.E.). We found no difference among treatments in maximum photosynthetic rates (Amax; umol m-2 s-1), dark
respiration (Rd) and the light saturation point (LSP) but we did find significant species differences. Betula had higher
Amax, Rd and LSP compared to the other species. We found no significant effect of treatment on survivorship after
one year, however, there way a trend toward higher survival in plots with reduced aboveground competition (e.g.
shrub removal and tieback) in Acer and Betula. In conclusion, we found no effect of competition on leaf physiology
but a significant effect on leaf morphology. This suggests that competition may be more evident when scaled to the
whole plant level. Future research on above and belowground biomass allocation is warranted.
link to view abstract with formatting and characters:
http://www.cnr.umn.edu/FR/people/facstaff/montgomery/index.html look for Kathryn Smith Abstract at bottom of this
page.
TRANSCRIPTION FACTOR FOXL2 REGULATION OF THE ANTIZYME GENE PROMOTER
Brianne R. Givens (Dr. Jean Escudero, and Dr. Kenneth Escudero), McNair Scholars Program, Department of
Biology, Texas A&M University-Kingsville, Kingsville, TX 78363
Certain mutations in the forkhead transcription factor FoxL2 have been shown to cause premature ovarian failure
(POF) in women. Previous studies aimed at identifying genes that are regulated by FoxL2 have suggested that the
ornithine decarboxylase antizyme gene is affected by FoxL2 expression. These studies utilized the herpes simplex
virus VP16 protein. VP16 is a very strong transactivator that enhances the transcription of genes. Fusion of VP16 to
FoxL2 creates a hybrid protein that theoretically activates genes regulated by FoxL2. Ornithine decarboxylase (ODC)
is a key enzyme in the biosynthesis of polyamines and is regulated by antizyme. Binding of antizyme to ornithine
decarboxylase promotes the dissociation of ornithine decarboxylase homodimers (the active form) and also promotes
degradation of ornithine decarboxylase. The aim of these studies is to determine if FoxL2 regulates the expression of
antizyme by using Real Time PCR analysis. The hypothesis is that FoxL2 regulates the expression of antizyme and
this will be tested by transfection of mammalian cells, total RNA extraction and Real Time PCR analysis. If FoxL2
acts as a repressor, over expression of FoxL2 will cause antizyme expression to decrease. If FoxL2 acts as an
activator, over expression of FoxL2 will cause antizyme expression to increase. The results of this study will increase
the understanding of gene regulation involved in ovarian function and lead to the elucidation of one of the causes of
POF (Premature Ovarian Failure).
40
BIOLOGY
ATRAZINE EXPOSURE RESULTS IN ESTROGEN-LIKE ALTERATION OF SPATIAL DISTRIBUTION IN
FRESHWATER MUSSELS
Tanya M. Spellman, Peter R. Warny, and Katherine Flynn (Katherine Flynn), Biology Department, Adelphi University,
Garden City, New York 11530 and Wildmetro Inc., New York, New York 10163
Atrazine is a commonly used herbicide and a suspected endocrine disrupter; it may have estrogen-like activity, which
could include behavioral effects. To evaluate this, laboratory-acclimated adult freshwater mussels (Elliptio
complanata) were exposed to atrazine at 1.5, 15, or 150 ug/L for 72 hours (n = 8/dose). These concentrations
correspond to 0.1, 1, and 10 times the U.S. Environmental Protection Agency’s allowable atrazine concentration for
aquatic ecosystems. Estradiol at 100 ug/L and 70% ethanol at less than 1 mL in 30 L were used as positive and
negative controls, respectively. Spatial distribution was evaluated every six hours by separating each tank into four
grids, starting with the same number of mussels in each (two per grid), and then recording the number of mussels in
each of the four grids at each time. Distribution index (DI), defined as the fraction of mussels in the most crowded
grid, was then calculated for each tank at each time. All treatments resulted in decreased distribution of mussels with
respect to control. The DI for estradiol was 25% smaller than control and the DI for all atrazine treatments was 21%
smaller than control. A one way analysis of variance (ANOVA) with treatment as the between groups variable
indicated significantly different DIs among treatment groups. The Games Howell post hoc test revealed significantly
lower DIs in estradiol, mid and high ATR treated animals (p<0.02). These data suggest that in freshwater mussels,
temporary exposure to ecologically-relevant concentrations of atrazine results in an estrogen-like decrease in the
number of animals per unit area.
VIRTUAL CHINCILLA
Keith Taschner (Dr. Ignatios Vakalis and Dr. Will Ray), Department of Mathematics, Computer Science, and Physics,
Capital University, Columbus, Ohio 43209
Otitis media (middle ear infection) is one of the most commonly diagnosed childhood illnesses. Not only does this
illness cause pain and suffering for many children and their parents, it is also the most common cause of hearing loss
in children, and has an estimated six billion dollar economic impact in the United States alone. Current research on
otitis media is performed using the chinchilla as an animal model. A chinchilla is infected with bacteria that cause an
ear infection, observed for a period of time as the infection develops, and then is sacrificed. The chinchilla is then
dissected to determine the state and site of infection when the animal died. This approach only provides accurate
data at one point in the process of the disease and the researchers have no data on where the bacteria were prior to
that point in time. Clearly, the full history of the infection might allow researchers to more quickly and accurately
determine causes of otitis media and discover ways to prevent it. We can generate data using a photon-counting
camera, but the challenge in obtaining reliable results is that the interior of a chinchilla head must be accurately
modeled in three dimensions, in order to accurately locate the infection from the data.
We pursued a method which involved dyeing a decalcified skull, followed by embedding that skull in opaque wax,
and destructively removing five thousandths of an inch thick layers using a high-precision commodity milling machine
(http://www.maxnc.com/). A picture was taken of the top of the block, after each layer was removed. The resulting
data set was cleaned up using computer programs to remove most of the white wax, and then the Insight
Segmentation and Registration Toolkit (ITK) was used to segment the skull. A three dimensional model was rendered
using the Visualization ToolKit (VTK).
41
BIOLOGY
REGULATION OF NF KAPPA B ACTIVITY BY INTERACTION WITH CBP AND p38 MAP KINASE
Rachael Sullivan (Dr. Brian Ashburner), Department of Biological Sciences, The University of Toledo, 2801 W.
Bancroft St., Toledo, OH, 43606
NF-&#954;B is a transcription factor that controls expression of genes that play a role in inflammatory and immune
responses, cell growth and differentiation, and protecting cells from programmed cell death. NF-&#954;B activity is
dysregulated in different types of cancer and chronic inflammatory disorders. The main form of NF-&#954;B is a
dimer of p50 and p65 proteins. NF-&#954;B is found in its inactive form in the cytoplasm, bound to the I&#954;B
inhibitory protein of the IKK complex. Stimuli such as viruses, UV light, inflammatory cytokines, and bacterial toxins
can induce NF-&#954;B activity by initiating degradation of the IKK complex. After degradation of the complex,
NF-&#954;B is freed and can translocate into the nucleus to activate transcription. Although NF-&#954;B activity is
largely controlled through its nuclear translocation, it is also controlled through its interaction with other coactivator
and corepressor proteins in the nucleus. My project focuses on the role of the p38 MAP kinase in mediating the
interaction between the p65 subunit of NF-&#954;B and the CBP coactivator. Chromatin immunoprecipitation assays
are being used to assess the recruitment of NF-&#954;B, CBP, and p38 to the NF-&#954;B-regulatory promoter in the
presence or absence of p38 inhibitors. Through these experiments, the lab hopes to gain insight into how p38
regulates NF-&#954;B activity.
AN EVALUATION OF PROMEGA’S DIFFEREX&#61652; EXTRACTION SYSTEM, WHICH SEPARATES
SPERM FROM EPITHELIAL CELLS FOR FORENSIC DNA ANALYSIS.
Angela N. Yoch (Ann Marie Gross/Presley Martin), DNA Section, Minnesota Bureau of Criminal Apprehension, , 1430
Maryland Avenue East, Saint Paul, MN 55106, , (Biology Department, Hamline University, 1536 Hewitt Avenue, Saint
Paul, MN 55104)
The differential extraction used to separate sperm from epithelial cells has always been a long, tedious, and time
consuming process. Recently, Promega developed the Differex system, which takes the current differential extraction
and simplifies it through the use of phase density and color to physically and visually separate the sperm and
non-sperm cell fractions. This method allows for fewer washes and centrifugation steps resulting in a less time
consuming extraction. The two main goals of this research are to determine if the Differex system works, and to look
at its efficiency in extracting small amounts of sperm, which Promega has not yet done, but is crucial to forensic
casework. Early experiments containing samples with low amounts of sperm following the Differex system protocol
yielded poor results in generating partial or full male profiles. As a result, changes were made in the protocol and
samples were prepared with larger amounts of sperm, although no significant changes in the total DNA yield have
been seen. In a comparative study between the current organic method and the Differex system, it is apparent that
the Differex system is not as efficient in obtaining full male profiles in which alleles from all sixteen Identifiler® loci
are being called.
42
BIOLOGY
HO-1 PLAYS AN IMPORTANT ROLE IN THE REGULATION OF COX-2 INDUCTION FOLLOWING
ENDOTOXIN EXPOSURE.
Elizabeth J. Saladin (Laura Fredenburgh, M.D., Mark Perrella, M.D.) (Tom Sequist, M.D., Four Directions Summer
Research Program; Brian Campbell, Ph.D., Oklahoma-Louis Stokes Alliance for Minority Participation). Brigham and
Women’s Hospital, Pulmonary Dep
Background: Sepsis is a severe infection characterized by the release of lipopolysaccharide (LPS) from the cell wall
of gram-negative bacteria. The body`s defense mechanism during sepsis involves the production of anti-inflammatory
mediators through the inducible enzymes heme oxygenase-1 (HO-1) and cyclooxygenase-2 (COX-2). HO-1
activation results in the production of carbon monoxide (CO) and bilirubin while induction of COX-2 leads to
prostanoid synthesis. Prior studies have shown that LPS induction of COX-2 is exaggerated in the absence of HO-1,
however the mechanism is not known.
Methods: We studied the mechanism by which COX-2 expression is enhanced in the absence of HO-1 during sepsis
using a mouse macrophage cell line (RAW). We used HO-1 byproducts of CO and bilirubin to model the expression
of HO-1. We treated RAW cells with increasing concentrations of either a CO releasing molecule (10 ?M, 50?M, or
100?M) or bilirubin (5?M, 10?M, 25?M, and 50?M) in the presence or absence of LPS. Following this exposure to
HO-1 byproducts and LPS, we extracted RNA at 2, 4, 6, 8, 12, and 24 hours. Northern blot analyses were performed
to quantify COX-2 expression levels among the exposed cells. Control groups received vehicle and did not receive
CO or bilirubin at each of the time points.
Results: Baseline COX-2 expression in RAW cells was low, however increased significantly following LPS exposure.
CO enhanced LPS-induction of COX-2 at low doses (50 ?M); however, at higher doses (100 ?M), CO exposure
appeared to decrease COX-2 induction as measured by Northern blot band intensity. Bilirubin demonstrated no effect
on LPS-induction of COX-2.
Conclusions: Regulation of COX-2 expression by CO in the presence of LPS is dependent on the dose of CO, but not
bilirubin. This provides evidence for a specific mechanism through which HO-1 regulates the induction of COX-2
during endotoxemia.
DISTRIBUTION OF GASTROPOD MOLLUSKS ON THE INTERTIDAL ZONE OF VANCOUVER ISLAND,
CANADA
Travis F. Mackenzie (Dr. Peter Kondrashov), Department of Biological Sciences, Northwest Missouri State University,
800 University Dr., Maryville, Mo 64468
Over time gastropod mollusk faunas have gone through significant changes caused by non-biotic and anthropogenic
factors. This study documents species diversity, ecological preferences, and intraspecific shell variation of native and
invasive species of gastropod mollusks in the intertidal zone along the coasts of Vancouver Island, Canada. I
collected samples of over 20 species representing eight orders and thirteen families of gastropod mollusks from eight
localities. In order to document the species distribution and shell variability, the samples were cleaned, sorted by
locality and then shells were identified and measured. An array of localities was sampled in order to understand how
the species diversity varied in different habitats. Extensive intraspecific shell variation was documented in three
species of gastropod mollusks: Littorina scutulata, L. sitkana and Nucella emarginata. Two species, Littorina scutulata
and L. sitkana, were the most abundant gastropod mollusks at all the localities. Both Littorinas showed variation in
color and sculpture. Nucella emarginata varied in color (including the presence or absence of spiral bands) and in the
development of spiral ribs which were often reduced. Along with the native species of gastropods I discovered an
invasive species, Batillara attramentaria (Japanese false cerith), in the intertidal mud flats of Comox harbor and
Royston beach. Originally this species was only found in Japan and the Western Pacific, but had been brought to
North America in the early 20th century with the import of the pacific oyster. Batillara attramentaria was abundant at
both localities. In contrary, the native representatives of Cerithiidae, which are usually found in the same type of
habitat, were extremely scarce or absent at these localities. As our study shows, the native gastropod mollusk fauna
of Vancouver Island, Canada, is very diverse and abundant, but an introduction of a non-native species could
jeopardize the stability of native mollusk populations.
43
BIOLOGY
THE EFFECTS OF INHIBITION OF SEROTONIN SYNTHESIS ON T CELL PROLIFERATION
Melinda Hexum, Audrey Geiger, Katelyn Ruter, (Dr. Jodi Goldberg), Biology Department, Hamline University, St. Paul,
Minnesota 55104
Previous research has indicated that serotonin (5-hydroxytryptamine, 5HT) is a signal necessary for human T cell
proliferation. The role of 5HT in T cell proliferation was investigated by examining the impact of p-chlorophenylalanine
(pCPA), a serotonin synthesis inhibitor, on the proliferation of purified human peripheral blood T cells activated by
accessory cell independent (anti-CD3/CD28 beads) and accessory cell dependent (phytohemagglutinin, PHA)
methods. T cell proliferation in response to either method of activation was inhibited by pCPA, though T cells
activated by PHA and accessory cells were inhibited at significantly lower doses than pure T cells activated with
antibody-coated beads. Inhibition of proliferation with pCPA was most effective when added upon initiation of T cell
activation, with inhibitory effects progressively reduced when pCPA was added on day 1 or 2 after activation.
Attempts were made to reverse the inhibitory effects of pCPA on T cell proliferation by providing pCPA treated cells
with exogenous 5HT or 5-Hydroxytryptophan (5HTP), but neither restored proliferation at any concentration tested.
These data suggest that T cells are inhibited by pCPA, but that non-T cells may be more sensitive to pCPA’s effects.
It remains to be determined if the inhibition occurs via a serotonin-dependent pathway.
USING ELECTRORETINOGRAPHY TO MEASURE SPECTRAL SENSITIVITY OF DIFFERENT
PHOTORECEPTORS
Laura G. Meloney, Leo Fleishman, Biology Department, Union College, Schenectady, NY 12308
The retina of most diurnal lizards contains four different classes of cone photoreceptors. The spectral response of
each cone class is
determined by its absorptive pigment and by an oil droplet within the cone which acts as a color filter. The spectral
responses of
individual cone classes have never been directly measured. The
purpose of this experiment was to determine the spectral sensitivity
of the different classes of cones of two species of lizards:
Anolis sagei and Platysaurus broadleyi.
We used electroretinography, in which an electrode is placed
on the surface of the cornea of an anesthetized animal and an
electrical response of the retina is recorded in response to brief
flashes of light. Electroretinogram (ERG) amplitude is proportional to the strength of summed response of the
photoreceptors. In order to determine spectral sensitivity, monochromatic light of different wavelengths (from 360 to
700 nm in 10 nm steps) was flashed into the eye. Four different intensities were used as stimuli for each wavelength.
For each wavelength, a plot of light intensity vs. response amplitude was created, and these were used to estimate
the stimulus intensity required to achieve a criterion response. In our first experiments we found that retinal spectral
sensitivity was dominated by a single class of cone which responds primarily to long wavelengths (the L cone). In
order to characterize the other cones in the retina, it was necessary to eliminate the response of the L-cone by
bleaching, which we achieved by shining the output from a 630 nm laser into the eye. We measured spectral
sensitivity of the retina after long wavelength bleaching and discovered multiple peaks in the spectral sensitivity
curve, which corresponded to the spectral sensitivities of the remaining three classes of cones. We have thus
demonstrated that electroretinography can be used to measure the spectral response of each class of cone in the
lizard retina.
44
BIOLOGY
UNDERSTANDING TAMOXIFEN RESISTANCE IN BREAST CANCER CELLS
Ashley L. McClellan and Lauren S. Kawabata and (Dr. Maggie C. Louie), Department of Natural Sciences and
Mathematics, Dominican University of California, San Rafael, California 94901
Breast cancer is one of the most common malignancies that occur in women in the United States and accounts for 1
in 4 female cancers. Breast cancer can exist as either ER positive or ER negative depending on the presence or
absence of the estrogen receptor (ER). The development of ER positive breast cancer is determined by the activity of
the ER and circulating levels of estrogen. Current treatments for ER positive breast cancer include using
anti-estrogens such as Tamoxifen. Although endocrine therapy with anti-estrogens has resulted in some success,
resistance is often seen with prolonged therapy. How this resistance develops is not clear. A better understanding of
tamoxifen resistance in breast cancer will provide valuable information for future therapeutics and improve the
success of current drug therapies. Our lab has developed a tamoxifen resistant breast cancer cell line (MCF7-TamR)
from the parental MCF7 cells (MCF7). In order to understand the mechanism of anti-estrogen resistance, we have
compared the growth and protein expression of the MCF7-TamR to the parental MCF7 cells. Our results demonstrate
that tamoxifen resistant cells under hormone-deprived conditions expressed higher levels of Cyclin E and Cdk2, two
important cell cycle proteins that regulate the G1/S transition. To further understand the mechanism of tamoxifen
resistance, we also treated the two cell lines with 10-8M tamoxifen. Under these conditions, the MCF7-TamR cells
displayed a faster growth rate than the MCF7 parental cells, which correlates with the higher Cyclin E and Cdk2
levels. Additionally, ACTR and TIF2, two coactivators for the ER, were also elevated in the resistant cell line and
may also contribute to anti-estrogen resistance.
RELATIONSHIP BETWEEN TEMPERAMENT AND BEHAVIORAL LATERALIZATION IN CYNOMOLGUS
MACAQUES (MACACA FASCICULARIS)
Emily C. Pearman (Dr. Allyson Bennett), Departments of Physiology and Pharmacologyand Pediatrics, Wake Forest
University School of Medicine, Winston-Salem, NC 27157
Previous studies have shown that temperament measures, such as impulsivity, can be used to determine risk
factors for underage drinking. Also lateralization measurements, such as handedness, have also been shown to
relate to temperament characteristics. This study, using 21 adolescent Cynomolgus Macaques (Macaca fascicularis),
focused on determining if dominant animals had a shorter latency response to a novel food, if dominant animals were
more likely to be left-handed, and if right-handed individuals had the longest latency to mouth a novel food. A piece of
cactus (novel food) was placed in an upper quadrant; a single subject was allowed to enter the quadrant and the
session recorded for 10 minutes. The sessions were coded for latency to approach, touch, and mouth the cactus.
Dominance status was determined by social interaction observations of subjects during the same period as the novel
food test. The findings demonstrate that dominant animals are have a stronger hand preference (p=0.057) but were
not more likely to have a particular hand preference direction (p=0.96), dominant animals were more likely to mouth a
novel object (p=0.16) than subordinate animals, and that both behavioral lateralization and social dominance rank
contribute to individual variation in response to novelty. However the link between neurobiological factors and
temperament measure has yet to be completely elucidated.
address for author:
Department of Biology, Guilford College, 5800 W. Friendly Ave, Greensboro, NC 27410
45
BIOLOGY
Activity of Lsm-11, ZFP100, and SLBP in the Pre-mRNA Processing of Human Histone Protein
Berkow A, Wagner EJ; Marzluff W; Biochemistry and Biophysics; University of North Carolina at Chapel Hill; Box
3280, Coker Hall, UNC-Chapel Hill, Chapel Hill, NC 27599-3280
Histones are a vital protein in DNA production and packaging. This project studies how proteins help the stem loop
binding protein (SLBP) regulate the production of histones. The proteins being investigated are Lsm11, ZFP100, and
SLBP, itself. It is believed that these proteins bind in a complex on the histone pre-mRNA and aid in pre-mRNA
processing. A reporter utilizing the MS2 RNA binding site, a green fluorescent protein, and the histone stem-loop was
created in order to determine the activity of proteins of interest with regards to histone pre-mRNA cleavage. The
gene for each protein of interest was cloned into a vector with the gene for the MS2 binding protein. In vivo, these
clones create a protein fusion of each protein of interest and the MS2 binding protein. The MS2 RNA binding site on
the reporter elicits recruitment of each protein to the reporter, thereby putting the protein of interest in close proximity
with the stem-loop structure. If cleavage is induced by that protein, the GFP message will be allowed to travel to the
cytoplasm where it can fluoresce green. It is in this way that each protein of interest will be tested for its activity in the
cleavage of the histone stem-loop structure.
Effect of Wounding on Gene Expression in Tomatoes: Analysis of Specifc Transcripts that are
increased in total mRNA population.
Taleria Alston, National Science Foundation, Botany Department, North Carolina State University, Raleigh NC 27695
Experiments were conducted which involved heat-wounding the next-to-youngest leaf (#3) of tomato plants for
specified times, which were as close as 2 seconds apart, and then harvesting the younger, distant leaf (leaf #4). This
tissue was frozen in liquid nitrogen to immediately stop all biologic activity. The frozen tissue was pulverized in liquid
nitrogen and used for isolation of polysomes, polysomal RNA and total RNA. RNA was incubated with the appropriate
primers in the presence of reverse transcriptase to make cDNA, and the cDNA was used for PCR. The PCR reactions
were electrophoresed to measure the size of the PCR products. The cDNA was also provided to a graduate student
who did real time PCR to find out which of these transcripts changed after heat-wounding. The polysome profiles of
all 19 samples show there was little consistent change in polysomes after wounding, but all the profiles showed the
polysomes (and thus their mRNA) were undegraded. The PCR reactions gave products of the expected size, while
the real time PCR reaction showed that one particular mRNA (PEP carboxylase kinase) increased at least 20-fold
within 2 minutes of wounding.
46
BIOLOGY
ENVIRONMENTAL POLLUTION AND AGRICULTURAL INCENTIVES IN US AND EU PROGRAMS
Tory Hodges (Dr. Jennie Popp), Department of Agricultural Economics and Agribusiness, University of Arkansas,
Fayetteville, Arkansas, 72701
The United Kingdom (UK) and the Arkansas/Oklahoma (AR/OK) region in the US have a long history of production
agriculture. Today, both face similar water quality struggles from non-point source pollution, and the US and UK are
using voluntary, incentives-based farm programs to enhance environmental stewardship. The purpose of this study is
to present an economic comparison of agricultural incentives programs in the Common Agriculture Policy (CAP) of
the European Union and voluntary green programs including the Conservation Security Program (CSP) in the US in
order to evaluate the applicability of these programs to water quality issues from agriculture in the AR/OK region.
First, the primary incentives of CAP and CSP are assessed for similarities and differences. Then using the same
criteria, these programs are analyzed to determine which programs, or aspects thereof, could be utilized in the US.
Measurable criteria include required tax dollars and length of funding, while qualitative measures consist of
farmers/residents perceptions regarding conservation programs. Research through Great Britain’s Department for
Environment, Food and Rural Affairs (DEFRA), Europe’s Linking Environment and Farming (LEAF), and the Dutch
National Environmental Research Institute provide statistical economic information needed for this comparison, while
the United States Department of Agriculture’s National Resource Conservation Service (NRCS) is the primary source
for information on U.S. programs. After defining economic criteria for environmental programs, they will be used to
conduct a comparison between the CAP Rural Development Program and the Conservation Security Program to
assess which program might be applicable to the Illinois River dispute in the AR/OK region.
It is expected that the CSP has the potential to economically serve the needs of business, agriculturists, and
conservationists in AR/OK; however, this program is currently underdeveloped. Also, the High Level option of the
Environmental Stewardship Program in the CAP Rural Development Program could be applied in part to the Illinois
River dispute, and portions of other programs within the Single Payment Scheme of the CAP could prove beneficial to
AR/OK, though no single program is likely to be an instant fix-it for water quality degradation.
“Elucidation of Amino Acids Necessary for Transcriptional Activation in ToxT, a Key Regulatory
Protein in Vibrio cholerae”
Melissa Castaneda, Senior, Biology, University of Texas at San Antonio Honors College, , Advisors:, Brandon M.
Childers, Gregor Weber, Melissa. M Castaneda and Karl E. Klose, Department of Biology, University of Texas at San
Antonio, San Antonio TX 78249
Vibrio cholerae, is a motile, gram-negative, rod-shaped bacterium and is the etiologic agent responsible for the
diarrheal disease cholera. Research into the pathogenesis of V. cholerae, which afflicts thousands of people
worldwide each year, is critical for the development of novel vaccines and therapeutics. ToxT is a key regulatory
protein in V. cholerae, which directly activates the transcription of the two major virulence gene clusters, ctx and tcp,
which encode cholera toxin (CT) and the toxin-co-regulated pilus (TCP), respectively. Previous data demonstrated
that the N-terminus of ToxT (aa1-153) functions in dimerization of the protein, whereas the C-terminus (aa154-276)
functions in DNA binding. Interestingly, the ToxT C-terminus dimerized by a heterologous dimerization domain is able
to bind DNA, however it is unable to activate transcription, suggesting that there are determinants in the N-terminus of
ToxT involved in transcription activation. We are interested in determining which amino acids of ToxT are necessary
for these various functions. More specifically, the purpose of my project is to determine which amino acids in the
N-terminus are necessary for ToxT dimerization. Scanning alanine mutagenesis studies have shown that there are
approximately 30 N-terminal (aa1-153) ToxT mutants defective for transcriptional activation. To test the effects of
these individual N-terminal mutations on dimerization, the mutations were reintroduced into a plasmid that expresses
a ToxTN-LexAN protein fusion. These plasmids will be transformed into a sulAp-lacZ reporter strain (JL1436) and
then assayed for _-galactosidase. We anticipate that we will have characterized all 30 N-terminal mutants by March.
It is expected that that N-terminal mutants with defects in dimerization will exhibit high _-galactosidase activity due to
their inability to dimerize. Elucidation of the amino acids that are necessary for ToxT dimerization will help to further
characterize virulence regulator ToxT and assist in the development of therapeutics against cholera.
47
BIOLOGY
Investigating the Role of 5HT1A in Human T Cell Activation
Audrey Geiger, Katelyn Ruter, Melinda Hexum (Dr. Jodi Goldberg), Hamline University Biology Department, 1536
Hewitt Avenue, Saint Paul, MN 55104
The neurotransmitter 5-hydroxytryptamine, 5HT, is a necessary signal for human T cell proliferation. Specifically, the
5HT1A receptor subtype has been implicated in the activation of T cells. The importance of the 5HT1A receptor in
human T cell activation was assessed by measuring 5HT1A receptor expression and by assessing the effects of
5HT1A antagonists on T cell activation. Using two PCR primer sets, 5HT1A mRNA was shown to be expressed in the
human Jurkat T cell line, confirming published results. Human peripheral blood T cells were then purified and their
5HT1A receptor expression pattern was assessed over a 3-day activation timecourse. Unlike previous published
results, 5HT1A mRNA was present in resting T cells. Expression was also detected on each day of the timecourse
with the lowest expression on day 3. Assays were then performed to determine T cell proliferation in the presence of
5HT1A receptor antagonists. Purified T cells were activated by accessory cell dependent (phytohemagglutinin) and
accessory cell independent (anti-CD3/CD28 beads) mechanisms. The nonspecific 5-HT1,6,7 receptor antagonist
methiothepin exhibited inhibition starting at concentrations of 2.5 uM. The specific 5HT1A receptor antagonist
nan-190 failed to inhibit proliferation. Reversal of methiothepin by the inclusion of 5HT was not achieved.
THE EFFECT OF OXYTOCIN ON mGnRHI AND cGnRHII RELEASE IN SALMONOIDS
Sarah Ellenberger (Michael Woller, PhD), Department of Biological Sciences, University of Wisconsin –
Whitewater,Whitewater, WI 53190
This study focuses on the regulation of Gonadotropin Releasing Hormone (GnRH) release in fish, specifically
salmonoids. A single species of fish expresses and releases three distinct isoforms of the GnRH molecule: seabream
GnRH (sbGnRH), chicken GnRH II (cGnRH II), and salmon GnRH (sGnRH). Each variant is believed to be released
from a different region of the brain. Oxytocin has been shown previously to impact the release of mammalian
mGnRH I (mGnRHI) in several model systems. Brook trout brain tissue was divided into three regions: nucleus
olfactoretinalis, preoptic area and midbrain and placed into an in vitro perifusion system to collect samples linearly.
This study involves two experiments: 1) measuring endogenous release of GnRH isoforms from the three brain
regions; 2) evaluating any effects of oxytocin on GnRH isoform release from the trout brain tissue. For the first
experiment, the brain regions were exposed to media and samples were collected linearly for 5 hours. For the second
experiment, the tissue was exposed to 1 hour of media followed by 10 minutes of oxytocin and 3 hours of media.
Samples were collected every 10 minutes and mGnRH I and cGnRH II levels were determined by radioimmunoassay.
Oxytocin showed no effect on the release of either GnRH isoform tested. Differential release from the various brain
regions was not observed. This study represents the first observation of GnRH release in salmonoids.
IDENTIFICATION OF CIS-ACTING DNA ELEMENTS THAT CONTROL TISSUE SPECIFIC MUA-1
EXPRESSION IN C. ELEGANS
Jennifer L. Quinlan (Dr. Plenefisch) Department of Biological Sciences, The University of Toledo, Toledo, OH 43606
Movement of Caenorhabditis elegans is dependant upon the way its muscle is attached to the outer cuticle. Many
mutants have been identified where, initially, their movements and muscle differentiation are normal, but then the
muscles somehow detach from the cuticle and paralysis occurs. These mutants identify a class of genes required for
growth and maintenance of functional muscle attachments. These genes are named mua, for muscle attachment
defective. I will be taking a closer look at mua-1, particularly, by identifying DNA elements that control its expression
in specific tissues during the development of C. elegans. This gene is expressed in three tissues – the pharynx, the
epidermis, and the uterus (during the larval to adult molt only). This project is designed to use different potential
enhancers of mua-1 in an attempt to determine which enhancers are expressed where in C. elegans. The polymerase
chain reaction (PCR) will be used to generate expression constructs. Since the nucleotide sequence of mua-1 is
known, fragments can be amplified directly using the PCR technique and ligated into a plasmid vector upstream of
GFP (green fluorescent protein) using standard molecular techniques. The resulting recombinant plasmids, all
carrying the identical genomic DNA segment, can then be microinjected into C. elegans or propagated in E. coli
bacteria. GFP will fluoresce wherever the amplified fragment turns on expression. The ultimate goal of my project is
to find out what is controlling mua-1 – how its expression is being controlled, why it is being turned on during such a
specific time in the uterus, and what makes sure it is turned on. It is hypothesized that mua-1 is an element important
for localization to the nucleus.
48
BIOLOGY
MICROSATELLITE ANALYSIS OF GENETIC DIVERSITY AND INCIDENCE OF HYBRIDIZATION IN THE
ADIRONACK PARK COYOTE (C. latrans) POPULATION
Brian Curry (Dr. Nancy Elwess and Mrs. Sandra Latourelle), Department of Biological Sciences, Plattsburgh State
University of New York, Plattsburgh, NY 12901
Since the mid-1900s, coyote (Canis latrans) populations have steadily expanded throughout the United States and
into Canada, with ecological ramifications that have yet to be fully appreciated. While other predatory animal
populations have dwindled in the northeastern United States, coyote populations currently exist at densities sufficient
to make consideration of coyote ecology vital to any responsible assessment of conservation priorities. This research
quantifies the genetic diversity of the coyote population within New York`s Adirondack Park, currently the largest
publicly protected area in the contiguous United States. In addition, this research may have application in
determining the impact of coyote-wolf hybridization on coyote population genetics and ecology. Genomic DNA
isolated from tongue tissue samples from over 40 coyotes within the Adirondack Park was analyzed for allelic
variation of several microsatellite loci. This variation indicates the degree of genetic diversity within the coyote
population of the Adirondack Park. Also, variation as a result of coyote-wolf hybridization may be identified, with
some indication of the degree to which hybridization has occurred. This data could assist with management decisions
regarding conservation priorities within the Adirondack Park, and the northeastern United States in general.
GENETICALLY DECREASED GONADOTROPHIN-RELEASING HORMONE SECRETION:
CONSEQUENCES ON FERTILITY AND OBESITY IN MALE TRANSGENIC RATS
Francesco DeGiacomo, Sara Batres, Ashley Fale, Andrew Lewis, Lisa Lima and Christopher Heiser (Mohammed El
Majdoubi), Department of Natural Sciences and Mathematics, Dominican University of California, San Rafael,
California 94901
The pulsatile release of Gonadotropin-Releasing Hormone (GnRH) is the driving force of reproductive function in
mammals. Our previous investigation of transgenic rats with decreased GnRH secretion showed that middle-aged
females are infertile and obese. In the present study, we asked whether middle-aged male transgenic rats display the
same reproductive and metabolic abnormalities observed in transgenic females of same age. Young (3 month-old)
and middle-aged (9 month-old) male transgenic and wild-type rats were euthanized. Body weights were measured and
abdominal fat dissected and weighed. Testes were also weighed and evaluated histologically. Plasma levels of
luteinizing hormone (LH), Follicle-stimulating hormone (FSH), and testosterone (ie., reproductive hormones) together
with levels of insulin and leptin (ie., metabolic hormones) were measured by radioimmunoassay. As expected, LH
levels were decreased in young and middle-aged transgenic whereas no differences were detected in FSH levels
between transgenic and wild-type rats. Moreover, although testicular weight was significantly lower in young and
middle-aged transgenic males compared to wild-types, testosterone levels and the histology of the testes were similar
in both groups. In middle-aged males, there was an increase in body weight and abdominal fat content associated
with elevated levels of insulin and leptin. However, these metabolic changes affected both transgenic and wild-types
to a similar extent. We report that, unlike female transgenic rats, decreased GnRH activity in middle-aged male
transgenic rats was not associated with infertility and obesity. This work reveals a major sexual dimorphism in the way
GnRH regulates reproductive and metabolic functions, and provides new insights into the etiology of reproductive and
metabolic abnormalities in males and females.
49
BIOLOGY
ALTERED GONADOTROPIN-RELEASING HORMONE SECRETION IS ASSOCIATED WITH
INFERTILITY AND OBESITY IN MIDDLE-AGED FEMALE TRANSGENIC RATS
Jacqueline Maravilla, Elena Romero, Ofek Bar-Ilan, Alex Plascencia and Eric Hinderleider (Mohammed El Majdoubi),
Department of Natural Sciences and Mathematics, Dominican University of California, San Rafael, California 94901
Gonadotropin-Releasing Hormone (GnRH) is the main hormone controlling reproduction in male and female
mammals. Female transgenic rats, in which GnRH secretion was genetically decreased, have diminished fertility and
altered metabolism. In the present study, we investigated whether the severity of the reproductive and metabolic
alterations in female transgenic rats is age-dependent. Young (3 month-old) and middle-aged (9 month-old) female
transgenic and wild-type rats were euthanized. Body weights were measured and abdominal fat dissected and
weighed. Ovaries were weighed and evaluated histologically. Plasma levels of luteinizing hormone (LH),
Follicle-stimulating hormone (FSH), and estradiol (ie., reproductive hormones) together with levels of insulin and
leptin (ie., metabolic hormones) were measured by radioimmunoassay. As expected, LH levels were decreased in
young and middle-aged transgenic females whereas no differences were detected in FSH and estradiol levels
between transgenic and wild-type females. On the other hand, ovarian weight was decreased in young and
middle-aged transgenic rats compared to wild-types. However, only middle-aged transgenic females were anovulatory
(absence of corpus luteum) and had polycystic ovaries. Unlike wild-types, middle-aged transgenic females had a
significant increase in abdominal fat content together with elevated levels of insulin and leptin. We report that
decreased GnRH secretion was associated with infertility and obesity in middle-aged but not young female transgenic
rats, and that the likeliness of reproductive and metabolic problems associated with alterations in GnRH secretion
increases with age.
PHYLYOGEOGRAPHIC ANALYSIS OF BYTHOTREPHES INVASIONS IN MINNESOTA LAKES
Jacob M. Cooner (Dr. Leif Hembre), Department of Biology, Hamline University, St. Paul, MN 55104
Bythotrephes longimanus (the spiny waterflea) is a predatory zooplankton native to Eurasia that invaded Lake
Superior and other Laurentian Great Lakes in the 1980s. The invasion of a non-indigenous species could potentially
be catastrophic to the longevity of a habitat, forever altering pre-existing food webs due to competitive pressure on
native species’ morphology, abundance and composition. Since 1990, with the invasion of Island Lake (near Duluth,
MN), the species has spread to more inland lakes in Minnesota. A recent survey of lakes in the Superior National
Forest and the Boundary Waters Canoe Area discovered that the invasion of inland lakes in Minnesota is progressing.
In this study we are comparing the genetic composition (using allozyme markers) of Bythotrephes populations in
several Minnesota lakes in an attempt to determine the hub lake(s) of the inland invasion. Genetic data will be used to
develop a dendogram (tree diagram) based on the genetic distance among populations. This will provide insight into
possible invasion corridors that are enabling non-indigenous species to invade inland Minnesota lakes and will
potentially aid in preserving natural aquatic food webs.
50
BIOLOGY
EXPRESSION OF ASIC2 MRNA IN BRAIN TUMORS
Brooke L. Goodman* ( Jennifer T. Thomas*, Ph.D., Catherine M Fuller†, Ph.D., Dale J. Benos†, Ph.D.) *Departments
of Biology and Chemistry, Belmont University, Nashville, TN, †Department of Physiology and Biophysics, University of
Alabama at Birmingham, Bir
GBMs (glioblastoma multiformes) are responsible for 30% of all brain tumors and are the most common primary brain
tumor in adults. Patients have a median survival rate of 12 months with treatment. In both normal astrocytes and
cultured GBM cells, there is a group of sodium ion channels known as ASICs (acid-sensing ion channels). In normal
astrocytes, protein subunits ASIC1 and ASIC2 are present at the cell membrane, resulting in inactivation of the
channel. In GBMs, however, ASIC2 is not present at the membrane and the channel is continuously active. This
increased Na+ channel activity may contribute to the ability of the GBM cells to invade surrounding brain tissue, a
hallmark of the rapid progression of GBMs. The purpose of this study, therefore, was to determine why ASIC2 is not
found at the cell membrane in GBMs. We chose to examine the levels of ASIC mRNA in normal brain samples and
GBM samples to see if the problem is at the transcriptional level. Our findings demonstrate that all of the normal
brain samples expressed both ASIC1 and ASIC2 mRNA, indicating proper transcription of the genes for ASIC1 and
ASIC2. In contrast, while all of the GBMs expressed mRNA for ASIC1, only 50% of the samples expressed the mRNA
for ASIC2. These results suggest that transcriptional silencing of ASIC2 may contribute to the lack of membrane
protein expression of this subunit.
This research was completed in the Summer in Biomedical Sciences (SIBS) program at UAB through an NIH grant
(NCI-CA71933) and was supported by an additional NIH grant (CA101952) to D.J.B in the Department of Physiology
and Biophysics at the University of Alabama at Birmingham.
RESPONSE OF A STRAIN OF BACILLUS MYCOIDES TO A CARBON DIOXIDE ENRICHED CULTURE
ATMOSPHERE
Kate Genty (Joe R. Wolf), Department of Biology, Peace College, Raleigh, NC 27604
Bacillus mycoides 6462 is the ATCC type strain of a common soil bacterium. Previous studies of this organism have
focused on the genetic determinants of its pronounced rhizoidal colony morphology. We are interested in the effect
of environmental influences on colony morphology and have noticed an intriguing response of this organism when
cultured in a CO2-enriched environment. We observe reproducibly that the surface area occupied by bacterial
colonies increases with atmospheric CO2 concentration. We can reverse the “colony spreading” effect by including
CO2 chemical sinks such as NaOH and KOH in partitioned plate cultures. In order to determine if the observed
difference in colony size is due to a difference in cell number or cell size, we homogenized colonies of B. mycoides
and performed serial dilutions of the homogenate followed by plate counts. There appears to be little difference in
cell number between the two colony types, suggesting that a difference in cell length plays a role in the response of
this organism to carbon dioxide. Electron microscopic observation of the cells within colonies from the different
culture environments supports this idea. Efforts are underway to characterize changes in cellular chemistry that occur
in response to a CO2-enriched culture environment and to understand how these connect to the observed changes in
cell and colony morphology.
51
BIOLOGY
A SURVEY OF MESOPREDATORS IN A STATE PARK NEAR A MAJOR METROPOLITAN AREA
Christopher J. Laliberte, (Dr. Stephen G. Mech)
We surveyed for mesopredators (medium-sized predators) in French Creek State Park, near Reading, PA for the
Pennsylvania IMAP (Important Mammal Area Project). IMAP seeks to conserve land that is important mammal
habitat due to either high mammal diversity or presence of mammals of special concern. As a large contiguous forest
patch near a major metropolitan center (Philadelphia), French Creek is an important natural area for conservation
purposes as well as recreational purposes. Part of our goal was to perform an initial survey of mammalian diversity in
the park. We used two survey methods: hair traps and camera traps supplemented by some live trapping. We
surveyed three habitat types (rocky outcropping, wetlands, and dry uplands) throughout the park. Over the course of
the summer, we observed only two species of mesopredators: raccoons (Procyon lotor) and Virginia opossum
(Didelphis virginiana). We determined that raccoons were the most common mesopredators in the area, but they
occurred in relatively dry habitat, which was not expected. These data serve as a preliminary step in testing the
mesopredator release hypothesis which hypothesizes that the absence of large predators leads to an increase in
mesopredator density. We also trapped small mammals to determine the food base for mesopredators as well as to
determine the habitat quality based on mass of the small mammals.
USING SNNAP (Simulator for Neural Networks and Action Potentials) TO MODEL THE BEHAVIOR OF
NETWORKED MORIS-LECAR CELLS
Michael A. Fierro, (Dr. James J. Watrous), Department of Biology, Saint Joseph’s University, Department of Biology,
5600 City Avenue, Philadelphia, PA 19131
Using SNNAP, a ‘patch’ network, consisting of dimensionless Moris-Lecar type cells, was constructed. The cells within
the network consist of three voltage dependent channels - calcium, potassium, and leak, each of which is controlled
by a unique set of parameters. According to Morris and Lecar’s 1981 journal article “Voltage Oscillations in the
Barnacle Giant Muscle Fiber,” muscle fibers subjected to a constant current of electrical stimulation will exhibit
oscillatory behavior. The model assumes the presence of two non-inactivating conductances in the form of calcium
outside of the cell and potassium on the inside. Cardiac cells, which have many similar characteristics to barnacle
muscle fiber, can be represented using the same set of equations set forth by Morris and Lecar. Nine-cell networks
were modeled using both patch and geometric cells. The electrical currents and behavior of the three channels in
each cell of the system were analyzed over a period of 1.0 ms. The time interval between 0.2 and 0.5 ms was the
focus of this research. Action potentials were generated in the geometric network that fire at 0.315 ms and reach
hyperpolarization at 0.348 ms. They had a range of voltages of -56.2 mV to 30.7 mV between depolarization and
hyperpolzarization. The action potentials in the patch network fire at 0.310 ms and reach hyperpolarization at 0.345
ms. The voltage range of their action potentials was -47.5 mV to 30.1 mV. Because the patch network and geometric
network had nearly identical voltage and frequency characteristics within their action potentials, the simulation can be
considered valid.
CHEMICAL DEFENSE AND THE POPULATION ECOLOGY OF PHYTOLACCA RIVINOIDES, A
TROPICAL PIONEER PLANT
Aaron Garoutte and Mitchell Plosz, (Drs. K. Greg Murray & William Mungall), Departments of Biology and Chemistry,
Hope College, 35 E. 12th Street. Holland MI 49423
A key feature of the life history of most tropical pioneer plants is the maintenance of a persistent bank of dormant
seeds in the soil, even in the face of threats from rodents, insects, and fungi. Our lab is investigating the roles played
by defensive chemicals in seed persistence, and my work examines the chemical basis of antifungal defenses in
Phytolacca rivinioides, a pioneer whose seeds can persist in the soil for decades. Other members of the family
Phytolaccaceae are known to deter herbivores and fungi with saponins, but neither P. rivinoides nor the seeds of
those other species have been examined specifically. Using a bioassay developed for this study, we have
determined that Phytolacca contains powerful antifungal compounds that rival commercial fungicides in their efficacy.
Using a variety of separation and identification techniques (HPLC, GC-MS, and LC-MS) we are currently narrowing
our focus to the active fractions of whole seed extracts in order to identify the specific chemical compounds involved
in defense against fungi.
52
BIOLOGY
CORRELATION OF MEMBRANE FUSION EVENTS WITH SECRETION IN THE GASTRIC MUCOSA.
Tanner L. Bartholow and Peter J. Patitsas. (Jeff R. Demarest) Department of Biology, Juniata College, Huntingdon PA
16652.
A single cell type, the oxyntic cell (OC), is responsible for both acid (HCl) and pepsinogen (PEP) secretion by the
gastric mucosa of all non-mammals. Using scanning electron microscopy, we have shown that the stimulation of HCl
and PEP secretion by in vitro Necturus maculosus gastric mucosa is accompanied by dramatically increased oxyntic
cell apical membrane area. These two separate secretory pathways contribute to the increased membrane area with
different temporal patterns. Secretion of H+, Cl- and PEP were followed in isolated mucosae mounted in Ussing
chambers. Exposure to histamine, which predominantly stimulates the HCl secretory pathway, results in a slow (about
60 minutes) continuous 4-5 fold increase in Cl- secretion (ClISC) measured as short circuit current in amiloride treated
mucosae. Whereas, carbachol, which predominately activates the PEP secretory pathway, produces a sharp 4-5 fold
transient peak in ClISC within 1-3 minutes of application followed by a sustained 5-6 fold increase by 30 minutes.
Transepithelial electrical resistance decreases in both cases in parallel with increased Cl- secretion. The responses to
histamine are consistent with the well known time course of membrane fusion leading to the insertion of H+/K+
-ATPase into the apical membrane of OCs during the activation of H+ secretion. The initial transient effect of
carbachol is consistent with the abrupt fusion of mature protein secretory granules during the activation of PEP
secretion. Both processes are accompanied by increased Cl- secretion. The dynamics of the membrane fusion and
retrieval events of the individual secretory pathways responsible for these changes in apical membrane area and
secretion have been examined in isolated oxyntic cells with fluorescent cytoskeletal and membrane probes using
confocal microscopy.
Link to special characters version - http://students.juniata.edu/barthtl3/NCUR.htm
INVESTIGATION OF THE MECHANISM OF CD8+ T CELL HOMEOSTASIS AFTER ACTIVATION BY C.
NEOFORMANS-LADEN MICROGLIAL CELLS
Jessica E. Nordin (Dr. Karen Aguirre), Department of Biology, Coastal Carolina University, Conway, South Carolina
29528
The immune system has evolved innate and adaptive responses to protect itself from invading microorganisms.
Immune function is relatively well understood in the periphery, but qualitative differences exist when infection occurs
in the central nervous system (CNS), and these differences are poorly understood. One cell activated in response to
CNS infection is the brain–resident macrophage, or microglial cell. Microglia interact with T lymphocytes, which bind
to the MHC class I-peptide combination. Past research has focused on microglia/CD8+ T cell interaction in viral
models. However, the pathogenic fungus Cryptococcus neoformans, and other non-viral facultative and obligate
intracellular pathogens represent a considerable portion of reported CNS infections. Such pathogens do not interfere
with host cell gene expression as do viral pathogens, and therefore must be controlled by different mechanisms.
I plan to observe the viability of CD8+ T cells after microglial activation in a non-viral system to tell whether CD8+ T
cells undergo activation-induced cell death (AICD). AICD is a homeostatic mechanism by which large clones of
activated lymphocytes are removed from the system after infection has been cleared, leaving only a small population
of memory cells. It is not known how CD8+ T cell homeostasis is maintained after CNS infection. Activation will be
induced by adding Cryptococcal antigen and SEA (Staphylococcus enterotoxin antigen) to microglial/CD8+ T cell
co-culture. After 48 and 120 hours, CD8+ T cell viability will be quantified by trypan-blue exclusion. If AICD has
occurred, the TUNEL assay will be performed to investigate if the observed death is a result of apoptosis (or
programmed cell death). If AICD has not occurred, further research is necessary to determine how the immune
response is being shut off. Overall, an enhanced understanding of the mechanism of CD8+ T cells is significant as it
is a rational basis for designing immunotherapy or vaccines based on CD8+ T cells.
53
BIOLOGY
BAC RECOMBINEERING OF HUMAN ANDROGEN RECEPTOR: A FUNCTIONAL GENETICS
APPROACH TO MODELING HUMAN PROSTATE CANCER
Scott M. Krummey (Kristin Kee, Ph.D., and Jeffrey E. Green, M.D.), Laboratory of Cell Regulation and
Carcinogenesis, National Cancer Institute/NIH, Bethesda, MD 20892
Mutations in the human androgen receptor (hAR) alter hormone-responsiveness and have been found to contribute to
growth in refractory prostate cancer. Previously, mutations in hAR cDNA have been studied and shown to cause
cancer in mice; however, functional and regulatory studies of hAR mutations have been limited. This project seeks to
create a humanized mouse model of the hAR that will mimic endogenous expression levels in vivo. To achieve this
goal, we are using a powerful chromosome engineering technique called recombineering, which manipulates DNA in
vivo using a phage-based E. coli homologous recombination system. We have devised a two-step recombineering
strategy to generate a bacterial artificial chromosome (BAC) which will contain the full-length hAR sequence with its
upstream and downstream regulatory elements. This BAC will mimic endogenous expression patterns in cell culture
and humanized mouse model systems. Once constructed and tested for functional rescue in hAR knockout cell lines,
the hAR BAC can be mutated to study specific hAR mutations with a role in prostate tumor development and
progression in mouse models. Potentially, these studies can provide insight into novel therapeutic approaches
against androgen-independent prostate cancer.
Colgate University, Hamilton, NY 13346
FEEDING HABITS OF JUVENILE BLACK DRUM (POGONIAS CROMIS) FROM SOUTH TEXAS
CREEKS
Krisan M. Kelley (Jennifer Purviance,and Dr. M. Andres Soto), McNair Scholars Program, Department of Biology,
Texas A&M University-Kingsville, Kingsville, TX 78363
Black drum (Pogonias cromis) are one of the most commercially and recreationally important finfish in Texas. They
have an estuarine dependent life cycle. Adults are found coastally and migrate into estuaries to spawn. Eggs, early
larvae, and juveniles are found in bays. Larval and juvenile black drum from the Laguna Madre, a hypersaline
lagoon, are commonly found in creeks. The objective of this study is to examine the food habits of juvenile Black
Drum from these creeks. Black drum were caught from two creeks, San Fernando Creek and Escondido Creek.
Juvenile black drum were obtained using a cast net, and then placed in 10 % formalin that fixes the fish and stopped
further digestion. Standard length (SL) was taken for each fish. During dissection, the stomach and intestine was
examined and contents were identified to lowest possible taxon. Ten fish have been examined. The size of the fish
examined ranged from 4.1 to 11.5 SL. Food items found were larval insects, gastropods, ostracods, amphipods, and
isopods.
INITIAL CHARACTERIZATION OF THE EXPRESSION AND FUNCTION OF THE CIRCADIAN GENE
NOCTURNIN DURING EARLY DEVELOPMENT IN XENOPUS LAEVIS
Rachel M. Yanke (Dr. Kristen L. Curran), Department of Biological Sciences, University of Wisconsin-Whitewater, 800
West Main Street, Whitewater, Wisconsin 53190
The purpose of this research project was to begin characterization of the function of the NOCTURNIN protein in the
early development of Xenopus laevis. Nocturnin is a gene that is expressed in a circadian manner in the adult. In the
developing embryo it is expressed in the somites and pronephros during the early tail bud stages. NOCTURNIN has
been characterized as a deadenylase, a protein that destabilizes mRNA in the cell. It does this by removing the poly
(A) tail, a sequence of adenine nucleotides, from mRNA. In order to test for a role for Nocturnin during
development, we injected one cell of a two celled embryo with a morpholino specific for Nocturnin. Morpholinos act
to inhibit translation of mRNA and therefore, NOCTURNIN protein levels will be less than normal on the injected side
(50% reduction in protein to date). Preliminary results suggest that Nocturnin may affect the timing of somitogenesis
or somite formation. We are conducting additional replicates of this experiment to confirm our results and analyzing
whether NOCTURNIN depletion affects development of the pronephros.
54
BIOLOGY
NEURONAL DEATH OCCURS IN THE BRAIN OF PRETERM LAMBS MANAGED BY CONVENTIONAL
VENTILATION FOR 3 DAYS.
J Alvord, MJ Dahl, A Shumway, DM Null, RH Lane, KH Albertine Pediatrics, University of Utah, Salt Lake City, UT.
Background: Preterm birth followed by prolonged mechanical ventilation often results in chronic lung disease (CLD)
of prematurity. Associated with lung injury is brain injury. The molecular pathways that are affected in the brain of
preterm neonates during the evolution of CLD are unclear. On the other hand, animal models of intrauterine growth
retardation have shown that apoptosis is greater in the brain of postnatal rat pups with intrauterine growth retardation
compared to controls (Ke et al, AJP - 2004). A marker of apoptosis is expression of downstream targets of p53, such
as caspase-3.
Hypothesis: Apoptosis of neuronal cells in the brain is enhanced during the evolution of CLD.
Methods: Preterm lambs (~132d gestation) were managed by conventional mechanical ventilation (CV) or nasal
continuous positive airway pressure (nCPAP) for 72h (n=4/group). We used nCPAP as the positive control
(gold-standard) for lung outcome in chronically ventilated preterm lambs (Albertine et al, Pediatr Res - 2004). Brain
tissue was analyzed for caspase-3 protein abundance (immunoblot analysis) and topographic distribution
(immunohistochemistry).
Results: Immunoblot analysis showed that caspase-3 protein abundance in brain tissue homogenates from preterm
lambs was greater in the CV group compared to the nCPAP group, at 72h. Immunohistochemistry showed that
caspase-3 protein was localized in more Purkinje cells in the cerebellum in the CV group compared to the nCPAP
group at 72h.
Conclusion: Our results show that markers of apoptosis (caspase-3 protein abundance and immunolocalization) are
more prevalent in the brains of preterm lambs that are managed by CV compared to nCPAP. We conclude that CV
contributes to attrition of neurons during the first few days of life support with a ventilator. [Supported by CHRC,
HL62875, HL56401, T35 HL07744]
TROPHIC ANALYSIS OF MYOSOTIS LAKE, RENSSELAERVILLE, NEW YORK
Amanda B. Stella (Peter Tobiessen), Department of Biology, Union College, Schenectady, New York 12308
The purpose of this study is to determine a possible biomanipulation strategy for Myosotis Lake, located on the
Edmund Niles Huyck Preserve in Rensselaerville, N.Y. This lake serves as a reservoir for its surrounding population
and is currently experiencing algal blooms. Biomanipulation is a promising means of reducing unwanted algal blooms,
because it is based on the presumption that a change in one trophic level will result in changes of all other trophic
levels. It is thus hoped that a change in fish predator types in the lake will increase the herbivorous zooplankton
populations, therefore decreasing algal biomass. We collected water samples approximately once a month, from May
2005 through October 2005 to identify and quantify zooplankton and phytoplankton populations in the aquatic
community. Lake samples were also collected for water chemistry and chlorophyll content analysis. Water
transparency, DO content, and water temperature at various depths were recorded on every sampling day. Based on
the data collected, a suitable biomanipulation strategy will be recommended for Myosotis Lake.
55
BIOLOGY
COMPARISON OF DIETARY PREFERENCES IN THE GYPSY MOTH (LYMANTRIA DISPAR) BETWEEN
UNPARASITIZED AND PARASITIZED (GLYPTAPANTHELES FLAVICOXIS) LARVAE
Cassondra A. Myers (Dr. Jennifer Koehl), Biology Department, Saint Vincent College, 300 Fraser Purchase Road,
Latrobe, PA 15650
This study’s purpose is to compare the effect of hemlock diet verses control diet on parasitized and unparasitized
gypsy moth larvae. Zoopharmacognostics believe that animals alter their diet to fit varying ailments. If this is true,
then there are boundless medicinal practices we could learn through modeling animal behaviors. The project allowed
parasitized and unparasitized caterpillars the choice of eating a 0% or experimental diets of 25%, 50%, or 75%
hemlock diet to determine if a change in preferences would occur depending on their parasitized state. Data was
collected on 32 larvae over 20 days. A 2-sample t-test indicates (p < 0.5) that when given the choice, the parasitized
larvae (M = 34.42%, SD = 21.72) significantly ate more of the 75% hemlock diet than did the unparasitized (M =
13.66%, SD = 7.47). Choice was then eliminated from larvae to study the medicinal benefit of the control and
hemlock (50%) diet. Unparasitized and parasitized larvae’s weight changes from both diets were compared. Data was
collected on 26 larvae for 22 days. The final results for this section are still being determined, and will be re-analyzed
after ambiguous larvae are dissected to verify their parasitic state. Preliminary data indicates through a two-sample
t-test that before parasites exit, the parasitic larvae eating hemlock diet (M = -0.02g, SD =0.03) have the same weight
gain as the parasitized larvae eating control diet (M = -0.02g, SD = 0.04), t(8) = 1.423, p > 0.05. A two sample t-test
(p > 0.05) shows that there is no difference in weight gain between unparasitized larvae eating control diet (M =
0.03g, SD = 0.02) and unparasitized larvae eating hemlock diet (M = 0.02g, SD = 0.01) with current results. This data
indicates that parasitized larvae will eat more hemlock diet when given the choice over control diet, but an increase in
weight gain does not accompany the increase in uptake.
What is the molecular basis of the pink muscle fiber type of sheepshead, a teleost fish?
Mahfuza Ahktar (Dr. David Coughlin), Department of Biology, Widener University, One University Place, Chester, PA
19013
Swimming in fishes is powered by red, white and pink skeletal muscle. Slow swimming is powered by the red (or
slow-twitch muscle), fast speeds are achieved by the white (fast-twitch) muscle and pink muscle apparently serves an
intermediate function. In recent years, the physiological properties and molecular composition of red (slow) and white
(fast) muscle fibers have been well studied, while the intermediate pink muscle, which falls in a thin sheet between
the superficial red muscle and deeper white muscle, has received little attention. The goal of this study is to
determine the molecular basis of pink muscle in a fish, sheepshead (Archosargus probatocephalus). Since myosin
heavy chain (MHC) is the most important structural protein found in the muscle fibers, the first objective of this work is
to characterize MHC expression in the three muscle fiber types of sheepshead. This is being done using protein gel
electrophoresis (SDS-PAGE) and immunoblotting (Western blots). PAGE gels suggest that white and pink muscle
have similar patterns of MHC composition and are distinct from the red muscle. Western blots employing anti-myosin
antibodies also suggest that the three muscle fibers differ in MHC isoforms, although identifying specific antibodies
that distinguish each fiber type in sheepshead has proven more difficult than in other species. Further research will
be conducted to find antibodies that show high affinity for fast vs. slow MHC isoforms to help us clarify our
understanding of the MHC composition of the three muscle fiber types of sheepshead.
56
BIOLOGY
Antimutagenic and Antioxidative Testings of Selected Plant Polyphenols
Christine Hauther and Patrudu S. Makena (Dr. King-Thom Chung), Department of Biology, The University of
Memphis, Memphis, Tennessee 38152
Plant polyphenols have been reported to have antioxidative, antimutagenic and anticarcinogenic properties. Green
tea, fruits, and vegetables are great sources of these plant antioxidants. By reducing the production of reactive
oxygen species (ROS) through the consumption of these polyphenols, the amount of DNA damage caused by known
mutagenic compounds would be significantly reduced. We plan to test the antimutagenic capabilities of some plant
polyphenols such as: gallic acid, propyl gallate, ellagic acid, (-)-gallochatechin, (-)-rhein, (-)-epigallochatechin gallate,
(-)-gallochatechin gallate, and (-)-epechatechin against suspected mutagenic tryptophan metabolites
(3-hydroxykynurenine and kynurenine). A ROS sensitive tester strain of Salmonella typhimurium TA102, will be used
in the Ames Salmonella/microsomal mutagenicity assay. Different doses of the polyphenols (1&#956;g, 10&#956;g,
50&#956;g and 100&#956;g) will be used and antimutagenic potentials will be calculated. The polyphenols will be
ranked according to whether they are: a great antimutagen, a fair antimutagen, a poor antimutagen, not an
antimutagen or a mutagen. Antioxidative activities of these polyphenols will also be assessed. We will then compare
the antimutagenic potentials with the antioxidative activities of the polyphenols. We will attempt to find the
relationships between the antimutagenic and antioxidative potentials. This approach may shed a light on the possible
usefulness of these polyphenols as preventative agents in the fight against cancer.
Monitoring the Outcome of Habitat Creation the Spotted Turtle, Clemmys guttata, a Species of
Special Concern in Massachusetts
Kerry A. Muldoon and Ian M. Francis (John C. Jahoda), Department of Biology, Bridgewater State College,
Bridgewater, Massachusetts, 02325
The spotted turtle, Clemmys guttata, is considered a species of special concern in Massachusetts, due to its
drastically declining population. Causes include capture for pets, predation, road kill, exploitation, and habitat loss.
The research goal was to study the effectiveness of a newly created nesting and aestivation habitat and to determine
if the area is being used. This is part of a five year study which is approved by the Massachusetts Natural Heritage
and Endangered Species Program and funded by Waste Management Corporation. Our objectives were to visibly
locate any gravid females, individually mark them by filing the shell, and track them, using a bobbin, to their possible
nesting location. Sixty days after the first eggs were laid, careful observations were made to locate hatchlings and
return them to the nearby wetland. At the end of the first year, fourteen adult turtles (five females) were marked and
released and all females were located in the nesting area. Two hatchlings were also recorded, which confirmed the
success of this created habitat for nesting. At the end of year two, ten new turtles were found (eight females) in both
the nesting area and aquatic traps. Predators were also found in the area, which negatively impacted hatchling
success. The long-term goal is to better understand the parameters needed to successfully create habitat for the
spotted turtle population with the hope that one day the outlook for this species will improve so that it will not be
considered a special concern in Massachusetts.
57
BIOLOGY
DIFFERENTIAL BAX AND BCL-2 PROTEIN EXPRESSION AND LOCALIZATION IN TGFa &#61472;AND
NORMAL MOUSE MAMMARY GLANDS AT DAY FOUR INVOLUTION
Drew Scribner (Dr. Teresa Rose-Hellekant), Department of Physiology and Pharmacology, University of
Minnesota-Duluth, Duluth, Minnesota 55812
Mice engineered to express transforming growth factor alpha (TGFa) in
the mammary gland have developmental abnormalities and acquire mammary
tumors. As involution requires program cell death signals to function normally, we tested the hypothesis that two
proteins known to regulate apoptosis, bcl-2 and bax, were differentially expressed and localized during involution in
TGFa versus normal mice. Bcl-2 and bax are anti-apoptotic and pro-apoptotic proteins respectively whose ratio
determines whether a cell will live
or go through apoptosis. Comparing the ratio of these two proteins in TGFa&#61472;and normal mice may lead to a
better understanding of the mechanisms which allow cells to avoid cell death signals. Assays were carried out in day
four involuting mammary glands collected from TGFa versus normal mice. First we quantified the number of
pyknotic cells, an indicator of cell death. In addition, we measured bcl-2 and bax expression immunohistochemically
and through Real Time-Polymerase Chain Reaction (RT-PCR). We found that bax was located in myoepithelial cells
in TGFa mammary glands and absent immunohistochemically in normal mammary glands. Evaluations of Bcl-2
staining are underway. Using quantitative real-time PCR (QPCR) analysis, we found that the ratio of bcl-2 to bax gene
expression didn’t differ in whole mammary gland extracts from TGFa vs. normal mice. In summary, we found that (1)
involution was significantly delayed in TGF&#61537; mice and apoptotic levels were lower than in normal mice, (2)
bax protein was detected in myoepithelial cells in TGF&#61537; glands (3) myoepithelial cells weren’t present and
bax protein couldn’t be detected in normal glands, and (4) bcl-2/bax mRNA levels in whole mammary extracts was
unchanged in TGFa mice compared to normal mice even though the bcl-2 containing myoepithelial cell population
was absent in normal glands. To shed more light on the involvement of bcl-2 and bax in reduced program cell death
associated with involution observed TGFa mice, we are currently determining bcl-2/bax protein levels in whole
mammary extracts.
Identification Key to the Scale Patterns of Guard Hairs of Indigenous Pennsylvania Mammals
Melissa Cicchella, Kellie L. Kremser, and (Carlos A. Iudica)
Mammals can be identified based on several elements, such as cranial and postcranial morphology, dentition, and
behavior, among others. Scale patterns on guard hairs are a distinguishing feature among mammals that have been
used only sporadically over the years. We noticed the current need of a standardized identification key based on
guard hairs for the mammals of PA. Guard hairs were obtained from various sources, such as road kills, donations
from trappers, hunters, veterinarians, and the mammal collection from The State Museum of Pennsylvania in
Harrisburg. The hairs were cleaned with ethanol for the preparation of their scale casts. Casts were made using
smears of corrector fluid on glass slides. Once the casts were finished, a photograph of each one was taken and
those photographs were compiled into a master database. Each file was then compared, and based on the patterns
of the scales we grouped the pictures. The final step was organizing the pictures of the scale casts into a
dichotomous identification key. We envision our final product as a key that could be used as a practical tool for both,
the amateur and the professional naturalist in PA.
58
BIOLOGY
RALOXIFENE DOSE REPONSE ON DEVELOPING ZEBRAFISH (DANIO RERIO)
Stephanie Mills (DR. James Turner), Biology Department, VMI, Lexington, VA 24450
Zebrafish have long been used in studies testing the effects of hormone replacement therapy on the nervous system.
Raloxifene itself is a SERM (a Selective Estrogen Receptor Modulator) that does not cause as many physical
complications when used in lieu of Estrogen for hormone replacement therapy. The use of Estrogen on zebrafish has
show positive effects on the developing nervous system as shown in previous studies. This experiment implements
the same methods to test if Raloxifene has any similar affects on the nervous system. I tested different
concentrations of Raloxifene on the zebrafish by exposing eggs in solutions as early as 24 hours after fertilization. By
doing this the embryos would receive certain amounts of Raloxifene at their earliest and most crucial stages of
nervous system development. All the experiments were run with double controls that used Estrogen, and regular egg
rearing solution to compare results. First I had to test the concentrations bases upon physiological and
pharmacological levels of estrogen, because of varying degrees of toxicity to the fish. The pharmacological
concentrations that exceeded 10-6 M proved to be toxic while the fish were able to survive in lower ones. This was
mirrored in the concentrations used with Raloxifene. Repeating this experiment without the toxic levels showed
discrepancies between the developments of fish exposed to Raloxifene versus Estrogen, and the regular egg rearing
solution. This was specific to ocular maturation, spinal curvature, cranial vault size and relative hyperactivity shown in
early stages of development. All of these differences still need to be examined further, but it leads to the idea that
there are significant differences in the molecular pathways and how Raloxifene affects the nervous system compared
to Estrogen.
ROLE OF LARGE HERBIVORES IN SPREADING NON-NATIVE INVASIVE PLANTS INTO NATURAL
AREAS OF CALIFORNIA
Monika Alas, Mark Orcholski, Alma Martinez, Desaree Williams (Bonnie Davis, Sibdas Ghosh, and Mietek Kolipinski),
Department of Natural Sciences and Mathematics, Dominican University of California, San Rafael, CA 94901;
National Park Service, Pacific West
There have long been comments, assumptions, hypotheses and anecdotal information that some seeds viably pass
through the horse gut. Once in horse manure, they ultimately sprout, grow and germinate. Supposedly, some of
which could be noxious or invasive weeds. We did not find any conclusive literature references to any scientific
programs that validate the long held hypothesis about noxious or invasive weeds that may come from horse manure.
Thus, Dominican University of California in collaboration with National Park Service investigated the role horse
manure on environmental spread of noxious/invasive weeds in natural areas of California. Forty five samples were
collected from selected locations throughout the San Francisco Bay Area. Each sample was divided into halves: one
of which was allowed to air dry while the other was refrigerated. After one week, both dry and wet samples were
weighed and planted individually in sterile 4 inch pots containing weed free soil. A random number selection process
for pot locations in trays was computer generated along with 10 control samples which contained no horse manure.
Of ninety pots, 34 plants germinated in 21 pots of which 12 represent dry and 9 represents wet samples respectively.
We report that toad rush (Juncus bufonius), Italian ryegrass (Lolium multiflorum), bur-clover (Medicago polymorpha),
summer mustard (Hirschfeldia incana), cudweed (Gnaphalium luteoalbum), purslane (Portulaca oleracea), dooryard
knotweed (Polygonum aviculare), cheeseweed (Malva niceaeensis). Although some of these plants are nonnative to
California, none are listed as CDFA (California Department of Food and Agriculture) Noxious Weed.
59
BIOLOGY
INVESTIGATION OF CD8+ T CELL CYTOTOXICITY IN CENTRAL NERVOUS SYSTEM CELLS
Sherry Kathleen Pittman (Dr. Karen Aguirre), Department of Biology, Coastal Carolina University, Conway, South
Carolina 29526
Immune response to pathogens in the central nervous system (CNS) differs from response in the periphery. CNS
immunity has been studied in viral systems, where pathogens interfere with normal host gene expression, but many
CNS infections involve intracellular pathogens such as bacteria and fungi that do not shut down host gene expression.
In these infections, the host cell may deploy antimicrobial weapons not available to the virus-laden macrophage.
Outside the CNS, CD8+ T cells typically lyse virus-infected macrophages and other cells by cellular cytoxicity (CTL).
The intent of this project is to determine whether CTL occurs when CD8+ T cells interact with brain macrophages
(microglial cells) laden with non-viral pathogens. Two cells lines, TK1 (a CD8+ T cell clone) and EOC (brain
macrophages), will be used in the study. EOC cells will be activated to ingest blue fluorescent beads. Subsequently,
TK1 cells will be incubated with microglia, capsular polysaccharide of Cryptococcus neoformans, a fungal pathogen,
and staphylococcal enterotoxin, (SEA), a superantigen which stablizes microglia/T cell interactions. If CTL occurs,
beads will be released from the lysed macrophages. Three experimental conditions will be investigated: co-culture
with fluorescent beads, co-culture with fluorescent beads and antigens, and co-culture with beads, antigen, and
antibody that neutralizes FASL. FASL is a protein required in the signal cascade attending CTL. The different setups
will then be analyzed by a fluorometer The amount of fluorescence liberated will be determined, with extensive
fluorescence indicating that T cells do in fact lyse microglia infected with fungi. In the event that CTL does not occur,
further investigation of other mechanisms of CD8+ T cell-mediated clearance of cryptococci will be indicated. This
experimental approach has never been utilized for CNS-derived cells. My research will increase knowledge of
pertaining to CD8+ T cell cytotoxic response to microglial cells laden with medically-important non-viral pathogens.
CULTURABLE HETEROTROPHIC BACTERIAL DIVERSITY BETWEEN THREE FORESTED
ECOSYSTEMS AS DETERMINED BY TWO NUTRIENT-LIMITED MEDIA
Emily A. York (Dr. Sean O’Connell), Department of Biology, Western Carolina University, Cullowhee, North Carolina
28723
Some bacteria from soils can survive with extremely limited nutrients, even though most soils have a high organic
carbon content. Studies have revealed great diversity in DNA sequence heterogeneity from soil, however only a few
oligotrophic bacteria from most phyla have been cultivated. Soils were sampled from three forested plots in Great
Smoky Mountains National Park; Albright Grove (old growth), Purchase Knob (second growth impacted by logging),
and Cataloochee (second growth impacted by Chestnut Blight). Samples were spread onto nutrient plates (DDNB;
0.008g nutrient broth/L) after 100-millionfold dilutions in sterile water were made and incubated for three months
(Experiment 1) or frozen soils were revived after three months (Experiment 2). Twenty-two and 23 species from the
first and second experiments, respectively, were isolated and identified using DNA sequences of their 16S rDNA
following PCR. The isolates were from six phyla of bacteria. In the first experiment, Albright Grove was dominated
by Firmicutes while the two disturbed sites were dominated by the Proteobacteria (?, ?, ?); all sites had
Cytophagales/Flavobacterium/Bacteroides (CFB) species. In Experiment 2, Albright Grove yielded only Firmicutes,
while Cataloochee was co-dominated by Firmicutes and Actinobacteria; Purchase Knob displayed the greatest
phylum evenness. Incubation in nutrient-deprived, aqueous conditions selected against Actinobacteria and for the
CFB group. Samples that were frozen and then plated onto DDNB selected for Firmicutes and Actinobacteria and
against CFB species. The greatest number of phyla encountered was in Cataloochee in Experiment 2 and the lowest
in Albright Grove from the same experiment. All species from Experiment 1 were more closely related to known
species (>90% similar based on DNA sequence) than in Experiment 2 where nine isolates were only 70-90% similar.
In order to assess diversity in a sample, it is clearly best to use multiple techniques; however, each method employed
in this study indicated that there was a difference between the microbial communities from old growth versus
disturbed forested ecosystems.
60
BIOLOGY
PHYLOGENETIC RELATIONSHIPS WITHIN THE ACHLOROPHYLLOUS FLOWERING PLANT GENUS
BARTONIA (GENTIANACEAE)
Emily York (Dr. Katherine Mathews), Department of Biology, Western Carolina University, Cullowhee, North Carolina
28723
Bartonia is a member of the flowering plant family Gentianaceae. It contains three species native to eastern North
America and one Texas endemic species. Some members of the Gentianaceae family are highly dependent on
mycorrhizal fungi, including Bartonia, which has very small, scale-like leaves and purplish stems, and its putative
sister genus Obolaria, which has reduced purplish leaves. The dependency of these plants on mycorrhizae and their
inability to produce sufficient amounts of chlorophyll suggest that these plants may be parasitic. The object of this
study is to determine the evolutionary relationships of three species of Bartonia: B. paniculata, B. virginica, and B.
verna, using sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (~730 bp). Based on
morphological characteristics, B. paniculata and B. virginica are hypothesized to be the most closely related. Due to
the genetic variability of the ITS regions, it is often used to determine the phylogeny of closely related species. ITS
sequences for B. paniculata and B. verna were obtained in the lab via total DNA extraction from herbarium specimens
and PCR amplification of the ITS region, while the ITS sequence of B. virginica was obtained from the NCBI
GenBank database. Sequences of Obolaria virginica and Gentianella quinquefolia, another closely-related member of
Gentianaceae, were used as outgroup samples for phylogenetic analysis. Findings of relationships among the
Bartonia species will be discussed as well as the utility of ITS for resolving relationships in this genus.
Analysis of Ubx function in crickets
Jennifer Stone, Undergraduate Research Dept, Biological Sciences, Wayne State University, 5155 Gullen Mall,
Detroit, MI 48202
The arthropods have a special place in the animal kingdom due to their great morphological diversity. They achieve
this by possessing a segmented body plan that is also highly modularized. Among the four major arthropod lineages
(insects, crustaceans, myriapods, and chelicerates), the insects exhibit the widest range of morphological
diversification. One of the key aspects of insect evolution is differential enlargement of hind legs. Previous research
in our lab revealed that this process is strongly associated with the presence of the homeotic gene Ultrabithorax (Ubx)
in hind leg segments. Based on these findings, we proposed that Ubx controls the differential enlargement of hind
legs in insects.
The role of Ubx was tested by carrying out a functional assay in the house cricket Acheta domesticus, a species
that is characterized by greatly enlarged hind legs (T3 legs). Functional assay was performed using a highly effective
technique known as RNA interference (RNAi), which inhibits Ubx function in developing embryos. Our analysis show
that the size of embryonic hind legs in Ubx-RNAi embryos is indeed greatly reduced when compared to corresponding
wildtype appendages. For example, between the wildtype and Ubx- crickets, there is approximately 60% and 50%
reduction in the size of femur and tibia, respectively. These findings show that Ubx is indeed required for differential
enlargement of hind legs in insects.
61
BIOLOGY
RESOURCE KIT FOR MANAGEMENT OF NON-NATIVE PLANTS IN NATIONAL PARKS OF
CALIFORNIA
Monika Foster, Asta Haman-Dicko, Cynthia Brooks, Monika Alas (Lauren Quinn, Sibdas Ghosh and Mietek
Kolipinski), Department of Natural Sciences and Mathematics, Dominican University of California, San Rafael, CA
94901; National Park Service, Pacific West R
Introduction of non-native plants is detrimental to ecosystems. Once established, invasive exotic plants alter native
habitats and populations of indigenous species. Eradication is costly and often impossible. Thus, it is best to prevent
invasions. Dominican University of California and National Park Service (NPS), have investigated sources and
dispersal methods of non-native plants into new habitats. We have synthesized information pertaining to a California
weed free feed and mulch initiative. As education is integral to this project, we in partnership with other federal and
state agencies, such US Forest Service, Bureau of Land Management, and California Department of Food and
Agriculture, have developed a prototype resource kit for use in California`s National Parks. The kit serves as a basis
for educating NPS employees and concessionaires as well as the public about use of best management practices to
prevent invasion and spread of non-native species. Components of the kit include: information about the NPS Units, a
list of non-native plants for each park, and an explanation of how weeds spread and negatively impact natural
habitats and landscapes. In addition, effective practices and procedures to prevent infestations will be illustrated in
the kit. The goal is to change existing practices, associated with stock feed, mulches, soils, and by human activities
within and around California NPS Units that inadvertently result in establishment of non-native plants.
A DOMINANT METHANOGEN INHABITS THE DIGESTIVE TRACTS OF ELK FROM GREAT SMOKY
MOUNTAINS NATIONAL PARK
Amanda L. Buchanan (Dr. Sean O’Connell), Department of Biology, Western Carolina University, Cullowhee, North
Carolina 28723
Ruminants require microbial symbionts within their digestive systems to provide them with nutrition from the plants
they consume. Elk fecal pellets were collected from eleven individuals in Great Smoky Mountains National Park over
10 months. These elk are part of an experimental herd identifiable by numbered tags; they represent unique
opportunities for repeated sampling of individuals from a wild ruminant population. Pellets from two males, #2 and
#67, were subjected to DNA extraction, PCR using archaea-specific 16S rDNA primers, and denaturing gradient gel
electrophoresis (DGGE). DGGE is used to distinguish species based on melting DNA and provides a measurement
of species richness and evenness. Preliminary results indicated that some differences in elk could be observed
based on season and individuals sampled. However, DGGE banding patterns were faint and PCR and DGGE
methods were optimized. A nested PCR approach manipulating magnesium concentration and annealing
temperature yielded higher quality PCR products that were better suited for DGGE screening of the archaeal
communities. One dominant DGGE band was observed for both animals and was present in all twelve samples; DNA
sequence analysis revealed this species to be closely related to Methanobrevibacter spp. associated with sheep
rumen in Australia. Two other species were detected in the DGGE analyses, with one band occurring only for spring
samples (also related to Methanobrevibacter spp. from Australia), while two less distinct bands were observed in all
samples at all times. These species also were aligned with Methanobrevibacter spp., with one band the same
sequence as the dominant species and the other a unique species most closely related to free-living methanogens
and methanogens endosymbiotic with protozoa from sheep rumens. The low diversity but ubiquitous nature of
methanogens in elk indicate their importance in the digestive processes of these animals. Their relatedness to
archaea in other ruminants deserves further study including more sequence analyses as well as attempts at
cultivation to understand their role in the elk rumen.
62
BIOLOGY
BLOCKADE OF SEROTONIN (5-HT) 2A, 2B, 2C RECEPTORS IN MAMMALIAN SPINAL CORDS
Katie Asp (Dr. Bradley Seebach), Department of Biology, The University of Wisconsin at La Crosse, La Crosse,
Wisconsin 54601
The purpose of this research is to determine the role of the serotonin 2 (5-HT2) class of receptors in generating an
alternating bilateral rhythm. Common lab rats from date of birth (P0) to postnatal day 5 (P5) are dissected for
removal of the spinal cord. The spinal cord is immersed in a cerebrospinal like saline solutions. Suction electrodes
are attached to a pair of ventral lumbar roots L2 through L5 to record the electrical activity of action potentials.
Neurons are stimulated via the application of 50uM serotonin. To differentiate the three 5-HT2 receptor types, an
antagonist, SB206553, was applied to block the effects of 2B and 2C receptors. The SB compound is added to the
saline wash to establish a receptor blockade. Following the blockade, a wash with 50uM serotonin and varying
amounts of SB are applied to the spinal cord. The data sets of serotonin application can be compared to the data of
serotonin and SB to determine if SB caused any alteration in the bilateral rhythm. Accrued data suggests a possible
organizing effect of the SB compound. Of the nine successful trials using SB, seven trials showed an organizing
affect with the application of either 50nm or 250nm SB. This organization was shown in stable rhythmic burst patterns
or in stable alternating patterns of the central pattern generator. Two trials with SB concentrations at 250 nm did not
show organization and appeared similar to the disorganized serotonin patterns. Four trials showed alternating
rhythms with serotonin alone and five trials displayed alternating bursts with the application of SB in both the 50 and
250 nm concentrations. With the blockade of 2B and 2C receptors, the basic pattern of the central pattern generator
was not significantly altered in frequency or amplitude. The 2B and 2C receptors do not appear to possess a pivotal
role in generating the alternating bilateral rhythm.
Analyzing C. Elegans Lacking Sia-1 Gene Function
Megan Theede, Caroline Martin, Jodie Deshler, and Scott Ballantyne , Biology Department, University of
Wisconsin-River Falls, 410 South Third Street River Falls, WI 54022-5001
Seven-in-absentia (sina) is a gene required for male fertility in mice (M. muluscus) and eye development in fruit
flies(D.melanogaster). The worm C. elegans is a powerful model organism for studying animal behavior and
development. Worms contain a gene, sia-1, that is homologous to sina. We have eliminated sia-1 gene function in
C. elegans. Here we describe the results of our experiments analyzing worms that lack sia-1.
C. elegans exist as either self-fertile hermaphrodites or males. We have determined whether sia-1 disruption altered
hermaphrodite fertility. We found that hermaphrodite worms lacking sia-1 remained fertile and had brood sizes
comparable to control worms. In mice the sina gene homolog only affects male fertility. We asked whether this is
also true in worms. We found that eliminating sia-1 gene function from males did not cause sterility. We conclude
that sia-1 is not required for worm reproduction and thus is functionally distinct from the mouse gene.
Worms may not have eyes – but they do have senses and sensory neurons. DIO is a fluorescent dye that specifically
stains a subset of active worm sensory neurons. We asked whether sia-1 disruption alters the DIO staining seen in
normal worms. Initial experiments revealed fewer DIO stained neurons in worms lacking sia-1. This result was less
dramatic in the second attempt and absent in the third. We have identified a potential explanation for this variablity
and are testing whether it is correct. Our hope is that this experiment will shed-light on whether sia-1 has an effect on
sensory neurons found in C. elegans.
63
BIOLOGY
RECOVERY OF NEURONS AND DENTATE GYRUS VOLUME IN ALZHEIMER’S APP TRANSGENIC
MICE BY MUTATION OF ASP664
Gurleen Sidhu, Danielle Crippen and Surita Banwait (Vernica Galvan ), Department of Natual Sciences and
Mathematics, Dominican University of California, San Rafael, CA 94901; Buck Institute, Novato, CA 94945
Alzheimer’s disease currently affects 4.5 million American citizens, but there is yet to be a cure for this detrimental
disease. The characteristics of Alzheimer’s disease (AD) are believed to partly result from the toxic effects of
ß-amyloid peptides, a set of amino acid fragments derived from the ß-amyloid precursor protein (APP). Recently it
was shown that APP is also cleaved intracytoplasmically at Asp664. However, whether this intracellular processing of
APP plays a role in the Alzheimer’s disease is unknown. Therefore, we compared dentate gyral atrophy and neuronal
degeneration in transgenic mice modelling AD that have or do not have a functional Asp664 cleavage site. We
found that dentate gyral atrophy and neuronal degeneration were recovered in animals carrying a mutation that
obliterated the cleavage site at Asp664. These results suggest that cleavage of APP at Asp664 plays a key role in the
generation of AD-like pathogenic changes in human APP transgenic mice
Electrophysiological Properties of the Girk2a and Girk2c Ion Channel
Kristin R. Mattox, (Dr. Paul Slesinger and the Phelps Scholarship), Peptide Biology, The Salk Institute of Biological
Sciences, 10010 North Torrey Pines Road, La Jolla, CA 92037
Potassium channels are critical protein complexes found in all organisms. A better understanding of their structures
and mechanics in nerve cells could lead to clearer insight into the function of the brain, neurological disorders such as
Andersens syndrome and drug addiction. This study focused on the physical properties of the Girk2a and Girk2c
potassium channels. My goal was to identify the electrical currents associated with Girk2a and Girk2c in various
physiological conditions and in the presence of a newly discovered channel binding protein, Mrt1b. Initially, the
Girk2a, Girk2c, and Mrt1b genes were cloned into the pGEMHE expression vector. From these constructs
messenger-RNA was synthesized. Oocytes were surgically removed from Xenopus laevis and injected with mRNA.
When the genes had expressed, each oocyte maintained a specific channel, the binding protein (when appropriate)
and various receptors needed to induce electrical currents. The basal and induced electrical currents through the
potassium channels were recorded using a Two Electrode Voltage Clamp. From the data analysis it was clear that
both the Girk2a and Girk2c channels were inducible under excitatory conditions, allowing electrical current to flow
through. More research is needed to determine if the Mrt1b binding protein modulates either channel. Future
experiments would entail increasing the concentration of Mrt1b and testing for endogenous binding proteins that could
affect the currents.
Exploration of Varying Phytochemical Concentrations on the Feeding Behavior of Gypsy Moth
Larvae, Lymantria dispar (L.)
Kristen P. Smith and Nicole S. Arnold, (Dr. Vonnie Shields), Department of Biological Sciences, Towson University,
Towson, MD 21252
The gypsy moth, Lymantria dispar, is one of the major defoliators of forest habitats in North America. These
polyphagous larvae feed on the foliage of hundreds of plants, but prefer leaves from deciduous hardwood trees, such
as sweet gum, maple, and oak. In this study, we investigated the feeding behavior of gypsy moth larvae to various
plant phytochemicals using feeding choice test bioassays. Plants contain a variety of phytochemicals, some of which
are insect feeding deterrents. Previously, Shields et al., (2003) established a feeding preference hierarchy and found
that red oak leaves were preferred significantly in fifth instar gypsy moth larvae. In this study, we tested the effects of
several phytochemicals, including a glycoside (e.g. salicin), terpenoid (e.g. picrotoxin), coumarin (e.g.
7-hydroxycoumarin), flavonoid (e.g. morin) and multiple alkaloids (e.g. caffeine, strychnine, sparteine, nicotine). We
tested these phytochemicals using six concentrations spanning four logarithmetic steps. We detected feeding
deterrent threshold effects for some of these phytochemicals at 0.1 mM and 1 mM. We observed increasing
dose-dependent deterrent effects for all of the phytochemicals tested. Our results suggest that the phytochemicals
used in this study are potentially useful in designing crop protection strategies from insect pests and improve our
understanding of insect feeding behavior.
Supported by a Bridges NIH grant 5R25GM058264-03 and NIH grant 1 R15 DC007609-01 to V.S.
64
BIOLOGY
ANTIMICROBIAL PROPERTIES OF A RECOMBINANT PROTEIN FROM THE BLACK LY, SIMULIUM
VITTATUM.
Rory D. Applegate, (Elizabeth M. Whitley, DVM, PhD), Department of Pathobiology, Auburn University, Auburn, AL
36849
A major concern in the medical profession is the successful treatment of wounds. A common barrier to successful
wound healing is infection caused by bacteria growing in the wound bed. Traditionally, wound infections have been
treated using synthetic antibiotics, but as infectious organisms increasingly have developed resistance to antibiotic
drugs, novel methods of killing bacteria that contaminate wounded tissues are urgently needed. Our research is
centered on evaluating a recombinant protein derived from the saliva of the Simulium vittatum for potential
antimicrobial properties. To examine the direct effects of the protein on bacterial growth, we have evaluated the in
vitro growth rate of cultures of Escherichia coli with the addition of varying concentrations of the protein. Current data
indicates that there is no effect of the protein on the growth curve of E. coli in culture. To evaluate for potential effects
of the protein on phagocytosis and killing of bacteria by macrophages, an important cell type involved in the innate
immune response to wounds, we have compared phagocytosis of FITC-labeled microbeads by cells of the RAW264.7
mouse macrophage cell line cultured with and without the protein using flow cytometry. Results indicate that the
protein does not have an effect on the phagocytosis of FITC-labeled microbeads by RAW264.7 cells. Effects of the
recombinant protein on the phagocytosis and killing of E. coli by RAW264.7 macrophages currently are being
evaluated.
DECAY DYNAMICS OF NYSTATIN CHANNELS IN THE PRESENCE OF STEROL MICROSTRUCTURES
ON MEMBRANES
Benjamin L. Yoder (Dr. Carl S. Helrich), Department of Physics, Goshen College, Goshen, IN 46526
Nystatin (nys) is an antifungal agent that preferentially forms ion channels in membranes containing the sterol
ergosterol (erg). The structure of the nystatin channel is not clear, although it is clear that multiple nystatin monomers
require a sterol-rich membrane for aggregation. When nys/erg containing vesicles are fused to a sterol-free
membrane, characteristic spike changes in membrane conductance are observed. The decay of a conductance spike
is generally step-wise linear and the time of decay is a strong function of [erg]. These data indicate interdependence
among the nys/erg channels. A model has been developed in which nys channels form at the boundary of a lipid/erg
superlattice rafts (SLR) and channel decay is determined by erg diffusion from the SLR. This model was tested using
Monte Carlo (MC) simulations for the diffusion current from the SLR. The MC simulations predict a constant diffusion
current from the SLR which results in a rate of loss of channels which is linear in time. The steps in the decay are
understood in terms of multiple SLRs and SLRs with fissions. The model then provides a complete understanding of
the decay scheme observed experimentally. This interpretation also predicts previously confusing data relating
conductance spike height to vesicle diameter.
65
BIOLOGY
ESTROGEN INDUCED SEXUAL ABNORMALITIES: A GENETIC MODEL FOR ASSESSING
ESTROGENIC POLLUTANTS IN THE ENVIRONMENT
Kendra K. Scudder (E. Katherine Miller, Ph.D.) Dept. of Biology, , University of Wisconsin-River Falls, River Falls,
Wisconsin 54022
Residual pharmaceuticals and industrial chemicals found in natural water systems and public water sources may
exhibit toxic effects on humans. Among these, estrogenic compounds are causing increasing concern due to effects
they are having on wild populations. In order to understand the effects of estrogenic exposure on complex organisms,
it is necessary to begin by studying the effects on a more simple system. The present study addresses the question
by assessing the multi-generation effects of two estrogenics, bisphenol A (BisA) and 17-beta-estradiol (E2), on the
him-5 strain of the model organism Caenorhabditis elegans. Fecundity, reproduction, and physical abnormalities are
used as toxicity endpoints. Initial data show that exposure of C. elegans to BisA decreases egg production relative to
the increasing concentration of BisA and the generational time of exposure. For first generation exposed C. elegans,
treatment effects were statistically significant at the p< 0.05 level (p-value = 0.019). For fourth generation exposed C.
elegans, treatment effects were statistically significant at the p< 0.10 level (p-value = 0.066). Several developmental
abnormalities also appear, including a protruding vulva and a retarded growth period for exposed larva. The most
significant of the findings, however, was the change in male to hermaphrodite population ratios, dropping from
45-50% male to as low as 9% male. This suggests that BisA may have caused some degree of infertility in the
population. Studies using E2 are currently in progress. RNAi technology is used to “knock-out” selected genes in an
effort to understand the mechanism by which these effects are caused. In understanding how they are caused,
science can address the relation and impact on humans more effectively.
DECREASED PERMEABILITY IN SOYBEAN CYST NEMATODE EGGS TREATED WITH
AZETIDINE-2-CARBOXYLIC ACID
Ravi M. Pujara (Dr. Paul M. Tefft) Biology Department, Saint Joseph’s University, Philadelphia Pennsylvania 19131
The soybean cyst nematode (SCN), Heterodera glycines, is an economically important pest of the soybean crop
in the United States causing up to 1.5 billion dollars worth of damage annually. SCN hatching can be affected by host
plant physiology. Azetidine-2-carboxylic acid (A-2-C), a compound found in Lilly of the Valley, Convallaria majalis, is
a chemical analog to the amino acid proline. A-2-C substitutes for proline during translation making the formed
protein non-functional, thereby providing protection to the plant from herbivory and parasitism. In a previous study this
analog was found to decrease hatching. Before hatching, SCN eggs undergo a significant permeability change. One
purpose of our study was to determine which of a selected group of ten histological stains would be the best for
determining this eggshell change following exposure to the hatching stimulant zinc chloride. The uptake of the stain
Nile Blue correlated well to this permeability change. The staining of the eggs also correlated well to hatching and
saved time when compared to hatching assays. The stain technique required only a few days for completion rather
than two weeks normally required for a hatching study. The second purpose of the study was to determine if A-2-C
inhibited egg hatching by blocking permeability or by causing detrimental effects later in the hatching process. In our
experiment, we used Nile Blue as an indicator of permeability and ran hatching studies as controls. Eggs stimulated to
hatch with zinc chloride and treated with A-2-C had a significant decrease in permeability (30%) compared to eggs
treated in the zinc chloride stimulant alone. These results suggest that protein synthesis maybe required for the
permeability changes that occur prior to hatching.
66
BIOLOGY
Anthocyanin accumulation and photoprotection in the evergreen understory herb, Galax urceolata
Evan Crutchfield (Dr. Jonathan Horton), Biology, UNC-Asheville, One University Heights, Asheville, NC 28804
Galax urceolata is an evergreen understory herb found throughout the southern Appalachian Mountains, and is
often harvested for its aesthetically pleasing leaves for use in floral arrangements. Leaves exposed to high light
during the winter buildup anthocyanin pigment molecules, the same pigments responsible for some fall colors in
senescing leaves and many red fruits. The function of anthocyanins is thought to be that of protection from oxidative
damage from excess light energy. These pigments serve as light attenuators to absorb excess blue-green light that
can not be converted to chemical energy in the cold temperature and high light environments experienced over
winter. The purpose of our research will be to determine the effects that anthocyanins have on the photochemistry in
Galax over the winter months and then the resulting carbon gain in the following spring. We hypothesize that
anthocyanin pigments protect the photosynthetic biochemistry in the leaves over the course of the winter season and
allow for greater photosynthetic capacity in the subsequent spring and summer. Our research will be conducted on
the Blue Ridge Parkway near Mt. Pisgah where there are many populations of Galax urceolata in the forest
understory. Three separate plots with six treatments each have been established. Within each plot, treatments will be
subjected to different light and shade treatments that create a mixed population of green and red leaves. We will
monitor fluorescence and photosynthetic parameters at least once a month from November to May (and possibly
longer) to document any photochemical damage over winter and resulting decreases in carbon gain in the following
growing season. Overall we expect our results to show that anthocyanins do serve a protective role in Galax
urceolata when exposed to adverse conditions allowing for higher photosynthetic carbon gain in the following season.
SIMULATING CARDIAC PACEMAKERS USING R15, EPSTEIN-MARDER (EM), AND MORRIS-LECAR
BURSTING NEURON MODELS
Marc A. Miller, (Dr. James J. Watrous), Department of Biology, Saint Joseph’s University, 5600 City Avenue,
Philadelphia, PA 19131
The purpose of the study was to determine which of three bursting neurons best simulated cardiac pacemaker
activity. The properties of pacemaker cells are determined by a minimum of three voltage dependent ion channels:
calcium, potassium and sodium. Depending on the properties of these channels the cells either fire individual or
multiple action potentials. Multiple action potentials or a volley are referred to as a burst. The bursting neurons
examined were the R15, Morris-Lecar, and Epstein-Marder (EM) cells. These neurons were observed alone and then
imported into a network comprised of a bursting neuron synapsed solely to a feeder cell from which a signal
progressed to the eight-cell network of Hodgkin-Huxley (HH) type cells. The kinetics of the ion pools and calcium
channels of the Epstein-Marder and the R15 were similar to one another but different than those of the Morris-Lecar.
This difference explains the ability of the R15 and the EM cells to burst without any external impulse which is required
by the Morris-Lecar model. We concluded from our results that the R15 cell was the best pacemaker model in this
experiment due to its stability in the network and ability to generate 1 HH action potential for every R15 action
potential. It also produced uniform action potential amplitude throughout the network. The Epstein-Marder model
cannot accomplish either of these properties the R15 exhibits. Work continues in observing how altering the
properties of the R15 effects larger cell networks.
67
BIOLOGY
Pak-1 Expression in Mammary Carcinogenesis induced by 7,12 dimethylbenz[a]anthracene
MICHAEL DONAWORTH, JONATHAN KOOP, CAITLIN MAYNARD, LARRY E. DOUGLASS, AND JULIA H.
CARTER, Wood Hudson Cancer Research Laboratory, Newport, KY 41071
Induction of mammary carcinomas in female rats by 7,12 dimethylbenz[&#61537;]anthracene (DMBA) is used as a
model for human breast cancer. DMBA-induced breast tumors are similar to those found in women. Factors affecting
tumor induction and growth have been studied in the DMBA model; however, cellular and molecular changes
occurring within days of carcinogen administration are not well characterized. DMBA alters tissue homeostasis in the
mammary gland within one week. Pak 1 is a serine/threonine kinase associated with cell growth, survival, and
motility. Our studies of human breast cancers linked Pak 1 with metastasis. Here we studied Pak 1 expression
during breast cancer development. Sprague Dawley rats (n=146) were sacrificed at 1 or 7 days and 1 to 6 months
after an intragastric dose of 20 mg DMBA or vehicle at 50 days of age. Some animals were pretreated with the
anti-estrogen tamoxifen before DMBA administration to determine its effect on DMBA-induced changes in mammary
glands. Some tumor bearing animals were ovariectomized to determine the effect of estrogen deprivation on Pak 1
expression in breast tumors. Tissues were fixed in formalin, and paraffin embedded. Pak 1 expression was
determined immunohistochemically in histologic sections of mammary glands and breast tumors. Pak 1 expression in
developing mammary glands was reduced by both the carcinogen DMBA and the anti-estrogen tamoxifen. Pak 1 was
expressed in breast carcinomas in intact, but not in ovariectomized, animals. These data imply that the established
role of estrogen in breast carcinogenesis may be mediated in part by Pak 1.
IDENTIFICATION OF SITES OF TISSUE FRAGILITY IN C. ELEGANS
Dannielle N. Valletta, (John Plenefisch), Department of Biological Sciences, University of Toledo, 2801 W. Bancroft
St. Toledo, Ohio 43606
The basis of this research was to identify the specific sites of muscle separation in Caenorhabditis elegans. This was
done through a series of genetic crosses using various strains of C. elegans with different gene array::GFP constructs
and mua animals. The mua animals have deficient cell attachments that result in partial or complete paralysis. The
gene arrays were complexed with green fluorescent protein (GFP), which serves as a fluorescent marker on the
specific proteins of that array. The purpose of using the different gene array::GFP animals was that it allowed
different cell-cell attachment proteins marked with GFP to be viewed using a fluorescent microscope. Some of the
various cell attachment proteins that were examined included: integrins, hemidesmosomes, and intermediate
filaments. A phalloidin stain was also used in conjunction with the GFP to view the tissue separation sites in the mua
mutants. Phalloidin is a toxin that stains the microtubules of the muscles red. Three phalloidin techniques were
tested and it was determined that the ethanol technique produced the most vibrant muscle staining without
compromising the vibrancy of the GFP. Photographs will be taken of the mua animals that have incorporated the
gene array::GFP complexes to show the specific sites of tissue separation. These photographs will clearly show the
effects of the decencies in the mua genes in C. elegans.
68
BIOLOGY
THE ROLE OF CONDUCTANCE IN LONG-TERM POTENTIATION OF A FIVE CELL NEURONAL
NETWORK
Melissa S. Oliver (Dr. James Watrous), Department of Biology, Saint Joseph’s University, Philadelphia, Pennsylvania
19131
Altering the synaptic strength of the signal sent between two neurons causes a change in the transmembrane
potential and the action potential that is produced by the postsynaptic cell. An increase in the conductance of the
pre-synaptic synapse can result in an over stimulation of the postsynaptic cell causing the cell to stop firing. A
decrease in the conductance of the pre-synaptic synapse may cause the postsynaptic cell to never reach its threshold
potential. Similarly, in a neuronal network where there is a group of interconnected neurons, an increase or decrease
in synaptic conductance of a pre-synaptic cell can cause the entire network to over stimulate and cease activity or
cause some neurons to never fire. In order to create a continuous network of neurons, the synapses between cells
need to be firing continuously as well. Long-term potentiation (LTP) is the long-lasting increase of strength in the
connection between nerve cells. To simulate the role that synaptic conductance plays in long-term potentiation, a five
cell Hodgkin-Huxley type network was created interconnected by excitatory chemical synapses, using the computer
program, SNNAP(Simulator for Nerve Networks and Action Potentials). An initial stimulus is applied to the first
neuronal cell to begin the network. By applying one or more brief, high frequency stimuli to a pre-synaptic cell or by
applying a cooperative weaker stimulation to many cells, the cells in the network will continue to fire for an extended
period. By altering and observing the changes in the conductance of the synapses, it is possible to simulate long-term
potentiation within the network.
Cloning and Analysis of Zebrafish (Danio rerio) Supervillin
Steve N. Duleh1, Aaron J. Cohen1, Jeannie T.B. Collins2, Robert K. Pope1*, , 1Indiana University South Bend, 1700
Mishawaka Avenue, South Bend, IN 46634, 2University of Southern Indiana, Evansville, IN 47712
Supervillin is a 205-kDa F-actin and myosin II binding protein initially identified as a tightly bound peripheral
membrane protein in bovine neutrophils. Here we report the cloning and analysis of a 1,763 amino acid (200-kDa)
zebrafish homolog of supervillin. Supervillin has been identified in human, bovine, mouse, rat, dog, chicken, and now
zebrafish tissues. Distribution of human supervillin mRNA has shown that this protein is expressed in most body
tissues, with the lowest levels in nervous tissue. Supervillin protein sequences contain a highly conserved
carboxy-terminal half homologous to the villin protein, and a novel amino-terminal half. The amino-terminal region of
zebrafish supervillin is well conserved, like all supervillins cloned to date, and the carboxy-terminal half is very similar
to human supervillin. One major difference between zebrafish supervillin and other supervillin proteins is the lack of
nuclear localizations signals normally present in the amino-terminal half. These signals have been shown to shuttle
fusion proteins into the nucleus. While the known nuclear localization signals are not present, a novel predicted
sequence exists that may function in a similar manner, suggesting that zebrafish supervillin is also a nucleic acid
binding protein. Zebrafish supervillin starts at the same amino acid as human, bovine, and mouse supervillin; other
identified supervillin proteins have additional sequence at the amino terminal end. Most of the proposed
phosphorylation sites conserved in the other species are missing in zebrafish supervillin, indicating its function and
regulation may be unique when compared to these homologs. Another difference between zebrafish supervillin and
all other villin-like proteins is the fact that zebrafish supervillin contains an extra insert sequence in the villin
head-piece at the carboxy-terminus. In this study, we have cloned the zebrafish supervillin gene and performed
protein modeling to acquire a proposed structure for this protein. Currently, we are constructing fusion proteins and
investigating the expression patterns of zebrafish supervillin. Ongoing research will help us understand the possible
roles of zebrafish supervillin, and provide insight into the function of motifs that are novel or absent in the zebrafish
homolog.
69
BIOLOGY
Memory Recall in Meadow Voles: Possible Episodic-like Memory
Amy M. Combs, Andrew A. Pierce, and Javier delBarco Trillo (Dr. Micheal H. Ferkin), University of Memphis,
Department of Biology
Episodic memory involves the ability to recall information the what, where, and when of a single past event. Although
episodic memory is commonplace in humans, recent work suggests that animals may also possess episodic-like
memory. My work investigates whether meadow voles (Microtus pennsylvanicus) possess episodic-like memory. To
test for episodic-like memory, we place males in an arena that contains two chambers. One chamber contains a
pregnant female (24 hours prepartum). The other chamber contains a sexually mature female that is not pregnant or
lactating, a reference female (REF). Males are allowed to explore the arena. However, the males cannot make
physical contact with the female due to a transparent barrier that also contains holes for odor stimuli. 24 hours after
the exposure, the males are placed in the arena, which is now empty and clean. I record the amount of time that the
male investigates the two chambers. At this time, the pregnant female would have delivered her pups and entered
postpartum estrus (PPE). PPE females are more attractive to males and mate more readily as compared to REF. If
males have episodic-like memory they will spend more time investigating the empty chamber that originally housed
the pregnant female (now a PPE female) as compared to the chamber that originally housed REF. The second
condition of my experiment is identical to above expect that there is a PPE female in one chamber and a REF in the
other chamber. 24 hours later, the PPE female would be lactating. A lactating female is equally attractive and mates
as readily as a reference female. The males should spend about equal amounts of time investigating both chambers.
For male voles an episodic-like memory for the reproductive condition of females would allow males to remember the
location of potential sexually receptive females and avoid interacting with females that will not readily mate.
Keeping an Eye on Charlie Brown: Tracking Eye Movements while Reading Comics
Karen Stringer, (Dr. Jay Hosler), Department of Biology, Juniata College, Huntingdon, PA 16652
This study used an eye-tracking device to trace individuals` eye movements while reading comics. We used four
Peanuts comic strips, two wordless strips and two strips that included text. Each strip had 4 panels. Eye movements
of subjects were tracked, and fixation times were analyzed to compare the time readers spent looking at text versus
the time spent looking at the illustration in each panel. There was no difference in image fixation times between
wordless comic strips and strips with text for panels 1 and 2 of each comic. However, results indicate that time spent
with the images in panels 3 and 4 for the wordless comic strips was greater than that in strips with text. This was due
to the subjects transitioning their gaze between panels 3 and 4 more often in the wordless comic strips. These
findings suggest that subjects may have had more difficulty constructing meaning from solely images in the wordless
comic strips compared to the comic strips that were also guided by verbal cues.
AN EXAMINATION OF THE INCUBATION PATTERNS AND MICROBIAL COLONIZATION OF EGGS IN
OPEN-CUP NESTING PASSERINES.
Ashley D. Fitzgerald (Morgan Wilson, PhD; Renee Godard, PhD; Amy Fenster, MS), Dept of Biology, Hollins
University, Roanoke, Va., 24020
Open-cup nesting passerines face many reproductive challenges, including finding suitable nesting habitats, avoiding
predators and nest parasites, maintaining egg viability, and raising nestlings. Of these challenges, maintaining egg
viability may be one of the most difficult to control because of the impacts of both abiotic (e.g. ambient temperature
and humidity) and biotic factors (e.g. invasion by pathogenic microbes). While several studies have shown that
prolonged exposure of eggs to ambient conditions and/or high humidity prior to incubation can reduce egg viability of
domestic fowl, few studies have investigated the effects of pre-incubation exposure on the viability of wild bird eggs.
We investigated the effects of incubation patterns (i.e. synchronous vs. asynchronous incubation) and microbial
colonization on nesting success of several open-cup nesting passerine species (Cardinalis cardinalis, Carpodacus
mexicanus, Dumetella carolinensis, Mimus polyglottos, Melospiza melodia, Turdus migratorius). Incubation was
monitored using wireless thermocouples and egg shell microbes were sampled at different intervals (<4, 24, 48, 72
hours) after laying. Our preliminary analyses suggest: 1) observed incubation patterns of several species were
different from those of previously published data and 2) an abundance of Gram-negative cocci, when compared to
Gram-positive rods - a trend that differs from published data on egg shell microbes in tropical species. Our findings,
thus far, indicate that patterns of incubation may be more variable than previously thought and that birds may adjust
their incubation behavior based upon environmental conditions.
70
BIOLOGY
3-ETHYLPYRIDINE, A CIGARETTE ADDITIVE, INHIBITS GROWTH OF THE HAMSTER CONCEPTUS IN
VIVO
Barbara Williams, Marcelis Morris, Gloria Lai (Dr. Talbot), Department of Cell Biology and Neuroscience, University of
California, Riverside, Riverside, CA 92521
Pyridine and pyrazine derivatives from cigarette smoke were previously shown to inhibit growth of chick
chorioallantoic membrane at picomolar doses (Reprod. Biol. Endoc. 2005 (3):52). The purpose of my experiment was
to test the hypothesis that in vivo administration of 3-ethylpyridine will affect fecundity, number of implantations, and
the growth of the conceptus in a mammalian model, the golden hamster (Mesocricetus auratus). Female hamsters
(8-week old) were injected intraperitoneally with light mineral oil (control) or with 25 mg/kg or 50 mg/kg of
3-ethylpyridine (10 females per group) for the first six days of pregnancy. They were sacrificed on the 7th day of
pregnancy, and the effects of 3-ethylpyridine on various reproductive parameters were determined. The statistical
significance of these results was evaluated using a one-way ANOVA to compare the mean of the control to the means
of the treated females (25mg/kg and 50 mg/kg) for all the parameters tested. Means were considered significantly
different for p< 0.05. 3-Ethylpyridine did not significantly affect the percent of implantations, uterine horn weight and
length, ovarian weight, oviductal weight, distance between implantation sites, or weight of maternal organs (kidneys
and liver). However, 3-ethylpyridine caused a statistically significant decrease in both implantation site length (4.9mm
± 0.9 control vs. 4.4mm ± 0.7 for 25 mg/kg and 4.6mm ± 0.7 for 50 mg/kg) and width (5.2mm ± 1.2 for control vs.
4.8mm ± 0.6 for 25 mg/kg and 4.9mm ± 0.8 for 50 mg/kg), which was interpreted to represent an inhibition in the
growth of the conceptus. These data extend our study on the chick and are the first to show an in vivo effect of
3-ethylpyridine on the development in a mammal. These data suggest that 3-ethylpyridine, which is on the Flavor and
Extract Manufactures Association list of chemicals that are generally regarded as safe (FEMA-GRAS), may not be as
safe as previously thought and should undergo further toxicological testing.
Aquaporin 0 and Alpha-crystallin Interaction During Thermal Stress
Joseph M. Fachini, Ajay Pillai (Dr. S. Swamy-Mruthinti), Department of Biology, Department of Chemistry, University
of West Georgia, Carrollton, GA 30118
Aquaporin 0 (AQP0) is a transmembrane protein. Its function is to transport water across the cell membrane of lens
fiber cells. Normal function of AQP0 is essential to maintain cellular homeostasis and lens clarity. Alpha-crystallin is
a molecular chaperone, protecting other proteins during stress-induced denaturation. Recent studies from our
laboratory have shown that alpha-crystallin protects AQP0 from heat-induced aggregation. This study aims to show
through immunochemical and HPLC analysis that alpha-crystallin specifically binds to AQP0 under thermal stress.
Immunochemical data shows that alpha-crystallin is the predominant lens cytosolic protein involved in the interaction
with AQP0 compared to beta and gamma crystallins. HPLC data shows the formation of AQP0-alpha crystallin
complex during the thermal stress. These studies suggest that the binding of alpha-crystallin to AQP0 and the
resulting protection of the transmembrane protein are a direct result of the chaperone-like function of alpha-crystallin.
The degeneration of the chaperone-like activity of alpha crystallin during aging may be directly associated with the
aggregation of AQP0 and the subsequent formation of cataracts.
71
BIOLOGY
NUMERICAL DISCRIMINATION IN WILD MOCKINGBIRDS
Hank Kerschen, Sirisha Manyam, Michael Horejs (Dr. George Farnsworth), Department of Biology, Xavier University,
Cincinnati, Ohio 45207
Previous studies have suggested the presence of numerical discrimination abilities in animals. In one recently
conducted experiment, five wild northern mockingbirds (Mimus polyglottos) were presented with a bird feeder that
released mealworms (larvae of Tenebrio beetles) as a reward. The mealworms were held on a plank suspended by
varying numbers of bamboo sticks which protruded from both sides of the feeder. Upon complete removal of sticks
from either end of the feeder, mockingbirds received mealworms. However, if the side with fewer sticks was chosen,
then the mealworms were more easily attained. Mockingbirds were found to have the ability to distinguish between
one and six sticks and two and five sticks, however failed to distinguish between three and four sticks. To further
explore the extent of numerical discrimination abilities in wild mockingbirds, a new experiment was begun using a
modified feeder. The new feeder was designed to release mealworms only when approached on the end containing
the least number of sticks, thus providing a greater incentive for mockingbirds to discriminate between numbers. This
experiment examined the ability of mockingbirds to distinguish between two and five, and three and four sticks. As a
control to ensure that the subjects were responding to the number of sticks, three sticks were placed on both sides of
the feeder. We hypothesized that wild northern mockingbirds would be able to distinguish between two and five, and
three and four sticks by repeatedly selecting the fewer number of sticks in order to receive the mealworm reward. In
the control trials, we predicted that subjects would choose either end of the feeder equally.
ALTERNATIVE SPLICING AND SEX-SPECIFIC EXPRESSION OF SOWUSCHEL IS ASSOCIATED WITH
SEXUAL DIMORPHISM IN DIOECIOUS SPINACIA OLERACEA
Madhu Jagannathan, D. Noah Sather, (Dr. Edward M. Golenberg) , Department of Biological Sciences, Wayne State
University, Detroit, MI 48201, U.S.A
In both animals and plants, cells that are highly undifferentiated and can readily divide to form new tissue and organs
are called stem cells. Because of their central role as a source for all tissue and organs, understanding the regulation
of maintenance of stem cells is a central problem in biology. Stem cells in plants are located in a restricted number of
structures. In the aboveground tissue they are organized in the shoot apical meristem (SAM), axillary meristems,
inflorescence meristems and floral meristems. Because all plant organs arise from the indeterminate growth of these
meristems, proper regulation of these stem cells is of paramount importance in plant development.
The major gene that regulates stem cell identity in Arabidopsis thaliana is WUSCHEL (WUS). Loss of WUS
expression results in rapid depletion of stem cells in the meristem and cessation of growth. In the floral meristem,
WUS is expressed in the center of the flower and upregulates the expression of the floral organ identity gene
AGAMOUS (AG). The AG protein, once expressed, inhibits WUS expression in the flower. The result is a
determinate structure due to the lack of stem cell maintenance.
In Spinacia oleracea (cultivated spinach), individual plants produce only male or female flowers (dioecious).
Females produce a single terminal carpel at the center of the flower, but lack stamens. Males produce four stamens
but the central portion of the flower is flattened due to lack of cell growth. Recent work in our laboratory has isolated
two transcripts of the spinach WUS homolog SoWUS in male flowers. One transcript that was only found in male
flowers, which appears to be the product of alternative splicing of SoWUS mRNA, has a deletion of 16 nucleotides
that produces a premature stop codon and an altered amino acid sequence. Our hypothesis is that the alternative
splicing of the SoWUS message is critical for the sexual dimorphism in spinach, possibly through the production of
the smaller (truncated) protein in males, which may interfere with normal WUSCHEL functions.
72
BIOLOGY
MODELING FISH MUSCLE DYNAMICS AND FORCE GENERATION
Jinjin Qian (Robert G. Root) Department of Mathematics, Lafayette College, Easton, PA 18042
This project constructs fish muscle models that can faithfully model active and passive force generation under
realistic, dynamic conditions of stimulation and deformation. The models focus on the generation of force at the
sarcomere level and in particular, the cross-bridge attachments in a single sarcomere. Since developed tension for a
single sarcomere depends on the proportion and lengths of attachments of cross-bridges, a scaled linear spring model
gives a good approximation to the developed tension within a certain physiological range of lengths. However, when
muscle is overly stretched or compressed, the effect of passive tension developed becomes significant. Because in
undulatory locomotion of fish, strains of 30% are documented by experiment, we add a piecewise function into the
model describing the passive tension. Moreover, time dependence of the “active state” is another important feature
of fish muscle. Unlike skeletal muscle that typically remains under tetanus once stimulated, fish muscle stimulation
turns on and off rapidly and often cyclically. The active state intensity is introduced explicitly in these models. They
are comprised of a system of integro-differential equations, to which we implement a numerical solution method using
Mathematica. We give two slightly different systems to describe the cross-bridge population; the first is when thin
filament sites are so sparse that at most one is within range of any given cross-bridge at any given time. The second
type is when thin filament sites is a continuum. It is interesting that we can actually obtain Hill’s force-velocity model
by treating the rate of detachment as a constant in the second type of system. Our models are based on ones
developed by C. Peskin for human cardiac muscle, and have been verified by comparison with muscle data collected
and modeled by J. Van Leeuwen.
This research is supported by National Science Foundation grant DBI-0442269.
XENOPUS LAEVIS EMBRYO EXPOSURE TO PERFLUROOCTANOIC ACID AND THE CAUSE AND
AFFECTS OF DEVELOPMENTAL DELAY
Jenna K. Gay (Dr. Kristen L. Curran), Biological Sciences Department, University of Wisconsin-Whitewater, 800 West
Main Street, Whitewater, Wisconsin 53190.
The purpose of this project was to begin to determine if Perflurooctanoic Acid (PFOA) affects early development of
the African clawed frog (Xenopus laevis). PFOA is a man-made compound that is of high concern because of its
toxicity, ability to bioaccumulate,, persistence in the environment, and global distribution in freshwater. It has major
uses in today’s society due to its heat stable and non-stick properties. In particular, it is a major component of the
Teflon commonly found on cooking pots, pans, and utensils. In order to examine the possible effects PFOA has on
development, I treated 1-2 cell stage frog embryos with concentrations of 125 micromolar, 250&#61472;micromolar,
and 500&#61472;micromolar PFOA and compared them to untreated embryos. Preliminary results suggest that
developmental delay first occurs in gastrula stages at concentrations of 250&#61472;micromolar and 500 micromolar.
This is a time during development where the three embryonic germ layers are being formed (ectoderm, endoderm,
and mesoderm). The concentrations of PFOA used in my preliminary trials are higher than those found in the
environment. But the levels that may be reached in the future due to PFOA continually being released into the
environment, such as the concentrations used in my experiment, shows a delay in development. This concern, along
with other affects such as liver damage, tumorigenesis, changing levels of hormones, and adverse affects on the
immune system are related to PFOA exposure. Future experiments include repeating the results and establishing a
link between developmental delay and a specific cellular process.
73
BIOLOGY
HABITAT FLEXIBILITY IN THE TOOTH-BILLED BOWERBIRD (SCENOPOEETES DENTIROSTRIS):
IMPLICATIONS FOR SPECIES SURVIVAL IN A CHANGING CLIMATE.
Breckheimer, Ian K.(1), and Amanda N.D. Freeman (2). (Dr. Amanda N.D. Freeman)(1)Department of Biology,
Guilford College, Greensboro NC 27410. (2)School for Field Studies Center for Rainforest Studies, Yungaburra, QLD
4884 Australia
The Tooth-billed Bowerbird (Scenopoeetes dentirostris) is endemic to rainforest uplands of North Queensland,
Australia, and is potentially threatened by global warming. The degree of this threat depends on the species’
flexibility when faced with changing habitat. During the breeding season, males clear a forest-floor display “court” and
decorate it with upturned leaves, near which they vocalize to attract mates. Aspects of this male breeding effort
(including calling time, court maintenance time, court leaf number, and leaf turnover) were used as indicators of
habitat suitability for males in two habitats: regrowth forest (RG) and mature rainforest (MR). Calling time (n=4; RG
S.E. ±1.84, MR S.E.± 4.54) court maintenance time (n=4; RG S.E.±0.263, MR S.E. ± 0.198), and court leaf number
(MR n=15, S.E.= ±4.58; RG n=8, S.E. ±7.89) were statistically similar in the two habitats at a p>0.05. Court leaf
turnover was significantly greater in regrowth (p=0.0289; RG n=4, S.E.±1.72; MR n=8, S.E.±1.37). The trials fail to
give evidence that breeding effort was greater in mature forest than in regrowth. This supports two conclusions: (i)
that the species exhibits some habitat flexibility and (ii) factors other than resource availability will drive changes in
the species’ distribution in a warmer world.
Relation of Smallmouth bass Otolith Chemistry to Natal River
Brant Priest (Dr. Robert Humston), Department of Biology, The Virginia Military Institute, Lexington, Virginia
24450-0304
We used otolith trace elemental chemistry to discriminate natal origins of young-of-year smallmouth bass
(Micropterus dolomieu) from the James River and its tributary Maury River. Fry aged 2-4 weeks old were collected
along the entire length of the Maury River and locations along the James River above and below its confluence with
the Maury. The otoliths were analyzed using inductively coupled plasma mass spectroscopy to determine their trace
elemental chemistry. Discriminate function analysis of the multivariate elemental data allowed us to determine the
classification accuracy of natal origins between the James and Maury Rivers as well as between collection sites.
Analysis of the data yielded up to 95% classification accuracy of fry back to their natal river and between 85-90%
accuracy in discriminating their natal origin between river reaches. Otolith trace elemental chemistry can be used as
a means of identifying the natal origins of individual smallmouth bass. Determining the natal origin of individual fish
has the potential to allow for tracking the dispersal of recruits and adults between rivers that may delineate
metapopulations.
The Effects of Aromatase Inhibitor (AI) 4-hydroxyAndrostenendione (4-OH-A) and
Aminoglutathiamide (AMGT), and Estrogen Replacement Therapy on the De
Author: Bryan Nelson, Sponsor: Doctor James E. Turner, Department: Department of Biology, Institution: Virginia
Military Institute, Address: VMI Box 80883, Lexington VA 24450
It has been shown that the steroid hormone estrogen plays an important role in the development of the central
nervous system. The purpose of this study was to determine the effect of estrogen on zebrafish sensory-motor
behavioral maturation in the central nervous system (CNS). In these experiments the aromatase inhibitor (AI),
4-hydroxyandrostenedione (4-OH-A), as well as aminoglutathiamide (AMGT), which block estrogen synthesis, were
used to diminish estrogens effects on zebrafish CNS maturation. During the treatments, the zebrafish were analyzed
for neurological deficits, including tactile responses, spontaneous swimming movements, vestibular behavior, pectoral
fin, and eye movements. Over a three to five day developmental time period, in response to the 4-OH-A and AMGT,
none of these sensory-motor behaviors were expressed except for the early appearing tactile response that
disappeared over a 24 hour period post treatment. Furthermore, when the AI was removed from the treatment
medium the sensory-motor behaviors began to be fully expressed over a two to three day time period. In addition,
when AI and physiological levels of estrogen were added at the same time, there was a stimulation of developmental
appearance in all neurological parameters measured indicating that exogenous estrogen could in part replace the
endogenous levels reduced by AI treatment. Therefore, these studies have demonstrated that the presence of
estrogen in the early developing zebrafish embryo is necessary for the proper expression of several critical nervous
system sensory-motor behaviors which presents a suitable model for the study of its effects on the development of
brainstem and spinal cord circuitry related to the maturation of these phenomena.
74
BIOLOGY
GENETIC ANALYSIS OF THE PLANT HORMONE AUXIN IN A MODEL PLANT
Yefim Yushvayev-Cavalier, (Dr. Lawrence Hobbie), Department of Biology, Adelphi University, 1 South Avenue,
Garden City, New York 11530
This project involves a genetic analysis of the signal transduction and transport pathways of the plant hormone auxin
using the model plant Arabidopsis thaliana. This signal transduction pathway includes auxin response factors (ARFs),
which mediate auxin induced changes in gene expression, and components of the ubiquitin pathway, which degrade
unstable AUX/IAA proteins, which normally repress ARFs in the absence of auxin. Many proteins within this pathway
have been identified through a genetic approach. To expand the analysis of this pathway, 156 putative mutants were
identified as showing altered auxin response in root growth. Based on initial characterization of their resistance to
natural and synthetic auxin, these mutants were classified into four groups. Further experiments, which included
phenotypic analysis such as visualization of vasculature, complementation tests to known mutants, and cross
fertilization to wild type plants, led us to focus on eight potential mutants that may represent novel loci involved in
auxin response. In our recent experiments, we have used molecular gene mapping to determine the chromosomal
locations of these genes. Based on the map positions, we will identify novel mutations and eventually characterize the
genes affected in these mutants. This research will contribute to a better understanding of plant physiology and the
molecular basis of plant hormone action.
MOLECULAR EXAMINATION OF THE DEVELOPMENTAL SHIFTS IN VARIOUS SUNFISH
Sean C. Dodson (Dr. Coughlin), Department of Biology, Widener University, Chester, Pennsylvania 19013
This study explores developmental shifts in the contractile properties and protein composition of a jaw muscle, the
sternohyoideus, of sunfish (F. Centrarchidae). In fishes, the sternohyoideus aids in movement of the lower jaw,
including respiration and suction feeding. Physiology experiments have shown developmental shifts in the
sternohyoideus. In bluegill, the sternohyoideus shows a typical pattern of the maximum shortening velocity (Vmax) of
muscle slowing with age. However, in largemouth bass, Vmax of the sternohyoideus is highest in middle size fish,
moderate in large fish and shows variation among small fish. In both species, the sternohyoideus appears to the eye
to be entirely composed of white or anaerobic, fast-twitch muscle. What is the molecular basis of these variations in
Vmax? One answer may be the myosin heavy chain (MHC) component of the myofibrillar proteins. MHC composition
is thought to determine the Vmax of muscle. My goal is to analyze developmental shifts in MHC composition of the
sternohyoideus in sunfish. SDS Page gel electrophoresis and Western blotting have been implemented to identify
slow and fast isoforms of MHC. Through electrophoresis, the slow MHC isoform (found in red muscle) has higher
molecular weight than the fast MHC isoform (found in epaxial muscle). Three antibodies, F8, F59, and EB165 have
successfully been used to identify MHC isoforms. F8 and F59 bind only to the slow isoform, while EB165 binds to
both the slow and fast isoform. The fast isoform is always expressed in the sternohyoideus of largemouth bass.
However, variations are observed among the juvenile largemouth bass. In some juveniles, the fast isoform is
expressed but in others the slow isoform is expressed. These data correspond with the physiological results that have
been obtained. For instance the sternohyoid muscles of young bass vary both in terms of MHC composition and
contractile properties.
COMPARISON OF IMAGE ANALYSIS SOFTWARE FOR DNA MICROARRAYS
Scott C. Couden and Hollie E. Flick (Dr. Jens Hemmingsen), Chemistry Department, Capital University, Columbus,
Ohio 43209
DNA microarrays are used to measure differences in gene expression profiles. An important step in the experiment is
the analysis of images produced by the microarray scanner. In this study, image analysis software packages were
evaluated for the ability to locate spots (gridding and segmentation), the determination of spot intensities, background
correction and ease of use. Image analysis software was freely available to academic users and included Magic Tool,
ScanAlyze, TIGR Spotfinder and UCSF Spot. Analysis features varied among the software packages. Gridding was
fully automated in UCSF Spot and partially manual in the other three programs. A variety of segmentation algorithms
were available in each analysis program including fixed or variable diameter circles and seeded region growing. All of
the programs included background correction except Magic Tool. Each software package could be learned quickly,
but the quality of documentation varied among analysis programs. Ultimately, each software package could be used
to obtain spot intensities from DNA microarrays. However, differences in program features resulted in variation in the
intensity data obtained.
75
BIOLOGY
Comparison of Mycorrhizal Colonization Frequency of Bromus tectorum and Native Grass Species
at Antelope Island State Park
Shaunna Anne Goldberry & (Ron Joseph Deckert) Department of Botany, 2504 University Circle, Weber State
University, Ogden Utah
Bromus tectorum—cheat grass, is a non-native species that affects ecosystems by altering plant community structure.
Though much is understood about the role of symbionts in the autecology of native species, little is known about the
role of symbionts and invasive species establishment. The objective of the study was to compare the colonization
frequency of root symbionts (arbuscular-mycorrhizal and dark septate endophytes) between B. tectorum and several
species of native grasses. Native species were chosen according to forage potential and the degree to which they are
impacted by cheat grass invasion. Inoculum potential of the soil from the test sites was analyzed by growing Zea
mays and the test species in collected soil from Antelope Island State Park—an island in the Great Salt Lake of
northern Utah. After twelve weeks, roots were cleared and stained with trypan blue in lactoglycerol. Fifty 10.0 mm
root sections from twenty individual grasses were collected from the five test sites. Samples were quantified for root
fungal structures using the line intersect method. Fungal identifications were made where possible. Preliminary
results show higher colonization frequency of an unidentified dark-septate endophyte (DSE) in B. tectorum relative to
native grass species. We found differential distribution of an unknown Chytridiomycete in the non-native species B.
tectorum, and other native grass species. Identification of the isolated DSEs from B. tectorum root and a Koch’s
postulate test is ongoing.
SMALL MAMMAL SPECIES COMPOSITION AND VARIANCE BETWEEN RIPARIAN WOODLAND AND
CLEAR-CUT AREAS AT JEFFREY ISLAND HABITAT AREA
Kristi A. Bauer (University of Nebraska at Kearney), Department of Biology, The University of Nebraska at Kearney,
Kearney, Nebraska 68849
The Central Nebraska Public Power and Irrigation District is managing the Jeffery Island Habitat Area for wildlife.
This 1,700 ha parcel lies in the midst of the Platte River between Lexington and Overton in Dawson County,
Nebraska. In 2001, Central clear-cut 97.5 ha of woodland habitat in an effort to create a more attractive environment
for migratory bird species of conservation concern. From May 2005 to July 2005, we used live-trapping methods to
assess the composition and relative abundance of small mammal populations in the clear-cut area relative to remnant
wooded areas to assess possible effects of woodland deforestation. Each of 2 wooded and 2 clear-cut sites were
trapped using a drift fence system (pitfall buckets and funnel traps) and a configuration of baited Sherman traps.
Individuals of 12 species were captured during the 10-week trapping period. The two most commonly caught species
were Peromyscus leucpous and Peromyscus maniculatus, with leucpous dominating in the wooded habitat. A total of
3 species were captured using the Sherman traps versus 8 species captured with the drift fence system. The species
caught utilizing the Sherman traps included: Peromyscus leucpous, Peromyscus maniculatus, and Dipodomus ordii.
Relatively high recapture rates were observed for both the Peromyscus leucpous and the Peromyscus maniculatus.
For the two Peromyscus species combined, an 88.7% recapture rate was observed in the wooded sites versus a
78.7% recapture rate at clear-cut sites. Sorex shrews were present at every site and were caught only with the drift
fence system. A similar study was done by a student in 2004 and the work was presented at the 2005 National
Conferences for Undergraduate Research. Data from both studies was utilized and compared to illustrate variance.
76
BIOLOGY
DENTAL BIOFILMS AND THE BACTERIA THAT COMPOSE THEM
Matt Sroka (Dr. Jennifer Koehl), Department of Biology, St. Vincent College, Latrobe, Pennsylvania, 15650
Dental plaque is something that will always be present. Oral plaque is actually a biofilm comprised of many types of
bacteria. A biofilm is a community of multiple bacteria that live and grow together on a common surface. The most
dominant of the oral bacteria in these biofilms are Streptococcus species, particularly Streptococcus viridans,
Streptococcus oralis, Streptococcus mutans, and Streptococcus oralis. The pattern of biofilm growth is an initial
attachment of bacteria to a solid surface, formation of microcolonies on the surface, and finally differentiation of
microcolonies into a polysaccharide-encased mature biofilm. It is believed that S. oralis and S. sanguis are the first to
attach and therefore the primary colonizers in oral bacteria. The purpose of this experiment was to determine
bacterial growth of the Streptococcus species individually and in combination with the other Streptococcus species.
In addition, different food and beverage solutions (containing different carbon sources) may have an influence on
biofilm growth. It was initially decided to utilize a glucose-defined media for all experiments; this media, however, did
not allow for adequate growth of the individual bacteria. A specific biofilm medium was then utilized but various
medium components would not dissolve and the bacteria were unable to grow. Since all bacteria utilized grew in the
complex medium brain heart infusion (BHI), BHI was utilized although not an optimal medium (the exact composition
of BHI is unknown and varies from batch to batch). Overall, it has been shown that individually S. viridans grows best
in a biofilm. In combination, S. oralis and S. viridans and S. viridans and S. mutans grow best together a combination.
The next step of this research is to determine if different food and beverage supplements encourage or discourage
oral biofilm growth.
DETERMINATION OF WINDOWS OF VULNERABILITY FOR THALIDOMIDE INDUCED
TERATOGENESIS IN SEA URCHIN EMBRYOS CULTURED IN VITRO
Katherine L. Plocharczyk, Cherysse E.D. Lanns, Steven Heverley (Dr. Jan Reichard-Brown), Department of Biology,
Susquehanna University, 514 University Avenue, Selinsgrove Pennsylvania 17870
Thalidomide is an anti-inflammatory, immuno-modulatory, anti-angiogenic drug which was widely administered to
pregnant women in the 1950s to alleviate morning sickness. After being linked to numerous cases of limb reduction
deformities and other congenital defects, this highly teratogenic drug was banned in 1962. Although the mechanism
of action for thalidomide induced teratogenicity in humans remains uncertain, thalidomide is being tested in clinical
trials and marketed as a therapeutic agent for a variety of diseases. Mammalian testing has proven to be inconsistent
and expensive. Previous work has shown that sea urchin embryos (Lytechinus pictus) exhibit abnormal development
in culture when exposed to thalidomide. The current studies focus upon identifying windows of vulnerability for
teratogenicity in L. pictus embryos. Preliminary results indicate several potential windows. There is evidence that
one of these windows occurs early in development while others indicate susceptibility post gastrulation. Continuing
experimentation will delineate the precise timeframes as well as further these studies in a different sea urchin species
(Strongylocentrotus purpuratus). Sea urchins also express growth factor-like molecules similar to those that are
postulated to play a role in thalidomide’s pharmacological action in humans (TGF, EGF, and PDGF). Later phases of
this research will examine the interaction between these similar growth factors and thalidomide in sea urchins.
Mapping Distributions of Invasive Plants and Their Effects on Forest Regeneration
Georgia Sawyer, Nina Kean, Biology, North Carolina A&T State University, 1601 E Market , JOSEPH COVINGTON
(Curator of the Museum Park, NC Museum of Art, Raleigh, NC 27699), , RITA HAGEVIK (NC A&T Biology
Department)
Native forest species exhibit a well-known range of ecological roles with respect to natural disturbance regimes, from
pioneer phase to mature phase, and they regenerate from a range of sources, including dormant seeds, seed rain,
pre-established juveniles, and resprouts from damaged adults. In contrast, the ecological roles of invasive,
non-indigenous species in forest communities after natural disturbances are not well understood. Some previous
studies of invasive species have emphasized their weedy nature and their ability to colonize anthropogenic
disturbances. To study the effects of invasive plants on indigenous forests, the location of large areas within the
ecosystem of the Museum Park at the NC Museum of Art of four common invasive plant species, Kudzu,
Microstegium, English Ivy, and Purple Nutgrass will be mapped. Then two pairs of approximately 30 x 60 m research
plots per site, consisting of one control plot and one restoration plot; will be studied using an aggressive management
program. This restoration project will be a part of the restoration plans of the Park and will serve as educational
demonstration site to the public on the national problem of invasive plant species in the United States.
77
BIOLOGY
THE EFFECT OF ACID STRESS ON ADULT RANA CLAMITANS (GREEN FROGS)
Bukola Ojo, Raymond J. Howanski (Dr. Marc Brodkin and Dr. Itzick Vatnick), Department of Biology, Widener
University, Chester, Pennsylvania 19013-5792
The massive decline in amphibian populations in recent years has raised concerns among ecologists and
herpetologists around the world. Possible causes for this decline include infectious diseases and environmental
stressors. Ranid frogs are one of the species declining in the northeastern United States. Brodkin et al. (2003) showed
that acid exposure acts as an immune disruptor in Rana pipiens, suppressing the thioglycollate-induced inflammatory
response and phagocytic efficiency of white blood cells (WBCs). The authors also showed that exposure to pH 5.5
resulted in colonization of the spleen by bacteria. We have extended our investigation to the effects of acid on the
immune response of Rana clamitans. Alford et al. (1999) showed that the local extinction rate of Rana clamitans was
quite low. In addition, R. clamitans have been shown by other studies to flourish in places with low pH. For this
reason, we hypothesized that a low pH would have no effect on the innate immune response of R. clamitans. The
number of peritoneal white blood cells after stimulation by thioglycollate, a substance used to experimentally induce
peritonitis, was used as a measure of the acute innate immune response of the frogs. We randomly allocated 18 frogs
into four experimental groups: acid-exposed with thioglycollate injection, acid- exposed injected with saline,
neutral-exposed with thioglycollate injection, and neutral-exposed injected with saline. We counted the number of
white blood cells per ml of peritoneal fluid. We also counted the splenic bacteria using the spread plate method on
T-soy agar. Our results show that exposure to acid does reduce the number of white blood cells in stimulated frogs,
and also increases the number of bacteria per ml of spleen cells. Our results also suggest that although a low pH does
not cause mortality of green frogs, it suppresses their host defense mechanism against bacterial infection.
CENTER OF MASS MOVEMENTS OF PRIMATES VS. NON-PRIMATES: ELUCIDATING
EVOLUTIONARY PATHWAYS TO HUMAN BIPEDALISM
Anita Pai (Dr. Daniel Schmitt), Department of Biological Anthropology and Anatomy, Duke University, Durham, North
Carolina, 27701
During walking, all mammals studied to date are characterized by up and down movements of the center of mass
(COM) which allow for the efficient exchange of potential energy (PE) and kinetic energy (KE). Videorecords have
suggested that unlike other mammals, primates use a smoother, non-oscillating gait as an adaptation for movement
and foraging on thin, flexible branches. Video data only provides a rough estimate of COM movements. No direct
measures of COM movement in primates have ever been collected and the efficiency of mechanical energy
exchange in primates has not been documented.
Ground reaction forces and video data were collected on Ring-tailed lemurs (Lemur catta ) (N=2) and domestic cats
(Felis catus) (N=1) walking across a force platform embedded in a runway or attached to a long, horizontal pole.
Force plate data were used to calculate accelerations, velocities, and displacements of the COM as well as
fluctuations in mechanical energy. In Lemur catta, KE and PE oscillations were consistently in-phase and of different
magnitudes allowing little exchange of potential and kinetic energy. The domestic cat exchanged energy more
effectively than the lemur but not at the level typical of nonprimate mammals.
These data provide empirical support for the long-held view that primate locomotion is different from that of other
animals. The differences could reflect a critical adaptive shift associated with the origin and evolution of primates in
an arboreal environment, where stealth and stability may be more important variables than energetic efficiency. The
re-emergence of the energetically-favorable gait in early humans suggests that this locomotor mechanism is
beneficial for long-distance terrestrial walking.
78
BIOLOGY
BEHAVIORAL, MORPHOLOGICAL, AND PHYSIOLOGICAL CHARACTERIZATION OF A GIANT
JUMPING STICK STIPHRA SP. (ORTHOPTERA: PROSCCOPIIDAE) FROM PERU.
Angela Caranci (Dr. Stephen Madigosky and Dr. Itzick Vatnick), Department of Biology, Widener University at
Chester, PA 19013
The particular species of jumping stick used in this study (Stiphra sp.) has yet to be taxonomically described. Our
animals originated from a single gravid female obtained under the proper permit granted to the Cincinnati Zoo and
Botanical Garden, Cincinnati, Ohio. Little is known of changes in body mass and metabolic rate among the stages of
development of many insects and this includes Stiphra sp. We conducted a series of experiments to characterize the
behavioral, morphological, and physiological changes that occur in Stiphra sp. during development. These data now
serve as a starting point for further research on aspects of jumping stick development. We monitored daily food
intake, determined the efficiency in assimilation of controlled diets, and quantitatively tested food preference among
several plant species. Food items were selected based upon previously reported behavioral preference. A total of
seven adult females were offered firethorn (Pyracantha coccinea), blackberry (Rubus sp.), rose (Rosa sp.), and red
mulberry (Morus rubra). Results indicate a high preference toward firethorn and blackberry. Adult females showed
little interest in red mulberry. We have measured the energy content of food and feces in order to determine net
energy intake and efficiency of usage of different food items. We are in the process of analyzing information gathered
from these experiments to ascertain the overall energy balance. Also documented were the changes in body mass
and specific metabolic rate that occur as development progresses from eggs to adulthood. These results indicate a
successive increase in metabolic rate over the progression of all instar stages.
IDENTIFICATION OF DOWNSTREAM EFFECTORS OF NEUROGENIN1 SIGNALING IN DEVELOPING
DORSAL ROOT GANGLIA
Ben Mire (Richard C. Murray), Biology Department, Hendrix College, Conway, Arkansas 72032
In the nervous system, pain is sensed by nociceptive neurons which reside in the dorsal root ganglia (DRG) that flank
the spinal cord. These neurons derive from migrating neural crest cells and are absent in mice deficient in
neurogenin1 (ngn1)(Ma et al., 1999 Genes Dev. 13:1717), but little else is known about the molecular mechanisms
that regulate their development. In order to better understand nociceptive neuron differentiation, we will identify genes
that act downstream of ngn1 by comparing gene expression in heterozygote and ngn1 null DRGs by microarray
analysis. A preliminary microarray analysis of mRNA isolated from dissected DRGs comparing heterozygote and
ngn1 null embryos at E12.5 revealed several genes that were over- or under-represented in mRNA from heterozygote
embryos relative to mRNA from the null ngn1 embryos. We are currently confirming these preliminary results by
performing additional microarray analyses and in situ hybridization on the individual candidate genes. The results of
this study will contribute to our understanding of neuronal growth and differentiation in the developing nervous
system. (Supported by NIH P20 RR-16460 from the INBRE program of NCRR.)
79
BIOLOGY
RESERVA AMAZÓNICA, PERU AMPHIBIAN MONITORING AND ECOTOURISM
Sarah A. Orlofske (Dr. Erik R. Wild) Biology Department, University of Wisconsin Stevens Point, Stevens Point,
Wisconsin 54481
Loss of biodiversity, particularly amphibians, is a global concern illustrated by numerous threatened species and the
importance of amphibians as indicators of ecosystem health (Duellman and Koechlin 1991). A lack of long-term
amphibian population monitoring, particularly in tropical rainforest, limits perspectives on natural population
fluctuations and impedes conservation efforts. Numerous causes are likely responsible for amphibian population
declines and extinctions including human development and the associated habitat loss and degradation (Global
Amphibian Assessment 2004). However, further development in many areas of Latin America is inevitable.
Sustainable development activities such as ecotourism may play an important role in supporting local community
development while conserving critical amphibian habitat and protecting vulnerable populations for scientific research.
Sustainable forms of ecotourism and scientific research on amphibian populations have been continuing at Reserva
Amazónica, Peru, according to the Neotropical Biological Diversity Program (BIOTROP) established in 1988. During
the most recent field season monitoring of amphibian species occurrence, abundance and local distribution was
conducted to gain a long-term perspective on amphibian population health. Sustainable development is necessary to
preserve Reserva Amazónica as a research area. Therefore, scientific data are being developed into educational
materials and multimedia presentations. Interactive research projects conducted by the tourists involving common
species, such as Ceratophrys cornuta, Epipedobates hahneli and Colostethus trilineatus are also planned. This
research demonstrates how scientific data can be used to support local economic development, conservation and
continued research.
IMMUNOCYTOCHEMICAL AND ELECTRON MICROSCOPIC SURVEY OF CULTURED RAT
HYPOTHALAMI LOOKING FOR GAP JUNCTIONS BETWEEN GONADOTROPIN RELEASING
HORMONE NEUR
Jean Wennlund (Dr. Michael Woller), Department of Biological Sciences, University of Wisconsin – Whitewater,
Whitewater, Wisconsin 53190
The purpose of this project was to determine whether or not gonadotropin releasing hormone (GnRH) neurons in a
dispersed cell preparation of rat hypothalamic tissue are reforming gap junctions with one another in vitro. GnRH is
the essential molecule of reproduction, controlling the release of luteinizing hormone and follicle-stimulating hormone
by anterior pituitary, and subsequently that of estrogen and testosterone by the gonads. Coordinated release of
GnRH from the hypothalamus is crucial for proper reproductive function. To date, research has suggested that gap
junction connections between GnRH neurons might be responsible for the synchronized release of GnRH. However,
other work has shown that these connections may not exist. Our present investigation involves measuring GnRH
release from 3 different tissue/cell preparations from rat hypothalamus in a perifusion culture system. After 3 hours of
sampling GnRH release every 10 min, the tissue/cell preparations are collected, fixed, and prepared for
immunocytochemical analysis and electron microscopy. GnRH release from all 3 preparations was pulsatile as
determined by radioimmunoassay and pulsar analysis. Tissue preparations were viewed at the light microscopy level
for isolation of GnRH neurons, and then targeted for further observation by electron microscopy to assess the
presence of gap junctions. This study represents an important step in determining how GnRH neurons communicate
with each other to coordinate exocytosis and form pulsatile release events.
80
BIOLOGY
AN OUTSIDE SHOT AT UNRAVELING THE MYSTERIES OF WALDENSTROM’S
MACROGLOBULINEMIA
Derrel V Walker (Ruhul H. Kuddus, Ph.D.), Utah Valley State College, Orem UT 84058
Waldenstrom’s Macroglobulinemia (WM) is a low grade non-hodgkins lymphoma characterized by bone-marrow
infiltration and detection of serum monoclonal protein of IgM type. WM patients experience blood hyperviscosity and
enlargement of liver, spleen and lymph nodes. WM constitutes approximately 2% of all hematological cancers
affecting 2.5-5 subjects/million people. While affecting mostly Caucasian males over 65 years of age, no
susceptibility risk factors have been identified. Although it is known that about half of WM patients have deletions on
chromosome 6 (6q21) and a small portion have translocations in the IgM heavy chain locus 14q32, the mechanism of
the disease is poorly understood. Multiple siblings of a family may be affected by the disease but it is unclear whether
WM is familial. WM research is slow due to the rarity of the disease and because patients respond to conventional
cancer treatments (plasmapheresis and chemotherapy, although prognosis is slow and complete recovery is
extremely rare). I studied an extended family with two siblings affected by WM in order to identify possible genetic
markers associated with the disease. DNA was extracted from oral mucosal cells of the two affected persons, their
parents, spouses and children (a total of 22 subjects). Purified DNA was digested with three different restriction
enzymes, resolved in agarose gels and then transferred onto nylon membrane. The blot was hybridized with labeled
DNA probes derived from IgM heavy chain gene cDNA. No detectable difference in band pattern was observed.
Reprobing of the blots is currently underway. Random Amplified Polymorphic DNA analysis was also performed. The
resulting band pattern was highly polymorphic and several possible markers are currently being investigated by
cloning and DNA sequencing.
THE ISOLATION OF MONOMER AND DIMER COMPLEXES OF ADENYLATE KINASE AT VARIOUS
CONCENTRATIONS
Joan B. Watson (Anita K. Williams, Ph.D., Percy J. Russell, Ph.D) (School of Medicine and UCSD Chancellor`s
Undergraduate Research), Biological Sciences, UCSD 9500 Gilman Drive MC 0690, La Jolla, CA 92093-0690
Previous experiments showed that the glycolytic enzyme Adenylate Kinase (AK) was inhibited by Ascorbate (Vitamin
C), but it was concentration dependent. It was found that AK is not inhibited by Ascorbate at high concentrations.
Additionally, AK becomes more sensitive to Ascorbate inhibition and also exhibits a loss in activity with dilution. A
hypothesis was developed that AS exists as a dimer at high concentrations and dissociates to a stabilized monomer
at low concentrations. The monomer form is either not active or less active that the dimer in addition to the increased
sensitivity to ascorbate inhibition. We designed experiments to separate the dimers (MW= 42,000 kD) from the
monomers (MW= 21,000 kD). Centrifugal filters with selective molecular weight cut-off (MWCO) values ranging from
10,000 to 100,000 kD were used for separations. We found that when concentrated AK (200nM) was centrifuged
through MWCO value of 50,000 kD (MWCO50), only 10% of the activity passed through the filter. At MWCO30, only
3% passed through. At a lower AK concentration of 20nM with MWCO50, approximately 90% passed through. At
20nM and MWCO 30, about 50% passed through. Using MWCO30 at 20nM AK, we found that the AK that passed
through the filter (monomer) was more sensitive to Ascorbate inhibition that the AK that did not pass through the filter
(dimer). We showed that the dimer is not inhibited by Ascorbate and the monomer is inhibited as predicted by the
hypothesis.
81
BIOLOGY
THE EFFECT OF ACRYLAMIDE ON HUMAN CELLS AND THEIR EXPRESSION OF TELOMERASE
David A. Dempsey, (Dr. Jennifer Koehl), Biology Department, Saint Vincent College, 300 Fraser Purchase Rd.
Latrobe, Pennsylvania 15650
The purpose of this project was to determine the affect that the chemical acrylamide, a known carcinogen. has on the
expression of telomerase on human lung cells. Telomerase is a chemical produced by the cells that controls the DNA
sequences in the cell that control how many times a cell can divide. It is believed that an increase in the expression
of telomerase will indicate if the acrylamide affected the cells in a way as to make them malignant. Two cell lines
were cultured, CCD-16 Lu, and HS 229.T. CCD-16 Lu is a normal (experimental) cell line, and HS 229.T is a
malignant (control) cell line. These two cultures were exposed to the chemical acrylamide in four concentrations, 20
µg/ml, 40µg/ml, 60µg/ml, and 120µg/ml for 24 to 48 hours, and observed for changes in cell appearance. The
exposure to each concentration at both time durations resulted in no change in cell appearance. After exposure to the
chemical acrylamide, the cells were tested for viability via the trypan blue exclusion test. This test uses a dye that will
stain dead cells only, thereby indicating if the acrylamide had a toxic affect on the cells. Both cell lines were tested
for the levels of telomerase expression via a telomeric repeat amplification protocol (TRAP) assay. The levels of
telomerase will be compared between the two cell lines, the control and experimental. It is hypothesized that the cells
exposed to acrylamide will express telomerase to a greater degree than the cells that were not exposed to the
carcinogen. By comparing the TRAP results, it can then be determined if the acrylamide had an affect on the
expression of telomerase in the experimental culture.
Effect of human trails on avifauna in a tropical ecosystem
Dan Hannen-Starr (Dr. Robin Tyser), UW-L SAH Dean’s Distinguished Fellowship, UW-L SAH Travel and Supplies
Grant, UW-L Research Grant, UW-L Provost office grant, Biology Department, University of Wisconsin-La Crosse, La
Crosse, Wisconsin, 54601
Because ecotourism is increasing in tropical rainforests, resource managers need to ensure that trails in tropical
bioreserves do not disrupt wildlife. Breeding habits and population distribution of birds in North America are known to
be negatively effected by human trails (Zande, 1984; Miller, 1998); however, little research has been done on the
effects of trails in tropical ecosystems. Between May and August of 2005 bird counts were performed in Tirimbina
Rainforest Center (TRC), Costa Rica, to determine if bird distribution patterns were affected by: 1) the presence of
recreational trails and 2) human activity on the trails. In the summer months approximately 30 to 100 people per day
hike on the TRC bioreserve trails in tour groups. Bird abundance was determined through census counts while
following tour groups along the TRC bioreserve trails. Bird species and abundance within 25 meters of either side of
the trail were determined while walking 25 meters behind tour groups. Each census was paired with a similar census
(same trail length/direction, time of day, and weather conditions) conducted when tour groups were not present on the
trails. Bird abundance was greater when tour groups were not present (paired t-test: P=0.0042), indicating that bird
abundance may have been altered by the presence of tour groups. Point counts 100 meters from the trails were also
executed to determine the affect of the presence of trails on bird abundance. A significant pattern was not found
connecting the presence of trails to the abundance of birds. A longer study may reveal a relationship between the
presence of trails and bird abundance. The results of this study indicate that TRC is managing its bioreserve with
minimal impact on its wildlife.
82
BIOLOGY
THE EFFECT OF OLFACTORY ENRICHMENT ON THE BLACK HOWLER MONKEYS (ALOUATTA
CARAYA) AT JOHN BALL PARK ZOO
Molly Q. Lien (Dr. Roehling and Dr. Winnett-Murray), Departments of Psychology and Biology (respectively), Hope
College at Holland, Michigan 49423
A goal of many zoos is to study wild animals and teach others an appreciation of biological diversity and the need to
conserve it. To better do so, zoo environments often explore how enrichment can increase the complexity and
variability of the animals’ environment. John Ball Park Zoo in Grand Rapids, Michigan has initiated many enrichment
programs in order to positively stimulate their primate community, but the Black Howler monkeys (Alouatta caraya)
have continued to present an enrichment challenge. Because recent research indicates that howler monkeys may be
more positively affected by olfactory stimulation than other primates, the objective of this study was to determine the
behavioral effects of olfactory enrichment on the howlers, using fruit, non-fruit, primate, and non-primate scents. The
scents were placed on pieces of bark in the same location within the howler exhibit for a period of 1 day (totaling three
observation sessions each day). It was found that the howlers became significantly less bored and more active with
the enrichment-specifically with food scents (as compared to control and baseline categories). Notably, the level of
active, exploratory behaviors not associated with the scented bark increased, in addition to exploration of the scented
bark itself, indicating that the enhancement in behavior associated with the presence of novel scents has positive
effects extending beyond the object alone. In addition, there were differences in the responses of individual howlers
across the enrichment treatments, and diurnal patterns associated with the level of response to olfactory cues.
These findings have significant implications for zoos to enhance the environment of their captive primates. Further
analysis on what specific types of scents are effective would be desirable.
DESIGN, PRODUCTION AND CHARACTARIZATION OF A NOVEL HSV-1 BASED VECTOR FOR GENE
THERAPY
William D. Brubaker, (Satya Dandekar Ph. D.), Medical Microbiology and Immunology, School of Medicine, One
Shields Avenue, Davis, CA 95616-8645
I have designed a multi-step molecular strategy to engineer a novel gene transfer vector derived from a Type 1
Herpes Simplex Virus (HSV-1) amplicon vector and the PhiC31 phage integrase. By combining the properties of a
HSV-1 vector with the efficient and reliable site-specific integration of the PhiC31 integrase, the resulting gene
delivery platform should be capable of delivering a large amount of genetic information to almost any dividing or
non-dividing cell type in the human body. In addition, it will also integrate site specifically into the host cell
chromosome, providing safe, long-term transgene persistence. After gaining experience with the existing amplicon
packaging protocol, additional molecular biology techniques, and using the confocal fluorescence microscope, I
began construction of the new amplicon plasmid. The novel amplicon plasmid and control plasmids are being
constructed from an existing amplicon plasmid utilizing splice overlap extension PCR and restriction sites to insert the
phage integrase system and a red fluorescent protein expression cassette that will be shut off upon integration. In
addition to this a loxP site is included in the new plasmids. This will allow easy addition of filler DNA in order to
ensure that only a single copy of the plasmid was delivered by a single vector particle and will also allow easy
insertion of a new transgene cassette along with the filler DNA. Using Cre recombinase and a filler BAC of
appropriate length, the amplicon vectors will be brought up to 152 kb in size. The experimental and control amplicon
plasmids will then be packaged using the helper BAC, plasmid, and Vero cells used by the original amplicon system.
The resulting vector stocks will be used to transfect fresh Vero cells and will be assayed using confocal fluorescent
microscopy for both integration and expression persistence in the transfected cells.
83
BIOLOGY
Effect of All-trans Retinoic Acid on Proliferation in Pregnant Human Myometrial Cells
Ihunanya Mbata and Holly L. Boettger,Ph.D. (Holly L. Boettger-Tong, Ph.D.), Department of Biology, Wesleyan
College, Macon, GA 31210
All-trans retinoic acid (ATRA) is a natural derivative of Vitamin A that plays an important role in proliferation and
differentiation in various cell types. Previous studies have shown that uterine stromal and myometrial cells are
responsive to ATRA. Specifically, previous research has shown that ATRA inhibits estrogen-induced uterine stromal
and myometrial cell proliferation by halting DNA synthesis and cell division in vivo. In vitro, primary cultures of human
myometrial cells also exhibit dose-dependent retinoid sensitivity. Data in this presentation will indicate that
immortalized pregnant human myometrial cells respond to ATRA differently than their non-pregnant counterparts.
Dosages as low as 10^-9 M ATRA resulted in cell death in the PHM-1 pregnant myometrial cell line; this dosage was
not lethal in primary cultures of non-pregnant myometrial cells. To determine if this difference is due to the method of
immortalization in the pregnant myometrial cells, the current study has compared PHM-1 retinoid responsiveness to
that of another cell line. The HM6-1 cell line, immortalized by a completely different mechanism than that used to
immortalize PHM-1 cells, exhibits similar retinoid sensitivity and analogous dose-response characteristics. The
current studies also address the mechanism of retinoid induced cell death. Additional experiments will be conducted
to compare retinoid responsiveness between non-pregnant primary cultures of human myometrial cells and the cell
lines derived from the pregnant uterus. Partial funding for the research was provided by the Munroe Sisters
Endowment Fund.
CHARACTERIZATION OF A PHOTOPERIOD-SENSITIVE MUTATION IN ARABIDOPSIS
Samuel E. Shopinski and Karen A. Hicks (Karen A. Hicks), Department of Biology, Kenyon College, Gambier, OH
43022
Many plants rely on day length, or photoperiod, to regulate the transition from vegetative to reproductive
development. Proper timing of this transition is necessary to maximize a plant’s reproductive success. Arabidopsis
thaliana is a facultative long-day plant, flowering earlier in long-day (LD) conditions than in short-day (SD) conditions.
By studying Arabidopsis, we may better understand floral induction in other flowering species, such as rice, corn, and
tomato, and may then develop better technologies for producing crops. Although some of the molecular pathways
and biochemical processes involved in Arabidopsis floral induction have been characterized, much remains unclear.
By characterizing flowering mutants, we hope to identify additional genes in the floral induction pathways of not only
Arabidopsis but also other flowering species.
We are studying photoperiod-sensitive suppressor of elf3-1 2 (pse2), an Arabidopsis suppressor mutation that has a
subtle, late-flowering phenotype relative to wild-type flowering. elf3-1 mutants flower earlier than wild-type plants and
at approximately the same time in both LD and SD conditions. Because elf3-1pse2 double mutants flower
significantly later in SD than in LD conditions, we concluded that the pse2 mutation restores photoperiod-sensitivity to
elf3-1 mutants. We are using positional cloning to isolate the PSE2 sequence and identify its predicted protein
product, in order to determine the biochemical function of PSE2 in regulating flowering time. Using progeny from a
polymorphic cross, we can differentiate DNA derived from different ecotypes with PCR-based genetic markers of
known locations in order to determine linkage relationships. We have identified linkage between the PSE2 locus and
chromosome II, specifically in a region that spans approximately 20 cM or 4 Mb. We are currently narrowing the
region containing the PSE2 sequence by expanding the mapping population and identifying individual recombination
events with the hope of isolating the PSE2 sequence.
84
BIOLOGY
THE EFFECTS OF CALCIUM CHLORIDE ON THE BACTERIAL GROWTH OF ESCHERICHIA COLI
MN294, PSEUDOMONAS AERUGINOSA 27853, AND STAPHYLOCOCCUS AUREUS RN450
William K. Kunkle, Jr. (Dr. Jennifer L. Koehl), Department of Biology, Saint Vincent College, 300 Fraser Purchase
Road, Latrobe, Pennsylvania 15650-2690
The purpose of this research project was to determine the effects of calcium chloride on the growth of various
bacteria. Calcium chloride is a major component of skeletal bones; therefore, the research completed here can be
related to bone infections and potentially bone diseases such as osteoporosis. Three optimal calcium chloride
concentrations were determined by plating each bacterium on tryptic soy agar (TSA) containing calcium chloride
concentrations as follow: 0.00 M (control), 0.25 M, 0.50 M, 0.75 M, 1.0 M, 1.5 M, and 2.0 M. Each bacterium, per
concentration, was plated three times to ensure that the results were consistent and reliable. The top three
concentrations (those which exhibited the best growth from an observational standpoint) were identified as 0.25 M,
0.50 M, and 0.75 M. These concentrations of calcium chloride were then added to the complex medium, tryptic soy
broth (TSB), and growth curves were run. Optical density readings (OD 600) were taken pre-inoculation and then
approximately every 50 minutes. Gram stains were done to ensure contamination did not occur and to observe any
cellular differences when grown in different concentrations of calcium chloride. Overall, it was noted that the bacteria
growing in the various concentrations of calcium chloride were prone to a relatively short lag phase and a log phase
which peaked at a high optical density in a short amount of time, followed by a quick decrease in optical density. This
was in contrast to the bacteria growing in TSB alone. These control bacteria exhibited a mild lag phase, followed by
an extended log phase without any decrease; stationary and death phases were not present.
REARING THE SMALL HIVE BEETLE (COLEOPTERA:NITIDULIDAE) TO EXPLORE ANTS AS
PREDATORS FOR SMALL HIVE BEETLE LARVAE
Falon A. Mihalic (Dr. Elzie McCord), Division of Natural Sciences, New College of Florida Sarastoa, FL 34243
The Small Hive Beetle, Aethina tumida Murray, has recently become a major pest of beehives in the United States.
The beetle lays eggs in the brood comb of the European Honey Bee (Apis mellifera, L.) hive, and the larvae consume
the bee brood. Larvae exit the beehive to pupate in the ground, and may be vulnerable to attack by ground foraging
arthropods. This study aims to explore the Red Imported Fire Ant (Solenopsis invicta, Buren) as a possible predator
for Small Hive Beetle larvae. Beetle larvae were reared in the laboratory for all experiments and rearing techniques
are summarized. Ant palatability for the Small Hive Beetle was tested with ants collected in the field. Beetle larvae
were exposed to different densities of Fire ants, and their interactions observed. Frequency of attack by the ants and
beetle pupation was recorded. Results of these experiments indicate that the Red Imported Fire Ant is not a viable
predator for Small Hive Beetle larvae.
Real Time PCR Analysis of Myosin Light Chain 2 Expression in Rainbow Trout Swimming Muscle
Megan E. Donato (Dr. Frances Weaver and Dr. David Coughlin), Department of Biology, Widener University, Chester,
PA 19013
Previous physiological experiments in our lab have shown that Rainbow trout (Oncorhynchus mykiss) display
developmental shifts in the contractile properties of their swimming musculature. The younger, smaller parr have
faster contractile properties than older, larger smolts for both their red (slow-twitch) and white (fast-twitch) muscle.
This shift in physiological properties is correlated with a change in myosin heavy chain (MHC) expression. We
predicted that a similar variation in expression of myosin light chain 2 (MLC2) would be observed in the red and white
muscle of the same juvenile stages. MLC2 (or regulatory light chain) reportedly modulates contractile properties and
we hypothesized that shifts in MLC2 expression would relate to developmental changes in contraction kinetics. In this
project, real-time, quantitative PCR (qPCR) with MLC2 isoform-specific primers was used to examine MLC2
expression levels in the red and white muscle of parr and smolt. Total RNA was extracted and single strand cDNA
was obtained. Real time qPCR with the isoform-specific primers permitted the quantification of mRNA expression
levels of fast and slow isoforms of MLC2 in muscle samples. Both red and white muscle expresses both MLC2
isoforms. Red muscle displayed a significant down-regulation of fast MLC2 expression at the transition from parr to
smolt, while white muscle showed a significant up-regulation in slow MCL2 expression in older fish. Both of these
results would correspond to the slower muscle kinetics of both red and white muscle observed in the older fish.
85
BIOLOGY
Timing and Location of the Elastic Extracellular Matrix Protein, MAGP, in Zebrafish Embryos
Justin Harper and Barbara Trask, Department of Zoology, Weber State University, 2505 University Circle, Ogden, UT
84408-2505
A specific set of proteins designed to function in tissues (such as blood vessels, lungs and skin) that undergo repeated
stretching and recoil is collectively called the &#8216;elastic extracellular matrix&#8217;. While the elastic
extracellular matrix is comprised of multiple components, the protein upon which this study focuses is known as
&#8216;microfibril associated glycoprotein&#8217;, or MAGP. This study investigates the translation, timing, and
deposition of MAGP in Danio rerio (zebrafish) during their embryonic development. Because very little is known
about MAGP production in zebrafish during this physiologic process, the information obtained during these studies will
likely contribute to the understanding of the assembly of this important protein mixture. To accomplish this we have
used RT-PCR, molecular cloning, in-situ hybridization, northern blotting, gel electrophoresis and other molecular
biology techniques. The project was initiated by extracting zebrafish RNA from whole embryos from different
developmental stages. Using RT-PCR with degenerate oligonucleotide primers, a 300 bp probe specific for Danio
MAGP was generated. The creation of these primers was accomplished by aligning known sequences of MAGP from
different organisms. Initially this probe was used to analyze relative levels of MAGP through embryonic development.
Whole mounted embryos were then used to assess the spatial deposition of this protein more specifically.
The Responses of Microglia to Deafferentation in the Adult Zebrafish Olfactory Bulb
Harmony M. Smith (Dr. Christine A. Byrd) Department of Biological Sciences, Western Michigan University,
Kalamazoo MI 49008
The body has many responses to injury. One such response is the activation of phagocytic cells, which are
responsible for removing cellular debris. In the central nervous system these cells are known as microglia, which
proliferate near the ventricles and then migrate throughout the brain. Under normal conditions, microglia have a
ramified (resting) morphology; however, they can up-regulate to an amoeboid (mobile) form in response to injury
and/or debris. In this state, the cells are called macrophages. Here we analyze the proliferation, up-regulation, and
migration of microglia in the adult zebrafish brain when responding to damage. Animals were deafferented and then
analyzed at time points varying from 2 hours to 3weeks. This analysis was accomplished using four staining protocols
including IsoB4 Lectin, anti-Ox-42, &#945;- naphthyl butyrate esterase and &#945;- naphthyl acetate esterase. IsoB4
Lectin, &#945;- naphthyl butyrate esterase and &#945;- naphthyl acetate esterase showed a positive staining in brain
sections. While the results indicated that there was neither a proliferation nor a migration of microglia to the area of
damage in response to injury, they did suggest a change in morphology of the cells following deafferentation. These
findings imply that the brain responds selectively to injury. The olfactory system is accustom to dealing with
regeneration, which involves cell death and removal of cellular debris. Thus, the proliferation of microglia often
associated with a wound response, may not be a necessary for to damage caused by injuries such as deafferentation,
where cell death and removal of cellular debris also are probable. Furthermore wound responses can have a
damaging effect on the brain, lack of a considerable response to injury may help the olfactory bulb recover from
wounding. This may have interesting implications for recovery from central nervous system injury and disease.
Westminster College
1300 E 1840 S
SLC UT 84105
86
BIOLOGY
CURCUMIN ENHANCES TRAIL-INDUCED APOPTOSIS IN CHEMORESISTANT OVARIAN CANCER
CELLS
Heather N. Wahl, Sarah K. Fogoros, Milheon Choi, (J. Rebecca Liu), Obstetrics/Gynecology, University of Michigan
Health System 1500 E. Medical Center Drive 4130 CCGC, Ann Arbor, MI 48109
Objective: Curcumin, the active component of tumeric (Curcuma longa), exhibits growth inhibitory activity against
prostate, colon, and breat cancer, however the effect of curcumin on ovarian cancer cells is not known. Curcumin
has been shown to inhibit NF-kB transcription factors. We hypothesized that curcumin could induce cell death in
ovarian cancer cells, and enhance apoptosis induced by tumor necrosis factor-related apoptosis inducing ligand
(TRAIL)/Apo2L.
Methods: Chemoresistant ovarian cancer cell lines SKOV3, ES-2, CaOV3, and OVCA432 were used. Dose response
experiments were performed with curcumin, TRAIL, and curcumin/TRAIL together. Growth inhibition and cell death
was determined by sulforhodamine assay. Apoptotic fraction was determined by staining with propidium iodide and
analyzing subGo content by flow cytometry. Caspase activation was determined by immunoblotting.
Results: Curcumin alone induced cell death in the majority of cell lines at 25 µM. Curcumin at low doses (5-15 µM) or
TRAIL alone was not significantly cytotoxic to the cell lines tested (10% cell death). Pre-incubating cells with
curcumin at low doses (5-15 µM) for 4-6 hours prior to treating with TRAIL resulted in markedly enhanced cell death
(50-70%). The combined treatment of curcumin and TRAIL resulted in activation of both the extrinsic, receptor
mediated apoptotic pathway (cleavage of caspases -8 and -3) and the intrinsic, mitochondria-mediated apoptotic
pathway (cleavage of caspases-9 and -3).
Conclusions: The majority of patients with epithelial ovarian cancer eventually die from chemoresistant, recurrent
disease. Most conventional chemotherapeutic agents activate the intrinsic, mitochondria-mediated pathway of
apoptosis. We have shown that inhibition of NF-kB with curcumin can sensitize ovarian cancer cells to TRAIL
induced apoptosis. Because curcumin and TRAIL in combination can activate both the extrinsic and intrinsic
pathways of apoptosis, they may circumvent chemoresistance caused by conventional chemotherapeutic agents.
A ROLE FOR CELL ADHESION IN THE DEVELOPMENT OF THE APICAL ECTODERMAL RIDGE IN
THE LIMB FORMATION OF CHICKEN EMBRYOS
Alis G. Heidar (Andrew T. Dudley), Department of Biochemistry, Cell Biology, and Molecular Biology, Northwestern
University, Evanston, Illinois 60208, North Central College, Naperville, Illinois 60540.
The apical ectodermal ridge (AER) is a specialized epithelial structure that forms at the distal margin of the limb bud,
and directs outgrowth of the limb. The AER initially forms as a flattened structure that progressively grows into a
prominent, pseudostratified columnar epithelium. Since epithelial morphogenesis involves changes in cell-cell and
cell-matrix interactions, we examined the expression of cell adhesion molecules during AER development.
Specifically, I performed both section and whole mount in-situ hybridizations of chick embryos at various stages in
development, using probes that detect expression of molecules of the cadherin and eph/ephrin families of cell-cell
adhesion molecules, the integrin family of cell-matrix adhesion molecules, and several extracellular matrix associated
proteins. These experiments demonstrated that the homophilic cell-cell adhesion molecule E-cadherin and the
extracellular matrix associated protein Del-1 are expressed at the earliest stage of AER development. Collectively,
these data demonstrate that adhesion molecules are dynamically expressed during AER development and that
specialization of the extracellular matrix may play an important role in formation of the AER.
87
BIOLOGY
The Effect of Beta Glucan on Neutrophil Chemotaxis: Role of PI-3-Kinase
Leon E. Cushenberry Jr. (Dr. Jonathan Reichner), Rhode Island Hospital Department of Surgical Research, Brown
University, Providence, RI 02912
Neutrophils are phagocytic cells that are a component of the immune system’s defense against injury or infection. In
response to chemical signals (chemoattractants) such as complement protein (C5a), N-formyl
methionine-leucine-phenylalanine (fMLP) peptide and host-derived Interleukin-8 (IL-8), neutrophils leave the blood
vasculature and are induced to move through tissue toward affected sites. Neutrophils migrate towards
chemoattractants from different origins like. Soluble beta glucan, a glucose polymer derived from the cell wall of the
yeast Saccharomyces cerevisiae, has recently been shown to enhance neutrophil migration. The present study tested
whether beta glucan could also affect neutrophil migration towards fMLP. Results showed that beta glucan similarly
increased migration towards this chemoattractant. The cellular mechanisms of how beta glucan mediates neutrophil
migration are not well understood. Phosphoinositide 3-kinases (PI3K), cytosolic signaling proteins found in
neutrophils, are involved in the generation of other cell activation signals. Past studies suggest a role for PI3K delta in
the directional but not random migration of neutrophils. Using a specific inhibitor of PI3K delta, preliminary data
shows a reduction in the beta glucan enhanced migration. These results suggest that PI3K delta signaling is important
in mediating the effects of beta glucan on neutrophil migration. Understanding this relationship might lead to
increased therapeutic modalities for treatment of inflammatory diseases.
Utilization of C. elegans for the Measurement of Cellular Respiration
Ashley Dozier, Nick Ragsdale, Department of Biology, Belmont University, Nashville TN 37212
Cellular respiration is commonly explored in both lecture and laboratory exercises of the biology courses offered to
majors. Students confuse much of the lecture material due to the abstract nature of the molecular interactions.
Lecturers attempt to alleviate the confusion through laboratory exercise reinforcement. Currently, many of these
laboratory exercises require the utilization of test organisms that either do not help solidify the cellular respiration
concepts and/or that require the use of expensive equipment. The purpose of this study was to determine the
effectiveness of Caenorhabditis elegans as a model organism for the study of cellular respiration. Vernier&#8482;
software and transducers were utilized to measure the consumption of dissolved oxygen. Results indicate that C.
elegans cellular respiration can be measured with inexpensive materials. Furthermore, the rate of cellular respiration
can be altered with simple manipulations. Specifically, the effects of heat and food source were investigated. C.
elegans is an easy and inexpensive organism to demonstrate cellular respiration.
ROYAL JELLY AND HONEY AS AN ANTIBACTERIAL SURFACE AGENT
Graham Harbison (Dr. Koehl), Biology Department, St. Vincent College, Latrobe, Pennsylvania 15650
The purpose of this research was to determine if Royal Jelly and honey is a effective surface antibacterial agent
against nine pathogenic bacteria. Royal Jelly is produced from the mandibular glands on top of the nurse bee’s head.
It is not as well known as the traditional honey produced by worker bees. Royal Jelly’s purpose serves as a food
source for the larva and the queen bee. Royal jelly is rich in proteins, vitamins B-1, B-2, B-6, C, E, niacin, pantothenic
acid, biotin, inositol and folic acid and has therefore become an attractive supplement. To test if royal jelly is suitable
for hindering bacterial growth, nine common bacteria were grown. Six of these bacteria were Gram-negative
(Psuedomonas aeruginosa, Escherichia coli, Proteus mirabalis, Serratia marcescens, Salmonella colerasuis, and
Enterobacter aerogenes) and three were Gram-positive (Staphylococcus aureus, Staphylococus aeruginosa, and
Bacillus subtilis). The bacteria were grown on solid complex media with sterile paper disks. The disks contained
different concentrations (100%, 75%, 50%, 25%, 10%, and 5%) of royal jelly, blossom honey (light), clover honey
(dark), and distilled water (control). The clearing zones around these disks were measured to determine effectiveness
of the agent and the optimum concentration. Royal jelly has shown partial clearing zones in the concentrations above
50% below that there is little to no clearing. The majority of these clearing zones are partial except about 1 mm
around the paper disk. This area has total clearing and is probably from the Royal jelly spreading out from placing the
disk on the media. The concentrations above and including 75% for the blossom and clover honey show signs of
partial clearing and total clearing where the honey spread out during application for most of the lower concentrations.
Most of the most significant clearing for both the Royal jelly and honeys was the in the Gram-positive bacteria.
88
BIOLOGY
Morphological Analysis of Bacillus Stearothermophilus, Under Thermal Stress Using Atomic Force
Microscopy
Kristen E. LaRue, Tekyia Lewis and Aisha Pitts (Dr. Phyllis Freeman-Junior, Mrs. Patricia McCarroll,, Ms. Tiffany
Crenshaw, Ufoma Akoroda and Dr. Weije Lu), Fisk University, Physics Department, 1000 17th Avenue North,
Nashville, TN 37208
Threats of biological warfare have increased interest in spore producing bacteria such as Bacillus Stearothermophilus
and Bacillus Anthracis. Left untreated, the inhalation of Bacillus anthracis spores by humans often leads to internal
bleeding, server damage to the tissue of major organs, respiratory failure and death. In this investigation, the effects
of temperature stress on the morphology of Rhodospirillum rubrum, Microccocus roseus and Bacillus
stearothermophilus were studied using Atomic Force Microscopy (AFM). In this report special attention is given to
the sporulation of Bacillus Stearothermophilus, under thermal variations. Additionally, Microccocus roseus and
Rhodospirillum rubrum were observed via AFM after atmospheric and nutritional variations, respectively. Results of
this study indicate that the induction of thermal stressors largely inhibited growth, altered structural morphology, and
influenced the germination of spores.
ASSAYING FOR PROTEIN-PROTEIN INTERACTIONS IN C. ELEGANS USING THE YEAST
TWO-HYBRID SYSTEM
Ria Saroop and Havilah Gates, (Casonya M. Johnson), Department of Biology, School of Computer, Mathematical,
and Natural Sciences, Morgan State University, 1700 E Coldspring Lane, Baltimore, MD 21251
The broad number of mammalian developmental events affected by bHLH proteins brings to light the importance of
understanding how these proteins function inside the cell. Specifically, the bHLH protein, NeuroD, is involved (1) in
the differentiation of neuronal cells, (2) in ensuring that these cells maintain their differentiated state, (3) in the
differentiation of endocrine cells, and (4) in the production of insulin. Mutations in the human NeuroD have been
linked to increased susceptibility to both Type I and Type II diabetes mellitus. Mammalian members of the bHLHB5
subfamily can inhibit NeuroD activity in cell culture. Recently, our laboratory reported the molecular characterization
of the C. elegans homolog ofo bHLHB5, HLH-17. A homologs of bHLHB5 has been identified in the soil nematode,
Caenorhabditis elegans, and the role of NeuroD in regulating neuronal differentiation and in maintaining the
differentiated state of neuronal cells implies that HLH-17 will have critical roles in neuronal development. A long term
goal of our laboratory is to more thoroughly characterize the functional role HLH-17. The first step in accomplishing
this goal, is to identify the bHLH protein(s) that is capable of dimerizing with HLH-17 to regulate transcription. We are
using the Gateway®-compatible yeast-two hybrid system to assay for dimerization between HLH-17 and other bHLH
proteins in C. elegans. To date, we have fused both the full-length protein and the bHLH domain of HLH-17 to the
Gal-4 DNA-binding domain. We have also generated fusions between the Gal-4 activation domain and 13 of the 39
bHLH proteins found in C. elegans. We report here on our cloning strategy to generate these fusion proteins and the
results of our control experiments which show that HLH-17 is not self-activating in the yeast two-hybrid system.
(supported by NSF grant #MCB0516355, NIH/RCMI grant #5G12RR017581 and NIH/RISE grant #R25GM058904)
89
BIOLOGY
DESIGN AND SYNTHESIS OF A Ga12-INTERACTING FUSION PROTEIN
David W. Culver (Ted Meigs), Department of Biology, The University of North Carolina at Asheville, Asheville, NC
28804
G proteins are a class of proteins that reside just inside the surface of cells. As receptor-coupled proteins, they
receive “information” from cell surface proteins that bind ligands outside the cell, and transmit signals into the cell to
cause changes in additional, “downstream” proteins inside the cell, known as effectors. These proteins, which are
found in essentially all eukaryotic cells, mediate a wide variety of cellular processes including cell proliferation and
cancer development, as well as cancer metastasis (the stage at which cancer spreads to other areas of the body).
The G protein trimer consists of a, b, and γ subunits. Each G protein is classified according to the identity of its a
subunit. Using such identification, G proteins are classified into four subfamilies, one of which is G12. The G12
subfamily contains two members, Ga12 and Ga13. Ga12 has been demonstrated to play a role in cell proliferation,
differentiation, and cancer development. Ga12 interacts with several other proteins, including cadherins, Protein
phosphatase-5, Bruton’s tyrosine kinase, p115RhoGEF, Heat shock protein-90, zona occludens-1, and radixin; as well
as AKAP-lbc, AKAP110, and PYK2. To understand the role that Ga12 plays in cancer, as well as other cellular
processes, the interactions between these various proteins and Ga12 must be understood in greater molecular detail.
However, in order to test these proteins for their interaction with Ga12, protein “fusions” containing the interacting
proteins must first be created. This was the purpose of my research this summer—engineering constructs containing
the Ga12-interacting protein AKAP-lbc. One of the most difficult parts of this research was simply procuring the DNA
required to create this interacting protein construct. After procuring this DNA from a laboratory in Switzerland, several
strategies for engineering the desired DNA construct were designed. This research is very close to having a construct
for immobilized AKAP-lbc in hand, and will use this DNA to produce protein molecules useful for further research.
This work will involve testing this protein for binding to mutants of Ga12. The results of these tests may shed light on
the processes of cancer development.
EAVESDROPPING BY THE DUETTING KATYDID AMBLYCORYPHA ALEXANDERI (OTHOPTERA,
PHANEROPTERINAE)
Kendra A. Dagg, and David Mabe (Timothy Forrest), Department of Biology, The University of North Carolina at
Asheville, Asheville, North Carolina 28804
Katydids in the genus Amblycorypha form duets during reproductive pair formation. Duets between males and
females are initiated by the male's calling song and females answer with ticks precisely timed within the male's song.
As the duet continues, one or both sexes moves toward the other. Mating follows. Because duets provide
information about the location of sexually receptive females, we hypothesized that other males might eavesdrop on a
duetting pair and respond by moving toward females. We tested our hypothesis in an indoor arena using duets of
Amblycorpha alexanderi. A speaker broadcasting the song of a male was randomly placed at 1 of 8 locations around
the arena and the female’s speaker was randomly assigned to 1of 2 positions at 90o relative to the male speaker. For
each trial (N=21), a single male was placed at the center of the arena and we marked the position (to the nearest 5°)
that he exited the arena. Most males exited the arena within 40° of the female speaker and the distribution of their
azimuths was significantly clumped around the female speaker. Rather than moving directly toward the female
signal, male A. alexanderi were more likely to exit between the two speakers. Eavesdropping by male A. alexanderi
is probably a behavioral tactic that allows males to locate and mate with females that are answering other
competitors.
90
BIOLOGY
PHOTOSYNTHETIC RESPONSES OF MISCANTHUS SINENSIS ANDERSSON, TO VARIABLE LIGHT
ENVIRONMENTS I. LIGHT RESPONSE CURVES
Renee Fortner, (Jonathan Horton), Department of Biology, The University of North Carolina at Asheville, Asheville, NC
28804
Miscanthus sinensis, Andersson, a C4 grass native to eastern Asia, is an emerging invasive species in the United
States. Most C4 plants generally thrive in warm climates with full sun and dry conditions and are often unable to
compete in low light environments. The focus of this study was to examine the photosynthetic responses of M.
sinensis growing along a light gradient, to assess its ability to adapt in low light environments. Plants adapted to
shaded environments exhibit photosynthetic and morphological characteristics that differ from those adapted to sunny
habitats. Shade species have reduced photosynthetic rates and become light saturated at lower light levels.
Quantum yield are generally higher for plants growing in low light. Plant leaves also tend to be thinner in shade,
resulting in lower dark respiration rates that cause lower light compensation points. There also tends to be a decrease
in chlorophyll a:b ratios as plants allocate more to light harvesting complexes. Plants of M. sinensis were sampled
growing under two natural and one artificial light gradients (ranging from 5% to 100% full sunlight). Photosynthetic
light response curves were constructed to examine photosynthetic characteristics along these light gradients.
Photosynthetic rates in all plants (grown in both high and low light levels) were very high and did not become light
saturated at full sunlight (2000 mmol m-2 s-1), a response typical of high-light adapted C4 plants. However,
photosynthetic quantum yield was higher in plants growing in lower light, typical of low light acclimated plants.
Specific leaf mass and dark respiration rates decreased slightly in plants growing in lower light, resulting in lower light
compensation points. However, there was no shift in the chlorophyll a:b ratio in any of our plants. While, M. sinensis
possesses some characteristics that will allow it to survive in shade, our results do not suggest that it is very shade
tolerant.
PHOTOSYNTHETIC RESPONSES OF THE INVASIVE EXOTIC, MISCANTHUS SINENSIS ANDERSSON,
TO VARIABLE LIGHT ENVIRONMENTS II. INDUCTION AND INDUCTION LOSS
Maya Goklany (Jonathan L. Horton), Biology Department, The University of North Carolina at Asheville, Asheville, NC
28804
Miscanthus sinensis Anderss. is a C4 grass that has spread throughout open, disturbed habitats in the Eastern United
States. This species has been documented in forest understories where it may alter ecosystems by out-competing
native species. The focus of this study was to characterize the photosynthetic induction responses of M. sinensis to
determine whether the C4 pathway limits the ability of this species to respond to sun flecks, an important resource in
forest understories. Plants were sampled along two natural and one artificial light gradients (ranging from 5% to
100% of full sunlight) on the Biltmore Estate in Asheville, North Carolina. Rates of photosynthetic induction and
stomatal opening in response to increasing light were measured, as were the rates of induction loss and stomatal
closure in response to decreasing light. Typically, shade-tolerant plants induce quickly and lose induction slowly, in
order to make efficient use of sunflecks. Upon exposure to high light, M. sinensis growing in higher light induced
more quickly (4 – 5 minutes to 50% maximum) than shade plants (6 – 8 minutes to 50% maximum). Plants growing
in lower light had slower stomatal opening (22 – 23 minutes to 90% maximum) compared to plants growing in higher
light (18 – 19 minutes to 90% maximum). There was no significant difference, however, for photosynthetic induction
up to 90% maximum between plants grown in high and low light. In addition, there was no significant difference
between the ability of high and low light plants to retain their induction state when exposed to decreasing light. Slow
stomatal responses in returning to high light following dark acclimation, and slow photosynthetic induction during
variable light indicates that the C4 pathway does limit the environmental conditions in which M. sinensis can grow.
91
BIOLOGY
DETERMINATION OF ANTI-CARCINOGENIC PROPERTIES OF CHINESE HERBS
Ryan M. Kelly (Thomas E. Meigs), Department of Biology, The University of North Carolina at Asheville, Asheville, NC
28804
Traditionally, many of the healing compounds utilized in pharmaceutical therapy have been identified first in their
natural botanical state. Many “modern” medicines such as aspirin (from the Willow tree) and taxol (from the Yew
tree) were first recognized as botanical supplements and were later synthesized in laboratories for legitimate medical
use. The purpose of this research is to treat various cancer cell lines with a variety of botanical extracts. The plants
used for this testing have been used for centuries in Traditional Chinese Medicine and are claimed to have
anti-cancer properties. The intention is to determine whether such extracts have any effect on specific types of
cancer cells grown in the laboratory. The procedure to be used for determining the success of the herbal treatments is
based on observation of the cell’s growth rate through quantifying the protein concentration with in the cells. If the
cells respond positively to the treatment, their growth rate should slow, and protein content will abate over time. This
procedure of quantifying protein is performed by creating a standard protein curve and a series of growth assays. The
process of measuring the success of cell growth in this experiment can be categorized into three sections. The first
portion deals with the actual growth of the cell lines, which are two epithelial cancer cell lines: one of breast origin
(K435) and the other of colon origin (SW480). The next part of the process involves creating a standard protein
assay. This is developed as a measuring tool to determine the protein content of the cell lines. The third portion of
the procedure involves creation of growth assays of the two cell lines with the botanical extracts applied. The cells
are harvested at consecutive time periods of 24-hour intervals and their protein content is quantified to determine the
success of the treatments.It is expected that at least one of these extracts will have an effect on the proliferative rate
of at least one of the cell lines. Promising combinations of cell line and extract source will be used to determine an
inhibitory constant that will allow comparison of the effectiveness of these different extracts
PHYSIOLOGICAL ADAPTATIONS OF THE UPPER LIMB TO THE BIOMECHANICAL ENVIRONMENT
OF PLAYING TENNIS
Natasha C. Lucki (Christopher Nicolay), Department of Biology, The University of North Carolina Asheville, Asheville,
NC 28804
Like many sports and occupations, tennis requires the relative overuse of one hand. Grip strength and endurance in
25 male and 24 female Division I tennis players was compared with non-athletes (17 males, 18 females) to examine
the physiological changes associated with extreme asymmetry in training that occurs in tennis players. Grip strength
and endurance were measured during three experiments: single-repetition maximum voluntary contraction, 30
consecutive repetitions, and a 30-second static hold. Tennis players had significant directional asymmetry in forearm
strength (force production), as well as significant increases in forearm circumference, wrist circumference, and palm
width of the dominant limb. The dominant hand of female tennis players produced about 22% more force than the
opposite hand, while the difference between hands of males was 15%. However, tennis players had no significant
differences between the hands in endurance (percent reduction in force output) in either the static (30-s hold) or the
dynamic (30 repetition) tests. In contrast to tennis players, non-athletes had no significant differences between hands
in forearm anthropometrics, grip strength, or any measure of endurance. These findings indicate that the training
involved with tennis promotes muscle mass development and absolute strength in the dominant arm, but does not
significantly alter muscular endurance as measured by these experiments. To further understand the biomechanical
environment experienced during tennis, we are using a specially-designed tennis racket that records the actual grip on
the racket during different shots (serve, forehand, backhand). This data will help use understand how the hand grips
the racket during the swing, and how these forces are related to our results.
92
BIOLOGY
COMPARATIVE ANALYSIS OF THE CRANIAL MORPHOLOGY OF VAMPIRE BATS
(PHYLLOSTOMIDAE: DESMODONTINAE)
Jillian Shank (Chris Nicolay), Department of Biology, The University of North Carolina at Asheville, Asheville, North
Carolina 28804 USA
Two primary factors that influence the morphology of the mammalian skull are diet and evolutionary constraints. Bats
of family Phyllostomidae exhibit extreme dietary diversity, with different species that specialize on fruit, nectar,
insects, and other vertebrates. However, the three species of subfamily Desmodontinae (true vampire bats) may be
most unique, as they feed exclusively on vertebrate blood. Desmodus rotundus feeds on mammalian blood, while
Diaemus youngi and Diphylla ecaudata require avian blood. Principle Component (PC) Analysis was conducted to (1)
examine how vampires differ from other phyllostomids, and (2) identify patterns of morphological variation within
three species of vampires. To quantify skull morphology, 13 cranial measurements were taken on each of the three
vampire species (Desmodus rotundus n=40, Diphylla ecaudata n=40, Diaemus youngi n=25) and 114 individuals from
8 diverse species of phyllostomids. The first PC Analysis demonstrated that vampires were morphologically distinct
from other phyllostomids. PC1 (which accounted for 59% of the variance and generally reflected size) and PC2
(which added 25% of the variance based on length and width of the rostrum and mandible) demonstrate that
vampires form a distinct group from other phyllostomids as small bats with short, blunt snouts, and short, wide
mandibles. The short faces and specialized anterior teeth of vampires reflect the bite required to obtain blood.
Because vampire species each feed on different hosts, a second PC analysis examined only these three species.
Each species has a distinct cranial morphology. PC1 (size) accounted for 57% of the variance in the sample, and
separated Diphylla ecaudata from the two larger species. Another 17% of the variance was explained by PC2, which
separated Diaemus youngi from the other vampires based on its shorter, wider skull and mandible, reduced canine
length, and taller coronoid process. These factors suggest that Diaemus youngi has a large temporalis muscle and a
particularly forceful bite. The distinct morphologies of the three species of vampire bats indicate that even within the
unusual, specialized niche of blood-feeding, species employ different strategies to meet varying feeding demands.
93
BIOLOGY: BOTANY
EVALUATION OF PRECURSORS ON GROWTH AND TAXANE PRODUCTION IN CALLUS CULTURES
OF Taxus media
Graham D Clinthorne (Robert A Smith), Department of Biological Sciences, University of the Sciences in Philadelphia,
Philadelphia, Pennsylvania 19104
This study has analyzed the effect of the precursors phenylalanine, benzamide, and sodium acetate on growth and
taxane production in callus cultures of Taxus media cv. Hicksii. Previous studies indicated introduction of these
precursors into the media may increase taxane production. We tested the effect of precursor feeding at various
concentrations with the goal of achieving sustainable growth of cultures and production of taxane compounds. Growth
was measured by increase in fresh weight. Taxane production was analyzed using HPLC. Compounds were extracted
from the solid media using two 24 hour methanol washes. Without precursors, growth occurred; however, no taxanes
were produced. Addition of all three precursors at 1.0 mM concentration stimulated production of one suspected
taxane compound; however, growth ceased. In experiments using only one precursor in each media, benzamide was
found to be responsible for production of taxane compounds. At concentrations of 1.0, 0.5, and 0.2 mM, but not at 0.1
mM, taxanes were produced. Growth also occurred at 0.2 mM. Thus, 0.2 mM benzamide was considered optimum for
growth and production of taxanes.
CANOPY LIGHT AND PHOTOSYNTHESIS PHYLOGENETIC RELATIONSHIPS IN SPHAGNUM
Lynn E. Aclander (Dr. Steven Rice), Department of Biological Sciences, Union College, Schenectady, New York,
12308
Globally, Sphagnum (the genus of peatmosses) stores more carbon than any other genus of plants. The need to
understand Sphagnum physiology and energy exchange is vital, especially within the canopy where energy absorption
occurs. The purpose of this experiment was to study how variation in canopy characteristics affects light attenuation
and photosynthesis in related species of Sphagnum. I used nine different species of Sphagnum moss (S.fallax,
S.magellanicum, S.recurvum, S.tenerum, S.portoricense, S.lescurii, S.contortum, S.squarrosum, and S.girgensohnia)
that encompass physiological variation in the genus. I measured light absorption in the top 3 cm of the canopy,
chlorophyll content and surface roughness and compared these with plant tissue bulk density, canopy biomass, and
photosynthesis (maximum gross photosynthesis and photochemical efficiency). The physiological characteristics
were analyzed using an independent contrast method to reduce the effects of phylogenetic history on the results. The
phylogenetic contrast method explores the evolutionary convergence of the structure-function relationships between
related species. The results show a significant positive relationship between gross photosynthesis and bulk density,
and a negative relationship between photochemical efficiency and canopy light absorbance, based on the
phylogenetic contrasts. The relationship between branches, leaves, and surface roughness to the whole canopy
function is important for understanding their integration in the Sphagnum canopies. These results show evidence of
evolutionary convergence of physiologically important characteristics in the genus Sphagnum.
94
BIOLOGY: BOTANY
EFFECTS OF CADMIUM ON PHOTOSYNTHETIC APPARATUS IN BRASSICA RAPA
David Ramirez III, Taylor Welty, and Jonathan Abesamis (Sibdas Ghosh), Department of Natural Sciences and
Mathematics, Dominican University of California, 50 Acacia Avenue, San Rafael, CA 94901
Cadmium (Cd 2+) may be present in soils naturally at levels of 0.01-7 ppm. Dispersion primarily through
anthropogenic activities concentrates Cd2+, which may become phytotoxic. Even though protective measures such
as liming of soils are established, examining Cd2+ induced effects on photosynthetic apparatus is of interest since
heavy metals elicit specific plant responses. Cd2+ influx into the chloroplast can occur via some calcium channels,
morphologically affecting the entire plant. Most evident is chlorosis of leaves, which is explained by the reduction in
magnesium uptake and overall photosynthetic pigment content. Seeds of Canola (Brassica rapa L. cv I-33) were
germinated in flats containing Peters Redi Earth potting mix and placed in a growth chamber at 25°C and 80%
relative humidity. Seedlings were supplied half strength Peters Nutrient solution (20:20:20) by the watering system
throughout experiment. Plants were grown under 24-h photoperiod with 53 &#61549;mol m-2 S-1 of mixed fluorescent
and incandescent light. Plants were harvested the 18th day after sowing and specimens frozen in liquid nitrogen and
stored at -18 °C. We investigated the effects of increased levels of CD2+ by monitoring the growth and development
of Brassica rapa. Quantification of pigments and total protein was performed and the levels of both large and small
subunits of ribulose 1, 5-bisposphate carboxylase were analyzed. The activity of ribulose 1, 5-bisphospate
carboxlayse declined in Cd2+ treated plants. The Cd2+ influx into the cytoplasm and/or chloroplast can occur via
some calcium channels, morphologically affecting the entire plants. These morphological effects include the
cholorosis of leaves, lowering photosynthetic rates especially, photosystem II (PSII) efficiency and a decline in the
activity of ribulose 1, 5-bisphosphate carboxylase.
An Investigation into the role of Hosta (Liliaceae) as a source of allelopathic chemicals
Elizabeth Klinke, Holy Family University, School of Arts and Sciences, 9701 Frankford Avenue, Philadelphia, PA
19114
The study of allelochemicals is an emerging field in plant biology. The possible use of naturally-produced chemicals
as herbicides would be desirable over the use of synthetic compounds that may damage non-pest plants or
thatdegrade slowly in the environment. The purpose of this study is to determine the effect of Hosta leaf extract on
three target plants: Rye (RY), Clover (C), and Radish (R). A known source of allelochemicals (Alfalfa - A) was also
used for comparison to Hosta. Seeds were germinated in Petri dishes and treated with either dH2O or a dH2O
extract from the source plants: Alfalfa or Hosta. Seed germination as judged by radicle emergence from the seed
coat (approximately 48 hours) were reduced by: 14% for RY-A and 19% for RY-H, 22% for C-A and 9% for the C-H,
18% for the R-A and 35% for the R-H as compared to controls. Mean fresh weights showed a similar decrease
ranging from -21% to a -79% for all target plants. Leaf lengths for RY decreased of 35% and 34% for both Alfalfa and
Hosta treated, respectively. These results show that Hosta leaf extract is at least as effective in inhibiting plant growth
and development, as is Alfalfa. Keywords: Allelochemicals, Hosta.
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BIOLOGY: BOTANY
ANALYSIS OF COLEUS BLUMEI FOR USE IN A QUANTITATIVE, INQUIRY-BASED FRESHMAN
PHOTOSYNTHESIS LABORATORY.
Orlando D. Hardrick (Dr. L.E. Wimmers), Department of Biological Sciences, Towson University, Towson, MD 21128
We evaluated Coleus blumei for use in a quantitative, inquiry-based freshman biology laboratory. In Biology 201
courses at Towson University students participate in four different laboratory modules. In each the students design,
execute and analyze their own experiments. In the photosynthesis module students are encouraged to pose logicallybased hypotheses about the relationships between photosynthetic rate and irradiance level or carbon dioxide level.
Students can also investigate the photosynthetic rates of varying species but unfortunately their content knowledge is
not adequate enough for them to make logical hypotheses in this area. This project introduces the opportunity for
students to develop a logical hypothesis concerning the correlation between photosynthetic rate and the pigments
chlorophyll and anthocyanin and to test that hypothesis using Coleus plants of various leaf pigmentation patterns
including all green (chlorophyll only), primarily white (no chlorophyll or anthocyanin), dark red (chlorophyll and
anthocyanin) and primarily light red (anthocynanin only. Photosynthesis was measured using a student-grade CO2
gas analyzer system. Photosynthesis was reliable and robust, reaching steady state levels quickly enough for students
to test several plants during one laboratory period. Variability between plants was low, permitting useful statistical
analysis. Photosynthetic rate was not strongly dependent on leaf age and logically predicted student hypotheses were
supported by photosynthetic rates in plants of different pigmentation.
Authors Home Address: Baltimore City Community College, Baltimore, MD 21215
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BIOLOGY: MICROBIOLOGY
SCREENING FUNGAL FRUITING BODIES FOR ANTI-MYCOBACTERIAL ACTIVITY
Cassandra Sleger (Kathleen Engelbrecht, Dr. William Schwan, and Dr. Marc Rott), Department of Microbiology, The
University of Wisconsin-La Crosse, La Crosse, Wisconsin 54601
One of the world’s most pressing health problems is tuberculosis. One third of the world’s population is believed to be
infected with tuberculosis and many of the causative agents are drug resistant. New drugs to treat tuberculosis are
needed. Because fungal fruiting bodies (mushrooms) growing in the wild share an ecological niche with saprophytic
mycobacteria, we hypothesize that to avoid being degraded, some fungal fruiting bodies may contain
anti-mycobacterial compounds. The objective of this study was to screen a library of extracts from the fruiting bodies
of 146 fungi for activities that inhibit the growth of Mycobacterium smegmatis, a non-pathogenic indicator strain for
the causative agents of tuberculosis. Fungal fruiting bodies were collected, identified (a reference sample was
removed and stored), dried, weighed, and extracted using standard protocols. The dried extracts at a standardized
concentration in dimethyl sulfoxide, were screened for anti-mycobacterial activity using a modified minimum inhibitory
concentration (MIC) procedure in 96-well microtiter plates. Briefly, extracts were serially diluted two fold in growth
medium and bacteria were added to each well. After incubation for two days, the lowest dilution of extract inhibiting
growth of the bacterium was defined as the MIC. Extracts showing activity in these initial screens were re-assayed in
a double blind test for reproducibility and to eliminate reader bias. Controls included wells with rifampin, no fungal
extract, and no bacteria. No growth inhibition was observed with concentrations of dimethyl sulfoxide at or below 5%.
MIC values for active crude extracts ranged from 12.8 mg/ml to 0.2 mg/ml. Eleven of 146 extracts had MIC values <
0.8 mg/ml. Selected extracts with MIC values < 3.2 mg/ml are being tested for activity against a variety of fast and
slow growing pathogenic mycobacterial species. In future work, we will attempt to purify and characterize molecules
responsible for the inhibition of growth from selected extracts. This research could lead to the discovery of novel
antibiotics for treating tuberculosis.
CLONING, EXPRESSION, AND PURIFICATION OF SERINE-RICH REPEAT PROTEINS OF GROUP B
STREPTOCCUS FOR DETERMINATION OF GLYCOSYLATION PATTERNS
Allyson L. Samselski*, Amanda J. Billings, and Paul R. Watson (Dr. Kyle Seifert), Department of Biology, James
Madison University, Harrisonburg, VA 22807
Group B streptococci are a leading cause of neonatal pneumonia, sepsis, and meningitis, as well as pathogens of
adults with underlying health conditions. Although all 9 serotypes have been isolated from infected individuals,
serotype III organisms account for greater than 90% of all meningitis cases. Serotype III organisms can be further
divided based on genetic techniques, which separates these organisms into 4 groups. One of these subgroups causes
over 90% of all cases of GBS disease caused by serotype III organisms. Genetic analysis has determined that this
more pathogenic, or “hypervirulent” lineage, has a unique surface serine-rich repeat glycoprotein (Srr-2) that is
different than a similar glycoprotein (Srr-1) that is found in the less pathogenic strains. This project focuses on
determining differences in glycosylation between the two serine-rich repeat glycoproteins. The serine-rich repeat
proteins have been PCR-amplified, cloned, and expressed in E. coli. Amino-terminal fragments have also been
cloned and expressed. Expressed proteins are soluble, and purified proteins will be used as immunogens in rabbits to
produce polyclonal antibodies so that the native glycoproteins can be purified and analyzed for carbohydrate content.
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BIOLOGY: MICROBIOLOGY
Characterization of Mycoplasma pneumoniae Transposon Mutants Using Complementation and
Western Blot Analysis
Bryan E. Hart, National Science Foundation, Department of Microbiology, University of Georgia,, Microbiology
Department, The University of Georgia, 527 Biological Sciences, Athens, GA 30602-2605
Mycoplasma species are among the smallest organisms capable of a self-replicating existence. Over the course of
their evolution, mycoplasmas have produced a streamlined genome with limited biosynthetic capabilities and the
necessity of symbiotic host interactions for survival. In the case of Mycoplasma pneumoniae, these symbiotic
interactions are potentially detrimental to the host, producing an atypical or walking pneumonia in
immunocompromised individuals. Acting as both an extracellular and intracellular pathogen, M. pneumoniae is an
important infectious agent to study, in attempts to more fully understand its virulence factors and modes of invasion
for novel diagnosis and treatment of this potentially lethal organism. This study involved culturing, Western blot
analysis, and genetic complementation of four different M. pneumoniae mutants in order to further understand the
nature of their deviations from the wildtype state. Gene knockouts were previously produced by transposon
mutagenesis using a modified Tn4001. Genetic complementation was conducted by amplifying wildtype MPN 104,
524, 444, 443, 442, 436, and 254 genes, followed by the transformation of knockout mutants with wildtype genes
ligated into the complementation vector, pKV304. These transformants were then compared to both wildtype M.
pneumoniae and clonal mutant cell growth on SP4 motility medium. Western blots were developed for the Duf-16
motif mutants as well as the MPN245 knockouts to investigate how attachment organelle and virulence associated
protein synthesis was affected by the disruption of each gene. The proteins investigated included HMW2, HMW3, P1,
B, p65, p41, p30 p28, and p24. While the Duf-16 and cinA complementations were inconclusive, the MPN442
complementations supported the possible presence of antisense RNA interference against MPN444 translation.
Western blotting revealed no effect on attachment organelle protein synthesis yielded by the Duf-16 and cinA
mutations. Therefore aberrant motility and cellular division, which may be the basis for these observed mutant
phenotypes, is likely to result from an alternative source than interference in tip protein production.
THE EFFECT OF CULTURE MEDIUM NUTRIENT COMPOSITION ON THE MORPHOLOGY,
SPORULATION, AND ANTIBIOTIC PRODUCTION BY A BACTERIUM ISOLATED FROM SOIL
Alexis D. Ulerich (Richard T. St. John, Ph.D.), Department of Biology, Widener University, Chester, PA 19013
The effect of the culture medium nutrient composition on the morphology, sporulation and amount of antibiotic
production by a filamentous bacterium was studied. The bacterium is a gram positive filamentous Actinomycete
isolated from soil. It produces a narrow spectrum antibiotic that inhibits gram positive bacteria but not gram negative
bacteria. We observed that the degree of branching in broth culture, the production of visible asexual conidiospores
on agar plates, and the amount of antibiotic produced was significantly affected by the nutrient composition of the
culture medium. Initially three different standard bacterial culture media were employed; Tryptic Soy medium,
Nutrient Broth/Agar medium, and King’s Medium. The results indicated that antibiotic production was greatest on the
King’s Medium but that this medium suppressed filamentous growth and completely blocked conidiospore production.
Cultures grown on Nutrient Agar plates produced standard filamentous colonies with visible conidiospores but
antibiotic was produced in reduced amounts in nutrient broth cultures. Tryptic Soy cultures produced intermediate
results for both filamentous growth and antibiotic production but did not show any production of conidiospores. Two
culture medium components, glycerol and glucose, are variables in the recipes for these three culture media. A series
of experiments are underway to determine the exact nutrient component of the culture medium responsible for the
observed differences and the nature of the regulation of sporulation and antibiotic production by this soil bacterium.
98
BIOLOGY: MICROBIOLOGY
EFFECTS OF BACTERIAL CELL WALL COMPONENT (LPS) AND BACTERIAL TOXIN (ALPHAHEMOLYSIN) ON RETINAL EPITHELIAL CELL CULTURES (CRL-2302) TO DETERMINE CE
Krystal L. Weissert (Dr. Jennifer Koehl), Department of Biology, Saint Vincent College, Latrobe, Pennsylvania 15650
Bacterial endophthalmitis, which commonly results in visual loss, is the inflammation of the interior part of the eye.
Inflammation is commonly caused by bacterial components such as Gram-positive and Gram-negative bacterial cell
walls and bacterial toxins. The purpose of this project was to use an in vitro retinal epithelial cell culture to look at the
effects of eye toxicity to better understand bacterial infections of the eye. Different bacterial toxins and cell wall
components decrease cell viability at different rates and severities. The bacterial components used in this experiment
were lipopolysaccaride (part of the outer membrane of Gram-negative bacteria cell wall) and alpha-hemolysin (a toxin
isolated from Gram-positive Staphylococcus. aureus). Cells were maintained utilizing cell culture techniques and then
specific concentrations of bacterial components were added (2, 5, and 10µl alpha-hemolysin, 2,5,10, and 20µl
lipopolysaccaride for 24 hours). Cell viability and proliferation of the retinal cell culture as well as mitochondrial
function were measured with the WST-1 test. This test measured the number of viable cells through the detection of
cleaved tetrazolium salts by mitochondrial dehydrogenase and was expressed as percentages of the optical density of
treated versus untreated controls. The data was obtained by an optical density microplate and analyzed using an
independent-samples t-test. The alpha-toxin treated cell viabilities were significantly lower than the control cell
viabilities (p&#8804;0.05; 5 trials). The LPS treated cell viabilities were not significantly different to the control cell
viabilities (p>0.05; 4 trials). This experiment helps to explain how bacteria and their components alter the human eye
system. It also develops different severities that bacterial components have on the retinal cell cultures. It was found
that small amounts of alpha-hemolysin caused detrimental effects on cell viability, while lipopolysaccaride did not
cause detrimental effects on cell viability even though they are both components of bacteria.
NEUROANATOMICAL AND STEREOLOGICAL ANALYSIS OF THE DORSAL ANTERIOR CINGULATE –
PREFRONTAL MICROCIRCUIT INVOLVED IN COGNITIVE CONTROL
Zeljko Dvanajscak (Dr. Robert Morecraft) Department of Neurological Sciences, University of South Dakota School of
Medicine, Vermillion, SD 57069
Neuroimaging, lesion, and anatomical studies have proposed that the dorsal Anterior Cingulate Cortex (dACC), a
midline, limbic, frontal-lobe brain structure crucially contributes to the control of cognitive functions that underlie the
ability to overcome stimuli-induced, reflexive reactions in order to direct behavior toward future goals. The cognitive
contribution of the dACC depends on its interaction with the Prefrontal Cortex (PFC), a brain region known as the
executive center in the control of behavior. Evidence that the dACC is reciprocally interconnected with the PFC
abound. However, the precise characterization of this cognitive microcircuit has never been undertaken. Three rhesus
monkeys (Macaca mulatta) were used to uncover the patterns of dACC innervation of the PFC. By the use of
anterograde neurotracers, localization of synaptic terminals originating from neurons of the various dACC
subdivisions within the PFC was possible. By analyzing the coronal sections under brightfield microscopy with the aid
of computer software, unique findings were uncovered. First, the manner by which dACC innervates the PFC is by
clearly demarcated, interdigiating, translaminar arrays of terminal synapse columns readily observable by the plotting
analysis. Second, the reconstructions of the lateral cortex in two dimensions and the localization of labeled terminal
synapses revealed a qualitative diagram that neatly describes the projectional density patterns. Third, the use of
unbiased stereology, a computerized statistical tool, exposed the strengths and patterns of this connection. The
estimates show that the dACC subdivisions 24a and 24b project about 2.5 – 3 million synapses each to the PFC,
while subdivision 24c projects at one-fifth the strength. Furthermore, subdivision 24a principally innervates
ventrolateral PFC area 46, while subdivision 24b mainly innervates dorsolateral PFC area 9, suggesting functional
specializations of the dACC subdivisions within subsets of cognitive function. As physiological studies continue to
progress towards a unifying theory of dACC function, our study provides the anatomical data essential for complete
understanding of this microcircuit`s contribution to human cognition and behavior.
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BIOLOGY: MICROBIOLOGY
COMPARISON OF ANTIBIOTIC RESISTANCE IN ENTEROCOCCUS SPP. ISOLATED ABOVE AND
BELOW SEWAGE TREATMENT PLANT OUTFLOWS.
Adam A. Aguiar, Lydia Horvath, and Michael Levandoski (J. H. Middleton), Department of Biology, Fairleigh Dickinson
University, Madison, NJ. 07940.
We compared the level of antibiotic resistance of Enterococcus spp. isolated from shallow surface waters at sites
above and below sewage treatment plants in Morris County, NJ. Sampling sites included the Whippany River
(Butterworth treatment plant), Loantoka Brook (Woodland) and a drainage culvert (Fairmont). Water samples were
collect aseptically in May and in September and enterococci were isolated and enumerated on mEI agar. Total
enterococcal counts were higher at sites above sewage outflows than below outflows. All Enterococcus spp. were
verified by standard techniques and identified biochemically. At each site the nosocomial pathogens E. faecalis and
E. faecium were present in higher concentrations below than above outflows. Twenty-four unique isolates were
selected from each sample and replica plated on 13 different antibiotic media. No enterococcal isolates exhibited
resistance to chloramphenicol, vancomycin (30 &#956;g/ml) or to high level gentamycin (120 &#956;g/ml). Multiple
antibiotic resistance (MAR) patterns were evaluated for all isolates. At all sites nonpigmented isolates (including E.
faecalis and E. faecium) exhibited significantly higher diversity in combined resistance (mean MAR = 17) than
pigmented (E. casseliflavus and E. mundtii) isolates (mean MAR = 2). For all sites examined total E. faecalis and E.
faecium isolates demonstrated higher levels of multiple antibiotic resistance at sites below than above outflows. Total
antibiotic resistance was higher in E. faecium than in E. faecalis.
EXAMINATION OF THE EFFECT OF CALCIUM ON BIOFILM DEVELOPMENT AND PROTEIN
EXPRESSION IN VIBRIO PARAHAEMOLYTICUS
Heather E. Pontasch (Dr. Jodi Enos-Berlage), Department of Biology, Luther College, Decorah, Iowa 52101
Although calcium is known to play a critical function in numerous eukaryotic cell processes, the role of calcium in
prokaryotes is not well understood. Vibrio parahaemolyticus, a gram-negative marine bacterium and seafood
pathogen, serves as a model system to investigate the role of calcium in bacteria. This organism occupies
environments that differ substantially in calcium content, ranging from saturating levels (10 mM) in seawater, to the
human gastrointestinal tract, where calcium levels are up to 50-fold lower. Significantly, recent work has indicated
that a number of V. parahaemolyticus phenotypes are altered in response to calcium, including the ability of this
organism to attach to surfaces and develop a biofilm. The focus of this work was to obtain quantitative data
supporting the macroscopic observations of the effects of calcium on biofilm development and explore the possibility
that calcium might mediate these effects through changes in gene expression. Quantitation of surface-attached cells
under various calcium conditions indicated that increased calcium levels resulted in a five-fold increase in biofilm
development. In addition, two-dimensional electrophoresis of total cellular protein from V. parahaemolyticus grown
under different calcium conditions revealed up to 17 differentially-expressed proteins. These data provide
quantitative and qualitative evidence that external calcium levels influence biofilm development and protein
expression in V. parahaemolyticus.
100
BIOLOGY: MICROBIOLOGY
IN SEARCH OF A FAST-GROWING HALOPHILE FROM THE GREAT SALT LAKE: A HEAVILY
POLLUTED NATURAL ECOSYSTEM
Jonathan D. Oakes, (David Jordan and Ruhul H. Kuddus Ph.D), Biology Department Utah Valley State College 800
West University Parkway Orem, UT 84058
The purpose of the project was to isolate a salt and pollution-tolerant bacterium that can be used for bioremediation of
polluted marine environments. The Great Salt Lake of Utah, a naturally saline lake polluted with heavy metals
(selenium, mercury) and industrial/agricultural pollutants was the site chosen to search for such a bacterium. We
employed both culture-independent and culture-dependent methods to survey the microbial biodiversity in the Great
Salt Lake. PCR-amplification of 16srDNA fragments using total DNA isolated from lake water and mud (using a
universal primer) followed by cloning and DNA sequencing revealed very low biodiversity (a mere 11 different
microbial species). Restriction Fragment Length Polymorphism analysis of crude amplification product compared to
amplification products generated from artificial mixture of genomic DNA of 5-17 different bacterial species also
indicated low microbial biodiversity at the lake. Such low biodiversity is typical for highly polluted harsh environments.
Various culture media were also developed and inoculated with water and mud sampled from the lake. Only three
different bacteria colony types grew in one of the media types. The colonies were grown to pure culture and later
identified as a Salinovibrio sp. and a Halomonas sp. The Halomonas sp. appeared to be the bacteria suitable for
bioremediation. Although the species is an obligate halophile, it grows vigorously under several different
environmental and nutritional conditions. Selection of rapidly growing and oil and pollutant-tolerant strains of this
species is currently underway.
Analysis of Nitrogen-Cycling Bacterial Communities Present in Soils Overlying the Centralia,
Pennsylvania Underground Anthracite Coalmine Fire
Kristina Brown (Dr. Tammy Tobin-Janzen), Department of Biology, Susquehanna University, Selinsgrove,
Pennsylvania, PA 17870
The underground coalmine fire in Centralia, Pennsylvania has caused dramatic environmental changes in the
surrounding area. Overlying soil temperatures can exceed 450°C, and increased concentrations (>100 ppm) of
ammonium and nitrate are common in these soils due to venting gases from the mine fire below. The presence of
these nutrients makes Centralia a potentially ideal location in which to investigate nitrogen-cycling bacteria, and
previous research has suggested that these bacteria do inhabit hot soils (up to 60°C) in our study site.
Nitrogen-cycling bacteria can have dramatic impacts on soil fertility and are often involved in the release of
greenhouse gases. Nevertheless, they remain largely unstudied in hot soil environments. In order to identify the
resident nitrogen-cycling bacterial species in Centralia, and to begin to characterize the microbial communities in
which they reside, surface soil samples (0-10 cm depth) were collected from eleven sites that ranged in temperature
from 49.2 – 60.1 °C and spanned an active vent. Genomic DNA was isolated from these soil samples using a MoBio
Kit, and 16S rRNA genes were amplified via the polymerase chain reaction using universal Domain Bacteria primers.
The resulting fragments were then used to generate clone libraries for sequencing. NCBI BLAST and RDP II analysis
of these clones has shown that thermophilic nitrogen metabolizing bacteria such as Geobacillus thermoleovorans and
Sphaerobacter thermophilus are common in the site, as are novel bacteria that resemble uncultured denitrifying and
nitrogen fixing bacteria. Thus, it appears that Centralia will be an important site in which to identify and study
thermophilic nitrogen-cycling bacteria.
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BIOLOGY: MICROBIOLOGY
Characterization of Factors Influencing Cell Shape in Campylobacter jejuni
Sarah Nayoung Kim and Jonathan Olson, Ph.D.,, Department of Microbiology, , North Carolina State University,,
Raleigh, NC 27695
Campyobacter jejuni is a gram negative spiral bacteria that is the major cause of bacterial food poisoning in the U.S.
While the physiology of C. jejuni experiencing rapid growth has been studied extensively, little effort has been
directed at studying the physiological processes that occur when the bacterium is found in sub-optimal growth
conditions. When C. jejuni senses environmental stress or starvation, it quickly transforms to a metabolically inert
form, marked by a loss of spiral cells and the appearance of non-motile coccoid cells. In order to study the molecular
basis of the transition, different methods of analysis were conducted. First, a culture was grown for an extended
period of 27 days, and the optical density and the number of colony forming units (CFU) were recorded daily. There
was a result of a steady decrease in CFU from day one, which correlated to a switch from the spiral to the coccoid
morphology. Proteomic analysis was used to identify proteins that are expressed under conditions that promote the
spiral to coccoid transmission. Two-dimensional gels were used to identify protein spots that are differentially
expressed under the various growth conditions.
INVESTIGATING MECHANISMS OF HIV-1 ENVELOPE GLYCOPROTEIN (ENV)-MEDIATED
MEMBRANE FUSION, USING ENV- AND CD4-EXPRESSING CELLS
Kevin M. Hassler, Ashley A. Wells, Penny C. John, Rachel L. Puckett, David C. Martinez, Tenzin Bhutin, (Dr. Nejat
Duzgunes), Natural Sciences and Mathematics, Dominican University of California, 50 Acacia Ave, San Rafael, CA
94901.
The human immunodeficiency virus type 1 (HIV-1) infects host cells by binding to cell surface CD4 molecules and
chemokine receptors, followed by fusion with the plasma membrane. Our research project is focused on observing
HIV-1 glycoprotein-mediated cell to cell fusion. Instead of live HIV-1, we are using a human lymphoid cell line (TF
228.1.16) stably expressing the HIV envelope glycoproteins (gp120/gp41) on their surface. These cells are allowed to
fuse with human epithelial cells (HeLa-CD4) expressing the primary HIV receptor CD4. To monitor fusion the TF
228.1.16 cells are loaded with Carboxyfluorescein–acetoxymethyl ester and the HeLa-CD4 cells with Cell Tracker
Red. Generation of a fusion pore between the cells results in syncytia with orange fluorescence. We are
investigating the effects of inhibitors of the various stages of membrane fusion, to gain insight into the molecular
steps involved in this reaction. These molecules include the peptide DP-178 (T-20) that inhibits the gp41 six-helix
bundle formation, and the bicyclam JM-2987 that binds to the chemokine receptor CXCR4, a co-receptor for HIV. We
are also exploring the inhibitory effects of anti-gp41 and anti-gp120 antibodies.
THE ANIT-MUTAGENIC EFFECT OF FLAVONE IN BACTERIA EXPOSED TO UV RADIATION.
Bethany S. Miracle (Dr. Phil Musich) Department of Biochemistry & Molecular Biology, Quillen College of Medicine,
ETSU. Johnson City, TN 37614
Ultraviolet (UV) radiation, a component of sunlight, is damaging to DNA. UV-C (100-290 nm) is the shorter
wavelength UV radiation shown to damage bacterial and human cells. UV-B (290-320 nm) radiation also is harmful to
DNA in bacteria. Flavone is an anti-oxidant phytochemical found in dark-colored fruits and vegetables. Its high
absorbance of UV-B and UV-C light suggests that it might provide strong ‘sunscreen’ protection against UV radiation.
In addition, there may be ‘metabolic’ protection due to flavone’s cellular uptake and metabolism prior to UV exposure,
consistent with the beneficial effects of diets rich in foods containing flavonoids and other polyphenolic compounds.
We hypothesize that flavone will decrease the number of toxic mutations due to UV light exposure through its
‘sunscreen’ and the ‘metabolic’ effects. Flavone’s ability to protect cellular DNA against UV damage was tested, and
UV-B and UV-C radiation were compared to determine against which flavone offered the most protection. A bacterial
plating assay was used to measure the viability of cells after UV exposure. It was determined that flavone does
provide ‘metabolic’ protection against UV-C radiation, and preliminary trials also determined that it provides
‘metabolic’ protection against UV-B radiation. Ongoing research also will determine flavone’s protection against the
third component of sunlight, UV-A (320-400 nm) radiation. This might be much less than that of UV-C and UV-B
because of decreased absorbance. Therefore, flavone’s strong protection against mutations caused by UV-C and
UV-B radiation indicates that it and other plant polyphenols may be useful as additives to sunscreen lotions and an
important component of a healthy diet.
102
BIOLOGY: MICROBIOLOGY
Effect of Pomacea canaliculata on H5N1 Strain of Avian Influenza Survival in Water
Professor Arunee Thitithanyanont, Sasha Rihter, Department of Microbiology, Faculty of Science Mahidol Univeristy
Bangkok, Thailand
Avian H5N1 influenza has infected almost 100 people worldwide since 2003 and caused 69 deaths. Almost 150
million birds have died as a result of the disease or efforts to contain it. With the impending threat of avian flu
emerging from Southeast Asia, this study attempts to determine a possible method of viral transfer from migratory
birds to local fowl. The Golden Apple snail is a ubiquitously found food source for several large populations of
migratory birds in northern Thailand. Our study chose to investigate the species as a possible intermediate connection
between virally infected waters and diseased birds because the snail carries out its life cycle in H5N1 infested waters.
We specifically investigated Pomacea canaliculata as a possible carrier, protecting the virus from environmental
dangers and therefore extending its infectivity. Experimental conditions were created to mimic an infected water
source. Viral particles counts were measured in water prior to and at several time points after addition of the snail.
Analysis of water samples was conducted using Reverse Transcription Real-Time Polymerase Chain Reaction
(RT-PCR) using primers specific for the H5N1 Avian influenza strain. Viral particles used during the experiment were
isolated from recent outbreaks from several northern Thailand provinces. Results showed the presence of Pomacea
canaliculata had an immediate effect on the number of H5N1 viral particles present in water upon introduction to the
experimental environment. Analysis not only showed a definite connection between the Golden Apple snail and the
H5N1 virus, but also gave new evidence as to the longevity of the viral survival in water alone. The outcomes of this
study can provide new methods of viral control with a more efficient and less lethal manner of managing the spread of
the avian flu amongst birds and therefore open new doors for containment of the virus away from human populations.
103
BIOLOGY: MOLECULAR BIOLOGY
A GENETIC SCREEN TO IDENTIFY GENES REQUIRED FOR C. ELEGANS GONADOGENESIS
Amber D. Rex (Dr. Jennifer Miskowski), Department of Biology, University of Wisconsin- La Crosse, 1725 State St La
Crosse, Wisconsin 54601
For an organism to develop properly, many processes must be properly executed and coordinated together.
Understanding the molecular basis of these processes will increase our understanding of development and might
provide insight into disease initiation, progression, and potential treatments. We are studying the role of the NUD-1
protein in the formation of the Caenorhabditis elegans gonad. Proteins related to NUD-1 are found in many different
organisms, including humans, but their exact function is unknown. RNA-mediated interference (RNAi) was used to
severely reduce NUD-1 levels in C. elegans and defects in gonad development were observed. The RNAi-treated
animals (called nud-1(RNAi)) were sterile and possessed gonads that lacked the normal organization of somatic
tissues and germ cells. The treated animals also had defects in cuticle and vulva formation resulting in an
amorphous shape with a distinctly protruding vulva. These phenotypes have formed the basis for a genetic screen
that has been initiated to identify genes that function in similar cellular processes as NUD-1, and hopefully a mutation
in the nud-1 gene itself. A heritable mutation in the nud-1 gene does not yet exist, but isolation of one will facilitate
analysis of the role of NUD-1 protein in C. elegans. The screen has been designed to identify mutations that reside
on LGIII, the chromosome that nud-1 is located on. Experimentally, over 2,000 C. elegans genotypes have been
screened for the nud-1(RNAi) phenotypes listed above. Our progress will be presented.
DETERMINING THE LOCALIZATION OF THE NUD-1 PROTEIN IN CAENORHABITIS ELEGANS
Rachel McCabe (Dr. Jennifer A. Miskowski), Department of Biology, University of Wisconsin- La Crosse, La Crosse,
Wisconsin 54601
The broad goal of our research is understand the molecular basis of gonad formation in the nematode Caenorhabditis
elegans. Specifically, this project is focused on the NUD-1 protein that has been highly conserved across evolution.
NUD-1/NudC proteins have been implicated in diverse processes such as asymmetrical cell division in yeast, nuclear
migration in fungi, and neuronal migration in humans. The NUD-1 protein has been shown to function in the formation
of the C. elegans gonad using RNA-mediated interference (RNAi), but its specific role is unknown. Determining in
what cells the NUD-1 protein is found, and also where in the cell it is located, will provide insight into the possible
functions of the protein. An antibody generated against the active version of mammalian NUDC has been found to
cross react in C. elegans and was used for indirect immunofluorescence studies. After optimizing the fixation and
staining methods, we found NUD-1 to be localized to the gonad in L4 and adult staged worms. In L4 animals, NUD-1
was observed in a perinuclear punctuate pattern in primary spermatocytes. Adults showed staining in the
spermathecae, where mature sperm are found, and surrounding the oocytes. Further experiments will be conducted
to determine if the staining is in the oocytes themselves or in the somatic sheath cells which encircle oocytes and
support their development. This work promises to provide insight into aspects of C. elegans gonad development not
well studied. It might also reveal mechanisms of organ formation that are conserved in higher animals.
ANALYSIS OF THE MEIOTIC PHENOTYPES OF THE CDC7 AND DBF4 MUTATIONS IN
SACCHAROMYCES CEREVISIAE
Kimberly Henderson (Dr. Anne Galbraith), Department of Biology, University of Wisconsin-La Crosse, La Crosse, WI,
54601
The CDC7 and DBF4 genes are required for S phase during the mitotic cell cycle. The CDC7 gene encodes a
cyclin-dependent kinase (CDK) and the DBF4 gene encodes its cyclin. Several labs have investigated the role of
these two genes in mitotic S phase, and it is clear that the Cdc7-Dbf4 kinase is necessary to initiate DNA replication.
We are interested in determining the role of these genes in the other cell division process, meiosis. In order to
analyze the meiotic role of these two genes, we examined temperature sensitive cdc7 and dbf4 mutants in meiosis.
We induced meiosis in wild-type, cdc7, and dbf4 diploids by starving them in sporulation media. Time points were
taken every hour for 12 hours and then again at 24 hours (the time needed for meiosis to be completed in these
strains). We analyzed these samples by a variety of methods to determine which step(s) of meiosis was affected by
the absence of CDC7 and/or DBF4 function compared to the wild-type control. The major meiotic events of DNA
replication, meiotic recombination, and the meiotic divisions were all analyzed. We also measured overall sporulation
efficiency and spore viability. The results from these experiments will be presented and a model proposed to explain
them.
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DETERMINING THE EXECUTION POINT OF THE CDC7 AND DBF4 GENES IN YEAST MEIOSIS
Bethany A. Volkmann (Anne M. Galbraith), Biology Department, University of Wisconsin-La Crosse, 1725 State St, La
Crosse, Wisconsin 54601
In yeast, humans, and other eukaryotic organisms, the CDC7 and DBF4 genes are required for meiosis, a type of cell
division, to occur properly. If errors occur during this cell division process due to mutations in either of these two
genes, results can include death of the cell, or disease of the organism. Our lab has also been testing the effects of
both dbf4 and cdc7 mutations on meiosis. We have shown that mutations in either of the two genes prevent meiosis
from occurring properly. In yeast, this means that the normal haploid products of meiosis, the spores, are not
produced. The objective of my research was to determine at what point during meiosis the CDC7 and DBF4 genes
are needed by examining the time that the cells are arrested in meiosis in mutants versus wild-type strains of yeast. I
constructed three different strains of yeast: wild-type, cdc7 mutant, and dbf4 mutant. I then took samples of these
strains at various times during meiosis and placed them at a high (restrictive) temperature that did not support
meiosis in the mutants. These cells were then analysed using flow cytometry, double strand break analysis, DAPI
staining, and fluorescence microscopy to determine what step of meiosis was affected. Results from these
experiments will be presented, along with a model for how these genes may function in yeast meiosis.
COMMENSAL MICROFLORA AND HEPATIC RESPONSES TO FASTING AND RE-FEEDING
Eboghoye O. Eluka, Rachael Rigby, Victoria Newton, Kay P. Lund , (Partnership for Minority Advancement in the
Biomolecular Sciences (PMABS)) , Department of Cell and Molecular Physiology, The University of North Carolina at
Chapel Hill, Chapel Hill, North
Obesity and metabolic syndrome are conditions that affect a high percentage of the western population. Recent
studies in germ-free (GF) mice that lack commensal bacteria in the intestine indicate that commensals may play a
role in adiposity (fat storage), but the relationship between bacteria and obesity is not yet known. We wanted to
examine the effects of commensal bacteria on nutritional status and uptake. This research examines body and liver
weight and hepatic Insulin-Like Growth factor 1 (IGF-1) levels, following fasting and re-feeding in mice. We predict
that commensal bacteria increase the availability of nutrient and promote energy storage. IGF-1 is a blood protein
that indicates growth hormone levels in tissues. It is mainly produced in the liver and functions to promote growth and
proliferation of cells. This study utilizes the C57/B6 mouse strain, raised under different bacterial conditions, Germ
Free (GF), Specific Pathogen Free (SPF) and Conventionalized (CONV). Body and Liver weight were measured at
sacrifice. Total mRNA was extracted from liver tissue and used in a Northern blot to determine IGF-1 gene expression
levels. Our findings show that there is a decrease in liver weight following fasting in SPF and CONV animals
compared to GF mice. There was also an increase in IGF-1 gene expression levels in GF animals following fasting
compared to CONV animals. These data indicate that the presence of commensal flora limit IGF-1 production,
possibly growth, and fat storage in times of nutrient deprivation.
Alterations in Caldesmon Expression Following Traumatic Brain Injury
Noah C. Morgan, Christian Kreipke PhD., Theodor Petrov PhD., Jose Rafols PhD., Detpartment of Anatomy and Cell
Biology, Wayne State University School of Medicine, Detroit, MI, 48201
Caldesmon (Cd) is associated with the contractile mechanisms regulating vascular tone. Modifications in caldesmon
expression are thought to regulate vessel contractility and hence, are associated with the regulation of blood flow.
Our lab has previously shown that following traumatic brain injury (TBI) there is a significant decrease in blood flow.
This study was aimed at drawing a causal association between changes in Cd expression and regulation of
neurovascular tone following TBI. Using the Marmarou’s model, used to simulate TBI, brain tissue was collected for
protein analysis. Double immunofluorescence, using antibodies raised against Cd, smooth muscle actin (SMA)(a
marker for smooth muscle cells (SM)), and lectin (a marker for the endothelial cells (En)) revealed that in control
animals Cd colocalized with SMA and with the marker for En. Cd expression significantly increased in SM at 4hours.
This increase was sustained throughout the duration of the experiment (48hours). Cd expression increased at 24
hours post TBI within En, to a level of nearly 400% that of control. However, this upregulation was not sustained at
48hours with a return to control levels. This data, in conjunction with MRI results showing decreased blood flow
following TBI, supports a causal association between modifications in Cd expression and the regulation of
neurovascular tone following neurotrauma. In conclusion, this study emphasizes the dynamic process of vessel
contractility and represents an initial investigation into the upstream events following TBI. Supported by NIH grant
NS039860.
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The physiological state of the Msi1p complex in Saccharomyces cerevisiae, does not change with
different carbon sources
Scott Halkyard (Dr. Stephen Johnston), Department of Biology, North Central College, 30 North Brainard Street,
Naperville, IL 60540
http://scott.halkyard.students.noctrl.edu/NCUR%20abstract.htm
Using Mitochondrial COI Gene Sequence to Evaluate Relatedness in the Genus Unionicola (Acari:
Unionicolidae)
Crystal M. Harmon, Kevin S. Myers, Dawn L. Scarlett, Bradley L. Stilger, (Dale D. Edwards and Brian R. Ernsting),
Department of Biology University of Evansville, Evansville, Indiana 47722
Water mites of the family Unionicolidae are common symbionts of freshwater mussels. Many species within
Unionicolidae are morphologically similar, and thus determining the degree of relatedness within subgenera is
especially challenging. This study compared the mitochondrial DNA Cytochrome Oxidase I (COI) locus among mite
species and host-associated populations from three subgenera of Unionicola: Parasitatax, Clarkatax, and
Unionicolides. DNA was extracted from approximately five mites for each study species or host-associated population
and PCR was preformed to amplify the COI gene. Gel electrophoresis was used to confirm amplification and PCR
products were sequenced. The sequences were assembled using CodonCode Aligner© and consensus sequences
were aligned using ClustalX©. Our research produced sequences from ten mite species and host-associated
populations. A neighbor joining tree grouped the members of the subgenera—Parasitatax, Clarkatax and
Unionicolides—closely together, further supporting the classification of the three different subgenera. These findings
support the morphological information available and this study is the first to fully examine the evolutionary
relatedness between the subgenera of Unionicola using molecular phylogeny. Future studies will amplify other gene
targets from current and additional species in the related subgenera to add support for the relatedness proposed in
this work.
The Use of Green Fluorescent Protein to Create Expression Plasmids for Two Salvage Pathway
Enzymes
Adaku F. Iwueze (Dr. Judith Wubah and Dr. Daniel Wubah), Department of Biology, James Madison University,
Harrisonburg, VA 22807.
In the United States, 4 million babies are born each year and approximately 150,000 of these are affected by
congenital malformations. It is not known whether the 4,000 neonates born each year that suffer from mitochondrial
DNA defects are included in this number. The mitochondrion, which is the energy generator in the cell, functions in
other metabolic processes. Deoxyguanosine kinase (dGK) and thymidine kinase2 (TK2) are nuclear genes, which
encode the rate-limiting enzymes in the mitochondrial salvage pathway, a process for generating deoxynucleotide
triphosphates. Mutations in dGK and TK2 lead to various forms of mtDNA depletion syndrome and early infantile
deaths. A novel isoform of dGK, dGK3 and TK2 are being analyzed with a long-term goal to determine their possible
role in birth defects. The objective of this study is to create expression plasmids of dGK3 and TK2 which will be used
to determine the intracellular location of these two genes. Using total RNA from adult mouse liver, full-length dGK3
and TK2 complementary DNA (cDNA) were amplified which contained EcoR1 sites at both ends. The cDNA products
were each cloned into pGEM-T vector and digested with EcoR1 enzyme to ensure incorporation of the sites. The
digested products were ligated with a digested pEGFP vector. Screening of several clones has yielded an expression
plasmid for dGK3 and three possible expression plasmids for TK2. The generated TK2-GFP expression plasmids will
be sequenced for confirmation. In summary, GFP expression plasmids will be confirmed by sequencing and used in
future transfection studies.
Funded by the Jefferres Memorial Trust Fund Grant # J699 and the NIH/Bridges Grant #R25GM063995.
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IDENTIFICATION OF CBP SITES PHOSPHORYLATED BY P38
Amber N. Young (Brian Ashburner), Department of Biological Sciences, The University of Toledo, Toledo, Ohio 43606
NF-&#954;B is a transcription factor that is involved in essential biological processes such as immune and
inflammatory responses, cell proliferation, and programmed cell death. Classical NF-&#954;B is a heterodimer of the
subunits p65 and p50. The ability of NF-&#954;B to activate transcription of genes is dependent on its interaction with
transcriptional coactivators, such as the CREB-binding protein (CBP). However, the mechanisms regulating
interaction between CBP and p65 is not completely understood. Therefore, the focus of my current research is to
gain a better understanding of how the interaction between the p65 subunit of NF-&#954;B and CBP is regulated.
Preliminary evidence has indicated that the p38 MAP kinase is involved in regulating the p65 and CBP interaction.
One potential mechanism is that p38 regulates the p65 and CBP interaction by interacting with and possibly
phosphorylating CBP. Our data indicates that p38 can phosphorylate CBP at multiple sites in in vitro kinase assays;
however, it is not clear if these sites are also phosphorylated in vivo. To test this, site directed mutagenesis will be
used to mutate potential p38 phosphorylation sites (serine or threonine) to alanines. We will then determine if these
mutant forms of CBP are still phosphorylated by p38 and will determine the effect of these mutations on the ability of
CBP to function as a coactivator for NF-&#954;B.
THE ROLE OF DALLY CORE PROTEIN IN DROSOPHILA DEVELOPMENT
Daniel M. Lercher (Scott Selleck, MD, PhD), Departments of Genetics, Cell Biology, and Development and
Department of Pediatrics, 321 Church St SE, 6-160 Jackson Hall, University of Minnesota, Minneapolis, Minnesota
55455
Dally is a Drosophila heparan sulfate proteoglycan (HSPG) that includes a core protein on the extracellular cell
surface anchored by a GPI linkage. Enzymes add heparan sulfate chains to the core. The purpose of this project
was to determine if the Dally core protein has biological function during Drosophila development. In one view of
HSPG function, the heparan sulfate side chains are thought to bind the ligand to promote signaling, while the core
protein only delivers the chains to the right cellular location. To characterize the function of the core protein, we
mutated the heparan sulfate addition sites by single point mutations, changing the three serine attachment sites to
alanines. This change prevented the addition of heparan sulfate. Using the UAS/GAL-4 system in Drosophila
melanogaster, we expressed this non-glycanated form of Dally in a dally homozygous mutant background to
determine the core protein’s function. In dally mutants, the L5 wing vein stops short of the wing margin and has fewer
chemosensory bristles at the margin. In addition, the fly loses two sets of bristles on the notum, the anterior
dorsocentral and posterior supraalar. Non-glycanated Dally substantially rescued the L5 wing vein defect,
chemosensory bristles, and anterior dorsocentral bristles, but not the posterior supraalar. Rescue of these
phenotypes shows that non-glycanated Dally still functions surprisingly well in Dpp and Wingless signaling, but not as
well as wildtype.
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DISRUPTION OF PHOSPHODIESTERASE 4D: IMPACT UPON MURINE CARDIAC CONTRACTION
RATE
Christine N. Chang (Dr. Andrew J. Patterson and Dr. Marco Conti ) Anesthesia, Stanford University, Stanford, CA
and Obstretrics and Gynecology, Stanford University, Stanford, CA
&#946;1 adrenergic receptors (&#946;1ARs) are primarily responsible for increasing heart rate during catecholamine
stimulation. &#946;2ARs are secondary in this regard. In vitro data suggest that compartmentalization of the rise in
cAMP after &#946;2AR stimulation helps to distinguish this receptor subtype from the &#946;1AR.
Phosphodiesterase 4D (PDE4D) has been shown to surround &#946;2ARs after agonist-induced activation and
prevent cAMP diffusion. &#946;1ARs are not surrounded by PDE4D and a global intracellular rise in cAMP occurs
after &#946;1AR activation. In cultured myocytes, PDE4D disruption leads to a global rise in cAMP after &#946;2AR
activation.
We hypothesized that mice with disruption of PDE4D will have higher baseline heart rates and detrimental myocardial
remodeling over time due to &#946;2ARs functioning like additional &#946;1ARs, as well as the absence of
&#946;2AR-mediated cardiac protection. We evaluated 19 resting awake male PDE4D KO, &#946;1/PDE4D Double
Knockout (DKO) as well as wild type (WT) mice. Heart rates were measured using a Visitech BP 2000 infrared tail
sensor device (Visitech Systems, Apex North Carolina). ECG-gated MRI (4.7T magnet) was used to determine left
ventricle end-diastolic volume (LVEDV), left ventricle end systolic volume (LVESV), and heart rate. Animals were
anesthetized using isoflurane during the analyses. Stroke volume (SV) and cardiac output (CO) were calculated.
From 2 to 6 months of age, DKO mice exhibited significantly lower heart rates than WT and PDE4D KO animals (both
p<0.001, one way ANOVA with Newman Keuls post-hoc analysis). The heart rates of PDE4D KO and WT mice were
similar. No significant differences in SV or CO were noted between WT and PDE4D KO or &#946;1/PDE4D DKO
animals at two months of age, or at four months of age. These results suggest that our hypothesis with regard to heart
rate may be incorrect. While data at four months of age revealed no significant structural or functional differences by
MRI assessment, changes may still occur as the animals age. Upcoming analyses at eight and ten months of age
should allow us to investigate this possibility.
COMPARING T-CELL RECEPTOR USAGE IN DIABETES-PRONE NOD MICE TO
DIABETES-RESISTANT NOD.IE TRANSGENIC MICE
Jonathan L. Hassel (Dr. Anthony Quinn), Department of Biological Sciences, The University of Toledo, 2801 W.
Bancroft St., Toledo, Ohio 43606
The earliest T-cells arising in the islets of Langerhans in the pancreas of NOD (Non-Obese Diabetic) mice, which
develop spontaneous Type I Diabetes (TID), are directed against GAD65 p530-543 through immunization. The
majority of the p530-specific T-cells in the NOD mice utilize V&#946;4 for the TCR (T-Cell Receptor) gene family.
The prevention of responses by the T-cells may be sufficient to prevent TID in NOD mice. There are two ways to
accomplish this: 1) Regulatory T-cells can be used to inhibit the response of the islet-invasive p530-43 specific
T-cells. 2) Alternatively, deletion of pathogenic clones would also be expected to provide protection. The addition of a
second Major Histocompatibility Complex (MHC) Class II molecule added via transgenesis protects NOD mice from
disease, as is seen in the NOD.IE mice. The MHC is made of a membrane glycoprotein and a presented antigen on
the surface of antigen-presenting cells (APC). This complex is recognized by T-cells specific for the antigen
presented. The samples were tested with fluorescence activated cell sorting (FACS) analysis, reverse-transcriptase
PCR (RT-PCR), and various other microbiology techniques. The data suggest the NOD.IE mice down-regulate or
delete the pathogenic clones specific for the p530-43 fragment, which utilize V&#946;4-J &#946;2.7 genes. NOD.IE
mice do mount a response to p530-543; however, they utilize a different repertoire. Therefore, the V&#946;4-J
&#946;2.7 TCR, seen in the NOD mice, can be linked to Type I Diabetes as a pathogenic receptor and the deletion of
this receptor could provide immunity in the NOD.IE mice.
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AAV MEDIATED ALPHA CAM-KINASE II SUPPRESSION AND UP REGULATION: BEHAVIORAL AND
WESTERN BLOT ANALYSIS
Deven D. Robinson, (Dr. Mike Babcock), Complex Biological Systems Summer REU, Montana State University, 1
Lewis Hall, Montana State University, Bozeman, Mt. 59717-3505, Home Institution: Carroll College, 1601 N. Benton
Avenue, Helena, Montana 59840
The goal of the present study was to inhibit the &#945; CaM-Kinase II (&#945; CaM-K II) enzyme, and to over express
the &#945; CaM-K II enzyme in order to observe if behavioral and learning problems occurred from induced memory
changes. An adeno-associated virus (AAV) with either sense or anti-sense RNA to the &#945; CaM-K II protein was
designed for the over expression and suppression of the &#945; CaM-K II protein. A Morris-water maze and an open
field maze were used to test for the effects of the differences in protein concentration. An increase in protein in the
sense rats, as apparent in the Western Blots, correlated with significantly greater probe trial times when compared to
the controls as expected if &#945; CaM-K II protein was involved in the establishment of memory. The sense group
(n=6) differed significantly (p=0.02) from the GFP control group (n=5). Concentrations of &#945; CaM-K II protein in
the hippocampi were determined 29 days after surgery. The sense bands demonstrate an increase in concentrations
of &#945; CaM-K II protein as compared to the control bands. A statistically significant difference (p<0.05) was
observed between sense (n=5) and control (n=5) conditions.
CADMIUM PROMOTES BREAST CANCER CELL PROLIFERATION
Pham H. Vu, Lauren S. Kawabata, Ashley L. McClellan, and (Dr. Maggie C. Louie), Department of Natural Sciences
and Mathematics, Dominican University of California, San Rafael, California 94901
Breast cancer is the most common non-skin cancer in the United States, leading to the second highest
cancer-associated deaths in women. Majority of breast cancers initially develop as hormone dependent, which
implies that estrogen can enhance the progression of the cancer. Hormone dependent breast cancers are typically
treated with hormone ablation therapy and/or anti-estrogen treatment. However, hormone responsive breast cancer
frequently develops into a more aggressive phenotype that is hormone independent. In many cases, the estrogen
receptor (ER) is still present. However, the role of ER in hormone refractory breast tumorigenesis and the underlying
mechanism is unclear. A potential mechanism may involve endocrine disruptors including heavy metals such as
cadmium. Cadmium is as an environmental contaminant that enters the body through diet or cigarette smoke.
Recent studies have suggested that cadmium can stimulate estrogen receptor (ER) activity and promote neoplastic
growth in mice. (Stoica et al. 2000, Martin et al., 2003). This suggests that increased incidences of breast cancer in
the US may be associated with human exposure to cadmium. Preliminary data from our lab also suggests that
cadmium can promote hormone independent breast cancer cell growth in a cell culture model. Additionally, we have
also shown that cadmium can induce the expression of various ER target genes (cyclin D1, cathepsin D, c-myc) and
non-ER target genes including cell cycle regulators, cyclin E, and cdk2. However, further studies are necessary to
better elucidate the role of cadmium in promoting breast cancer cell growth.
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COMPARISON OF FIVE COMMERCIAL DNA EXTRACTION KITS FOR THE RECOVERY OF
FRANCISELLA TULARENSIS DNA FROM SPIKED SOIL SAMPLES
Hannah E. Hottel, (Dr. Chris A. Whitehouse and Dr. Susan M. Ensel), U. S. Army Medical Research Institute of
Infectious Diseases, Fort Detrick, MD
Francisella tularensis is the etiologic agent of the zoonotic disease tularemia and is found throughout the Northern
Hemisphere. F. tularensis is highly infectious with exposure to less than 10 organisms being able to cause disease.
The highly infectious nature of the organism has made it attractive as a bioweapon in historical biological warfare
research programs and has prompted the U.S. Centers for Disease Control and Prevention (CDC) to list it as a
Category A bioterrorism agent. To gain a better understanding of the ecology and epidemiology of Francisella, it will
be important to further explore its distribution in the environment. As a prerequisite to these studies, methods must
be established to efficiently extract Francisella-specific DNA from the soil and be able to eliminate potential PCR
inhibitors. Thus, we evaluated five commercial DNA extraction kits for their ability to recover F. tularensis-specific
DNA from soil samples and eliminate potential PCR inhibitors. Soil samples were spiked with gamma-irradiated F.
tularensis SHU-4 strain [corresponding to a range from 1 to 105 colony-forming units (CFU)]. Spiked samples were
extracted with each kit according to manufacturer’s instructions and evaluated using both a F. tularensis-specific
real-time PCR assay, which targets the fopA gene, and an internal positive control (IPC) assay that measures the
presence of potential PCR inhibitors. Comparisons of these kits will include processing time, cost, and effectiveness
when utilizing multiple soil types.
Home institution: Department of Chemistry and Physics, Hood College, Frederick, MD
EFFECTS OF SPROUTY1, AN INTRINSIC INHIBITOR OF THE ERK SIGNALING CASCADE, IN THE
HUMAN HEART
Sean P. Polster, Nathan J. Charles, Robert C. Huebert, Neeta Adhikari, Ami Mariash, Mohammed-Karim Ezzat,
James Rider, Leslie W. Miller, (Jennifer L. Hall), Department of Medicine, Cardiovascular Division, The University of
Minnesota, Minneapolis, MN, USA,
Heart failure is the second major cause of death in the world behind infectious disease, and the prevalence continues
to rise. Mechanical unloading of the failing human heart with a ventricular assist device (VAD) has proven to partially
reverse, halt, or inhibit left ventricular wall stress and the accompanying structural decompensation. To dissect the
critical genes and networks governing this reverse remodeling process, we compared paired gene expression profiles
from 19 patients at the time of implant and explant of the VAD and identified a significant up-regulation of Sprouty1 in
post VAD samples. Subsequent studies confirmed this finding with RTQPCR and identified a parallel up-regulation in
protein expression. Sprouty1 is an evolutionarily conserved gene that was described as a potent inhibitor of the ERK
cascade. We reconfirmed previous reports demonstrating that ERK is down-regulated following mechanical unloading
with the VAD. Our hypothesis is that Sprouty1 is an intrinsic rate-determining mediator that permits ERK inhibition and
alters the process of myocardial remodeling. Sprouty1 up-regulation inhibited bFGF activation in Cos and HL-1 cells.
We have recently begun to characterize a new line of &#945;-MHC Sprouty1 transgenic mice to critically test the
hypothesis that Sprouty1 alters remodeling. In conclusion, Sprouty1 is a novel endogenous inhibitor of the ERK
cascade and the transgenic model should help us fully define the role of Sprouty1 in heart failure.
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JUVENILE HORMONE REGULATION OF YP1, YP2 AND YP3 GENE EXPRESSION IN FAT BODY AND
OVARIES IN DROSOPHILA MELANOGASTER.
Emily E. Bowling (David S. Richard). Department of Biology, Susquehanna University, Selinsgrove, PA 17870.
Dipteran insects are significant agricultural pests and disease vectors making control methods valuable.
Investigations with the fruitfly Drosophila melanogaster may provide insight into the endocrine regulation of
vitellogenesis, the production and uptake of yolk proteins by the oocyte, a process required for female fertility.
Juvenile hormones (JH), ecdysteroids and insulin-signaling are involved in regulating vitellogenesis. The role of
juvenile hormone in ovarian gene expression was studied by microarray analysis using Affymetrix EST Drosophila
chips on apterous56f (ap56f) mutant females which are deficient in JH. ap56f females were treated with either 1µg
JH III in 1µl acetone (shown previously to rescue the flies from their usual 6 hour vitellogenic delay) or 1µl acetone
(negative control) at 6 hours post-eclosion and their ovaries were dissected at 12 hours post-eclosion. This treatment
elicited up-regulation of more than 80 genes and down-regulation of 20-50 genes in the ovary. The up-regulation
included an increase in gene expression of YP1 and YP3. YP2 expression did not increase to the same extent. All
three YP genes are found on the X chromosome. It is assumed that abdomen yolk proteins come from the fat body
(late vitellogenesis); ovary yolk proteins come from the follicle cells (early vitellogenesis). Semi-quantitative RT- PCR
analysis has shown that YP1, YP2, and YP3 gene expression in the ovary and abdomen vary across developmental
series. Therefore, these studies may provide insight into the endocrine and molecular control of reproductive
processes in insects. Supported in part by NIH grant #GM/OD54905 to DSR.
INSULIN-SIGNALING AND DYNAMIN MAY BE INVOLVED IN THE REGULATION OF
RECEPTOR-MEDIATED ENDOCYTOSIS IN DROSOPHILA MELANOGASTER.
Kristen Shilling (David S. Richard). Department of Biology, Susquehanna University, Selinsgrove, PA 17870.
Many insects are agricultural pests or disease vectors so rational methods of control are being sought. A model
organism, the fruitfly Drosophila melanogaster, may provide insight into the endocrine regulation of vitellogenesis (the
production and uptake by developing eggs of yolk proteins: YPs) a process required for female fertility.
Receptor-mediated endocytosis by nurse cells in developing oocytes utilizes non-specific proteins such as clathrin
and adaptin as well as specific receptors to sequester YPs from the hemolymph. Ring canals connect adjacent nurse
cells and allow for cell communication and cytoplasmic streaming during egg maturation. YP is therefore trafficked
from the hemolymph, via nurse cells, into the developing oocyte. Females deficient in the insulin-signaling pathway
protein CHICO, are non-vitellogenic, and receptor-mediated endocytosis appears to be disrupted (Richard et al.,
2005). Dynamin (shibire) is a GTPase critical for receptor mediated endocytosis. Dynamin may function in the
separation of the endocytic vesicles from the nurse cell plasma membrane, therefore internalizing YPs to be
trafficked into the oocytes. The specific role of dynamin in this process as well as the production localization of
dynamin remains to be determined, but is of potential importance for the coordination of this vitellogenic process.
Supported in part by NIH grant #GM/OD54905 to DSR.
Richard,D.S., Rybczynski,R., Wilson, T.G., Wang,Y., Wayne,M.L., Zhou,Y., Kroll,T.R., Partridge,L. and
Harshman,L.G. (2005) Insulin signaling is necessary for vitellogenesis in Drosophila melanogaster irrespective of the
roles of juvenile hormone and ecdysteroids.. Journal of Insect Physiology. 51, 455-464.
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RAB-7 MAY BE INVOLVED IN RECEPTOR-MEDIATED ENDOCYTOSIS OF YOLK PROTEINS BY
OOCYTES IN DROSOPHILA MELANOGASTER.
Andrea Huntley (David S. Richard) Department of Biology, Susquehanna University, Selinsgrove, PA 17870
Drosophila melanogaster (Diptera) is a model organism useful in studying aspects of insect disease vectors and
agricultural pests that cost millions of dollars every year. Thus insect population control is important and one
mechanism of control may be via vitellogenesis (the synthesis and uptake of the yolk proteins (YP) into the oocyte).
YP uptake by oocytes is by receptor-mediated endocytosis involving clathrin coated vesicle formation in follicle cells
and nurse cells. Control of the process of vitellogenesis appears to be under the control of the ecdysteroids, the
juvenile hormones (JH) and the insulin-signaling pathway. Rab-7 is a small GTPase that is involved in endosome
formation in endocytic cells. Microarray analysis of ovarian mRNA levels from, JH-deficient females treated with
topical application of JH, and diapausing females treated by ecdysteroid injection, show that Rab-7 expression is
elevated, indicating that the endocrine system regulates this facet of reproductive development. By establishing
Rab-7 transcript levels in a developmental series in wild-type ovaries, we may understand better the role of Rab-7 in
vitellogenesis. Semi-quantitative RT-PCR, with two different Rab-7 primer sets, using mRNA from a post-eclosion
developmental series indicate that expression of this gene is developmentally regulated. This information may lead
to further insight into the role of Rab-7 in receptor-mediated endocytosis of YPs in Drosophila ovaries. Supported in
part by NIH grant #GM/OD54905 to DSR.
CLONING AND EXPRESSION OF LUCIOLA ITALICA LUCIFERASE GENE AND PRELIMINARY
PROTEIN CHARACTERIZATION
Jennifer P. DeAngelis (Dr. Bruce R. Branchini and Ms. Tara L. Southworth), Department of Chemistry, Connecticut
College 06320
The purpose of this project was to clone the Luciola italica luciferase gene; and to express, purify and characterize the
corresponding bioluminescence-catalyzing enzyme. Fireflies were collected in the hills surrounding Bologna, Italy
and transported to the laboratory of our Italian collaborator Professor Roda where they were flash frozen in liquid
nitrogen. Total RNA was then isolated from the firefly lanterns and the L. italica luciferase cDNA was cloned by
RT-PCR using a gene-specific primer set based on the DNA sequence of the Eastern European Luciola mingrelica
luciferase. The gene was ligated into the pGEX-6P2 plasmid, which is designed to produce proteins as N-terminal
glutathione-S-transferase-fusion products. A highly specific protease can then be used to cleave the protein of
interest away from the GST and linker peptide thereby facilitating protein purification. The L. italica gene was
sequenced and determined to be 1647 base pairs in length with an open reading frame of 548 amino acids. Prior to
expressing the protein, it first was necessary to realign the reading frame of the cDNA in the pGEX-6P-2 plasmid by
PCR-based mutagenesis. The L. italica gene was then expressed as the GST-fusion protein in E. coli BL21 cells and
purified to homogeneity by affinity chromatography. Initial characterization of the enzyme showed that L. italica
protein exhibits bioluminescent activity similar in intensity to the common North American Photinus pyralis luciferase;
however; it produces light that is slightly red-shifted (maximum emission at 565 nm vs 557 nm). By steady state
kinetics analysis, the L. italica Km for LH2 was found to be 0.095mM, approximately 6-fold greater than that of the P.
pyralis enzyme. Interestingly, both enzymes had similar Km values for Mg-ATP (0.160 mM). Phylogenetic analysis
showed that L. italica luciferase was similar to that of other fireflies in the family Lampyridae, subfamily Luciolinae
with 95.8% and 95.6% homology to the Hotaria unmunsana (Korea) and Hotaria parvula (Japan) luciferases,
respectively.
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IDENTIFICATION OF PYRUVATE CARBOXYLASE AS A GLUCOSE-REGULATED TARGET GENE OF
CHREBP
Luke N. Robinson (Howard C. Towle), Department of Biochemistry, Molecular Biology and Biophysics, University of
Minnesota, Minneapolis, Minnesota, 55455
Lipogenesis is the process by which simple carbohydrates, such as glucose, are converted to triglycerides, the
preferred energy storage molecule in mammals. When mammals are fed a high carbohydrate diet, the process of
lipogenesis is accelerated in the liver, in part due to increased gene expression of enzymes involved in lipogenesis.
ChREBP (Carbohydrate Response Element Binding Protein) is a recently discovered transcription factor that
regulates the expression of lipogenic enzyme genes in response to glucose. ChREBP acts by binding a regulatory
DNA sequence, the carbohydrate response element (ChoRE). To date, functional ChoRE sequences have been
characterized in the promoters of L-type pyruvate kinase, S14, fatty acid synthase, and acetyl-CoA carboxylase
genes. Using the sequences of these ChoREs in a promoter database search, a putative ChoRE (-429/-413) was
identified in the distal promoter of the mouse pyruvate carboxylase (PC) gene. PC catalyzes the ATP-dependent
carboxylation of pyruvate to oxaloacetate, playing a crucial role in the metabolic processes of gluconeogenesis,
lipogenesis, and anaplerosis. Comparison of the sequences of PC distal promoters demonstrated that the putative
ChoRE was conserved in several mammalian species. Primary rat hepatocytes cultured in varying glucose
conditions were used to further investigate the putative PC ChoRE. As determined by real time polymerase chain
reaction, mRNA transcripts specific to the PC distal promoter increased 2-fold in response to high glucose.
Transfection of a reporter vector with two copies of the putative PC ChoRE into hepatocytes resulted in a 20- to
50-fold induction in response to high glucose conditions. Using the electromobility shift assay (EMSA), ChREBP was
found to bind to an oligonucleotide containing the putative PC ChoRE. Thus, when taken together, these results
suggest the PC ChoRE functions as a glucose-responsive regulatory element that confers an induction of PC
transcripts under conditions favoring lipogenesis.
MOLECULAR ANALYSIS OF A TRANSCRIPTIONAL SILENCER IN DROSOPHILA MELANOGASTER
Christopher Chamberlain (Dr. Presley Martin), Department of Biology, Hamline University, St. Paul, Minnesota 55104
The MM-50 transgenic line of Drosophila melanogaster contains a D. melanogaster Alcohol dehydrogenase (Adh)
transgene inserted at position 25C of chromosome 2. Previous analysis has shown that expression of this Adh
transgene is inhibited by a silencing activity located in an 1180 b.p. region near the 5` end of the inserted gene. The
sequence of the silencer region revealed the existence of two 20 b.p. homologies, which have been shown to inhibit
Adh expression by 50%. The objective of this investigation was to determine what other sequences within the
silencing region are required to produce the near 100% inhibition of expression observed when the whole region is
present. Seven unique fragments of the silencer sequence were cloned using PCR and purified using standard
procedures. Two fragments contain the first homology of the sequence, one fragment contains the second homology,
and 4 fragments contain neither of the homologies. Each of the seven fragments were transformed into plasmids
containing a functional Adh gene, thus yielding seven complete plasmids each containing one of the fragments and
one copy of the Adh gene. Each plasmid will be injected into Drosophila embryos and the larvae will be assayed for
the level of Adh gene expression.
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ANALYSIS OF RAD51 GENE EXDPRESSION DURING DIFFERENTIATION AND AFTER DNA DAMAGE
IN THE CELLULAR SLIME MOLD, DICTYOSTELIUM DISCOIDEUM
Robert M. Ostrowski, Jr. (Dr. Stanley K. Mauldin), Department of Natural Sciences, Holy Family University,
Philadelphia, PA 19114
The formation of double-strand breaks (DSB) in DNA is one type of damage that causes genomic instability. The
RAD51 protein is essential to the repair of these breaks. Many studies have used dividing cells to understand the
mechanisms of DSB repair. Few studies have explored the repair of DSBs in differentiated cells. The cellular slime
mold, Dictyostelium discoideum, has been widely used as a model system for studying differentiation and
development. The purpose of this study was to determine the expression of the D. discoideum homolog of RAD51
during differentiation and after DNA damage with ionizing radiation. Using a BLAST search of the DictyBase genomic
database (www. DictyBase.org), a gene was located that is homologous to the RAD51 gene (rhp51) of
Schizosaccharomyces pombe. A 211 bp fragment of the D. discoideum RAD51 homolog was cloned using primers
from sequence information obtained from the DictyBase website. This fragment was used as a probe to analyze the
expression of RAD51 after ionizing radiation and during differentiation. Initial experiments in actively growing D.
discoideum AX3K cells showed approximately a ten-fold increase in the amount of mRNA within one hour of exposure
to 40 krads of ionizing radiation. Further experiments are planned to analyze the expression of the RAD51 gene
during the various stages of development and differentiation both with and without ionizing radiation.
Plant PK-GAD Interactions: Sequence Specific or Generic?
Heather Fields (Dr. Robert Locy), Biological Sciences, Auburn University, 101 Rouse Life Sciences Building, Auburn
University, AL 36849
Occurring ubiquitously in organisms from bacteria to mammals, glutamate decarboxylase (GAD) is an enzyme
responsible for catalyzing the formation of 4-aminobutyric acid (GABA) from glutamate. GAD, however, is implicated
in various functions in cells including stress stimulated metabolism in plants and neurotransmitter production in
mammals. In order to function, GAD requires the binding of the cofactor, pyridoxal phosphate (PLP), and in many
organisms, especially mammals, PLP insertion into the active site of the enzyme is facilitated by pyridoxal kinase
(PK) which can bind directly to GAD. In this way interaction between PK and GAD may play an important role in
regulating GAD activity.
The purpose of this project was to determine how and to what degree different GAD and PK proteins from Arabidopsis
thaliana, a representative plant species, interact. There are four GAD and one PK genes capable of isolation in this
species; however the PK gene produces two splice variants. These genes were obtained and cloned using the pGEM
T Easy vector cloning system. The genes were then placed in the pET 23 vector for protein production. Each of the
six proteins was collected and purified using nickel-resin affinity chromatography. The proteins are now being used
for fluorescence polarization, which will quantify the interactions between the various proteins and establish whether
the PK-GAD interaction is sequence specific between enzymes from closely related organisms or generic in nature
and unrelated to the sequence of the interacting proteins. This work serves to shed light on a relatively
uncharacterized yet vital process in the stress response in plants. This work could also provide a possible option for
improving a plant`s response to stress by increasing the efficiency of this critical stress related pathway making it
possible to improve plant productivity during environmental stress.
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Telomere length control in Aspergillus nidulans
Megan E. Campbell (Prof. Steven James), Department of Biology, Gettysburg College, Gettysburg, PA 17325
Our laboratory focuses on the control of DNA synthesis in the fungus Aspergillus nidulans. Recently, we have
become interested in possible links between control mechanisms at the telomere and those involved in bulk
chromosomal DNA synthesis. Surprisingly, the telomeres in Aspergillus are unlike those of animals, protists, and the
budding and fission yeasts in that they are both very short and have discrete lengths. These observations imply that
telomere length control mechanisms, which are well known in other systems, must exert very stringent control to
maintain the exceptionally short, fixed-length telomeres in Aspergillus. For this reason, we are examining telomere
length control (TLC) in Aspergillus by using a Southern blot assay, and have thus far shown that mutations in several
key regulators of chromosomal replication do not alter telomere length. Currently, we are deleting orthologs of
several known TLC genes, loss of which in budding or fission yeast lengthens telomeres substantially. We have
deleted the apparent orthologs of budding yeast Rap1 and Tbf1, and fission yeast Taz1 (equivalent to human
TRF1/TRF2). These deleted strains, along with several others, are being tested for dysregulation of TLC.
Characterization of snoAp, an inhibitor of DNA synthesis in Aspergillus nidulans
Kalin V. Vasilev (Prof. Steven James), Department of Biology, Gettysburg College, Gettysburg, PA 17325
In the filamentous fungus Aspergillus nidulans, nimO-Dbf4 and cdc7 encode regulatory and catalytic subunits of the
conserved Dbf4-dependent kinase (DDK). DDK initiates DNA synthesis by phosphorylating components of the
replicative DNA helicase to trigger origin unwinding. A temperature sensitive nimO mutation, nimO18, can be
suppressed by mutations in two genes, snoA and snoB. Semi-dominant snoB mutations lie within the cdc7 gene, and
suppress by stabilizing the association of cdc7p with nimO18p. Recessive snoA suppressors not only rescue nimO18
ts-lethality, but they also rescue strains hypomorphic for expression of nimO+. However, snoA suppressors cannot
bypass the complete loss of nimO. These observations reveal that snoA suppression is not allele-specific, and that
snoA appears to act by inhibiting the stability or activity of nimOp. We used a positional cloning approach to localize
the snoA gene within a 340,000 bp interval of the Aspergillus genome. snoA was identified by deletion of candidate
ORFs, one of which phenocopied the nimO18 suppressor phenotype of snoA mutants. In this study, two-way fusion
PCR is being used to insert three copies of the hemagglutinin (HA) epitope at the snoA C-terminus. One-step gene
replacement will be used to generate a strain carrying snoA::HA3 as the sole copy of the snoA gene. Western blotting
will then be applied to characterize this tagged snoA protein. (Supported by NSF-RUI #01-14446 to SJ)
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TRANSCRIPTOMIC CHANGES IN SKELETAL MUSCLE IN RESPONSE TO DISUSE:
CHARACTERIZATION OF MICROGRAVITY AND SUSPENSION MODELS OF MUSCLE ATROPHY
Seiha Thorng, Jeremy L. Barth (Dr. W. Scott Argraves Laboratory, MUSC SURP 2005, NASA), Department of Cell
Biology and Anatomy, Medical University of South Carolina, Charleston, South Carolina 29425
It has been established that immobilization and disuse cause changes in physiological systems that are detrimental to
human health. Skeletal muscle in particular undergoes rapid degradation in response to disuse. With the help of
DNA microarray technology and bioinformatics tools, we are able to investigate the transcriptomic mechanisms
underlying the physiological responses to disuse. Here we have performed DNA microarray analysis of two models of
skeletal muscle disuse (microgravity and a ground based hindlimb Suspension). Genes differentially expressed in
response to disuse versus control were identified, hierarchical clustering and heat mapping of expression patterns was
performed, and biological processes significantly represented among differentially expressed genes were identified.
Our results show that gene expression levels change following 12 days of disuse. In response to microgravity 332
genes were differentially expressed (fold change above 1.5 and t-test p-value < 0.05) in comparison to ground control
samples. QPCR analysis of 6 differentially expressed genes confirmed changes in response to space flight. In
response to suspension 52 genes were differentially expressed compared to control samples. QPCR analysis
validated the differential expression of 3 of these genes. In muscle samples reloaded for 4 hr following hindlimb
suspension 57 genes were differentially expressed in comparison to suspended samples. QPCR analysis confirmed
the differential expression of 3 of these genes. Among the genes differentially expressed in each disuse model,
several functional categories were found to be overly represented, particularly: 1) induction of transcription factors in
the microgravity model, 2) nerve ensheathment/maturation and fatty acid/lipid metabolism in the suspension model,
and 3) stress response in the reloaded suspension samples. Future experiments will clarify how these biological
processes are related to the physiological aspects of skeletal muscle atrophy, either as a function of disuse or
disease.
Dominican University of California
50 Acacia Avenue
San Rafael, CA 94901
Nitric Oxide as a Neurotransmitter in the Salivary Glands of Manduca Sexta
Karly A. Burke, (Dr. Harry Itagaki), Department of Biology, Kenyon College, Gambier, OH 43022
Organisms must have finely tuned exocrine secretion pathways in order to regulate protein, fluid, and ion levels. One
means of regulation is through neurotransmitter release. Manduca sexta salivary glands have been little studied, yet
offer a simple cellular system in which to study neurotransmitter release, signaling, and downstream effects. The
paired glands are tubular structures and are composed of a single-cell layer. Though five discrete cellular regions are
found in the Manduca salivary gland, only the fluid-secreting region is innervated. Therefore, it is of interest to
determine other neurotransmitters and signaling pathways involved in the remaining four regions of the salivary
gland. Nitric oxide, the product of the nitric oxide synthase (NOS) enzymatic reaction, is a gaseous neurotransmitter
found in the central nervous system of invertebrates. Preliminary immunocytochemistry experiments indicate the
presence of nitric oxide synthase in Manduca salivary glands. Polyclonal anti-NOS antibodies were used to localize
nitric oxide synthase in cross-sections of salivary glands from 5th instar caterpillars. Staining patterns suggest that
the enzyme may be distributed evenly across the organ. Semi-quantitative RT-PCR experiments show that NOS and
soluble guanylyl cyclase (sGC), a common target of nitric oxide, is expressed throughout the salivary gland. These
results suggest that Manduca salivary glands may use the NOS-sGC signaling pathway to communicate between
cells. These data correspond with known work on the NOS-sGC pathway in the Manduca olfactory system in the
antennal lobe.
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CHARACTERIZATION OF A MOUSE MODEL FOR ALTERED INTEGRIN FUNCTION IN BONE
Sam Frankel, Jennifer Saasaki & Darilis Suarez. (Dr. Jonathan Phillips and Dr. Ruth Globus), Bone & Signaling Lab,
NASA Ames Research Center, Mountain View, CA 94040.
One of the challenges facing astronauts is the effect of a weightless environment on the skeletal system. The specific
mechanisms that translate mechanical information, such as the pressure of gravity, into a biological response are
unknown. Cell culture experiments suggest that mechanical stimuli are transmitted by a specific form of integrin
receptor on a type of bone cell called osteoblasts. Integrins are a class of transmembrane receptors that mediate the
cell’s adherence to extra-cellular matrix (ECM) proteins. The term “ß1-integrin” refers to an integrin receptor
containing the ß1 subunit paired with one of several complementary ? subunits. How ß1-integrins influence bone
homeostasis in vivo and biological reaction to mechanical stimuli are major questions in cell biology. In order to study
the function of ß1-integrins, we maintained a transgenic mouse line with a conditional knockout of the ß1-subunit
specific to osteoblasts. ß1-integrins are functionally distinct in their ability to bind an important ECM-protein called
collagen. To characterize the function of ß1-integrins, differences between ß1-deleted and control mice were
examined after use of the hindlimb-unloading procedure. Hindlimb unloading, a well established model of
microgravity, removes mechanical stimuli from the rear limbs. Collected tissues are to be analyzed upon reaching an
appropriate sample size for each genotype. We also refined an assay based on the binding of fluorescent
“microspheres” to cells. The targets of the assay were primary osteoblasts, dissociated and cultured directly from
animal tissue. Because all the forms of integrin receptor that bind collagen contain the ß1-subunit, the relative binding
ratio of collagen as compared to other ECM-coatings may be altered in ß1-deleted cells.
Positional Cloning of a C. elegans Supressor Mutation of a Neuronal &#945;-tubulin Defect
Lester Laddaran (Dr. Renee Baran), Biology Department, Occidental College, Los Angeles, CA 90041
The Baran laboratory is using the nematode Caenorhabditis elegans to study the molecular mechanisms that regulate
synapse formation and stabilization. Microtubules play multiple functions in neurons including axon formation,
transport of vesicles and organelles, and regulating synapse growth. We focus on tba-1, a gene that codes for the
&#945;-tubulin subunit of microtubules. Using a molecular genetic approach, we work with ju89, a tba-1 missense
mutation that alters the exterior of tubulin polymers. We hypothesize that this mutation disrupts critical residues to
which microtubule associated proteins bind. My project is concerned with mapping sup51, an unlinked mutation that
compensates for the ju89 defect. Using a high-throughput SNP (single nucleotide polymorphism) mapping strategy
and PCR (polymerase chain reaction), three candidate regions in two chromosomes have been identified. When the
correct region has been identified, we will use cosmids and transformation rescue to further narrow down the region.
To further study the regulation of tba-1, I am also creating a transgene that will fuse a flp-13 promoter and a red
fluorescent protein gene to the 3’ end of the tba-1 gene, which will be introduced into the C. elegans genome via a
standard injection protocol. The promoter will direct expression of the transgene to the six C. elegans motor neurons
that synapse into the dorsal muscles, enabling us to visualize these sites with a fluorescence microscope.
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HOW TO CONTROL AN ESSENTIAL GENE: BUILDING A CONSTRUCT TO FIND THE FUNCTION OF A
NOVEL ACINETOBACTER BAYLYI GENE
Natalie J. Funk (Dr. Leslie Gregg-Jolly and Dr. James Swartz), Department of Biology, Grinnell College, Grinnell, IA
50112
Cancer is a widespread disease and in the pursuit of a cure, we have discovered that it is often caused by the
unchecked proliferation of damaged DNA. Cells possess the ability to repair such damage with the help of specific
DNA repair genes, some of which are conserved in many species (1). A new gene, termed LGJ14, was recently
discovered in the naturally transformable bacterium Acinetobacter baylyi (2, 3). The LGJ14 gene is hypothesized to
be involved in DNA repair because its expression is increased specifically in response to DNA damage (2). The goal
was to determine the function of LGJ14, but the gene also appears to play an essential role in cell growth.
Conventional methods of study such as mutational insertions were impossible because they caused cell death (2).
The project thus involved engineering a genetic construct in which the LGJ14 gene could be manipulated without
causing lethality. The ultimate goal was to place the LGJ14 gene under the pobA promoter where its level of activity
could be incrementally turned up or down, thus allowing its function to be determined. The process of engineering this
construct contains many steps; the gene has been PCR amplified, inserted into a pCR® 2.1-TOPO vector, and its
orientation determined within the vector. The LGJ14 gene is now ready to ligate into the pobA gene and transform
into the bacterium. This construct will help determine the gene’s functions both in cell growth and DNA damage
response. Once the functions are understood of LGJ14 and other genes involved in repairing damaged DNA, this
knowledge could greatly aid the search for new cancer treatments.
(1) Walker, G. 2000. Understanding the Complexity of an Organism’s Response to DNA Damage. Cold Spring Harbor
Symposia on Quantitative Biology.
(2) Gregg-Jolly and Hare, personal communication.
(3) Metzgar, D. et al. 2004. Acinetobacter sp. ADP1: an ideal model organism for genetic analysis and genome
engineering. Nucleic Acids Research 32: 5780-90.
The Zinc-Finger Mutation, DDS, disrupts Wilms` Tumor 1 (WT1) Regulation of Vascular Endothelial
Growth Factor (VEGF) Promoter in LNCaP Cells
Jacquelyn Gorman, Julie Hanson, Jennifer Reese, Yasmeen Tandom, Kylie Graham (Gail Fraizer), Biological
Sciences, Kent State University, Kent, OH 44242
Prostate Cancer is the second leading cause of death in men and the number of incidence is on the rise. Our
objective was to determine whether the mutant, WT1-DDS, transcriptionally regulates VEGF. In previous studies, we
demonstrated that both WT1-LNCaP and WT1-DDS LNCaP cells were growth suppressed in mice compared to
vector control-LNCaP cells. In order to identify WT1-regulated pathways crucial for growth suppression of prostate
cancer, we conducted cDNA microarray analyses of vector control-LNCaP, WT1-DDS LNCaP, and WT1-LNCaP cell
lines. We observed differential vascular endothelial growth factor (VEGF) expression in WT1-LNCaP and WT1-DDS
LNCaP cells and validated these results with quantitative real-time PCR. To understand how the structure of WT1
determines it’s function as a transcription factor, we examined the activity of the zinc-finger mutant, WT1-DDS. We
transfected wildtype WT1 and WT1-DDS into LNCap cells and compared luciferase activity of VEGF luciferase
promoter constructs. While WT1 transcriptionally regulates the VEGF promoter, the DDS mutant remains inactive. In
conclusion, the WT1-DDS mutation caused a loss of function compared to wildtype WT1, thus the zinc finger
structure of WT1 is important in determining it’s function. Given that angiogenesis and tumorigenicity is correlated
with VEGF expression, a potential mechanism of prostate cancer growth suppression is the regulation of VEGF
expression by WT1. Supported by NIH (CA113360), Ohio Board of Regents and KSU-University Research Council.
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EXPLOITING THE GLYCOLYTIC PHENOTYPE IN CANCER CELLS
Jenny Y. Huang, Milheon Choi, Lijun Tan and J. Rebecca Liu, Department of Obstetrics and Gynecology, University
of Michigan, Ann Arbor, MI 48109
Objectives: In mammalian cells, autophagy (“self-eating”) is an adaptive response to nutrient starvation. The
breakdown of macromolecules by autophagy produces amino acids and other elements needed for intermediate
metabolism and biosynthetic pathways. Eventually, if nutrient deprivation is sustained, the cell will undergo
autophagic cell death. Both amino acid deprivation and glucose deprivation can induce autophagic cell death. In this
study, we sought to determine whether cancer cells were preferentially sensitive to glucose deprivation or amino acid
deprivation induced cell death.
Methods: A panel of cancer cell lines was screened for sensitivity to 2-deoxyglucose, (2-DOG, a specific glucose
inhibitor) using the sulforhodamine assay. Cell lines were incubated in media containing both amino acids and
glucose, amino acids without glucose, 25 mM glucose without amino acids, 5mM glucose without amino acids, or in
the absence of both amino acids and glucose. Cells were monitored for a 48 hour time period and viability was
determined at every 12 hour intervals by sulforhodamine assay.
Results: Sensitivity to 2-DOG varied among cancer cell lines. Sensitivity to 2-DOG did not correlate with cell growth.
In all cell lines tested, including the normal human fibroblasts, deprivation of both amino acids and glucose, or amino
acids alone resulted in the rapid induction of cell death (by 12 hours). These results were seen in the presence of both
25 mM glucose and 5 mM glucose, indicating that amino acids are essential for the survival of all cell types. In
contrast, glucose deprivation resulted in cell death in all cancer cell lines by 48 hours as compared to normal human
fibroblasts which remained viable.
Conclusions: Cancer cells exhibit increased rates of glycolysis as compared to non-transformed cells. Many patients
with solid tumors develop resistance to conventional chemotherapeutic agents. Chemoresistance is associated with
alterations in the apoptotic pathway. Because cancer cells are dependent upon glucose for survival, agents that
inhibit glycolysis, such as glucose analogs, may have broad clinical benefit to cancer patients.
THE ROLE OF NUCLEAR RECEPTORS IN THE DAUER MOLT OF CAENORHABDITIS ELEGANS
Theresa Buggs, Mark Corkins and Shannon McCarragher (Dr. Kirsten Crossgrove), Department of Biology, The
University of Wisconsin-Whitewater, Whitewater, Wisconsin, 53190
The purpose of this project is to determine whether nuclear receptors play a role in the dauer molt of Caenorhabditis
elegans. C. elegans, a small free living soil nematode, serves as useful genetic model due to its easy maintenance,
short life cycle, and fully sequenced genome. As they develop, C. elegans go through a series of molts (shedding of
the outer cuticle to allow growth) culminating in an adult hermaphrodite. However, if conditions are undesirable, the
worms may take an alternate route in the life cycle by becoming dauers instead of normal L3 larvae. Dauer larvae are
designed to survive harsh conditions as they await a more favorable environment. When optimal conditions are
detected, the worms molt and continue their lifecycle into an adult. Parasitic nematodes also have an arrested stage
in their life cycle that prohibits further development until appropriate environmental cues, such as the entrance into a
host, is perceived. We are interested in the genetic control of these developmental transitions. Members of the
nuclear receptor superfamily of transcription factors have been implicated in control of molting. Through the use of
temperature-sensitive dauer constitutive mutants, or daf-c worms, which form dauers regardless of environmental
conditions, the role of nuclear receptors can be tested. The daf-c mutants are exposed to double stranded RNA
(dsRNA) specific for nuclear receptor genes through consumption of Escherichia coli bacteria engineered to express
the dsRNA. After ingesting the transformed bacteria containing the dsRNA, the targeted genes are silenced in the
worms, which allows for the observation of loss of function phenotypes. We will present data showing that nhr-23 is
required for the entry into dauer, while nhr-85 plays a less vital role. We are also testing several nuclear receptors
whose microarray expression pattern suggests a role in exit from dauer. Our results demonstrate a role of nuclear
receptors in the dauer molt of C. elegans.
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Generation of Cul5 mutants in Drosophila
Ashley N. Salaz (Patrick J. Reynolds, Dr. Robert J. Duronio), Department of Biology, UNC-Chapel Hill, Chapel Hill, NC
Regulated protein degradation is an important process in the cell cycle. Several diseases including Alzheimer’s,
Parkinson’s, and some cancers may be caused when proteins are not properly degraded. For example, inefficient
ubiquitination of cell cycle machinery can cause cells to over proliferate. The ubiquitin proteasome pathway, which
consists of E1, E2 and E3 enzymes, targets proteins to be destroyed by the 26S proteasome. Many E3 ligases
contain a Cullin protein that forms a scaffold on which the rest of the complex assembles. Lethal mutants have been
identified for Cullins 1-4. The main objective of my research is to create a cullin 5 (Cul 5) mutation in Drosophila
melanogaster and determine the phenotype of the mutant. I am doing this by excising a P-element located just
upstream of Cul5 and looking for imprecise excision events that remove part of or the entire gene. With my research
so far I have created seventy-seven possible Cul 5 excision lines.
ROLE OF CLU1P IN MITOCHONDRIAL GENOME STABILITY AND MORPHOLOGY IN BUDDING
YEAST
Crystal R. Allen, Christine Hochmuth, and (Rey A. Sia) Department of Biological Sciences, State University of New
York at Brockport, Brockport, NY 14420
Maintenance of the mitochondrial genome is critical for normal cellular respiration. Mutations in the genome have
resulted in loss of mitochondrial function. Damage to the genome leads to premature aging and heart, lung, and
neurodegenerative diseases. Little is known about the pathways governing mitochondrial genome stability.
Saccharomyes cerevisiae (budding yeast) is a facultative anaerobe that can grow in the absence of respiration when
supplied with a fermentable carbon source. The lab has used a yeast two-hybrid assay with the known mitochondrial
protein, Ilv5p, to isolate genes whose products may be involved in the organization, repair, and recombination of
mitochondrial DNA (mtDNA). The CLU1 gene was identified in this screen. Clu1p was found to be essential for
proper mitochondrial morphology and distribution (1). CLU1 in S. cerevisiae was found to be homologous to the cluA
gene of Dictyostelium discoideum.
Our data provides evidence that Clu1p plays a role in mitochondrial genome stability and morphology. The lab
has shown that a clu1&#8710; strain displays a 9.5 fold increased rate of respiration loss as evidenced from increased
petite formation. Increased petite formation may arise from DNA polymerase slippage events at microsatellite tracts
or from an increase in mtDNA point mutations. Using clu1&#916;::URA3 reporter strains, the lab has found a 1.6 fold
increase in the rate of alterations in microsatellite tracts and a 2.1 fold increase in the rate at which mtDNA point
mutations form. Using a recently developed direct-repeat-mediated deletion reporter strain, loss of Clu1p results in a
5.3 fold decrease in the rate of deletion formation relative to that of wild-type. Microscopic imaging supports an
earlier report indicating that a clu1&#8710; strain displays a “clustering” phenotype (1). We found that the
clu1&#916;:URA3 strain exhibits a branched mitochondrial network that is localized or “clustered” to one side of the
yeast cell.
1. . Fields SD, Conrad MN, Clarke M. (1998) J Cell Sci. 111 ( Pt 12):1717-27.
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LPL PvuII Polymorphism is Associated with Performance in Endurance Running
Rosalyn Gonzàlez Universidad de Puerto Rico en Cayey, Avenida Antonio R. Barceló Cayey, P.R. 00736, Stavros
Kavouras, Harokopio University, Athens176-71, Greece Costas Anastasiou, Harokopio University, Athens176-71,
Greece, Labros Sidossis, Harokopio Univ
Lipoprotein Lipase (LPL) is a capillary bound enzyme that breaks down triglycerides in chylomicrons providing free
fatty acids for cells as substrates. In skeletal muscle, LPL mRNA is increased significantly in response to even a
single bout of endurance exercise. Studies with rats showed significant linkage between regions of chromosome 16,
orthologous to chromosome 8 in humans, and endurance running capacity. Chromosome 8 harbors various genes
related to fat metabolism including LPL at 8q22. PURPOSE: To examine the hypothesis of association between a
polymorphism on the LPL gene (PvuII C to T) and endurance running performance. METHODS: DNA was extracted
from white blood cells of 80 volunteers, participants of the 2005 Olympus Mountain Marathon; a 44Km footrace over
the 2900m high Mt. Olympus from Dion to Litochoro in Greece. All subjects are Caucasian and 90% are Greek. A
430bp fragment of the LPL gene was amplified by PCR and then digested with 10U of PvuII restriction enzyme; which
recognizes and digests the C allele. Subjects were placed in one of two groups of 40 according to their finishing times
(top 40 (T40) vs. lower 40 (L40)). Genotypic frequencies were compared and analysed by X2. RESULTS: For the
whole group of 80 runners allele (C= 52% T= 48%) and genotype (CC= 29%, CT= 46%, and TT= 25%) frequencies
were not significantly different from the expected frequency for the general Greek population based on previous
results from the literature. However, when compared by subgroups, genotype frequency for T40 (35%CC, 50%CT and
15%TT) was significantly different from L40 (23%CC, 42%CT and 35%TT) (X2 = 12.9, df=2, P=0.02). The difference
was even stronger when comparing the top 20 (30%CC, 65%CT and 5%TT) vs lower 20 (15%CC, 50%CT and
35%TT) (X2 = 38.2, df=2, P<0.001). In both cases, the TT genotype is less frequent among top performers. Mean
finishing time for the C allele carriers (CC + CT) was significantly lower (P=0.025) than for the TT subjects (C carriers
= 521.9 +/- 100.5 min vs. TT = 582.2 +/- 95.1 min). CONCLUSION: The results support the hypothesis that LPL gene
sequence variation is associated with endurance running performance among Greeks.
Expression and Purification of Leghemoglobin from Partridge Pea
Lawnie S. Woolsey and Frank A. Kovacs, Department of Chemistry, University of Nebraska at Kearney, Kearney, NE
68849
Chamaecrista fasciculata also known as partridge pea is a prairie legume whose biochemistry remains largely
uncharacterized. As a legume, it maintains a symbiotic relationship with a nitrogen fixing bacterium that infects the
roots of the plant and cause the formation of nodules. Like other legumes, partridge pea produces leghemoglobin
(Lb), which is a small heme binding protein found in its root nodules. Lb functions as an oxygen forager and
transporter in bacterial nitrogen fixation. The purpose of this study was to express and purify Lb from partridge pea.
This work represents the first step towards our long term goal of studying nitrogen fixation in partridge pea and
comparing it to other leguminous plants. The gene for Lb was obtained from a complimentary DNA library from
partridge pea root nodules. The gene was cloned into a vector containing a 6xHistidine tag. Lb was then expressed
in E.coli and samples were purified by Ni-NTA purification. The purified protein was then analyzed by SDS-PAGE for
purity and BCA assay for concentration and yield.
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GENERATING INSERTIONAL DELETIONS IN TYPE III SECRETORY GENES OF THE ERWINIA
CAROTOVORA CHROMOSOME.
Meghan B. Moll (Dr. John Pritchard), Department of Biological Sciences, The University of Wisconsin-Whitewater, 800
W. Main St., Whitewater, Wisconsin 53190
The gram-negative bacteria Erwinia carotovora infects many significant crops through the Type III Secretion System
(TTSS). The TTSS inserts bacterial proteins into host cells to weaken the host’s immune system. The TTSS is a
requirement for E. cartovora to successfully infect a host plant. Sorting through the library of insertional mutants we
can locate and characterize genes involved in the TTSS pathogenicity. Our efforts to use a mini-Tn5 system to
generate insertional mutations in the genome of E. carotovora suggest that this method is effective in creating TTSS
mutant clones. Following phenotypic screening of TTSS mutants, we will use PCR to amplify the genomic insertions
along with the sequences of the flanking regions and then map the insertion sites using the already published E.
carotovora genome. This project is being conducted jointly with other researchers to develop the mutant library, but
the specific project reported on here concerns the identification, mapping, and characterization of different genes
responsible for unique phenotypes. Further characterization of the TTSS genes, their regulation, and the protein
products will offer clues for the prevention and treatment strategies of crop diseases.
ISOLATION AND CHARACTERIZATION OF METHYL BUTENOL SYNTHASE FROM PONDEROSA PINE
Kathryn R. Amerell (Dr. Eric Singsaas and Dr. Ed Gasque), Department of Biology, University of Wisconsin-- Stevens
Point, Stevens Point WI, 54481
The purpose of this project is to identify, isolate, and sequence the enzyme methyl butenol synthase (MBS)
responsible for making methyl butenol (2-methyl-3-buten-2-ol) from Ponderosa pine (Pinus ponderosa L). This will be
the groundwork for isolating the gene for MBS which can be transformed into E. coli bacteria for production of butenol
-based biofuel. The immediate goals of this project include the induction of methylbutenol production through heat
treatments in Ponderosa pine seedlings, confirmation of the synthesis of methyl butenol using gas chromatography,
extraction and then isolation of the protein using 2-D gel electrophoresis, and sequencing of the protein at the
biotechnology center at the University of Wisconsin-Madison. The long-term outcomes of this project will be to
transform MBS into E. coli for production of butenol-based biofuel could offer a practical biomass-based fuel source
useful for transportation that will be superior to corn-based ethanol production because the feedstock does not
compete with food production, there is substantial energy savings when the feedstock comes form waste products
such as corn stover or paper mill waste, and methyl butenol’s higher energy density
A New Model to Study Adult Skeletal Muscle Gene Function
Peter S. Wulff (Dr. Karyn Esser), Physiology Department, University of Kentucky College of Medicine, 800 Rose
Street Lexington, KY 40536
In order to study gene function it is important to observe the gene of interest in a physiologically relevant, spatial,
temporal and quantitative manner. My project is to create a new mouse model that will enable us to study gene
function only in adult mouse skeletal muscle cells. My project is to create a double-transgenic mouse via
cross-breeding.
First, I will take a mouse that produces Cre-recombinase under control of the Tetracycline Operon promoter (TetO)
and breed it against a transgenic mouse carrying a skeletal muscle specific promoter driving the reverse
tetracycline-controlled transactivator (rtTA). When the double transgenic mouse is given doxycycline, this binds to the
rtTA molecule which activates the TetO promoter and drives expression of Cre-recombinase only in skeletal muscle.
Cre-recombinase is an enzyme that will cut genomic DNA at loxP sites to induce a recombination event. To test
whether the double transgenic mouse actually maintains proper skeletal muscle specific expression of
Cre-recombinase in response to doxycycline, I will cross it with the ROSA mouse. The ROSA mouse contains a stop
sequence flanked by LoxP sites upstream from the LacZ gene. When the doxycycline is fed to the offspring of the
cross, the Cre-recombinase will cut out the stop sequence and enable expression of &#946;-galactosidase. I will
collect different types of tissue and stain them with X-gal to identify the cells expressing &#946;-galactosidase, which
will only be skeletal muscle. This new mouse model will enable me to study gene function by inducing skeletal
specific gene knockouts.
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EXPRESSION OF KERATINOCYTE TRANSGLUTAMINASE (TGASE1) IN HUMAN CERVICAL AND
VAGINAL CELL LINES
Manira Rayamajhi (Dr. Wanda T. Schroeder), Department of Biology, Wesleyan College, Macon, Georgia 31210
Keratinocyte transglutaminase (TGase1), essential for the normal differentiation of epidermal cells, is usually
expressed during the latter stages of keratinocyte differentiation. Previous studies, performed in ovariectomized rats,
examining KTG mRNA levels at varying time points after administration of exogenous estrogen, have demonstrated
that TGase1 mRNA is differentially regulated in uterine and vaginal epithelia in response to estrogen. Recent in vivo
studies in mice, examining the expression of TGase1 protein in mouse uterus and vagina at different stages of the
estrous cycle, have demonstrated that the TGase1 protein is also differentially expressed in estrogen-responsive
uterine and vaginal epithelia. Surprisingly, a marked difference in TGase1 protein expression has been observed
between uterine horn and cervical tissue. To more closely examine the expression of TGase1 in human vaginal and
cervical epithelia and to determine if there are differences in TGase1 protein expression within the human uterus,
three immortalized normal human cell lines have been obtained from ATCC, corresponding to vaginal mucosa
(CRL-2616), ectocervix (CRL-2614) and endocervix (CRL 2615). Immunohistochemistry experiments performed on
these cell lines utilizing BC1 human TGase1-specific monoclonal antibody have determined the expression pattern of
TGase1 protein expression in these estrogen-responsive epithelia in humans. These studies may be useful in
understanding control of differentiation in both vaginal and transitional cervical epithelia.
A MOLECULAR GENETIC SCREEN FOR XENOPUS PROTEINS THAT BIND TO HERMES REVEALS A
NOVEL INTERACTION WITH THE CELL CYCLE CONTROL PROTEIN, CYCLIN A1.
Charles J. Packard (Dr. Scott M. Ballantyne), Department of Biology, University of Wisconsin – River Falls, River
Falls, Wisconsin 54022
Hermes is an RNA binding protein that is needed for the development of the frog Xenopus laevis. Hermes controls
cell division in the early embryo1 and is also required for proper heart formation during later development2. We have
performed a yeast two hybrid screen to search for frog proteins that bind to Hermes. We obtained ~70 yeast colonies
that contain candidate Hermes binding proteins. We are isolating and sequencing the cDNA that encode these
proteins. By identifying the proteins that interact with Hermes, we hope to better understand how Hermes controls
development.
We have now obtained partial cDNA sequences for three of the candidate Hermes binding proteins. Bioinformatic
analyses indicate that one cDNA encodes the protein, cyclin A1. Cyclin A1 is required for M-phase progression during
the cell cycle. Cyclin A1 normally associates with and regulates a cyclin dependent kinase. Our results raise the
intriguing possibility that cyclin A1 may also act in concert with the RNA binding protein Hermes to control early
embryonic cell divisions. We are currently further characterizing the Hermes:cyclin A1 interaction as well as
continuing to isolate and sequence cDNA that encode candidate Hermes binding proteins.
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DIFFERENTIAL EXPRESSION OF KERATINOCYTE TRANSGLUTAMINASE (TGASE1) IN ESTROGEN
RESPONSIVE EPITHELIA DURING MOUSE ESTROUS
Abena Redwood, Yo-Leigh Gardner, and GayeLeon Williams (Dr. Wanda T. Schroeder), Department of Biology,
Wesleyan College, Macon, Georgia 31210.
Expression of keratinocyte transglutaminase (TGase1) occurs in differentiated layers of epidermal tissue and
influences the cornification of the outermost layers of keratinocytes. While TGase1 expression has been thoroughly
investigated in epidermis, little is known of its expression pattern in hormone-responsive epithelia including uterine
and vaginal tissues. Previous studies, performed in ovariectomized rats, examining KTG mRNA levels at varying
time points after administration of exogenous estrogen, have demonstrated that TGase1 mRNA is differentially
regulated in uterine and vaginal epithelia in response to estrogen. This study was designed to determine the
expression pattern of TGase1 protein in mouse uterine and vaginal epithelium at different stages of the estrous cycle
including proestrus, estrus, metestrus, and diestrus. Histological staining of cells flushed from the reproductive tract
of cycling female mice was used to stage mice during the estrous cycle. Mice were sacrificed at different stages of
the cycle and immunohistochemical staining of frozen sections of uterus and vagina were performed utilizing a rat
monoclonal antibody specific for TGase1. These studies demonstrated that while TGase1 protein was expressed in
mouse vaginal epithelium during all stages of the cycle, there was no expression of TGase1 protein in the body and
horn of mouse uterus. However, TGase1 protein was expressed in cervical epithelium at estrus when estrogen levels
are at peak concentration and there is cell proliferation. Thus, TGase1 protein is not only differentially expressed in
different estrogen-responsive tissues during estrus, but is differentially expressed in different areas of the uterus as
well. This study provides valuable information on differentiation pathways in estrogen-responsive epithelia.
Determination of the Protein Level of Site-Specific Mutants of GerE During Sporulation in Bacillus
subtilis
Travis R. Russell (Dr. Dinene Crater), Departments of Chemistry and Biology, High Point University, 833 Montlieu
Ave., High Point, NC 27262
GerE is a transcription factor produced by Bacillus subtilis during the late stages of sporulation. It is the smallest
transcription activator from the LuxR-FixJ family. Past research has found that the F6A mutant of GerE exhibited a
lower level of transcription from the cotC promoter, but had no affect on transcription of the cotX promoter. Both the
L67A, and L74A mutants showed a lowered transcription in the cotX promoter, and had no affect on the cotC
promoter. Our future goals are to test for the presence of the mutant forms of GerE in the mutant strains. After
determining that GerE is in fact present in the mutant strains, we will examine the concentration of mutant GerE that
is present in cells compared to the control wild type strain. We are examining the presence of GerE over time during
the sporulation phase of B. subtilis. By using SDS-PAGE we are able to separate the proteins, then we use Western
Blot to transfer the proteins to a PVDF membrane. By utilizing Chromatogenic Immuno-Blot assay (BioRad) we are
able to visualize the presence of GerE using anti-GerE antibodies. There are three GerE mutant strains we will be
testing, F6A, L67A, and L74A. Our research will determine if the lowered transcription of cotC, and cotX is due to a
reduced amount of GerE present in the mutant strains or if it is the result of a lowered affinity of the GerE to sigma K
RNA polymerase.
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TRANSACTIVATION ACTIVITY OF XENOPUS LAEVIS ARYL HYDRCARBON RECEPTORS:
AHR1-ALPHA AND AHR1-BETA EXHIBIT LOW RESPONSIVENESS TO STRUCTURALLY DIVERSE L
Megan A. Ely (Dr. Wade H. Powell) Biology Department, Kenyon College, Gambier, OH USA
Dioxin (2,3,7,8 tetrachlorodibenzo-p-dioxin; TCDD) is a widespread environmental contaminant. It is a potent
developmental toxicant in most vertebrates. TCDD toxicity is mediated by the aryl hydrocarbon receptor (AHR), a
ligand activated transcription factor. Frogs are remarkably insensitive to TCDD, especially at early life stages.
Recent studies in Xenopus laevis, the African clawed frog, demonstrate that two aryl hydrocarbon receptors,
AHR1-alpha and AHR1-beta, bind TCDD with at least 20-fold lower affinity than the mouse AHR, a property that likely
plays an important role in frog TCDD insensitivity. In this study we sought to determine the relative potency of a
range of potential agonists of X. laevis AHRs in assays using COS-7 cells transfected with a single receptor and a
luciferase reporter gene driven by a portion of the mouse CYP1A1 promoter. Few PCB congeners elicited a
significant response, although the dioxin-like PCB 126 exhibited an EC50 greater than 40 nM with each receptor.
Similarly, beta-naphthoflavone, a potent agonist of mammalian and fish AHRs, elicited no response, even at high
concentrations. These data are consistent with the low toxicity of polychlorinated biphenyls and other aromatic
hydrocarbons in frogs. Indigo, a dye compound found in human urine, is a potent agonist for the mouse AHR. In
conjunction with the frog AHRs, indigo exhibited a dose response comparable to TCDD (EC50 of 50-80 nM) but was
nonetheless substantially less potent than with mouse AHR. Taken together, these data suggest that the low potency
of TCDD is not a ligand-specific property of frog AHRs. Rather, it extends to a structurally diverse range of
compounds that are strong ligands for AHRs from species more sensitive to TCDD toxicity. The presence of an
endogenous, high-affinity ligand in frogs remains a possibility. [Support: NIH R15 ES011130]
THE ROLE OF ALPHA V BETA 3 INTEGRIN IN THE ANGIOTENSIN II STIMULATED INCREASE IN
FOCAL ADHESION ABUNDANCE IN CARDIAC FIBROBLASTS
Alissa M. Stull (Dr. Thomas Peeler), Department of Biology, Susquehanna University, Selinsgrove, PA 17870
The vasoconstrictive peptide angiotensin II (AII), which induces hypertension and eventually cardiac hypertrophy, also
stimulates an increase in the abundance of focal adhesions in cardiac fibroblasts in culture. Focal adhesions are
cytoplasmic structures that serve as connections between the cytoskeleton and integrins. Integrins are
transmembrane proteins that bind to extracellular matrix (ECM) proteins and act as signal transducers by relaying
ECM structural information to the cytoskeleton. Previous research has shown that AII increases &#945;v&#946;3
integrin expression at the mRNA level in both neonatal and adult rat cardiac fibroblasts. We tested the hypothesis that
the AII stimulated increase in focal adhesions was specifically due to an increased level of &#945;v&#946;3 integrin
protein, and that the &#945;v&#946;3 integrin would be found in the newly formed focal adhesions. Cardiac
fibroblasts were collected from 1-3 day old neonatal rats, isolated, and cultured. The cells were treated with 10-7 M
AII, and after incubation the cells were fixed. Antibodies to the &#945;v&#946;3 integrin were used in order to
visualize the integrins by fluorescence microscopy. Our results show the AII stimulated increase in focal adhesions is
due specifically to an increase in the abundance of &#945;v&#946;3 integrin, and that the &#945;v&#946;3 integrin
was preferentially localized in focal adhesions.
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ROLE OF QUORUM SENSING IN THE EXPRESSION OF ANTIGEN I/II IN STREPTOCOCCUS MUTANS
Marquicia Pierce and Zhiyun Chen (Dr. Richard Gregory), Indiana University School of Dentistry, Indiana
University-Purdue University Indianapolis, Indianapolis, IN 46202-5107
Streptococcus mutans has been proven to have a major role in the development of dental caries (cavities). This
bacterium adheres to the surface of the tooth and produces harmful acids. Antigen I/II is a cell-surface adhesin that
aids in this binding process. It was observed that antigen I/II expression in S. mutans was regulated along its growth
curve. From early-to-late log phase, antigen I/II expression level decreased by at least two folds. Previous research
has indicated that neither pH nor metabolic products alone have effect on antigen I/II expression. However, when the
cell density was altered the expression of antigen I/II changed. This leads us to suspect that quorum sensing plays a
role in antigen I/II expression. Quorum sensing enables a bacterial population to regulate gene expression
collectively. Quorum sensing in S. mutans is controlled by a number of genes including the comCDE and X.
Therefore, different comCDE and comX knockout mutants were used to identify the role of the com genes in this
alteration of antigen I/II expression. Western blotting experiments indicated that the comC, comD, comE and comX
mutants did produce antigen I/II at all cell densities and suggested that the com system is not essential for antigen I/II
expression. Further research will be done to see if this specific genes control regulation instead. This would indicate
that quorum sensing genes may regulate antigen I/II expression in S. mutans and play a role in the virulence of this
pathogen. In the future antimicrobial agents may be developed to inhibit this quorum sensing process.
Exploring Biochemical Interactions Between Histone H3 and the Transcription Factor Spt16
J. Ira Miller (Dr. Andrea Duina) Department of Biology, Hendrix College, 1600 Washington Ave. Conway, AR 72032
The vast length of DNA that composes the genome of any eukaryotic organism, from yeast to humans, is sequestered
within the small confines of the cell nucleus. In order to facilitate this compaction, DNA associates with a number of
proteins to form a highly condensed structure known as chromatin. The basic unit of chromatin is the nucleosome, a
particle formed by the association of DNA with a complex of proteins known as histones. In addition to facilitating
DNA compaction, the histone proteins are also involved in the regulation of gene expression through their interaction
with other proteins generally referred to as transcription factors. The focus of our project is to gain novel insights into
the functional and physical interactions that occur between a specific histone protein, histone H3, and the highly
conserved transcription elongation factor Spt16. We describe an experimental system designed to identify domains of
histone H3 that play important roles in the proper function of Spt16. In addition to providing novel insights into
histone H3-Spt16 interactions, this system could also be adapted to identify other factors that regulate the functions of
Spt16. Our studies are carried out using the yeast Saccharomyces cerevisiae, a widely used model organism that
allows us to utilize a combination of genetic, molecular biological, and biochemical experimental approaches. Since
the factors and processes we are investigating are highly conserved from yeast to human cells, our results are likely
to also be relevant to our understanding of gene expression mechanisms in human cells.
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A Screen for Genes Necessary for Drosophila Midline Cell Formation and CNS Development
Warren M. Perry, II, Yi Zheng (Dr. Patricia Estes), North Carolina State University Genetics Department, The
Graduate School, 212C Peele Hall, Box 7102, Raleigh, North Carolina 27695-7102
People all over the world are affected by baffling neurological diseases such as Huntington’s, Alzheimer’s, and
Parkinson’s. In addition to these neuro-degenerative diseases, spinal chord injuries and the natural aging of the
human body are both acompanied by neuron loss. Because of these reasons, the understanding of how the nervous
system forms and operates is essential.
The goal of our study is to identify new genes that control CNS midline cell development in the fruit fly, Drosophila
melanogaster, and subsequently determine if the genes identified play similar roles in mammalian development. The
midline is the important structure in the nervous system that divides the embryonic CNS in half. Midline cells produce
signaling molecules that play a vital role in CNS morphogenesis and axon guidance because many neurons cross the
midline in route to their final destination in the embryo. In our study, we are using the embryonic midline of
Drosophila to help us uncover new genes that affect CNS development.
To identify genes required for normal CNS midline cell development, male Drosophila containing a midline gene
marker that allows us to easily visualize midline cells during development are treated with a mutagen, ethyl methane
sulfonate (EMS), at a concetrantrion that produces appoximately two or three mutations per chromosome. Next, a
series of genetic crosses are used to produce embryos homozygous for the induced mutation and the midline gene
marker. The embryos are then screened for an abnormal midline phenotype using fluorescent microscopy.
Our early results include the screening of 412 mutations and the identification of fifteen mutations that disrupt midline
cell development. These mutations have been mapped to discrete regions of the Drosophila genome. From here, we
are fine mapping the mutations to one gene. We will characterize each gene by describing their functions within the
CNS and looking for the expression patterns at both the RNA and protein level.
Our study of CNS development at a genetic level is designed to understand mechanisms of neuron growth and
pathfinding which can ultimately be used to design treaments for patients with nerve damage caused by spinal chord
injuries or neuro-degenerative disorders.
PARTICIPATION OF ATM PROTEIN KINASE IN THE DEVELOPMENT OF INSULIN RESISTANCE IN
RATS AND IN INSULIN-REGULATED GLUT4 TRANSLOCATION IN L6 MUSCLE CELLS
Stephanie Traversie*, Jinghua He, and Jody C. Hibma (Da-Qing Yang), Division of Basic Biomedical Sciences, The
University of South Dakota, School of Medicine, Vermillion, South Dakota, 57069, * Dakota Wesleyan University
Ataxia-telangiectasia (AT) is an autosomal recessive disorder characterized by cerebellar ataxia and oculocutaneous
telangiectasias. Patients with A-T also have high incidence of type 2 diabetes mellitus. The gene mutated in this
disease, ATM (A-T,mutated), encodes a 370-kD protein kinase. To study the functional link between ATM and type 2
diabetes, we induced rats with insulin resistance by feeding them with a high-fat diet. Significant differences in insulin
levels and QUICKI values between rats on the high-fat diet and control rats on a regular-diet indicate insulin
resistance was induced. We then compared the expression of the ATM protein in muscle tissues of both groups of
rats. The results showed that the rats fed a high-fat diet over a three month period had dramatically reduced ATM
levels as compared to regular chow-fed controls. Next, we did a colorimetric (OPD) assay of the cell surface glucose
transporter 4 (GLUT4) after co-transfecting L6 myoblasts with plasmids encoding GLUT4 and wild-type(WT) or
kinase-dead (KD) ATM.
o-Phenylenediamine Dihydrochloride (OPD) assay showed that in cells transfected with WT-ATM and GLUT4, insulin
caused an increase of OPD activity similar to those seen in cells transfected with only GLUT4 plasmid. However,
cells transfected with KD-ATM and GLUT4 significantly inhibited the OPD activity. Moreover, we performed an
immunofluorescence assay. Confocal microscopy analysis showed that KD-ATM, in contrast to WT-ATM, also
significantly inhibited the translocation of GLUT4 to the cell surface by insulin. Our results provide new insights into
the role of ATM in the pathogenesis of type 2 diabetes.
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PHOTOPROTECTION IN HALOPHILIC ARCHAEA: CORRELATION OF CAROTENOIDS AND DNA
DAMAGE
Ashlee D. Allred (Dr. Bonnie Baxter), Department of Biology, Westminster College, Salt Lake City, Utah 84105
Halophilic Archaea are much more resistant to ultraviolet (UV) light damage than Escherichia coli and other species
of Bacteria. These extreme halophilic organisms use light-driven ion pumps to maintain an acceptable internal
environment despite living in water that has salt concentrations up to 30%. This need for light requires that the
organisms experience more UV exposure than most other known microorganisms. For this reason, they have
developed survival mechanisms, including efficient DNA repair processes. We propose that halophilic Archaea also
employ photoprotective mechanisms such as low adenine-thymine ratios to avoid thymine dimers and pigmentation to
protect from UV damage.
Several strains of Archaea, including unidentified isolates from the Cargill salterns near Great Salt Lake, Utah were
examined for sensitivity to UV light. UV survival experiments showed that the LD50 (lethal dose for 50% of the cells)
for the Cargill isolate A is 24 times that of E. coli. Other halophilic Archaea strains give similar results. Data will also
be presented from strains such as Cargill isolate B, which is deficient in pigment syntheses and supports the notion of
photoprotection by these carotenoid compounds. Methods of analysis included thymine dimer detection as well as
isolation and identification of carotenoid compounds using HPLC.
Analyzing differential gene expression in photoperiod mutants of Arabidopsis thaliana: the search
for a cryptic pathway
Authors: Emily A Vaughn, Karen A Hicks , Research Sponsor: Karen A Hicks, Institution and Institutional Address:
Department of Biology, Kenyon College, Gambier, Ohio 43022
A better understanding of what gene pathways cause plants to transition from a vegetative to a reproductive state is
of great scientific and agricultural interest. One of the pathways responsible for inducing the change is the
photoperiod pathway, which senses the seasonably variable number of hours of light that a plant receives per day.
Many plants, including rice and soybeans, rely upon information from the photoperiod pathway order to maximize
reproductive success. The floral regulation gene pathways of the photoperiod sensitive plant Arabidopsis thaliana are
particularly well characterized, as the plant is often used as a model system for plant genetics studies. Surprisingly,
certain A. thaliana double mutants whose photoperiod and autonomous pathways have been disrupted nonetheless
show sensitivity to photoperiod. This finding leads us to believe that the current model of floral induction is
incomplete. We propose that the double mutants display photoperiod sensitivity due to the action of an
uncharacterized cryptic day-length pathway composed of genes or gene functions not previously identified as
regulators of floral induction.
In order to identify novel genes involved in floral regulation and build support for our cryptic pathway hypothesis, we
will assay single and double mutant A. thaliana plants using whole-genome microarrays. The assays will reveal
differences in mRNA levels of autonomous pathway, photoperiod pathway, and double mutant plants. This
information will allow us to explore the possibility and composition of a cryptic day-length pathway, identify novel
genes involved in floral induction, and further characterize known floral induction pathways.
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THIOREDOXIN REDUCTASE IN ARTEMIA AND ITS ROLE IN SELENIUM DETOXIFICATION
Rylan Larsen (Dr. Brian Avery), Department of Biology, Westminster College, Salt Lake City, Utah 84105
The interaction between selenium and the life in the Great Salt Lake is of great current interest due to the potential
ecological impacts of increased selenium levels in the Great Salt Lake. Selenium, a vital trace element in mammalian
cells, has long been associated with toxicity in organisms exposed to high selenite concentrations. The multifaceted
thioredoxin reductase enzyme family has been implicated in the detoxification of selenium in a wide variety of
organisms. In addition to the detoxification of selenium, thioredoxin reductase plays important roles in the synthesis of
Deoxyribonucleotides, cell growth, apoptosis, and cell signaling systems. In addition, thioredoxin reductase may also
have connections to diseases such as Alzheimer’s Disease and cancer.
While there is some data on the Selenium levels in the lake water, sediments, birds, and brine shrimp (Artemia),
little is known about the effect of selenium on the brine shrimp themselves. Our goal is to identify and characterize
the Artemia thioredoxin reductase gene and study its regulation in the attempt to provide insights into the reaction of
brine shrimp to selenium in their environment. As thioredoxin reductase is so highly conserved among other
organisms, it is believed that it may play a role in selenium detoxification in Artemia. This expectation is based on
evidence from yeast that selenium treatment induces the expression of the yeast thioredoxin reductase (TRR-1) gene
and that the function of thioredoxin reductase(TrxR) is conserved in Drosophila. To identify TrxR in Artemia, we have
employed Reverse Transcriptase PCR with degenerate primers targeted to conserved sequences in order to isolate
the Artemia thioredoxin reductase. PCR results have demonstrated a potential TrxR fragment and currently the
sequence of this band is being investigated.
The role of extracellular matrix in angiotensin II stimulation of focal adhesion abundance in cardiac
fibroblasts.
Daisy E. Conduah (Dr. Thomas Peeler), Department of Biology, Susquehanna University, Selinsgrove, PA 17870
Angiotensin II (AII) is an eight amino acid peptide that regulates blood pressure and fluid volume in animals. AII also
has direct effects on the growth of cardiac cells, stimulating cardiomyocytes to increase in size, and stimulating
cardiac fibroblasts to divide. Previous work in our lab has shown that AII stimulation results in an increase in the
abundance of focal adhesions prior to cell division in cardiac fibroblasts grown in culture. To test the hypothesis that
this effect is regulated by the extracellular matrix (ECM) environment of the cells, we grew neonatal rat cardiac
fibroblasts on laminin, fibronectin, collagen I, and collagen IV, and treated the fibroblasts with 10-8 M AII. At various
times after stimulation, the cells were fixed, and fluorescence microscopy was used to visualize focal adhesion
abundance. Our results indicate that the number of focal adhesions per cell varies with the ECM present, but that AII
stimulation results in an increase in the number of focal adhesions in all cases. These results show that AII
stimulation results in an increase in focal adhesion abundance regardless of the ECM environment.
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The Effects Of Calcineurin Inhibition On Cocaine-Seeking Behavior In The Mouse Model
Neeti S. Sasi1,2, Adeola R. Davis2 and Danny G. Winder2 , 1Belmont University, Nashville TN 37212, , 2Department
of Molecular Physiology and Biophysics, , Vanderbilt University Medical Center, Nashville, TN 37232
Cocaine addiction involves profound changes in behavior and brain morphology. Addiction to cocaine is characterized
by a progressive elevation in cocaine self-administration and compulsive pre-occupation with the drug. Previous
research indicates that the dorsal striatum of the brain is a major target for cocaine and contributes to habit-learning
(Gerdeman et al. 2003). Calcineurin (PP2B), a calcium-dependent protein phosphatase, is also highly expressed in
the dorsal striatum, and its activity has been proposed to be involved in the actions of cocaine. Double transgenic
mice were generated to overexpress the autoinhibitory domain of calcineurin, thereby reducing calcineurin activity.
This study investigates the molecular mechanism of calcineurin inhibition in the dorsal striatum and its relation to
cocaine-seeking behavior, utilizing biochemical methods and the behavioral paradigm of Conditioned Place
Preference (CPP). The separate effects of both calcineurin and cocaine have been extensively studied in other brain
regions in mice. In this study, we used the cocaine CPP paradigm to assess behavioral phenotypes in the
calcineurin-inhibited transgenic mice. Our results indicate that the calcineurin-inhibited mice (E211/tTA) exhibit a
trend towards a decrease in CPP compared to wild type and E211 littermates, although additional control experiments
will be necessary. Previous research has shown that sex and estrous cycle influence cocaine-seeking behavior
(Kippin et al.). Therefore in addition to testing for behavioral differences between wild type and transgenic mice, we
also tested such differences between male and female transgenic mice with CPP. Collectively, our study indicates
that calcineurin may play an important role in drug reward circuitry and that sex differences may not influence
calcineurin-directed cocaine-seeking behavior.
Estrogen and Testosterone Protect Against Camptothecin-Induced Apoptotic Cell Death in the
Zebrafish (Danio rerio)
Andrew Holt (Dr. James E. Turner), Biology, Virginia Military Institute, VMI/Lexington, VA 24450
Estrogen and Testosterone are sex hormones that are synthesized in the brain in both males and females. These
hormones are believed to play not only a major role in protecting the brain, spinal cord, and retina against disease
and trauma whose end result is nervous tissue death by the process of apoptosis in not only humans, but also in the
fish speces, Danio rerio. This study demonstrated that estrogen used in low physiological levels (10-8 M) protected
the zebrafish from camptothecin-induced apoptotic cell death (125 uM). Estrogen co-treated zebrafish showed a
85-100% survival rate compared to the camptothecin exposed zebrafish who showed only a 30-50% survival rate
after a three day assay. Preliminary studies with testosterone and estrogen may lead to the development of effective
strategies for the treatment of neurodegenerative diseases.
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Is Estradiol Sufficient to Restore Sexual Behaviors in Female Meadow Voles After Six Hours of Food
Deprivation?
Izu Iwueke (Andrew Pierce and Dr. Michael Ferkin), Department of Biology, University of Memphis, Memphis,
Tennessee 38152
Food deprivation inhibits sex behaviors in female meadow voles. Estradiol is a mechanism that influences both
feeding behavior and sexual behavior in mammals. Our objective was to determine whether estradiol treatment was
necessary and sufficient to restore odor preferences and receptive behavior in ovariectomized (OVX) food-deprived
female meadow voles. We used 48 female voles; 20 voles were intact control animals and twenty-eight OVX females.
We treated 16 of the OVX females with estradiol (E2) capsules and 12 with empty capsules. Half of the females in
each treatment group were ad lib-fed and half were food deprived for 6 hours at the time of testing. Intact, ad lib-fed
females showed preference for odors of opposite-sex conspecific vs. same-sex conspecifics, and sexual receptivity at
a 60% rate. In contrast, when food-deprived, intact females showed no preference for same or opposite-sex odors;
these voles also showed no sexual receptivity. Ad lib-fed and food deprived OVX+E2 females had odor preferences
and receptivity rates similar to those of ad lib-fed intact females. Ad lib-fed or food deprived OVX females without
estradiol, showed lack of odor preferences and receptivity, similar to intact food-deprived female voles. Estradiol
treatment was necessary and sufficient to restore odor preferences and sexual receptivity in food-deprived female
meadow voles.
DIET ANALYSIS OF LITTLE BROWN BAT (MYOTIS LUCIFUGUS) FEMALES DURING THE
REPRODUCTIVE SEASON: A COMPARISON WITH INSECT AVAILABILITY
LaKeisha A. Hall (Dr. Karen A. Campbell), Department of Biology, Albright College, Reading, Pennsylvania 19612
Little Brown Bats (Myotis lucifugus) were studied for a ten week period in the summer of 2005 at two summer
maternity colonies less than ten miles apart near Reading, Pennsylvania. The diet of these insectivorous bats was
compared in order to determine if they are dietary specialists or generalists, and whether their diet preferences
change over the course of the reproductive season, as insect availability changes. This was accomplished by
capturing over 150 bats of known age, sex, and reproductive condition. The bats were captured using several
techniques, and were placed in individual containers until guano samples were excreted. Insects were also captured
using a variety of techniques in order to obtain the broadest sample possible each night before the bats were captured
for guano collection. Insect samples were collected for the purpose of determining what was available for the bats to
ingest the previous night, and were also used to aid in the identification of the insect remains in the guano samples.
Diet analysis, when compared to insects captured, indicates that the bats eat what is most readily available, in
high quantities, and what might be considered to be the most energetically favorable orders based upon body size,
such as Lepidoptera, Coleoptera, and Tricoptera. Diptera were found in a small proportion of the guano samples from
bats at only one of the sites despite similar availability of Diptera at both sites. This research sheds some light on the
foraging behavior of insectivorous bats as they meet the high energy demands associated with reproduction and
growth.
Lodge-site Fidelity of the North American Beaver (Castor canadensis) in Northern Minnesota,
2003-2005
Christopher L. Hewitt, Janna R. Willoughby (Dr. Timothy Lewis and Rosemarie Burley), Department of Biology,
Wittenberg University, Springfield, Ohio 45501
The North American beaver, Castor canadensis, plays an integral role within the transitional boreal forest area of
northeastern Minnesota. Beavers construct lodges within critically selected habitats in order to provide optimal shelter
and protection for a colony. We compared recorded lodge-sites within the Boundary Waters Canoe Area Wilderness
over a 2 year period to determine lodge-site fidelity. The continued presence of prime habitat conditions and food
sources allowed us to hypothesize that lodge-site positions would remain unchanged. We examined the periphery of
lakes, ponds, rivers, and streams for lodges via canoe and compared the position of lodges discovered to those that
had been recorded two years previous in 2003. A total of 36 lodges were found in our study compared to 34 in 2003.
Of the 36 lodges, only 19 had been present two years earlier, 15 lodge-sites had been terminated, and 17 new lodges
constructed. The loss of 44.1% of the lodges is most likely attributed to the active trapping of beaver within the
Boundary Waters Canoe Area Wilderness. The newly constructed lodges provided evidence that the vegetation and
habitat within the study area was still favorable in terms of beaver settlement, and the migratory patterns effectively
fulfilled the necessity of recolonization in order for a population to persist.
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DOES PREY AVAILABILITY DETERMINE WETLAND PREFERENCE OF BLANDING’S TURTLES?
Samantha Foster, Michelle L. Jonovic and Kristen A. Smith (Dr. Mary Linton), Department of Biological Sciences,
University of Wisconsin-Whitewater, Whitewater, WI 53190
A major challenge for conservation biologists is prioritizing which habitats to preserve for imperiled species. The
Blanding`s turtle (Emydoidea blandingii), a threatened turtle in the Midwestern USA, is a particular challenge because
it spends much more time in terrestrial habitats than do other semi-aquatic turtles. This is due to it’s migration
between wetlands and the female’s annual trek to suitable nesting areas. We studied Blanding’s turtle habitat usage
in Southern Wisconsin to identify those habitats essential for preservation of this species. At our study site,
Blanding`s tracked via radio telemetry consistently use a small subset of available wetlands. Characteristics of
wetlands that might account for this preference are hydroperiod, plant density, food availability, or proximity of good
nesting areas. We monitored Blanding`s movements from April to November, and compared food availability in used
vs. unused wetlands. Turtle food was sampled in the summer using minnow traps and stovepipe samplers. Food
from minnow traps differed between high and low use wetlands (MANOVA p = 0.018), with Odonate and amphibian
larvae, and giant water bugs accounting for most of the difference. Food from stovepipe samples also showed
differences with preferred ponds showing a greater abundance of prey in most categories. This suggests that
Blanding`s are showing preference for ponds that have more prey from typically consumed categories. Other
characteristics may still be of overwhelming importance - especially hydroperiod. It was found that the radio tagged
Blanding’s spent more time in wetlands that maintained a longer hydroperiod. The greatest numbers of turtles were
found in open wetlands. Plant type and density did not appear to be a significant factor between high and low use
wetlands. We will continue monitoring turtle movement and prey availability during 2006 Spring migrations, and
begin collecting hydroperiod data to assess patterns of seasonal change in hydrology characteristics. The data
gathered would be used to determine prey preference and to assess hydrologic determined turtle movement.
COMBINING FOCAL MALE AND TRANSECT DATA TO INVESTIGATE COLOR POLYMORPHISM
MAINTENANCE IN THE TROPICAL FISH TELMATHERINA ANTONIAE
Samuel F. Miller (Dr. Jeffrey S. McKinnon), UW-Whitewater McNair program, UW-W Undergraduate Research
Program, Department of Biological Sciences, University of Wisconsin-Whitewater, Whitewater Wisconsin,
53190-1790.
Male Color Polymorphisms (MCPs) are frequently hypothesized to play a key role in speciation, particularly in the
explosive divergence of several model fish systems. The study of MCPs are important both to understanding such
explosive speciation events and to link the processes maintaining variation within species with those causing
reproductive isolation. Our lab is studying the factors maintaining an MCP in the Indonesian freshwater fish
Telmatherina antoniae, which possesses distinct blue and yellow color morphs. We have previously reported on three
years of observational data that suggested trade-offs in some fitness components and more intense interactions
between males of the same morph. Here we add an additional year of data to these analyses and combine the
behavioral data sets with new transect data on morph abundance and pairing rates. The new combined data sets
allow us to evaluate more definitively the relationship between time of day and mating success for each morph, which
is of particular interest because relative T. antoniae morph abundance varies with time (McKinnon et al. unpublished).
The combined data also allow assessment of relationships between components of fitness and density. Finally, we are
also investigating a hypothesized relationship between morph-specific mating success and the presence or absence
of cloud cover, which affects lighting conditions.
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HELMINTH PARASITES IN THE INVASIVE GAMBUSIA AFFINIS: THE EFFECTS OF THE
DISTRIBUTION OF MOSQUITOFISH IN RESPONSE TO WEST NILE VIRUS
Jan Marie Cheng, Marielle Discipulo, Nicole Fronteras, Nenna Olumba, Caitlin Sullivan, Joyce Valencia (James
Cunningham, Sibdas Ghosh, Mietek Kolipinski, Anindo Choudhury), Department of Natural Sciences and
Mathematics, Dominican University of California,
The mosquitofish, Gambusia affinis, has been distributed throughout the San Francisco Bay area in an effort to
control mosquitoes responsible for spreading West Nile Virus. Although this introduction is thought to be beneficial in
reducing the risk of spreading the virus, the nearly unmonitored dispersal of the nonnative species raises concerns
about the translocation and establishment of exotic, potentially pathogenic parasites. A parasite survey was
conducted on mosquitofish to identify parasites that may have been introduced with this fish species. Samples of G.
affinis were collected from two counties around the San Francisco Bay Area, including Point Reyes National
Seashore, to determine if there are invasive parasites that are detrimental to freshwater fish species. Dissections of
75 G. affinis reveal a total of 13 internal symbiotes. Of the sample population, 17.33% of the species are infected.
The average length of infected fish is 34mm. Three species of parasites were found: Clinostomatidae gen. sp.
(metacercaria), Clinostomum sp. (metacercaria) and Octospiniferoides chandleri (Acanthocephala). Potentially
harmful parasites (clinostomatids) that are found may include those having life cycle involving certain bird species,
the specific type of which has yet to be determined.
THE EFFECTS OF SPIDER CUES ON COLORADO POTATO BEETLE (LEPTINOTARSA
DECEMLINEATA) FORAGING BEHAVIOR.
Jeffrey J. Garofalo, Steven R. Heverley, (Matthew H. Persons), Biology Department, Susquehanna University,
Selinsgrove, Pennsylvania. 17870
Colorado potato beetles, Leptinotarsa decemlineata, are chemically-protected crop pests on economically important
solanaceous plants. We measured predation on potato beetles by four species of spider: the crab spider,
Misumenoides formipes, the long-jawed orb weaver, Tetragnatha laboriosa, and the wolf spiders, Hogna helluo and
Pardosa milvina. We also measured leaf damage on eggplants, Solanum melongena, in the presence of two of the
four spider species (H. helluo & P. milvina) as well as leaf preference or avoidance for leaves previously walked on by
one of the four spider species for 24 h. We found no predation on beetles by any of the four species of spider during a
4 h test period (n = 24/spider species). We found significant reductions in herbivory in the presence of Hogna helluo
but not the smaller wolf spider, Pardosa milvina (n = 48/spider species). When given a simultaneous choice between
eggplant leaves with spider cues and those without, beetles showed a significant initial avoidance for leaves
previously occupied by H. helluo, but beetles showed no significant initial avoidance of spider-cued leaves of any of
the other three spider species (n = 48/spider species). In the absence of live spiders, beetles showed significantly
lower herbivory levels on leaves previously occupied by H. helluo, P. milvina, or T. laboriosa, compared to leaves
without spider cues, but beetles showed no reduction in herbivory on leaves previously occupied by M. formipes (n =
48/spider species). Our results suggest that even chemically-protected herbivores show reduced feeding in the
presence of some generalist predators and that chemical cues from predators (silk and excreta) may also reduce
herbivory even when these predators present no predation risk.
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HABITAT PREFERENCE AND MOVEMENT OF THE WOLF SPIDER PARDOSA MILVINA IN A STRIP
CROP SYSTEM
Amanda M. Sangillo, Sarah N. Kiemle (Matthew H. Persons), Biology Department, Susquehanna University,
Selinsgrove, Pennsylvania 17870
We measured habitat preference of the wolf spider Pardosa milvina across different developmental stages (juveniles,
subadult males, subadult females, adult females, adult females with egg sacs, adult females w/ spiderlings) within
different crops (tilled corn, no-till corn, no-till soybean, and a fallow field). We used three methods to measure
preference: field censuses, laboratory habitat choice tests, and direct observations of spider movement within
different tillage and crop types. Within a 10.5 ha strip crop system, we censused P. milvina within 0.5m2 rings from
May 30-July 26, 2002 (N = 654). We also sampled at 0, 5, 10, 25, 50, and 100 meters from a hedgerow and
quantified litter depth, plant density and meteorological variables. Spiders of all developmental stages preferentially
avoided hedgerows. We found significant differences in distance from hedgerow, crop type, and tillage practice in
spider density. Spiders at different developmental stages also exhibited significant differences in habitat preference.
We also measured time spent in one of two habitat types for all developmental stages and both sexes under
laboratory conditions. All developmental stages and both sexes preferentially avoided bare soil, but the strength of
preference was significantly different by age and sex (N = 438). Nine 2 m2 plots were constructed containing three
different substrate types; bare soil, grass, and no-till soil with corn stubble. Flags were placed every 20 cm forming a
grid within each plot. Wild male, female, adult and subadult, spiders were caught, marked, and placed within the
center of each plot (N = 180). We then measured time to leave the plot, speed, distance traveled, and space use.
We found a significant difference is preference by substrate type, as well as a difference in movement depending on
sex and age of the spider. Results indicate that P. milvina habitat preference varies by sex, age and reproductive
status of the spider and that tillage practices greatly effects spider density and movement.
ECOLOGY, BODY SIZE, AND SPECIATION: BEHAVIORAL MECHANISMS IN MALE STICKLEBACKS
Nicole Frey, Nick Hamele, and Jess Greene (Dr. Jeffrey McKinnon), Department of Biological Sciences, University of
Wisconsin-Whitewater, Whitewater, Wisconsin 53190
The purpose of this study was to determine if male courtship preferences might contribute to ecological speciation in
threespine sticklebacks (Gasterosteous aculeatus). Previously, our laboratory has shown that body size plays a
central role in the evolution of reproductive isolation between large, ancestral anadromous threespine sticklebacks
and smaller, derived stream-resident sticklebacks. Here we investigate behavioral aspects of reproductive isolation in
detail. Using males of both ecotypes presented with females of both ecotypes manipulated to large or small sizes, we
test (1) the hypothesis that male courtship behavior will reflect a preference for females of similar size; (2) the
alternative hypothesis that ecotype similarity determines male preferences. From videotapes of experiments
conducted in our laboratory, we collected data on several male behaviors, with special attention given to bites per
minute and nest works per minute. Data for anadromous and stream resident males suggest that although they
perform similar behaviors, the pattern of performance is very different. There is a significantly positive correlation
between nest works/min and bites/min for anadromous males, which suggests an increase in aggression when an
anadromous male is ready to mate. On the other hand, stream males tend to bite fewer times per minute when their
nest works per minute increase, suggesting a less aggressive approach to courtship. Both types of male show higher
rates of nest work with females of their own size: large for anadromous males, small for stream males. Both male
types bit large females more, but we suspect for different reasons: anadromous males to court them, stream males to
reject them. We found little evidence that males altered their behavior based on female ecotype. Thus the size
hypothesis is best supported and male courtship preferences may contribute to ecological speciation.
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CORRELATIONS BETWEEN TESTOSTERONE AND MALE COURTSHIP BEHAVIOR AND
MORPHOLOGY IN MALLARDS (Anas platyrhynchos)
Brian Darkow, Rachel Rugen, Michelle Toman (Dr. Ellen Davis), Department of Biological Sciences, University of
Wisconsin-Whitewater, Whitewater, WI 53190
Pairing of males and females in many waterfowl species occurs in the fall with breeding occurring the following spring.
After pairing in fall, male mallards mate-guard throughout the winter and spring, which allows females to forage more
efficiently and increases their reproductive output. Recent evidence suggests that females choose males in the fall
that will be good mate-guarders, but, it is still unknown what cues a female uses to choose a male in the fall that will
predict his ability to mate-guard in the spring. One logical link could be testosterone, which has been shown recently
to positively correlate with male mate-guarding success. It is possible that testosterone may also influence male
characteristics that seem to be important in female choice during the fall, such as courtship, dominance rank,
plumage or male molt order. We collected blood samples, in August, from 21 males before they molted (baseline
blood sample), and then separated the males and females. We then recorded the order in which males molted and
the quality of their plumage. Blood samples were collected again in early November before placing the males and
females back together. We are currently collecting data on male courtship displays as well as dominance rank data
and other morphological characteristics. Starting in January, we will begin collecting male mate-guarding data. We
will look for correlations between testosterone and these behavioral and morphological characteristics during the fall
as well as with male mate guarding behavior in the spring to determine whether these fall characteristics predict mate
behavior in the spring and whether testosterone may thus serve as a physiological link between fall and spring
characteristics.
EFFECTS OF DOLPHIN ACTIVITY ON SEATROUT SPAWNING BEHAVIOR AT AGGREGATION SITES
Dena R. Hodges (Dr. Gorka Sancho), Department of Biology, College of Charleston, Charleston, SC 29424
Atlantic spotted seatrout, Cynoscion nebulosus, spawn within estuaries from late April to early September in South
Carolina. During this season seatrout form dense aggregations everyday, from the late afternoon and into nighttime,
when males produce loud courtship sounds. The formation of sonic fish aggregations is predicted to attract the
attention of bottlenose dolphins, known predators of sciaenid fishes. I studied the possible influence of dolphin
predation on spotted seatrout spawning aggregations in Charleston Harbor, SC. Throughout the summer of 2005,
during peak seatrout spawning, dolphin activity was monitored through hour-long observations carried out twice a day,
once in the morning and once in the evening. Dolphin activity was recorded from land close to a known seatrout
spawning site, while seatrout spawning activity was continuously monitored by an acoustic recording device deployed
near the fish aggregation site, recording male seatrout callings. Dolphin presence and activity levels were identical
during non-spawning morning and spawning evening surveys. Tides seemed to have no effect on either dolphin
presence or seatrout spawning activities. Daily male seatrout sound production was extremely regular in time,
seemingly unaffected by dolphin presence or tidal cycles. Bottlenose dolphins seem to not selectively feed on spotted
seatrout while forming spawning aggregations.
SINGING BEHAVIOR OF THE NEW ZEALAND GREY WARBLER (GERYGONE IGATA)
Bianca S. Gabriel, Shabrina Azim, and Mercedes Pacheco (Dr. James B. Cunningham), Department of Natural
Sciences and Mathematics, Dominican University of California, San Rafael, CA 94901
This study was undertaken to investigate the singing behavior of male Grey Warblers. The Grey Warbler is a small
native New Zealand song bird that occupies both native as well as non-native forests. Only males of the species sing.
The songs used in this study were recorded in Kowhai Bush, a small patch of native forest near the town of Kaikoura
on the east coast of the South Island, New Zealand. Each male recorded was color marked so that individual males
could be recognized and re-recorded on different days. The recordings, on reel-to-reel tapes, were digitized and
analyzed using the program Raven. Resulting sound spectrographs were printed out and studied. Each male has a
long (up to 3 minutes) warbling song, the pitch of which is between 4 and 5 kHz. A bout of singing consisted of a
series of simple introductory notes followed by two complex phrases (phase A and phrase B) which were alternated
several times until the male stopped singing. Each male sang a highly stereotyped song. The songs of neighboring
males were very similar in structure but were individually distinctive. We compared the singing behavior of the Grey
Warbler with other passerines and with other members of the genus Gerygone.
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A COMPARISON OF SEA CUCUMBER POPULATION DYNAMICS IN PROTECTED AND
UNPROTECTED HABITATS NEAR CHUMBE ISLAND, ZANZIBAR, TANZANIA
Jennifer M. Blaine, Christopher A. Muhando, James M. Welch; Institute of Marine Sciences, University of
Dar-es-Salaam, P.O. Box 568, Stone Town, Zanzibar, Tanzania; Department of Biology, Wittenberg University, P.O.
Box 720, Springfield, OH 45504
Chumbe Island is located 10 km southwest of Stone Town, Zanzibar, off the coast of Tanzania. The west side is a
protected reserve; the east side is subject to commercial fishing pressure. In order to examine the population
dynamics of sea cucumbers in the protected and unprotected waters of Chumbe Island, an 18-day study was
conducted between 14 November and 1 December 2004. A total of 138 belt transects, each with an area of 80m2,
were conducted in four habitats of each side of the island: intertidal zones, reef flats, seagrass beds, and reefs. In
each habitat, distribution, size, density and diversity of the sea cucumbers were determined. Of 858 total individuals,
87% were found in the protected habitats. Twenty-eight species of sea cucumbers were identified in the protected
area, while only 8 of these were present in the unprotected area. Although H. cinerascens was the most abundant
species (>200 individuals, but most in one transect), H. atra and H. leucospilota were the most widely distributed. In
seagrass beds, the average length of H. atra was significantly larger in the unprotected area. With the exception of
the intertidal areas, the populations in the protected habitats were significantly denser than in the unprotected (t-test,
p<0.001). Also with the exception of the intertidal areas, the protected habitats were more diverse than the
unprotected. The most diverse habitat was the protected reef (H`=2.16) and the least was the unprotected seagrass
beds (H`=0.95). The differing population dynamics between the protected and unprotected habitats indicates that sea
cucumbers are most likely being overharvested on the unprotected side of Chumbe.
COMPARISON OF FEMALE REPRODUCTIVE CYCLES BETWEEN URBAN, SUBURBAN, AND RURAL
BLACKNOSE DACE (RHINICHTHYS ATRATULUS) POPULATIONS
Aaron H. Basler (Ms. Amanda Richardson and Dr. Joel Snodgrass), Department of Biological Sciences, Towson
University, 8000 York Road, Towson, Maryland 21252
As urbanization of watersheds proceeds at a rapid pace, the resultant increase in impervious surface cover causes
major changes in stream hydrology, channel morphology, and ultimately, overall habitat quality. It remains unclear
what ecological factors allow a few tolerant species to persist in degraded habitats and whether they become adapted
to conditions in urban streams. Blacknose dace (Rhinichthys atratulus) are stream minnows that are able to persist in
highly degraded urban streams. To investigate the potential influence of watershed urbanization on blacknose dace
reproductive cycles, we examined the ovaries of 437 female dace from eight populations occurring across an
urban-rural gradient. The reproductive condition of each set of ovaries was assessed based on stage of maturity
according to an established classification scheme. Overall, ripe ovaries were found from April through June in all
populations and the number of females with ripe ovaries declined throughout this reproductive season. Following the
reproductive season, from July through February, the frequency of ovaries containing eggs in later stages of
maturation increased. When urban, suburban, and rural dace collected during winter months (November-February)
were compared, a portion of small dace from suburban, and to a greater extent, urban streams contained ovaries with
eggs in early stages of maturation. The recruitment of small size dace into suburban and urban breeding populations
during winter months suggests these individuals are maturing within their first year of life. Taken together, our results
suggest that in urban streams, female dace are allocating more energy to reproduction in their first year of life.
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EVIDENCE FOR A TREMATODE PARASITE CAUSING SEVERE LIMB MALFORMATIONS IN
AMPHIBIANS
Donald J. Larson (Dr. Daniel Sutherland), Department of Biology and Rivers Studies Center, University of
Wisconsin-La Crosse, La Crosse, WI
Ribeiroia ondatrae is a parasitic trematode that has been shown to cause malformations in amphibians. R. ondatrae’s
life cycle includes three hosts: a predatory bird or mammal definitive host, a planorbid snail first intermediate host,
and an amphibian second intermediate host. Eggs are shed in the predator’s feces, and R. ondatrae emerge as
miracidia. The miracidium penetrates a snail host and develops into sporocysts. Asexual reproduction within the
snail produces rediae and cercariae. Cercariae then burrow out of the snail and seek out amphibians. . Some
cercariae access the gill cavity. Most cercariae enter the anus and penetrate the cloacal mucosa. Metacercariae
encyst near the tadpole’s limb buds and may interfere with limb development and cause malformations. Increasing
the shedding of cercariae from planorbid snails should increase the mean intensity (average infection per infected
amphibian) leading to more malformations. Duck Pond in Eden Prairie, MN was found to be a malformation hotspot
in July 2003 with a malformation frequency of 49% in Rana pipiens. Tadpoles and metamorphs of Rana pipiens, Hyla
chrysoscelis, Bufo americanus, and planorbid snails were collected every two weeks from Duck Pond in 2003
(July-October) and 2004 (April-October) and monthly in 2005 (April-October). Analysis indicated mean intensity of R.
ondatrae infections in amphibians had no correlation with prevalence in snails but did positively correlate with snail
shedding. Mean intensity also decreased over the three year period in R. pipiens and B. americanus, but not
significantly in H. chrysoscelis. Malformation rates also declined dramatically over the last two summers. Only two
minor malformations were identified in R. pipiens during all of 2005. As parasite shedding in snails remained constant
or increased, mean intensity in amphibians increased. Declines in shedding snails caused declines in mean intensity
for amphibians.
BIODIVERSITY AND ECOLOGY OF BEES (HYMENOPTERA) IN CENTRAL CALIFORNIA NATIONAL
PARKS
Paula Mancilla, Jessica M. Van Den Berg, and Gregory Cameron (Mietek Kolipinski and Sibdas Ghosh), Department
of Natural Sciences and Mathematics, Dominican University of California, 50 Acacia Ave, San Rafael, CA 94901; ,
National Park Service, Senior Scien
Of over 100,000 species known in the Order, Hymenoptera (bees, wasps, ants, etc) 4,000 occur in the United States.
We intend to understand and explain what role bees play in naturally functioning ecosystems, as those found in
national parks, such as Pinnacles National Monument (PINN) and Sequoia and Kings Canyon National Parks (SEKI).
Also, we are examining relationships between native and non-native bees and how they relate to occurrence and
distribution of native and non-native plant species. Our analysis of bee surveys found that only forty species of bees
were documented in SEKI, while PINN, a much smaller park, has an amazingly high bee diversity of nearly 400
reported species. Interestingly, these two parks have twelve species in common, six of which belong to the genus,
Lassioglossum. Several National Park Service (NPS) Units in California have developed lists of Hymenoptera that
occur within their boundaries. An analysis of these databases is in progress which will provide for further insight into
regional distribution patterns. Our study aims to assist NPS managers in determining how to track long-term
population trends, so they can identify potential problems quickly.
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Captive Florida Manatee Behavioral Response to Public Viewing
Michelle L. Latham, Katie M. Rivera, Michael S. Wehmann, Tony Zak, Sean Kirby, Brid McDonnell,Adam McHenry,
Amanda Hemmer (Charles J.Grossman, Ph.D.) , Department of Biology, Xavier University, Cincinnati, Ohio 45207.
Behavioral responses of captive Florida manatees (Trichechus manatus latirostris) at the Cincinnati Zoo were studied
during different times of the day to determine if certain behaviors, specifically swimming activity, of the manatees
were being affected when visitors were present for public viewing. The two manatees (Slip and Little Joe) were males
of approximately the same age and were recent arrivals to the Manatee Springs exhibit, which houses only two
manatees at one time. Because the construction of the exhibit allows visitors to approach the windows of the tank,
the manatees can clearly see and hear people through the glass. Activity data was collected for each manatee when
there were people present in the exhibit hall, and this was compared to activity data collected for each manatee when
there were no people present in the exhibit hall. When people were present, there was an increase in Slip’s activity
up to &#8776;58% and an increase in Little Joe’s activity up to &#8776;50% as compared to the data when there were
no people present. Our findings strongly suggest a relationship between the behavioral responses of captive
manatees, and the presence of people in the exhibit hall. Further investigation may help determine if the increase in
manatee activity is stress-related, which would lead to a better understanding of how to maintain a healthy
environment for manatees in captivity
PREDATION RISK OF MALES AND FEMALES OF THE WOLF SPIDER PARDOSA MILVINA DURING
COURTSHIP AND COPULATION.
Bryant T. Upton (Matthew H. Persons), Biology Department, Susquehanna University, Selinsgrove, Pennsylvania
17870
Courtship and mating may impose substantial predation risks to wolf spiders due to reduced vigilance, conspicuous
courtship displays, and lack of mobility during copulation; however, the level of risk may differ between males and
females. The wolf spider, Pardosa milvina, is known to modify their courtship and mating behavior in the presence of
silk cues from a larger co-occurring predatory wolf spider, Hogna helluo. We compared predation risk by H. helluo of
male and female P. milvina during courtship interactions and copulation as well as differences in male and female
behavior with and without silk cues from Hogna helluo. We measured predation and mating success, predation
latency, courtship latency, copulation latency, courtship intensity, and copulation duration for both P. milvina sexes.
We compared four treatments: 1) introduction of H. helluo during male courtship toward a female P. milvina, 2)
introduction of the predator, H. helluo to copulating pairs of P. milvina, 3) introduction of H. helluo to courting P.
milvina with access to silk cues from H. helluo, 4) introduction of H. helluo to copulating pairs of P. milvina with
access to silk cues from H. helluo. We predicted that male predation risk would be higher both during courtship and
copulation. Male courtship displays could attract predators such as H. helluo and males are positioned dorsally
relative to the female during copulation and are therefore more prone to predatory attacks. We also predicted that
both male and female predation would be reduced when silk cues were present from H. helluo. Preliminary results
suggest that copulation rates are reduced under predation risk but that males are not prone to more predation than
females during courtship or mating interactions.
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THE EFFECTS OF HABITAT TYPE ON THE DISTRIBUTION OF FROGS IN VERNAL PONDS OF
HARTLEY NATURE AREA, DULUTH MINNESOTA
Nicole R Barg (Lyle Shannon), Biology Department, University of Minnesota Duluth, 257 JISSB Duluth, MN 55812
Vernal Pools are temporary bodies of water that fill each spring as a result of spring rains and snow melt. These small
ponds provide critical breeding habitat for many amphibian species. This study investigated 4 ponds in Hartley Nature
Area (Duluth, Minnesota) to determine which frog species used them for spring breeding. These ponds were selected
to encompass a range of habitat types, differing in area, depth, vegetative cover, and duration of their wet period.
Breeding frogs were censused using frogloggers constructed from Sony Minidisk recorders fitted with stereo
microphones. These devices were used to record frog calls over a 24-hour period once a week at each site. Water
and air temperatures and light intensity were continuously recorded on all sites using HOBO data loggers. Data was
analyzed by listening to the first five minutes of every half hour to determine the species present and the intensity of
their calling (scored on a scale from 0 to 3).
Four different species of frogs were recorded during the mid-April through May sampling period. Wood Frogs (Rana
sylvatica) were the most widely distributed and were found at all four ponds. These animals were more active in
mid-April and the beginning of May. The other species were more narrowly distributed. Spring Peepers (Pseudacris
crucifer) were only found at two sites. Chorus Frogs (Pseudacris ornate) were found at one site, and were very active
from the end of April to mid-May. Gray Tree Frogs (Hyla versicolor) were found at a single site in late May. These
differences in distribution were primarily related to differences in pond vegetation and depth.
For most species, calling intensity was related to light intensity, and typically increased in the evening and dropped off
when the sun came up. Chorus frogs, however; did not show a nocturnal peak and typically called throughout the day.
DISTRIBUTION AND ABUNDANCE OF INVERTEBRATES IN VERNAL PONDS OF HARTLEY NATURE
AREA, DULUTH MINNESOTA
Brittany R Rowland (Lyle Shannon) , Biology Department, University of Minnesota Duluth, 257 JISSB Duluth MN
55812
Vernal pools are shallow, intermittently flooded wetlands that experience seasonal drying. This unique hydrology
limits their biotic community to species adapted to withstand stresses associated with periodic desiccation. This study
examined the role of physical and chemical characteristics of such wetlands in determining the distribution and
abundance of macro and microinvertebrate communities. Four study ponds were located in the Hartley Nature Area
in Duluth, Minnesota, all within a mile radius of one another. Three of these were vernal pools which flood in the
spring and dry annually in late summer or fall. The fourth pond was added for comparative purposes and had
continuous standing water and supported populations of brook sticklebacks (Culea inconstans) and mud minnows
(Umbra limi). The ponds differed in area, depth, vegetative cover, conductivity, and duration of the wet period.
This study was conducted between April 17 and May 23, 2005. Physical and biological measurements were made
weekly at each site. Water and air temperature and light intensity were continuously recorded using Hobo© data
loggers. Invertebrate communities were sampled using funnel traps evenly spaced in a line traversing each pond
which were left for 24 hours before collection. Specimens were identified in the lab to the lowest practical taxonomic
level.
Each pond had a similar biological community, although these were differences in the relative abundance of each
taxon. A Kruskal-Wallis One Way Analysis of Variance showed that densities of mosquito larvae, chironomid
larvae, a snail (Gyraulus sp.), female calanoid copepods, and ostracods differed significantly among ponds.
Macroinvertibrate density was positively correlated with emergent woody vegetation and negatively correlated with
the presence of fish. Microinvertebrate density was highest in the pond containing fish. Diversity among ponds was
similar, ranging from 18-20 distinct taxa.
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BIOLOGY: ZOOLOGY
SILK DRAGLINE DEPOSITION AND CONSPECIFIC COMMUNICATION IN THE WOLF SPIDER HOGNA
HELLUO.
Christopher A. Latanich (Matthew H. Persons), Biology Department, Susquehanna University, Selinsgrove,
Pennsylvania 17870
Spiders produce silk threads (draglines) as they move through the environment. Adult female draglines are known to
convey information to adult male spiders about the maturity and mating status of females, but whether or not females
vary the quantity or quality of the draglines in the presence of males remains unknown. Furthermore the dragline
function among males remains unknown. We tested the function of dragline silk in intersexual communication among
adult males and adult females of the wolf spider, Hogna helluo. We placed adult male and female H. helluo on
grid-bearing 80 mm dia. paper disks for four hours within transparent plastic containers. Spiders were then allowed to
observe a conspecific adult female, adult male, juvenile, or no spider through the transparent container. We then
quantified the amount, type, and pattern of silk and excreta deposition for each spider on the grids. Preliminary results
suggest that male silk deposition patterns remained unchanged in the presence of conspecific spiders, however there
was a non-significant trend toward increased deposition of attachment disk silk in the presence of mature females and
mature males compared to juveniles and the blank control. Fine gauge silk deposition occurs almost exclusively
among mature females suggesting a sex-specific function of this silk type.
The contractile properties of the circular mantle muscles of cuttlefishes (Cephalopod molluscs)
change during growth.
Joshua H. Brody and Lisa M. Crescenti (Dr. Joseph Thompson), Department of Biology, Saint Joseph`s University,
Philadelphia, PA 19131
Squids and cuttlefishes are versatile swimmers, having the ability to hover in one spot, change direction or orientation
rapidly with apparent ease, and ascend/descend almost vertically. Although they have fins and arms that are used to
varying degrees for propulsion, stability, and maneuverability, jetting is the foundation of the locomotive system. Jets
in squids and cuttlefishes, which differ from the more familiar undulatory locomotion of fishes, are generated by
alternately filling an internal mantle cavity with water and ejecting that water by powerful mantle contractions through
a maneuverable funnel.
Thick filament length in the mantle muscles that provide power for jet locomotion (i.e., the circular muscles) increases
significantly during the growth of some species of squids and cuttlefishes. This increase in thick filament length may
affect the mechanical properties of the circular muscles. The contractile properties of striated muscles, such as the
obliquely striated circular muscles of the mantle, depend on the lengths of the thick filaments and sarcomeres, the
load on the muscle, and the rate of cross-bridge cycling (e.g., Josephson, 1975). Thick filament length is inversely
proportional to shortening velocity and is directly proportional to peak isometric tension (e.g., Josephson, 1975).
Assuming all else is equal, we predict that the circular muscles of newly hatched animals will produce lower peak
isometric tension than the circular muscles of juveniles and adults. We performed a set of preliminary muscle
mechanics experiments at 20ºC on a species of cuttlefish (Sepia officinalis). The tests performed consisted of both
length-tension and slack step methods (e.g. Edman, 1979). We found peak isometric tension (brief tetani, 50Hz,
100ms), as well as an average contracting velocity of 4.87 L/s (N= 7, range: 1.11 L/s – 12.56 L/s). The preliminary
results are consistent with the prediction that peak isometric tension should increase during ontogeny in the species
we studied.
140
BIOLOGY: ZOOLOGY
White muscle creatine kinase activity and sprint swimming performance in blacknose dace
(Rhinichthys atratulus)
Angela Amato (Dr. Jay A. Nelson), Department of Biological Sciences, Towson University, 8000 York Road, Towson,
MD 21252
Locomotor performance of feral animals is of considerable interest from management, physiological, environmental,
ecological and evolutionary perspectives. Flow characteristics of streams in the Baltimore area have been measured
and are significantly different between streams in urban and rural areas. These differences in flow of the organism’s
natural environment have been shown to cause variation of locomotor performances in dace. Both endurance and
sprint swimming performances have been shown to vary along an urban/rural gradient.
The differences in sprinting ability between fish from different streams could be due to a variety of factors. One
possibility is that the animals` ability to supply energy to contracting muscle is different. Examination of the
differences in metabolic enzymes in the muscle of these individuals could help determine the underlying reason for
differences in sprint swimming performance. Creatine kinase is an enzyme responsible for the supply of immediate
energy to the contractile apparatus after depletion of ATP occurs. Creatine kinase catalyzes a reaction where in
creatine phosphate donates inorganic phosphate to ADP, which produces ATP, the immediate energy source for
contraction. Thus, this enzyme is a good candidate for contributing to differences in sprint swimming performance.
The purpose of this project is to evaluate whether the activity of this metabolic enzyme is related to sprint swimming
performance differences in blacknose dace. The activity of creatine kinase is coupled to the hexokinase reaction,
ultimately causing an increase in NADPH. The concentration of NADPH is measured using spectrophotometry in
order to determine activity level differences within fish of known sprinting ability.
Thus far, creatine kinase activity seems to be linked to sprinting ability; a higher enzyme activity is present in fish
from urban streams when compared with fish from rural streams. This increased ability of urban fish to rapidly
regenerate ATP in maximally contracting muscle could explain the observed differences in sprinting ability.
THE EFFECTS OF CYCLIC AMP AS AN INTRACELLULAR SIGNAL MOLECULE INVOLVED IN GnRH
RELEASE
Melissa G. Williams (Dr. Michael Woller and Dr. John Wu), Department of Biological Sciences, The University of
Wisconsin-Whitewater, Whitewater, Wisconsin 53190
Cyclic adenosine monophosphate (cAMP) is an intracellular signal molecule involved in GnRH release. GnRH is a
decapeptide that is released from neuroterminals in the hypothalamus in discrete pulses to the median eminence.
After GnRH is released it is degraded by a neuropeptidase, EP24.15, which breaks GnRH between fifth and sixth
amino acid, leaving GnRH 1-5 and GnRH 6-10 metabolites. GnRH 1-5 metabolite has been shown to be a positive
feedback mechanism to the hypothalamus. Treatment with GnRH 1-5 metabolite increases GnRH and cAMP
production. We are investigating the communication between neurons that may be conducted by cAMP. Cyclic AMP
is a second messenger hormone that may be used to signal the release of GnRH to other neurons. GnRH binding to
its receptor causes the receptor to bind to a complex of proteins (the G proteins). Through many steps ATP may bind
to a calcium channel which may add in the conversion of ATP to cAMP. Increased cAMP can lead to an increase in
GnRH release. We plan on testing the affects of GnRH 1-5 on the production of cAMP in the cell and the release of
GnRH decapeptide by running an Enzyme-linked immunosorbent assay (ELISA) to measure cAMP levels and a
Radioimmunoassay (RIA) to measure GnRH levels concurrently. Tissue will be sent to a collaborating lab to run a
superarray analysis. They will extract mRNA to make a cDNA library. This will allow us to look at every gene that is
being up-regulated and down-regulated in our samples to further explain what is happening in the GnRH neurons in
response to GnRH 1-5 stimulation.
141
BIOLOGY: ZOOLOGY
EFFECT OF HUMAN DEVELOPMENT ON SOIL MACROINVERTEBRATE RICHNESS IN A SMALL
SWAMP
Marian Howse (Dr. Jill Wicknick), Department of Biology, Chemistry, and Mathematics, University of Montevallo,
Montevallo, AL 35115
Macroinvertebrates found in the soil contribute to the soil ecosystem by cycling nutrients through the ground and
creating and changing soil structure. Soil macroinvertebrates are also good indicators of soil and water quality
because of their varying sensitivities to pollutants. I examined soil invertebrate richness at sites nearer to and further
from human development. I hypothesized that pollution sensitive invertebrates would be more abundant in sites
further away from development and that pollution tolerant invertebrates would be more abundant in sites closer to
development. Soil samples were collected from Ebenezer Swamp, Montevallo, Alabama, from June 22-October 5,
2005. The samples were collected from two sets of sites, each set including sites close to human development and
sites further away from development. In the lab, samples were placed in Baermann funnels, and macroinvertebrates
identified, usually to family. Community similarity analyses indicated that the sites nearer to human development
were 56-64% similar to the sites further away from development. There were, however, some differences in the
distribution of pollution-indicator species. Midges (Order Diptera, Family Chironomidae), which are pollution tolerant,
were more abundant in sites closer to human development while black fly larvae (Order Diptera, Family Simuliidae),
which are pollution sensitive, were more abundant in the sites less affected by development. Most of the pollution
sensitive animals were
found in sites further from human development and all of the pollution tolerant animals were found in sites closer to
human development. These data provide support for my hypothesis. As the area surrounding this 65-acre swamp is
currently experiencing rapid development, further impacts on the soil invertebrate community are expected.
COMPARISON OF VARIANCE PARTITIONING TECHNIQUES TO ESTABLISH CLUTCH SIZE
VARIANCE ATTRIBUTABLE TO SPECIFIC ENVIRONMENTAL FACTORS IN TUNDRA SWANS
Kezia R. Manlove (Saint Olaf College Center for Interdisciplinary Research, NSF Program to Enhance the
Mathematical Science Workshop NSF0354308), Department of Mathematics, Statistics, and Computer Science, Saint
Olaf College, 1500 Saint Olaf Avenue, Nort
In this study, we compare three different methods for quantifying variance in a phenotypic trait associated with tundra
swan productivity. Using data obtained through an eight-year mark-recapture study of tundra swans (Cygnus
columbianus), we obtained several different estimates of repeatability on clutch size (the number of eggs associated
with a nest). Repeatability measures the proportion of total variance attributable to genetic and general
environmental factors; it lets wildlife managers formulate predictions about future population growth, and helps them
assess the impact of specific events on population productivity. The first repeatability estimate we generated relied
on the accepted formula for repeatability. However, while repeatability estimates have traditionally been used on
normally distributed response variables, such as length or weight, and operate under the assumption that response
variables are normally distributed, clutch size is not normally distributed. In tundra swans, it follows a Poisson
distribution, so the repeatability estimate should not be based on the normal distribution; it should utilize the gamma
distribution. Therefore, the second estimate we generated used a gamma distribution and treated the clutch size as a
manifestation of several explanatory variables, each with its own variance, but a universal lambda value. This model
did not account for correlation between clutches in the same location or clutches produced by the same hen.
Therefore, the third estimate we generated employed regression models with random effects for hens and locations.
The random effects models correct for consistently large or small clutches being produced in the same location or by
the same hen. We complete the study by comparing the three estimates and assessing the theoretical and practical
advantages of each method.
142
BIOLOGY: ZOOLOGY
THE EFFECTS OF DIETARY RESTRICTION ON ORGANISMS THAT LIVE LONG
Jung-Won Soh (Robert Arking), Wayne State University Honors Department and the Department of Biological
Sciences, Wayne State University, Detroit, MI 48202
Dietary restriction (DR) is a regime used to increase healthy life span in treated organisms by reducing the food
consumption compared to an organism that is allowed to eat freely. The food-stress induced gains in life span have
been observed in many organisms and demonstrate the conserved nature of DR throughout evolution. Organisms
that live long can be artificially selected from normal aged organisms. The effects of dietary restriction on organisms
artificially selected to live long are unknown. DR has been demonstrated to be inducible in normal-lived organisms. It
is possible that long-lived organisms use a constitutive form of the DR mechanism to enhance their life span. To
critically test this idea, a traditional DR experiment was performed using various strains of the fruit fly, Drosophila
melanogaster. Five strains were tested, including w1118 Oregon R, our lab’s robust but normal-lived (Ra), and our
lab’s long-lived (La) strains of flies. A DR-induced increase in median lifespan was observed in the two control strains
(w1118 and the Ra flies), most notably in the males. The La male flies showed no significant additive effect of DR in
their already increased longevity. DR enhances longevity of normal-lived fruit flies. However, the non-additive effect
of DR on the La longevity leads us to conclude that the La flies constitutively express the DR effect. Our observation
offers us a vehicle for future analysis of the cell signaling pathways responsible for the expression of the DR effect.
BODY SIZE CHARACTERISTICS OF SPOTTED TURTLES, CLEMMYS GUTTATA FROM 1889-2005 IN
OHIO
Rebecca M. Stewart, Timothy L. Lewis, Department of Biology, Wittenberg University, Springfield, OH 45501
Spotted turtle populations have greatly declined because of loss of wetlands. Changes in body size in a population
are reflected in habitat quality. Ohio is on the southwest edge of spotted turtle range in North America. This study
measured the physical characteristics of the spotted turtle, Clemmys guttata, in Ohio from 1889-2005. Data collected
on turtles from before 1980 was obtained from preserved specimens at the Ohio Biological Survey, while recent turtle
data were collected by us periodically from various counties in Ohio. We found no significant change in plastron or
carapace length or weight over the past hundred years between sex or latitude. Mean female weight (120 g, n = 130)
was slightly greater than males (112 g, n = 74). However the average carapace length for spotted turtles was greater
in males (9.1 cm, n=74) than females (8.8 cm, n = 130). Also no steady increase in carapace size was found over the
twelve degree range from South Carolina to Ontario. Overall the physical characteristics of spotted turtles in Ohio do
not seem different from spotted turtles elsewhere in its range. Even though Ohio’s spotted turtle population has
greatly declined due to the destruction of their habitat, it seems as though there is little change in the physical
characteristics of the spotted turtles that are left.
143
BIOLOGY: ZOOLOGY
TERRESTRIAL DISPERSAL OF JUVENILE ENDANGERED SALAMANDERS: THE LAST
CORNERSTONE
Emilio Gabbai-Saldate (Dr. H. Bradley Shaffer and Dr. Peter C. Trenham), Department of Evolution and Ecology,
University of California at Davis, Davis, California 95616
Amphibian species extinction and population decline is a worldwide problem. A study by Trenham and Shaffer (2005)
on the federally endangered salamander, Ambystoma californiense (CTS) effectively described the dispersal of adult
salamanders; however immature juvenile dispersal was not well characterized. Because juveniles comprise most of a
given population in CTS, best management practices must include upland habitat use of both juvenile and adult life
stages. This study describes the dispersal and masses of juvenile salamanders, during their first two months as
metamorphosed terrestrial subadults. Using a trap array from the waterline up to 400 meters away, upland dispersal
of the juveniles was tracked by capturing, individually tagging, releasing and recapturing salamanders. We trapped
animals emerging from two adjacent vernal pools with different larval densities. From May 11th to July 27th 2005, we
accumulated 4,474 juvenile salamander captures; of these, we toe-clipped 233 animals with cohort-specific clip
patterns and individually marked 1,097 salamanders with visible, subcutaneous tags. The mean number of captures
per trap-night was 94 salamanders at Olcott lake and 8 salamanders at Round pond, confirming that different larval
densities produce different numbers of recruits. Based on captures of individually marked CTS at Olcott lake (97) and
total captures at Round pond (260), the majority of emerged CTS juveniles remained between 10m and 300m of the
waterline. Average masses of emerging CTS were 10.55 g at Olcott lake and 15.20 g at Round pond; at both sites,
mean masses of emerging juveniles gradually declined during the trapping period. The results suggest that the
largest, and potentially most fit, juveniles emerge first from the ponds, and a minimum area of 300m from the
shoreline around a pond is necessary to protect emerging CTS. Differences in larval density do not appear to affect
dispersal, but are correlated with mass at emergence. The study was continued through winter of 2005-2006 in order
to describe and analyze later juvenile CTS movements.
144
BIOTECHNOLOGY
The Structure of Filamentous Bacteriophages via Dynamic Light Scattering
Amy Serfis, Jay Newman, Physics Department, Schenectady, NY, 12308
The purpose of this project was to determine the overall structure of filamentous bacteriophages and see if they curl,
a phenomenon in which the backbone of the phage spirals. For many years, filamentous bacteriophages, consisting of
a single closed loop of DNA covered by thousands of identical coat proteins, have appeared slender and fairly
straight, having an average diameter of approximately 7 nm and lengths of over 700 nm. Recently, scanning
transmission electron micrographs and liquid crystal studies have found evidence for curling in a large family of these
phages, those which have a non-integral number of coat proteins per DNA base. In this project, dynamic light
scattering is used to detect the different degrees of curling with and without the addition of silver ions, known to
interact and lengthen the DNA. Different concentrations of X phage 2.2 mg/mL were produced, specifically, 0.1
mg/mL, 0.2 mg/mL, and 0.4 mg/mL. Each concentration was examined with and without added silver. The sample
was placed in a glass cuvette which was inserted in a temperature regulating water bath. An argon ion laser beam
having a wavelength of 514 nm was then focused on the sample and the light scattered at various angles was
collected by a phototube. Fluctuations of the scattered light intensity at each angle were analyzed by a digital
autocorrelator that was then able to extract the diffusion coefficient, D, related to the size and shape of the phage. D
was about 11% lower for all concentrations with saturated amounts of silver than without, suggesting that the addition
of silver, which causes an increase in DNA pitch, results in an increase in the phage’s overall length. These
differences in detected diffusion coefficients for X phage with and without the presence of silver ions implies that
curling occurs
CLONING AND EXPRESSION OF THE Yop Q/K PROTEIN OF Yersinia pestis FOR THE
DEVELOPMENT OF A RECOMBINANT VACCINE AGAINST THE PLAGUE
Zach Bradley, (Joseph Kimsey, Ronald Raab, Ph.D.) College of Integrated Science and Technology, James Madison
University, Harrisonburg, VA. (Captain Jeremy Goodin, Ph.D.), United States Army Medical Research Institute for
Infectious Diseases, Ft. Detrick
Bioterrorism is a threat in the world of today. The Plague, Yersinia pestis, is naturally occurring and is still present in
animal reservoirs around the world, including the United States. The plague is readily available from these natural
reservoirs and has been weaponized and used by countries in the past. There is currently a plague vaccine in clinical
trials for the plague. The United States Army Medical Research Institute for Infectious Diseases (USAMRIID) is
studying other possible vaccine candidates that may prove more effective, and serve as an alternate vaccine. The
Yop Q/K protein of the Yersinia pestis genome has been shown to be an important protein involved in the
development of a systemic infection. Using recombinant DNA technology and the IMPACT-CN system of New
England Biolabs, the Yop Q/K gene was cloned, expressed, and purified from E. coli. The end product of the research
yielded a significant quantity of the full length Yop Q/K protein. The YopQ/K protein was sent to USAMRIID for further
analysis, and will be tested in animal models for specific antibody production as part of an immune response to
against a plague infection.
145
BIOTECHNOLOGY
Inhibitory Effects of Minocycline on Gliosis in the Hydrocephalic H-Tx Rat.
Alexander G. Shanku, Janet M. Miller, PhD; James "Pat" McAllister, PhD, Department of Neurosurgery, Anatomy and
Cell Biology, Physiology, Wayne State University School of Medicine , Childrens Hospital of Michigan, Detroit,
Michigan 48201
Introduction: Persistent gliosis, if present in the hydrocephalic brain, has the potential to alter biomechanical
properties, impair cerebral perfusion, and impede neuronal regeneration and plasticity. The purpose of this study was
to determine the ability of minocycline, a specific inhibitor of glial reactivity, to reduce glial scar formation in the H-Tx
rat model of congenital hydrocephalus. Methods: Minocycline (45mg/kg/day i.p. in 5% sucrose at a concentration of
5-10mg/ml) was administered to two groups of hydrocephalic H-Tx rats from postnatal days 5-21 and 15-21. Treated
animals were compared to aged-matched untreated hydrocephalic littermates. White and gray matter of the
neocortex was processed for immunohistochemistry (Isolectin B4 for microglia and Glial Fibrillary Acidic Protein for
astrocytes) and analyzed light microscopically and stereologically. Results: Preliminary data suggest that minocycline
administration significantly reduced astrocyte density from 54,270 +/-19,800 cells/mm3 (mean +/-sdev) in untreated
hydrocephalic rats to 18,540 +/-12,000 cells/mm3 in the treated hydrocephalic animals. Additionally, minocycline
appeared effective in reducing the relative amount of microglia. The reactive morphology of microglia was reduced,
especially in the periventricular white matter. There was no apparent effect on ventricular size or cortical mantle
thickness. Conclusions: Overall, our data suggest that minocycline treatment may be effective in reducing the gliosis
that accompanies ventriculomegaly, and thus may provide an added benefit when used as a supplement to
ventricular shunting.
Creating Transgenic Drosophila melanogaster Using Site Specific Recombination
Polina Frolov (Dr. Victoria Meller), Department of Biological Sciences, Wayne State University, 5155 Gullen Mall,
Detroit, MI 48202
The overall goal of this study is to determine the function of non-coding roX1 and roX2 RNAs in dosage
compensation of the male Drosophila melanogaster. Dosage compensation is a process that enables each sex to
produce similar amounts of X-linked products from different numbers of sex chromosomes. I wish to be able to use a
newly developed site-specific intergrase system for the purpose of creating transgenic flies carrying roX1. In order to
utilize this system I need several integrase target sites on the X chromosome. Stocks containing an insertions of the
target site in a P element on the second chromosome where obtained from the Drosophila Stock Center. I mobilized
this P element and collected flies carrying hops to other chromosomes. New insertions have been mapped to
chromosome. A total of two hundred and nine hops were recovered, seventeen of these were to the X chromosome.
In order to determine the exact insertion of the P element on the X chromosome, in situ hybridization to chromosome
squashes is now being performed.
SURFACE RADIATION EFFECTS IN LASER ASSISTED CANCER IMMUNOTHERAPY
Hsin-Wei Chen, Josh Wilson (Dr. Pradip Bandyopadhyay), Department of Physics, Hendrix College, 1600 Washington
Avenue, Conway, AR 72032
Current methods for the treatment of cancer include surgery, chemotherapy, and radiation. Laser Assisted Cancer
Immunotherapy(LACI) is a method that shows promise for the treatment of solid tumors. LACI involves laser radiation
of a tumor that has been injected with an immunoadjuvent(Indocyanine Green) and a light absorbing dye(Glycated
Chitosan). The radiation causes tissue damage(necrosis) which excites an immune response that is enhanced by the
immunoadjuvant. The tumors are grown between the skin and muscle layers on mice until they reach a size of 5-7
mm in diameter. A 805 nm diode laser irradiates the tumor using a silica fiber tip. Temperature measurements were
performed with microthermocouples placed inside the tumor. Surface temperatures were measured with an infrared
detector. The tumor damage profile using the surface irradiation mode will be compared with that using interstitial
irradiation. This study aims to investigate the efficacy and amount of necrosis resulting from surface laser radiation .
146
BUSINESS
Perceived value to employers of MBA degrees depending on what country they are obtained in.
Marine Eberhardt, Nancy Miller, Marketing Research, Berry college, Martha Berry HWY Rome GA 30149
This research is used to find out how employers value different MBA degrees depending on the country in which they
are obtained. The belief is that certain countries have different values in the eyes of employers, and the goal is to
find out how much of that opinion is placed on facts versus simple "brand name" of a country.
An Analysis of Mergers and Acquisitions in the Telecommunications Industry
Ekaterina Trepalina, Senior, Department of Finance and Honors College, , Dr. Lalatendu Misra, Thesis Advisor
This study examines mergers and acquisitions in the telecommunications industry which changed from a regulated
monopoly to a competitive oligopolistic structure starting with the breakup of AT&T. Subsequent waves of mergers
and acquisitions have resulted in a consolidated industry. I analyze the structural evolution of the industry, and
evaluate the stock price performance of parties in the merger activity over the last two decades. The merger activity
before the 1984 AT&T break-up is not taken into account, as AT&T controlled 85% of the market share. I find that
targets gain approximately 12.5% and acquirers lose 3.5% during the two-day announcement period. Although my
finding suports previous academic research, the acquirer loss is substantially lower than that found in mergers in other
industries. The changes in synergies and gains to each party over time as the industry consolidates are analyzed by
conducting a multiple regression of the gains against independent variables. Synergistic losses to acquirer are
negatively related to the relative size (equity market capitalization) of acquirer to the target, and the time variable.
This suggests that relatively larger acquisitions in this industry have lowered the acquirer abnormal return and that
earlier acquisitions in the industry brought higher returns to the telecom acquirers.
WORK-FAMILY ENRICHMENT
Misti M. Stoddard (Dr. Susan Madsen), Department of Business Management, Utah Valley State College, 800 West
University Parkway, Orem, UT 84058
Work-family conflict has been under the microscopes of researchers over the past few decades. Attention has mainly
focused on the time-, stress-, and behavior-based spillover between these roles as well as the work to family and
family to work interference (Carlson, Kacmar, & Williams, 2000; Greenhaus & Buetell, 1985). Several instruments
have been developed to measure the degree of conflict and stress associated with work and family life (e.g., Carlson
et al., 2000; Stephens & Sommer, 1996). Interestingly, decades of research has focused on the negative impact
multiple roles have had on workplaces and homes, while little attention has been given to the individual benefits that
may result from simultaneous participation in these roles. Fortunately, Greenhaus and Powell (in press) have recently
provided a conceptual foundation for the work-family enrichment construct and have defined it as “the extent to which
experiences in one role improve the quality of life in the other role” (p. 2). Also recently, Carlson, Kacmar, Wayne,
and Grzywacz (2005) published a piece on the development and validation of a work-family facilitation scale to
measure the positive side of the work-family interface. The purpose of this current study is threefold: 1) to examine
the degree to which employees at retail establishments perceive work-family enrichment in their lives; 2) to contrast
these with participants’ perceptions of their own mental and physical health; and 3) to investigate the relationship
between work enrichment perceptions and the following employee demographics: gender, number of children, marital
status, length of employment, employee age, and educational level. This study, distributed to 150 retail employees,
utilized the newly developed 18-item Carlson et al. (2005) work-family enrichment scale with choices ranging from “1”
being “strongly disagree” to “5” being “strongly agree.” This research provides important insights into the level of
interdependency between work and family roles as well as the relationships between work-family enrichment, health,
and the previously listed demographics.
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WOMEN ENTREPRENEURS: ONCE JUST “ROCKED THE CRADLE”—NOW ROCKING THE WORLD
OF BUSINESS!
Ruth Davis (Dr. Susan Madsen), Department of Business Management, Utah Valley State College, 800 West
University Parkway, Orem, UT 84058
During the past few decades it has become clear that more women are now starting their own businesses. According
to the U.S. Bureau of Census in 2004, women held the majority of privately owned business (51 percent) (Center for
Women’s Business Research, 2004). In the last five years the profit margin from these women entrepreneurs
increased from 75 to 700 million which was an increase of 39 percent (Prospero & Michael, 2005). Between 1997 and
2004, the three fastest growing states housing women entrepreneurs were 1) Utah, 2) Arizona, and 3) Nevada. The
Center for Women’s Business Research (2004) noted that in Utah alone approximately half of privately held
businesses are now owned by women. Hence, the hands that once just “rocked the cradle” are now rocking the world
of business. Women interested in business, creativity, and profits are realizing their own dreams and aspirations, and
they are making money in the process. The purpose of this qualitative research study is to explore the reasons
women have decided to start their own businesses. The study focuses on women business entrepreneurs within the
state of Utah. Ten women entrepreneurs will be interviewed to investigate the reasons behind their decisions to create
and continue operating their own businesses. It will also explore the benefits and barriers they have experienced
throughout their journeys. A list of women Utah Entrepreneurs was obtained, and the women on this list within two
specific counties were contacted by email and/or phone. The first ten who agreed to participate were interviewed for
30 to 60 minutes. The interviews were taped, transcribed, and analyzed using theme generation techniques. This
research provides important insights into the motivation, benefits, and barriers perceived by Utah women
entrepreneurs.
Finding it Hard to Swallow: Consumer Perceptions and the Marketing of Genetically Modified Food
Products
John E. Brazel, Jr. (Dr. Nancy D. Albers-Miller), Department of Marketing - CSOB, Berry College, P. O. Box 495024,
Mount Berry, GA, 30149
Recently, genetically modified foods (GMOs) have been portrayed by the media in a very negative light. Horror
stories about the potential harms to consumers have been circulated both in the press and from consumer to
consumer. Documented benefits to nutritional value, to human health, to the environment, to crop productivity, and to
spoilage reduction rarely receive the same media and Internet coverage. Despite many potential benefits from
enhancing and modifying food products, there is a growing body of evidence that consumers are increasing skeptical
about genetically modified foods. This study examined consumer perceptions of GMOs from multiple perspectives.
First, the degree to which consumers were concerned about buying and eating GMOs was measured. In the second
part of this study, respondents were randomly sorted across control and treatment groups to test various advertising
messages for GMOs. Variation in the use of terms describing the type of modification (none, genetically modified,
engineered, scientifically improved), variations in the documented benefits (none, increased nutritional benefit, plants
- reduced use of pesticides/animal - reduced use of growth hormones) were experimental manipulated for two product
categories (tomatoes and beef). Therefore, a total of 24 hypothetical advertisements were created. Respondents
were randomly sorted across treatment groups and were shown only one of the 24 advertisements. Respondents
answered the same set of questions. The results offer insight which should prove valuable to consumers, producers
of food products and to retailers. Public policy and managerial implications are provided.
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Preparing Young Minds for the Future: Exploring Internships in a Novel Setting
LaToya J. Taylor (Research Faculty: Justin Vidovic and Gwen Gorzelsky, Department of English, Wayne State
University), Honors Department, Wayne State University, Detroit, MI 48202
This research project is a qualitative study on the internship program at University Preparatory High School (UPHS),
an urban charter school in Metro Detroit. UPHS employs an internship program based on Deweyan ideals of a
democratic education that integrates hands-on work with academic analysis. All students enrolled must find and
participate in several internships as a requirement for graduation. Fishman and McCarthy draw attention to the fact in
John Dewey and the Challenge of Classroom Practice that while the practical applicability of Dewey’s ideas has long
been contested, there has been little or no empirical research done on schools that employ Deweyan ideas. In an
effort to research the effects of a Deweyan internship pedagogy, this study endeavors to understand how internships
help students develop academically, how internships help students develop “life skills” such as responsibility,
conscientiousness, increased confidence and self-esteem, and any future benefits students get from participation in
internship programs.
This qualitative study will employ the ethnographic methods outlined by Emerson, Fretz, and Shaw in Taking
Ethnographic Fieldnotes in response to calls by educational researcher John Szwed for more ethnographic studies of
educational practice. I will be observing internship sites and reviewing past projects and portfolios of participating
students. In addition to participant-observation, I will conduct interviews with all persons involved in the UPHS
community including students, faculty, the internship coordinator, parents and the mentors. The results of this study
are significant for several reasons. First, it is an opportunity to learn whether the Deweyan theories that are the
foundation behind the concept of the school actually work in a practical setting. Second, many research studies have
been conducted in an effort to assess the future benefits to students who participate in internships while few have
focused on discovering what, if any current benefits students gain.
HMONG PROFESSIONALS IN WISCONSIN: THEIR CONNECTIONS AND PATHWAYS
Mai Chue Xiong (Susan Huss-Lederman), Department of Language & Literature, University of Wisconsin-Whitewater,
800 W. Main Street, Whitewater, WI 53190
By the end of 2005, there were over 35,000 Hmong people in the state of Wisconsin. Hmong professionals are
increasing in number and venturing into new professional fields. Yet, the stories of their struggles and successes
have never been adequately documented. The experiences of these pioneers are important for Hmong youth and
educators to understand as the United States population continues to grow more diverse. This study addresses three
open-ended research questions. First, what factors influence the choice of fields that Hmong professionals enter?
Second, how do incumbent professionals create pathways for future professionals? Third, what is important in
guiding or assisting Hmong youth to choose a particular career? This qualitative study utilizes individual interviews
since the nature of the research entails different fields of professionalism. Fifteen to twenty Hmong professionals are
interviewed in the study: educators; lawyers; doctors; social workers; business owners; artists; musicians; politicians;
political activists. Preliminary results suggest that interviewees’ reflections demonstrate the importance of mentorship
by teachers and family members. The importance of mentorship was also borne out in this researcher’s previous
study, “Hmong American College Student Speak Out,” in which university students expressed the importance of
guidance from parents, siblings, and teachers in pursuing higher education.
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Credit Risk Reduction in the Auto Industry: A Case Study of Derivative Securities
Gabriela Peeva(Dr. Ralph Trecartin), Department of Business Administration & Economics, The State University of
New York - Brockport, Brockport, New York 14420
Recently, debt rating downgrades and bankruptcy cases have been flooding the news: names like Refco, Delphi,
Delta, GM and Ford resonate in the financial press. This is especially evident in the automotive industry where giant
companies like GM and Ford have faltered and their bondholders have suffered catastrophic losses. This
environment presents the perfect opportunity for instruments like credit derivatives and options to be used for risk
reduction activities. The purpose of this study is to document credit derivative usage by companies in the automotive
industry and by their investors. Industry and individual company data is summarized and analyzed from a variety of
sources including: COMPUSTAT, The Value Line Investment Survey, Analyst Research Reports and the Wall Street
Journal. A brief outline of credit derivative security basics is covered along with pricing mechanisms for default
insurance on loans and bonds. A major thrust of the study is to illustrate the application of credit default swaps,
spreads, and put options to the current credit risk scenario. Actual usage of these securities will be compared with
alternative strategies in an attempt to quantify the difference between actual losses and the hypothetical losses that
would have been incurred if derivative securities were to be used.
The Effect of Positive and Negative Nutritional Elements in Food Product Decisions
Author-Jessica Bennett, Sponsor-Steven Cox, Business Administration, Meredith College, 3800 Hillsborough Street
Raleigh, NC 27607
The Nutrition Labeling and Education Act of 1990 required that all prepared foods carry a prescribed label identifying
specific nutritional elements. These labels contain attractive items; vitamins, minerals, protein, and fiber, and
negative items; calories, carbohydrates, fats, sodium, and sugar. Employing approach-avoidance theory, this study
examined nutritional label usage. Over 150 female nutritional label users evaluated from least important to most
important, a standard set of items listed on a nutrition label. Results showed that the participants are more persuaded
by the avoidance items than the approach items on the label. These results confirm earlier studies which
demonstrated that concern for negative consequences is more compelling in food product selection than the value of
positive outcomes.
Regression Towards the Mean and Investment Portfolios
Michael D. Canning (Colonel H. Francis Bush), Department of Business and Economics, The Virginia Military Institute,
Lexington, Virginia 24450
This research tested the statistical property of regression towards the mean against the efficient market hypothesis of
capital markets. If markets are not efficient, then a portfolio of under-performing stocks, even though some stocks
will ultimately fail, will give a higher return on investment than a high-performing portfolio. The mean return of the
high-performing portfolio will fall towards the mean return of the population of stocks, and the mean return of the
low-performing portfolio will rise towards the mean return of the population stocks. Two portfolios of ten stocks each
were gathered from Fortune 500 magazine for each of the years 1983 to 2004. To assess the success of each
portfolio, the return on investment was calculated. The research was conducted to show the practical application of
regression towards the mean in investing. The results were surprising; the efficient market hypothesis was not
rejected. Over the 20 years from which data was taken, the low-performing portfolio outperformed the high-performing
portfolio by only 1.4% of the years. These results reject the efficient market hypothesis, but are not statistically
significant enough to reject it outright.
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When Advertisers Miss the Mark: Off-Target Consumer Responses to Sex-Role Values in
Advertising
James E. Colvin (Dr. Nancy D. Albers-Miller), Marketing, Berry College, P. O. Box 495024, Mount Berry, GA 30149
When advertisers use mass forms of communication to reach prospective customers, they are often in the position of
reaching a multitude of individuals who are not in their target audience. While advertisers are obviously concerned
with the ability to appropriately appeal to their designated target market, they must do so without offending others who
are not in the target market. Countless studies have looked into how gender affects cognition, behavior and values.
For decades, social science researchers, conducting various studies, have shown that the genders are sex-typed or
perhaps biologically wired towards differences in values. The majority of this research has indicated that traditional
feminine values include values such as being cooperative, caring, empathetic, while traditional male values include
values of achievement, success, and justice. Over the years, these studies have shaped how advertisers market to
men and women. Despite the richness of the previous research in to gender roles, there appears to be a significant
gap in the advertising literature. Marketers must be concerned not only with how feminine viewers respond to
feminine appeals and how masculine viewers respond to masculine appeals, but also how off-target masculine
viewers respond to feminine appeals and feminine viewers respond to masculine appeals. The purpose of this study
was to experimentally validate the reaction of off-target consumers facing appeals which are not salient or even which
violate their value systems. The results of this study provide important managerial implications for advertisers and
marketing managers.
Subjugated Students: The Negative Consequences of Loss of Control on Consumer Evaluations of
Product and Service
A. Daniel Margrave (Dr. Nancy D. Albers-Miller) Marketing, Berry College, P. O. Box 495024, Mount Berry, GA 30149
Student consumers are often required to buy without deciding. Many items, including textbooks, course supplies, etc.,
are selected by someone else. Students are expected to buy items regardless of inconvenience, price, or quality.
These situations form a unique type of captive consumption. When a student is required to live on campus and/or
purchase a meal plan, they are held captive for life basics. Research into captive consumption has documented
unfair treatment and disproportionate economic burdens (Rotfeld 2003, Krieger 1995, Savage 1993). Consumers in
an involuntary captive setting, unable to leave and facing few options are not loyal and are often unhappy (Moreno
2005, Pan, Lo and Hsiao 2004, Turley and Shannon 2000). The negative reactions to captivity may impact the
students` evaluation of the product and services. The purpose of this study was to quasi-experimentally evaluate the
degree to which there is a transfer effect of negative feelings of captivity to the evaluation of the quality of product
and service. The study examined evaluations of the quality of food and service at the campus dining facility at a
private college. Evaluations of students who ate at the dining facility using their required meal plan were compared
to evaluations of students who ate at the dining facility voluntarily (e.g. they did not have a meal plan and yet
selected, paid for and ate meals at the dining facility). The results of this study have important implications for
commercial suppliers of food services on college campuses. Additionally, administrative implications are provided.
Corporate Social Responsibility and its Effects on Shareholder Wealth
Kristina Lollman, Economics, Hartwick College, 200H Bresee Hall, Oneonta, New York
Corporate Social Responsibility (CSR). The neoclassical economic perspective states that a company should only
focus its efforts on expenditures that only enhance shareholder wealth, whereas the recently-emerged stakeholder
theory states that a company that recognizes its responsibility to other stakeholders instead of shareholders alone will
increase a company’s reputation and led to increased shareholder wealth. Data was collected from 10 different
companies over a 10 year period. The benchmark used for CSR was participation in the Global Reporting Initiative
(GRI). An independent variable of Selling, General, and Administrative Expenses (SGA) was collected to judge the
effects of advertising on a company’s shareholder wealth. The results show that participation in GRI has a negative
effect on shareholder wealth (p-value=0.037); a dummy interaction variable between GRI and SGA indicates that the
negative effect is decreased by 0.625 for every dollar spent on effective advertising of a company’s CSR activities
(p-value=0.264).
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Enterprise Service Level Improvement
Ryan Hergrueter & Tadas Mankevicius are the authors, Professor Barry Cobb is the sponsor, Department of
Economics and Business, Virginia Military Institute , 335 Scott Shipp Hall , Lexington, VA 24450
The measurement of customer service levels is essential in today’s competitive market place if internal supply
chains are to be aligned to, and supportive of, an organization’s profit goals. Further, customers are increasingly
seeing customer service measures as a sign of “best practice” and some demand them as a condition of trade. This
project will examine the recent service level improvement project of a $2.1 billion dollar privately owned global
cosmetic and beauty company. Historically, this particular company has measured service levels by manually
compiling data in programs such as Excel. Beginning in September of 2005, this company made the transition to a
fully automated, internet based method of measuring service levels through a product called Stratum Ezviewer.
Specifically, this project will use service level measurements over a 30-day period and apply quality control and
decision analysis tools to determine the overall performance of organizational processes. If performance deficiencies
or areas of superior performance are found, these may be presented to the company, along with recommended
solutions. This project will connect theoretical operations management models with business practice. Additionally,
this research aims to guide the sample company with diverse recommendations and various proposals throughout the
first months of this service level project.
EXPLORING THE LIMITS OF GLOBAL MARKET INTEGRATION AND CROSS-CONTINENTAL
ARBITRAGE: THE CASE OF AMERICAN DEPOSITORY RECEIPTS ISSUED BY RUSSIAN FIRMS
Ilian Draganov (Dr. Ufuk Ince), Business Program, University of Washington, Bothell, Box 358533, 18115 Campus
Way NE, Bothell, WA 98011-8246
In this study we investigate the validity of financial market efficiency hypothesis in global financial markets. We
examine statistical and time-series properties of the American Depository Receipts (ADRs) issued by a pool of
Russian firms on New York Stock Exchange (NYSE) with the characteristics of the corresponding securities in their
native economic environment in Russia. Out of the forty Russian ADRs in the US we focus on six firms that are
trading on NYSE and have extensive regulatory filing requirements with the Securities and Exchange Commission.
Using daily stock market and exchange rate data, we identify possible arbitrage opportunities trading the twin
securities across the American and Russian financial markets after taking into account the transaction costs in the
stock and currency exchange markets. We use correlation analysis as well as time series regressions with varying
lags to measure the degree of price and return divergence. In addition, we provide possible explanations for the
discrepancy in security price process in the two markets: market sentiment, microeconomic conditions, regulatory
ownership restrictions, and trading hour overlap. The findings enable us to answer the question of how correlated,
therefore efficient, the pricing of Russian ADRs trading on NYSE are. We expect the conclusions from this study to
have implications for analyzing market efficiency and integration in other emerging financial markets.
An Exploratory Investigation of the Roll of College Professors in Facilitating International Student
Transition
Lewis Chidziva, Jair Cortazar (Letty Workman), Business Management, Utah Valley State College, 800 West
University Parkway, Orem, UT 84058
As the primary contact between international students and the new and often challenging environment in which these
students find themselves, college professors represent a valuable lens for examining the social, cultural and
economic perceptions and expectations of international students. As the International student population is
decreasing(Institute of International Education, 2004), the teacher-student relationship represents an important
student touch point in developing effective methods for recruiting and retaining international students in the United
States, as well as facilitating transnational migration, adaptation and integration (Horton and Owens, 2004). The
purpose of this research is to: 1) understand professors’ perceptions of international students; 2) identify common
attitudes and behavior patterns of international students; 3) develop and implement better methods for integrating
international students with their new surroundings. This study will employ qualitative methods in exploring teachers’
perceptions of international students in a transnational community through: 1) a diverse focus group of Ph.D.
professors and international students, and 2) a comprehensive questionnaire, based on the newly developed
contextual methods of Flores and Clark (2005).
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PERSONAL BIAS IN STUDENT PEER ASSESSMENT
Suzanne Britton, (Dr. Jane E. Barnes) School of Business, Meredith College, Raleigh, North Carolina 27607
This study considers whether a personal relationship affects students’ abilities to grade each other objectively and
expands on the literature of peer evaluations. Peer assessments have become a popular and effective tool in higher
education; however, there are concerns regarding evaluations made by acquaintances and friends (Brown and Knight,
1994; Nilson, 2005). Data is from eight different classes from 2003 – 2005, and consists of 3199 paired observations
from 161 students who made evaluations of their fellow students’ oral presentations in two different business
management courses. The same professor gave similar evaluation instructions to each class. For the purpose of this
study, we define “friends” as those who sit next to each other in class. (After the first week of class, each student sat
in the same location in the classroom.) Demographic data on each student was collected from the Registrar. The
study hypothesizes that students who are friends will grade each other differently than those who are not. Previous
research suggested that we control for age, race, course level, and admission status (e.g., transfer student). For the
analysis, we performed correlations and multiple regressions. The results indicate that there is not a significant
difference between the grades of friends and those students who were not acquainted with each other. However, the
study does provide other interesting results. The tests suggest that older students and those in the higher level
course tend to give significantly lower grades than their younger counterparts or those in the lower level course,
respectively. We also tested race effects initially using four different categories (Caucasian, African American,
Hispanic and Other) but because of small sample sizes the results were not significant. When non-Whites are
combined into an “ethnic” variable, however, the results indicate that minorities tend to give lower grades to other
students.
Negative Effects of Poker
Tyler Carman, Jon Littlefield, Marketing, Berry College, 186 Berry College, Mt Berry, GA 30149
Negative Effects of Participation in the Poker Culture: An Ethnographic Investigation
Abstract:
Poker playing has experienced an upswing in today’s culture reaching levels of popularity never before seen for a
card game (Packham 2005), with players participating in such contexts as casinos, local “poker nights,” and online
play. Much of poker’s newfound popularity can be attributed to media portrayals such as the televised World Series
of Poker (WSOP), first televised in 1998. The “poker night” ritual has both financial and social repercussions to
players that carry over to other aspects of the players’ lives. In this study, I use ethnographic methods to examine the
connections among the media’s portrayals of poker players, the players’ actions in their individual playing groups, and
the solitary online behavior exhibited by many social poker players. Specifically, I conducted depth interviews with
poker players, and used participant-observation to document conduct during weekly poker nights.
Findings relate to negative effects of participation, including both social effects and players’ incorrect risk perceptions.
It seems that poker players evaluate and accept risk in unique ways during play. For instance, the level of social risk
to the player is much greater than players are willing to admit. My findings also support previous studies in the
gambling literature in suggesting that poker players do not correctly evaluate outcome probabilities, although the
current research focuses on the game (rather than the individual poker hand) as the unit of analysis for making these
evaluations. While physical and financial effects of poker playing are identified (consistent with previous research),
emotional effects are seen as a new contribution of this study. One emotional finding, for example, is players’ refusal
to recognize negative effects of poker, frequently making short-term attributions for long-term trends.
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Antecedents and outcomes of trust in peer learning groups: Is enhancing member satisfaction with
peer learning groups possible?
Adrienne D. Bumgarner, Paul H. Jacques, Ph.D., and John W. Garger, Western Carolina University, Management &
International Business Dept., College of Business, Cullowhee, NC, 28723
The objective of this research is to determine the antecedents of trust and nature of the relationship between trust and
satisfaction with subjects’ peer learning groups. Subjects in this research consisted of 426 upper-level undergraduate
students that were placed in various semester-long assignments with peer learning groups with key deliverables due
throughout the semester. Surveys were administered to 24 course sections across various courses within various
academic colleges and departments at a mid-size university located in the southeast region of the United States. The
hypothesized relationships in this research include antecedents to trust such as benevolence, integrity, ability,
trustworthiness, propensity to trust, and satisfaction with group. Findings generally support key hypothesized
relationships. Major findings for antecedent constructs include the relatedness of integrity and trust (p<.001) and
between benevolence and trust (p=.10). Also, study findings support the notion that there is a strong and positive
relationship between trust and subject’s self-reports of satisfaction with their peer learning group (p <.001). The
impact of group size on scale means of all non-trait based measures in this research was particularly noteworthy. The
size of the peer learning groups varied from two to seven students and means of benevolence, trustworthiness, trust,
benevolence, and subject’s satisfaction with their groups decreased as the size of the group increased. As expected,
self-ratings of a trait-based construct, propensity to trust, remained stable regardless of group size. In addition, the
relationship between group size and satisfaction with group was significantly negative. One implication of this study is
that educators who employ small groups in their course design should also be willing to take an active role to
contribute to the development of trust in these settings. Secondly, these findings suggest that the fundamental
relationships necessary for the development of trust within peer learning groups and subsequent satisfaction with
those groups deteriorate markedly as group size grows. Groups larger than 4 students appear particularly at risk.
Additional implications, recommendations, and directions for future research are discussed in the paper.
Title: Customer Relationship Management (CRM) and Market Segmentation
Robert Tiegs, Kira Lakner, Megan Halverson, Buddy Seiner, Kariline Rogers, Amy Brommer, , University of South
Dakota School of Business, 414 East Clark, Patteson Hall, Vermillion SD 57069
According to relationship marketing literature, building a long-term customer relationship increases a firm’s
profitability. Therefore it is important to create and maintain good rapport with its customer base. However, many
firms attempt to create this relationship without understanding the heterogeneous segments within the market. Our
research aims to explore the importance of market segmentation in Customer Relationship Management (CRM) to
enhance and build better relationships between firms and their customers. Specifically, this research is intended to
test one of the CRM theories proposed by Boulding, Staelin, Ehret, and Johnson (2005)*. We surveyed existing and
future customers of a health fitness service provider. We used a convenience sampling technique (N = 49) and
offered a cash drawing opportunity to compensate their time for participation. We measured consumer reaction to
service offerings (e.g., attitude, purchase intention, importance of service attributes) as well as segmentation bases
(e.g., frequency of usage). Our results showed significant differences in purchase intention as well as consumer
evaluation of key decision factors such as breadth of services, atmosphere, price, and sales promotions, depending
upon the type of market segments. In particular, our findings suggest that in regard to heavy exercisers versus that of
light exercisers, coupons tend to be a more effective sales promotion. These results support the CRM theory asserted
by Boulding and his colleagues (2005) that consumer heterogeneity should play a significant role in building long-term
customer relationships. Theoretical and practical implications in the relationship marketing area will be further
discussed.
* Boulding, William, Richard Staelin, Michael Ehret, and Wesley J. Johnston (2005), “A Customer Relationship
Management Roadmap: What is Known, Potential Pitfalls, and Where to Go,” Journal of Marketing 69(October),
155-166.
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BUSINESS
The Flat Tax: From Post-Cold War Europe to Steve Forbes
Nicholas Stinson (Dr. Karen Oxner and Mr. Stephen Kerr), Department of Accountancy, Hendrix College, Conway,
Arkansas 72032
The purpose of this project is to debate the potential effects of the Hall-Rabushka flat tax on the contemporary United
States of America by examining its evolution from Post-Cold War Estonia to the failed presidential candidacy of
Steve Forbes in 1996. As the flat tax grew in popularity throughout the 1990s it has been adapted by theorists and
later political leaders to reflect the economic needs and wants of the culture in which they live. The various
manifestations of the flat tax from theory to practice reflect different historical periods and political environments. An
examination of the flat tax over time and across geography offers a unique insight into the political vacuum created
by the fall of the Soviet Union. The flat tax of 1994 Estonia is not the flat tax proposed by Steve Forbes in 1996. This
project illustrates how different situations in the two countries resulted in drastically different images of the
Hall-Rabushka flat tax. Ultimately this project establishes that the economic success of the flat tax in Estonia is not
achievable in the contemporary United States of America.
THE SARBANES OXLEY ACT OF 2002: HOW BUSINESS EDUCATION IS RESPONDING
Lindsey Longmire (Dr. Sarah Stanwick). School of Accountancy, Auburn University, Auburn, Alabama 36849
Few pieces of legislation will impact the accounting profession and the business community as profoundly as the
Sarbanes-Oxley Act of 2002 (SOX, the Act). The Act is government’s response to the increasing number corporate
scandals and was created to increase internal controls and the quality of financial reporting. Accounting firms and
corporations alike have been forced to comply with significant changes brought about by the Act. The real question
that persists is “How is Business Education addressing this issue in the current curriculum?” This study addresses
current business education program approaches to incorporating the legislation into their existing curriculum.
Accounting department chairs were surveyed in an attempt to capture a glimpse of how accounting professors are
currently covering the Sarbanes-Oxley Act in their courses, their future plans for addressing the Act, and how they
believe SOX should be incorporated into business curriculums. The study also includes the results of a survey of
Certified Public Accountants regarding what they believe students should be taught about SOX before graduation.
The paper will compare the results of both surveys to provide insight on how accounting programs should be
educating future accountants and business professionals about the Sarbanes-Oxley Act.
BRAND LOYALTY IN THE NASCAR COMMUNITY
Ross Boner (Jon Littlefield), Department of Marketing, Berry College, Mount Berry, GA 30149
NASCAR is a significant and growing consumption phenomenon. Each of thirty-six annual races attracts anywhere
from 80,000 to 150,000 fans at ticket costs that reach into the hundreds of dollars. In addition, Sunday races are
viewed by many fans not attending games, a level of sport viewing second only to the National Football League in
sporting events. In all, NASCAR’s 75 million fans exhibit a high degree of loyalty both to the individual drivers and
the companies that sponsor them, and to other members of the NASCAR fan community. For example, loyalty is
exhibited publicly through membership displays including personal vehicle customization. This study examines the
relationships between fan preferences and driver/brand loyalty.
Qualitative ethnographic methods were used to provide understanding of these relationships. These methods
included depth interviews with fifteen NASCAR fans, with members of driver organizations, and with driver sponsors.
In addition, participant-observation was conducted by attending NASCAR events. Findings suggest that the NASCAR
experience results in a temporary virtual community that uses unique means of serving needs of community
members. In addition to this “micro-level” consumption connection, a “macro-level” connection also exists through
which brand relationships mediated by driver sponsorships fulfill important social membership needs of members of
the fan community.
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Student Entertainment and Restaurant Habits (A Marketing Study)
Adam Woodward, Jennifer Allen, Everett Smith, Kevin Perez (Dr. Beheruz N. Sethna and Ms. Melanie Hildebrant)
Department of Marketing and Real Estate, The University of West Georgia, Carrollton, Georgia 30117
The Maple Street Mansion Project was a research initiative designed to determine how a local restaurant and
historical landmark, The Maple Street Mansion, could maximize its business with UWG students. A one-page
questionnaire was constructed with questions relating to students’ choice in restaurants, frequency of restaurant visits,
types of advertising, favorite foods and attractions, and pricing for cover charges. The questionnaire was administered
to students through en event called a “Marketing Carnival”, in which games and prizes were used to entice students to
fill out questionnaires. Additional questionnaires were also collected from several classes around campus, bringing
the grand total to 442.
After coding the completed responses, which consisted of both open-ended nominal and ratio data questions and
Likert scale interval questions, statistical tests and analyses were performed on the data. Chi-Square tests and data
provided from the University’s office of Institutional Research and Planning were used to determine whether or not the
observed frequencies of the different genders in our sample were statistically different from the true population. Our
percentages were 45% males and 55% females, compared with 40% males and 60% females for the total population.
A statistical test of one proportion showed that the data had a gender breakdown that is representative of the
population of West Georgia at a .010 level.
Correlations were used to determining the characteristics of several segments, including gender, class, and age.
For evaluating the successfulness of some proposed future attractions, students were asked to rate a list of possible
attractions with a score from 1-10. Among the majority of attractions, there was no significant gender segment, except
in the cases of Karaoke and Televised Sporting Events. A Chi-Square test on both variables revealed that females
preferred Karaoke almost 20% more often than males, at the .014 level. Also the same was true for Males when the
variable was changed to Televised Sporting event, showing a similar preference at the <.05 level. From the results, it
was concluded that targeting different gender segments (possibly through advertising to the Greek system) for
Karaoke and sporting events respectively, would be the most beneficial.
156
BUSINESS: ACCOUNTING
EXECUTIVE COMPENSATION PACKAGES AND FINANCIAL RESTATEMENTS
Andrea Herndon (Dr. Steven Hall), Department of Accounting/Finance, University of Nebraska-Kearney, West Center
Kearney, NE 68849
In the past several years there has been a rash of corporations with accounting irregularities. Most official response
has been to strengthen enforcement of existing regulations. However, an alternative approach is to investigate the
incentives managers have to misstate earnings. The goal for my research is to analyze the relationship between
executive compensation packages and firms that report accounting irregularities. I hypothesize that there is a
positive correlation between those firms that use a bonus or a paricular form of bonus as part of their compensation
package and those firms who have been asked to restate their financial statements by an external auditor. This
hypothesis is justified because a bonus may give managers incentive to inflate earnings to increase compensation. I
will gather compensation data of a sample of firms that have been asked to restate their financial statements and
correlate these findings against a control sample. Compensation plans are described in proxy statements filed with
the Securities and Exchange Commission and are available from online resources. The results of the study will be
beneficial for corporate compensation committes in deciding whether a bonus is an appropriate way to compensate
executives of the firm. It will also be useful to policy makers who might consider an incentive change in addition to
strict enforcement to combat the incidence of fraud and other accounting irregularities.
157
BUSINESS: FINANCE
DEVELOPING AN UNDERGRADUATE STUDENT PERSONAL FINANCE GUIDE: AN APPLIED
RESEARCH PROJECT
Maylee Xiong (Rosita Chang, University of Hawaii-Manoa), Finance Department, University of Wisconsin-Whitewater,
800 W. Main Street, Whitewater, WI 53190
According to Nellie Mae (2004), over 2/3 of students began the academic year with at least one credit card, with
average outstanding balance of $1,585. The percentage of students that have at least one credit card substantially
increased to over 90 percent. Students have an average total credit card debt balance of $3,864 by the end of
undergraduate study. The Nellie Mae study also reported that 74 percent of undergraduates use credit cards for
school supplies, while 71% students use credit cards to pay off charged expenses. Although this study indicated that
students are extensively using credit card to pay off some of their school-related fee, only 21 percent reported paying
off account balance in full each month. Approximately 11% said they make less than the required minimum payment
each month, while 44% make a little more than the minimum payment but carry a balance forward. By making only
the minimum balance payment, students will end up paying far more the purchased amount. However, the ability to
control credit card using can be done through proper budgeting and planning. The purpose of this study was to
develop a personal finance guide that will enable college students to better understand how to manage their finances.
Utilizing an analysis of secondary literature, data was gathered to provide a step by step process for students to follow
in reaching their financial goals. This guide includes five steps: setting financial goals; learning how to budget;
selecting the right financial institutions; becoming aware of the types of student loans available; making wise
decisions when applying for credit cards. Finally, this applied research process accomplishes its ultimate objective of
providing financial knowledge in a simple manner in order to enrich students’ understanding of the financial system.
CHARACTERISTICS OF ONLINE IDENTITY THEFT VICTIMS
Whitney Price (Diversity Scholars Research Program), Center for Research and Learning, Indiana University-Purdue
University Indianapolis, 755 W. Michigan St., Indianapolis, IN, 46202
Over 9.3 million individuals were identity theft victims in 2004 (Wall, 2005). The number of identity theft victims is
continuously increasing, because of a relatively new identity theft scam called phishing. Phishing is the act of sending
out emails to lure Internet users to imitated websites to steal their financial and personal information (dictionary.com,
2004). The purpose of this study is to identify certain habits, behaviors, and knowledge that leads individuals to
become phishing victims and to see which characteristics could help in the decline of phishing attacks. This study will
test several characteristics including knowledge about phishing and behaviors and habits online and offline of 100
randomly selected IUPUI Internet users about their reactions to a phishing scam. The subjects will view and evaluate
3 emails, in which some are fake and some are real, and their responses will classify each of them as potential
identity theft victims and non-potential identity theft victims. Next, the subjects will be asked personal questions
about their habits, knowledge, and behaviors. Based on information we gather, we expect to find that the individuals
who would classify as an identity theft victim would exhibit certain behaviors and habits and would be less
knowledgeable about phishing than the individuals who could correcting identify a fake email.
158
BUSINESS: MANAGEMENT
LEADERSHIP DEVELOPMENT NEEDS ASSESSMENT FOR HISPANIC CHAMBER OF COMMERCE IN
MARIN COUNTY CALIFORNIA
Roger N. Forte (Dr. Christopher Leeds and Dr. Denise Lucy), The Institute for Leadership Studies and School of
Business, Dominican University of California, 50 Acacia Avenue, San Rafael, California 94901
The purpose of this research is to develop and test a practical and effective needs assessment tool to determine the
need for leadership development in small businesses in Marin County, with particular emphasis on small business
members of the Hispanic Chamber of Commerce. The results of this research will be used to better facilitate the
creation of a new leadership education program at Dominican University of California focused on aiding small
businesses in achieving a greater degree of overall business performance through leadership development and
training as well as raising overall awareness of the value added by these same disciplines. Research will also be
aimed at assessing the current unmet needs of small business proprietors and entrepreneurs for leadership
development and assessing currently available resources in the greater San Francisco Bay Area. The creation of the
needs assessment tool was guided by current leadership best practices in business and a comprehensive review of
existing needs assessment tools for small businesses, with particular focus on California and Marin County.
Assessment of current leadership needs is being determined through the use of surveys, focus groups and
strategically targeted interviews with small businesses and entrepreneurs in Marin County. Each member of the Marin
County Hispanic Chamber of Commerce is being electronically surveyed with a questionnaire designed to assess
current leadership training/development needs. Following the surveying, four focus groups will be randomly selected
from the respondents, each consisting of six to eight individuals. In addition, 15 randomly selected respondents will
complete in-depth interviews. Recognized needs will be used to help pilot a new leadership development program,
including the curriculum and pedagogy for courses, within the Institute for Leadership Studies at Dominican University
of California.
159
BUSINESS: MARKETING
ROLE OF APPEARANCE ON SALES PERFORMANCE VIA STATE OF MIND
Lindsey LaShell (Dr. Sri Seshadri), Department of Marketing & MIS, University of Nebraska at Kearney, Kearney,
Nebraska 68849
The purpose of this study was to show how an improved appearance (through dress and grooming) leads to better
performance through changes in self-confidence, mood and anxiety. A pre and post test methodology was used to
assess the influence of appearance on task performance. Participants were college students in an upper level sales
management class (8 males and 9 females). On Tuesdays students were allowed to wear whatever they liked;
Thursdays students dressed in “business attire” to receive extra participation points. A Tuesday and a Thursday were
selected for the students to give their sales presentations. Students self-reported their appearance, mood,
self-confidence and anxiety levels each day before they gave their presentations. Two external judges assessed
subject’s performance of the sales presentation (performance task). The average scores were found for each
construct by summating the items scores for each individual and then dividing by the number of items in that
particular scale. To find the final score for performance, the average performance scores from each item (from two
judges) for a given participant were first averaged and then using the summated score for each student, the average
performance score was computed. The average mood, self-confidence, anxiety, appearance and performance were
calculated for both Tuesday’s and Thursday’s sales presentations. Results show that the correlation between
appearance and performance is established through mood and self-confidence. By improving their appearance
(dressing up and grooming), participants experienced increased levels of self-confidence, and reported being in a
better mood. Because of the changes in self-confidence and mood, performance of the participants improved and
they received higher scores on the evaluations of their sales presentations. Surprisingly, in this study appearance did
not have a significant impact on anxiety, nor did anxiety have a significant direct impact on performance.
WHAT`S IN A NAME?: A CONTENT ANALYSIS OF JOB TITLES IN AUTOMOBILE SALES AND THEIR
EFFECT ON CONSUMER PERCEPTIONS
Zachary C. Ames (Dr. Kenneth R. Bartkus and Dr. Stacey Hills), Department of Business Administration, Utah State
University, Logan, Utah 84322
Marketing scholars and professionals generally believe that when job titles project a dated, inappropriate, or negative
image, they should be changed. Examples include “stewardesses” (now called flight attendants) and “mailmen” (now
called postal or mail carriers). One job title that appears to have a negative connotation is that of the automobile
salesperson. Since there is no known data outlining the extent to which this job title has changed, this research
sought to examine this issue. First, content analysis of business cards from 92 separate new car dealerships was
used to determine the amount of variance in job titles. The results indicate that the most common job title used was
“consultant” (34.8%), the second most used title was a departmental designation such as “sales” or “sales and leasing”
(32.6%), and the third most common title was simply no title at all (17.4%). No other job title accounted for more than
5 percent of the total. The next step was to then examine the extent to which the use of the most common title, sales
consultant, would elicit more positive attitudes about automobile salespeople than the traditional title of “salesperson”.
The results of the experimental design indicate that substituting the term “sale consultant” for “salesperson” does not
result in more favorable attitudes. That major implication seems to be that if the intent of the change is to create
more favorable impressions on the part of public, the strategy may be ill-advised.
160
BUSINESS: MARKETING
ASSISTING BATH & BODY WORKS IN ITS DESIRE TO APPEAL TO THE COLLEGE STUDENT
MARKET (MARKETING RESEARCH)
Emily Briegel, Jessica Henry, and Brannen Morris (Dr. Beheruz N. Sethna), Department of Marketing and Real Estate,
University of West Georgia, 1601 Maple Street, Carrollton, Georgia 30118.
The objective of this project was to provide Bath & Body Works of Carrollton, Georgia assistance in designing appeals
that would be effective in attracting the college/university student market. The data was gathered using an innovative
“Marketing Carnival” method; it is a way of making the collection of data appealing to the respondent. The Carnival
approach helped attract and engage respondents through games and prizes. Respondents filled out a questionnaire
and played games that corresponded with questions. The questionnaire was constructed in conjunction with Bath &
Body Works’ management; the goal being to find the most relevant data to answer the client’s questions. Several
issues that were to be researched included: compelling factors for customers; the level of influence an attractive
display has; whether leaving store doors open attracts customers; frequency of visits and the influence that has on
other variables; and for whom customers normally buy products. The data was compiled and analyzed using SPSS. In
addition to meaningful descriptive statistics, statistical tests were done of differences between means to determine the
existence of relevant segments. In addition, correlations and Chi-Square / contingency table tests tested for
relationships between variables. A few important findings – all of which were significant at the 0.000 level – were:
Females constitute the primary heavy user segment; customers who shop for themselves visit significantly more than
those who buy for others; and Bath & Body Works’ tactics of keeping their doors open and using attractive displays
positively influence customers. The research and conclusions were given to Bath & Body Works to help improve their
overall store efficiency—with specific regard to future marketing campaigns.
Light Activated Materials
Chris Rider (Author), Dan Engebretson (Research Sponsor), University of South Dakota, 414 Clark St. Vermilion, SD
57069
The purpose of this project was developing a source of economic growth within the state of South Dakota by using
new technologies based on light activated materials. These materials (called photo-catalysts) are titanium dioxide
based substances that, when exposed to ultra-violet light and water, react with organic matter in their surroundings.
During this reaction the surface to which these chemicals has been applied is cleaned, sanitized, and deodorized in
an eco-friendly manner. In order to create an economic generator we had to determine how to introduce these
products into the market place. Through market research we determined that because we are based in the rural
Midwest a good market for us to capture would be that of the cleansing of livestock waste ponds. Our chemistry
department is developing the floating photo catalyst that will eliminate ammonia, methane, bacteria, and other toxic
substances in these ponds. Through funds won in a recent idea competition on campus, we are researching and
determining the proper market niche and size, pricing, and distribution. The development of this company and
technology is a large project that will take at least a year to complete; however we have made strong strides toward
our goal in this first month of our operation and I see no reason that our research and development of bringing these
products to market would be impeded.
161
CHEMISTRY
PROGRESS TOWARD THE SYNTHESIS OF A NOVEL, NATURALLY OCCURING ANTIMICROBIAL
AGENT
William Walkowicz (Dr. Aaron Monte), Department of Chemistry, University of Wisconsin-La Crosse, 1725 State St,
La Crosse, WI 54601
The ability of microbes to evolve rapidly and the continued misuse of antibiotics have rendered many of the standard
treatment regimens for infectious diseases ineffective. Current drug discovery methods employed by the larger
pharmaceutical companies in recent decades have yielded few new lead compounds in the area of anti-infectives.
Thus, our research efforts have been directed toward the discovery of novel antimicrobial agents from nature using
more traditional research methods. In this work, a unique Gram-positive-selective antimicrobial agent was discovered
in the leaves of the woody fern Comptonia peregrina. The agent was found to be a substituted phenoxystyrene. We
report here our initial preparations of the unsubstituted scaffold structure, as well as additional analogs of the natural
product, using a combination of modified procedures found in the literature. In general, styrene oxides are combined
with phenols and then dehydrated under acidic conditions. Our approaches to the preparation of various substituted
styrene oxides as starting materials will also be discussed.
SYNTHESIS OF SELECTIVE SEROTONIN 5-HT2A AND 5-HT2C RECEPTOR AGONIST
Dani M. Schultz (Dr. Aaron Monte), Department of Chemistry, University of Wisconsin-La Crosse, La Crosse, WI,
54601
Serotonin (5-hydroxytryptamine, 5-HT), a major neurotransmitter of the central nervous system, is involved in a wide
range of neuronal processes. Some of these include the regulation of mood, sleep cycles, awareness/consciousness,
and memory. To initiate specific responses, serotonin typically binds to an appropriate 5-HT receptor population and
initiates a chain of biochemical events that alters the brain`s physiology in some way. The 5-HT2 receptor is one type
of 5-HT receptor and consists of at least three distinct subclasses, 5-HT2A, 5-HT2B, and 5-HT2C. In previous work in
this laboratory, a serotonin agonist was prepared that showed high affinity for 5-HT2A/2C receptors but did not show
selectivity for either one of the two. We present here the synthesis of two new agonist ligands that may have
differential affinities for the 5-HT2A or 5-HT2C receptor subtypes. The eight-step synthesis was begun with
commercially available para-methoxyphenol, which was modified by introducing bromoethyl and bromopropyl groups
to the aromatic oxygen atoms. A tandem ring closure using n-butyllithium was then performed to create the key
tricyclic intermediate. Formylation of this asymmetric intermediate gave two regioisomers, which were separated
chromatographically and carried through a parallel sequence allowing final elaboration of the desired alkylamine
sidechains. Subsequent pharmacological analysis of the two target molecules will provide a more detailed depiction of
the 5-HT2A/2C agonist binding site.
ONIOM CALCULATIONS OF GRAPHITE SURFACES AND ROLLED GRAPHITE SURFACES
Ellen Brown (George Heard), Department of Chemistry, The University of North Carolina at Asheville, Asheville, NC
28804
Nanotubes have proved valuable in the rising field of nanotechnology because they can be used as an electron
source in portable X-rays, or possibly for the electronic wiring in a microcomputer. The research here will aid in
determining how to overcome the defects which currently hinder the control and growth of these nanotubes. In the
current field of nanotechnology, defects in single-walled carbon nanotubes affect their growth and termination
patterns. The intent of this project was to research graphite surfaces and rolled surfaces which would represent a
defect free nanotube, using computer programs to build models, and then optimize the geometries of these models.
Gaussian 03 was used to build a graphite surface, and a spreadsheet of coordinates was transferred into the
programs to build a tube of one hundred and sixty carbons. The ONIOM technique allowed very large calculations to
be run in less time as the carbon structures were divided into three layers of high, medium, and low levels of theory.
Using the ONIOM theory with Hartree Fock, PM3, and Semi-Empirical methods, respectively for high, medium, and
low, enabled a more versatile scrutiny of the size-able carbon structures. From these calculations, strict patterns of
angles and bond lengths were observed as the position of the bond or angle was closer or further from the edge of the
structure.
162
CHEMISTRY
EFFECTS OF 3`-DANGLING NUCLEOTIDES ON THE STABILITY OF SIRNA-LIKE MODELS
Jennifer L. McGinnis (Dr. Maria Nagan), Division of Science, Truman State University, Kirksville, MO 63501
RNA interference (RNAi) has become an extremely important tool for studying gene function. RNAi depends upon
small duplexes with two 3´-dangling bases on either end of the RNA duplex (Hannon, G. Nature, 2002, 418, 244).
Recently thermodynamic data has been collected that suggests that these 3’ overhanging nucleotides significantly
affect the stability of siRNA-like molecules (O’Toole et al., RNA, 2005, 11, 512). When the first dangling base is a
purine, it increases stability of the duplex regardless of the nature of the second base. However, when the second
single-stranded base is a purine, the duplex is less stable than if it was a pyrimidine. To rationalize these experimental
findings, molecular dynamics studies comparing structural and dynamic properties of duplexes containing various
3´-dangling ends were conducted using AMBER 8.0. Molecular dynamics simulations of ten models with GGCC cores
and varying purine and pyrimidine combinations of 3´-dangling bases were performed. All simulations were conducted
in octahedral boxes of explicit water with sodium ions to neutralize the system. Production runs of 4.0 ns were
obtained for all ten models. Convergence of simulations was evaluated by root mean square deviations from starting
structures and by monitoring various helical parameters. The data generated has been analyzed using root mean
square distances, 2-dimensional base stacking plots, and free energy parameters. Initial data indicates that there are
interactions occurring between the two dangling bases as well as interactions between the second base and the
phosphate background.
PSEUDOROTATION OF TRIGONAL BIPYRAMIDAL STRUCTURES FORMED DURING THE
HYDROPEROXIDOLYSIS OF A VX-LIK
Ashley N. Jay (Dr. Eric V. Patterson), Division of Science, Truman State University, Kirksville, Missouri 63501
VX, O-ethyl-S-(2-(diisopropylamino)ethyl) methylphosphonothiolate, is a highly toxic nerve agent that irreversibly
inhibits acetylcholinesterase. VX is a V-series nerve agent, which indicates that it is a non-volatile liquid and also
relatively unreactive. This means that VX is difficult to neutralize, but neutralization is achieved if the P-S bond is
cleaved. When VX is attacked by a nucleophile, a trigonal bipyramidal (TBP) structure is formed and P-O or P-S
bond cleavage occurs. The use of hydroxide is known to result in both P-S and P-O bond cleavage, with the product
from P-O cleavage remaining toxic. However, the use of hydroperoxide, an alpha nucleophile, leads to exclusive P-S
cleavage and complete neutralization. Attack by either nucleophile leads to the formation of a TBP with the P-O bond
apical. Since cleavage will preferentially occur from an apical position, a pseudorotation must occur prior to P-S bond
cleavage. The pseudorotational potential energy surface of a VX model compound (O,S-dimethyl
methylphosphonothiolate) reacting with hydroperoxide has been explored at the MP2/6-31+G*//MPW1K/MIDI! level of
theory. The energies were corrected using vibrational frequency calculations and energies of solvation. Several
pathways for P-S cleavage were found, but no competitive pathway for P-O cleavage was found.
COMPUTATIONAL CHEMICAL STUDIES OF THE ALKALINE HYDROLYSIS OF VX
Kelly Daniel (Dr. Eric V. Patterson), Division of Science, Truman State University, Kirksville, Missouri, 63501.
O-ethyl S-(2-(diisopropylamino)ethyl) methylphosphonothiolate (VX), is a nonvolatile liquid chemical warfare agent
that is unusually persistent in the environment. VX is a potent irreversible acetylcholinesterase inhibitor with an LD50
of only 3.5 mg/70 kg man. Exposure to a lethal dose of VX results in convulsions, loss of muscle control and death
through asphyxiation within 15 minutes. Neutralization of this deadly nerve agent requires cleavage of the P-S bond.
Hydrolysis in alkaline solution results in both P-S (87%) and P-O (13%) bond cleavage, with the product from P-O
bond cleavage remaining highly toxic. Previous studies on a model compound (O,S-dimethyl
methylphosphonothiolate) reveal that attack of hydroxide initially yields a trigonal bipyramidal (TBP) intermediate with
the P-O bond in an apical position. Since it is known that bond cleavage from phosphorus TBPs occurs preferentially
from the apical position and the major hydrolysis product results from P-S cleavage, there must be a process that
allows P-S bond cleavage to compete favorably with P-O bond cleavage. The potential energy surface for the
reaction of hydroxide with VX has been computed at the MP2/6-31+G(d)//MPW1K/MIDI! level of theory, with
corrections added for thermodynamic and solvation effects. Results indicate that hydroxide attack opposite the P-O
bond is slightly favored, but that processes leading to P-S bond cleavage are more favorable than direct P-O bond
cleavage.
163
CHEMISTRY
MOLECULAR DYNAMICS SIMULATIONS OF ESCHERICHIA COLI TRNAPHE
Bryan D. Sitzmann (Dr. Maria C. Nagan), Division of Science, Truman State University, Kirksville, Missouri 63501
Modifications of the four primary nitrogenous bases are quite common in ribonucleic acid (RNA). Nonstandard
nucleic acid bases occur in all organisms. Many people are interested in determining the role of these modifications in
transfer RNA (tRNA) structure and function. Escherichia coli tRNAPhe requires two modified bases,
2-thiomethyl-N6-dimethylallyl adenosine (ms2i6A) and pseudouridine for accurate translation of the genetic code. The
preliminary nuclear magnetic spectroscopy (NMR) structure of the fully modified and unmodified E. coli tRNAPhe
anticodon stem loop has been solved (Cabello-Villegas J. Mol. Biol. 2002, 319. 1015-1034). The purpose of this
project is to observe how E. coli tRNAPhe behaves in solution with and without its modified bases. Four molecular
dynamics simulations of E. coli tRNAPhe with various modified bases have been conducted in explict solvent, the
Parm98 force field and AMBER 8.0. Charges of the modified bases ms2i6A and pseudouridine have been derived.
Production runs of 4.0 ns each have been collected. Convergence of simulations and preliminary analysis of tRNA
structural parameters are evaluated. These studies will provide a better understanding of how nucleic acid base
modifications affect tRNA structure and therefore accurate translation.
COMPUTATIONAL SIMULATION OF HUMAN tRNALYS,3UUU WITH MS2T6A37, MCM5S2U34, AND
PSEUDOURIDINE 39 MODI
Nina E. McCrate, Mychel E. Varner, Katie L. Schembri, and Amanda E. Combs (Dr. Maria C. Nagan), Division of
Science, Truman State University, Kirksville, Missouri 63501
Accuracy in translation of the genetic code into proteins depends upon correct recognition of their cognate messenger
RNA (mRNA) codons by transfer RNA (tRNA) anticodons. Within tRNAs, incorporation of posttranscriptionally
modified bases is common; however, the roles of these chemical modifications are poorly understood. In human
tRNALys,3UUU three modified bases are present—2-methylthio-6-threonylcarbamoyladenosine at position 37
(ms2t6A37), 5-methyoxycarbonylmethyl-2-thiouridine at position 34 (mcm5s2U34), and pseudouridine (&#61561;) at
position 39—two of which, ms2t6A37 and mcm5s2U34, are required to achieve wild-type binding activity of wild-type
human tRNALys,3UUU (Yarian et al., Biochemistry, 2000, 39, 13390). To elucidate the function of each modified
base in the human tRNALys,3UUU anticodon stem loop, molecular dynamics simulations of eight tRNA anticodon
stem loops with different combinations of nonstandard bases were performed. RESP charges for nonstandard bases
ms2t6A, mcm5s2U, and &#61561; were derived and incorporated into the Cornell et al. (J. Am. Chem. Soc., 1995,
117, 5179) force field. From these simulations it was found that ms2t6A37 enhances stacking between itself and U36
and also prevents U36, the anticodon reading the first codon, from flipping out into solution.
COMPUTATIONAL SIMULATIONS OF THE HUMAN IMMUNODEFICIENCY VIRUS REV-RRE COMPLEX
Peri W. Herman (Dr. Maria C. Nagan), Division of Science, Truman State University, Kirksville, Missouri 63501
Many drugs fight Acquired Immunodeficiency Syndrome but due to its high mutation rate, new and different drugs are
needed, requiring a better understanding of how the Human Immunodeficiency Virus works. One area to study is the
involvement of the Rev-rev response element (RRE) complex. Rev, which contains an arginine-rich peptide, binds to
the RRE sequence of mRNA signaling transport of unspliced and partially spliced RNA out of the nucleus. It is
necessary to characterize the arginine side chain dynamics to understand how Rev-RRE works in vivo. To accomplish
this, three models of the complex, were simulated with explicit water and counter ions using molecular dynamics in
the program AMBER and visualized with the program VMD. All simulations were begun from NMR structures
(Battiste, J.L. et al., Science, 1996, 273, 1547) 4.0 ns of data have been collected for all three models. Root mean
square deviation of each model from its starting structure and the average structure indicate simulation convergence.
Atomic fluctuations have been calculated on all three models indicating that the system is fairly dynamic. Water
analysis is currently being run on all three models to find out if ion binding is involved. This research will hopefully
serve as a starting place for the next researcher. Moberly Area community College, Moberly, MO. 65270
164
CHEMISTRY
EXPRESSION, PURIFICATION AND INITIAL CHARACTERIZATION OF HALOBACTERIUM PROLINE
DEHYDROGENASE
Jeremy J. West and Tommi A. White (Dr. John J. Tanner), Department of Chemistry, University of Missouri Columbia, Columbia, Missouri 65211
Nature recycles proline by converting it to glutamate. This 4-electron oxidation process is catalyzed by two catabolic
enzymes, proline dehydrogenase (PRODH) and &#916;1-pyrroline-5-carboxylate dehydrogenase (P5CDH). Inborn
defects in PRODH and P5CDH result in the disorders hyperprolinemia I & II, respectively. These conditions are often
associated with mental retardation, convulsions, and brain disorders. PRODH has also been implicated in
schizophrenia susceptibility, cancer and p53-mediated apoptosis. Despite their importance in human health and
disease, these enzymes have not been extensively studied. Thus, the goal of this research is to characterize the
structure and function of a newly discovered homologue of PRODH found in Archaea, which we identified by
bioinformatics analysis of genome sequence data. Archaea are also genetically more closely related to eukaryotes
than bacteria, so study of their proteins may provide insights into homologous eukaryotic enzymes. The PRODH
researched here is from the Halobacterium (salt-loving), which can be found in places like the Dead Sea and Great
Salt Lake. Preliminary results include testing the expression of Halobacterium PRODH (YusM) in two different E. coli
expression systems, BL21(DE3)pLysS and Rosetta2. The latter strain was used to account for rare codon usage by
Halobacterium. Parameters varied in these tests included time and temperature of induction as well as
isopropyl-beta-D-thiogalactopyranoside concentration. YusM was found to be largely associated with the cell pellet;
therefore protein purification under denaturing conditions was investigated using urea as a denaturing reagent. Once
the protein was renatured it showed improved kinetic activity. We believe the improved activity is due to disruption of
improperly folded protein by the denaturant, followed by re-folding into the native, or near-native, state. Future
studies will include expression of YusM as a fusion protein with maltose-binding protein in order to improve solubility.
Truman State University
THE SYNTHESIS OF META-CHLORO- AND META-BROMO- ANALOGS OF CAPSAICIN: POTENTIAL
NEW ANALGESIC AGENTS
Katherine M. Phalen, (David B. Rusterholz), Department of Chemistry, University of Wisconsin-River Falls, 410 S.
Third St., River Falls, WI 54022
Capsaicin is the substance in hot chili peppers that is responsible for causing the burning pain sensation for which
these peppers are so well known. When a capsaicin molecule comes in contact with one of these sensory nerves, it
first attaches to a receptor molecule called TRPV1. The goal of this research project was to create several new
substances that have affinity for the TRPV1 receptor, but do not activate it. It is hoped that biological testing will
reveal that these compounds have the qualities needed in a useful pain-relieving drug. The target structures were
based on the capsaicin structure with several modifications. Most importantly the target structures contained atoms of
bromine or chlorine in the position where capsaicin has a methoxy group. This change was expected to enhance the
pain-relieving qualities of the resulting compounds. The synthesis of the target compounds was accomplished by a
convergent synthetic strategy. The final step involved the formation of a thiourea linkage between an isothiocyanate
precursor bearing the hydrophobic C region and a benzylamine bearing the halogen-modified A region. Details of the
synthesis will be presented and discussed.
165
CHEMISTRY
NEW BUILDING BLOCKS FOR COLLOID-BASED MATERIALS BY IMPRINTING PEANUT SHAPE
Stephanie F. Jones 1,2 (Chekesha M. Liddell 2) 1Depatment of Biological and Chemical Sciences, Wells College,
Aurora, NY 13026, and 2Department of Materials Science and Engineering, Cornell University, Ithaca, NY 14853.
Self-assembly techniques for creating photonic crystals with the ability to affect the propagation of visible light have
become highly attractive. While research on spherical colloids with narrow size distribution has matured, methods
resulting in monodispersed nonspherical particles appropriate for photonic crystals are limited. For this work,
nonspherical colloids were synthesized by the controlled growth of silica coatings on iron oxide peanut-shaped
colloidal templates via sol-gel reaction. Hollow silica shells that maintained the peanut shape were obtained upon
dissolution of the core. By varying the amount of silicon-containing reactant, tetraethylorthosilicate, the thickness of
the generated silica shells was controlled up to 100 nm. Calculations show that nonspherical colloids such as the ones
we have synthesized, when packed in a face-centered cubic structure, form photonic crystals that would produce
complete photonic bandgaps. The bandgaps generated by these nonspherical particles are more tolerant to disorder
arising from self-assembly processes than those associated with crystals composed of spherical building blocks.
Assemblies of these non-spherical particles will provide tunability of the bandgap width and the spectral position. Both
depend on the volume fraction of silica in the photonic crystal, partially determined by the shell thickness, a
parameter that we have been able to control.
DESIGN AND SYNTHESIS OF TRPV1 ANTAGONISTS; PROBING THE D-REGION BINDING SITE
Victoria D. Held (David Rusterholz), Department of Chemistry, University of Wisconsin - River Falls, 410 S. Third St.,
River Falls, WI 54022
TRPV1 (transient receptor potential vanilloid subfamily 1) is a membrane bound ion channel protein that mediates the
pain response elicited by capsaicin, resiniferatoxin, and similar drugs. Structure activity studies of compounds that
bind to TRPV1 indicate several binding sites, namely: 1) an aromatic “A” region, 2) a polar “B” region, and 3) a
hydrophobic “C” region. Recently a fourth region has been proposed based on the structure of resiniferatoxin. This
fourth “D” region is thought to be responsible for the high potency exhibited by resiniferatoxin. A new series of
potential TRPV1 antagonists was designed that incorporates molecular features intended to interact with the known A,
B, and C binding sites. In order to investigate the interaction of the D binding region, amidoalkyl groups were
incorporated into the target structures. The synthesis of several of members of the target series was accomplished
using a convergent synthetic strategy. Ultimately these compounds will be tested for their effectiveness as TRPV1
antagonists.
SYNTHESIS AND CHARACTERIZATION OF BIODEGRADABLE POLYMER MICROCAPSULES WITH
VARIABLE C/N/P RATIOS
Ashley E. Van Genderen (Dr. Paul Miller), Department of Chemistry, The University of Wisconsin-La Crosse, La
Crosse, Wisconsin 54601
The nutritional requirements of aquatic suspension-feeding organisms, such as the caddisfly larva Brachycentrus
occidentalis, comprise an important area of study because of the significant role these benthic organisms occupy
within the river food web. In their natural habitat, Brachycentrus larvae feed on detritus, algae, diatoms, and
microparticles, which range from 30 to 200 microns in diameter. Due to the highly variable composition of the
particles that make up the natural diet of these organisms, it is desirable to have synthetic microparticles which have
been prepared under carefully-controlled conditions. Biodegradable polymer microcapsules with variable carbon,
nitrogen, and phosphorus ratios were synthesized using an interfacial dispersion polymerization process which
consisted of an aqueous carbohydrate phase (dextrin in a 1 M aqueous NaOH solution) dispersed in a
cyclohexane/dichloromethane phase. The dispersion was stabilized using soy lecithin and the particles were
cross-linked using 1, 3, 5-benzenetricarboxylic acid chloride. Microcapsule size was determined by analyzing digital
microphotographs with the aid of a commercial computer program that measured the cross-sectional area of the
particles. Microcapsule size was determined as a function of the wt % of dextrin present in the aqueous phase,
cross-linking density, and the amount of stabilizer present in the polymerization system. Microcapsules synthesized
ranged in size from 72 to 190 microns in diameter. In general, an increase in the amount of cross-linking agent led to
a decrease in microcapsule size. In contrast, an increase in the amount of dextrin in the aqueous phase resulted in
microcapsules with larger diameters. Increasing stabilizer concentration led to an increase in particle size, up to a
maximum, after which additional stabilizer resulted in smaller microcapsules.
166
CHEMISTRY
DETERMINING THE MANGANESE CONTENT IN AQUATIC PLANTS USING EPR
Megan J. Gootee (Bryan Lynch, Donald Batema) Department of Chemistry, University of Evansville, Evansville,
Indiana 47722
Aquatic plants can absorb heavy metals and have been suggested as a possible way to manage heavy-metal toxicity
in aquatic media. Determining the amount of heavy metals absorbed by aquatic plants is usually performed with
atomic absorption, or similar spectroscopies. We have used Electron Paramagnetic Resonance (EPR) spectroscopy
to examine the manganese (II) ion content in aquatic plants in a local wetlands. We have found EPR to be a much
easier method for determining the manganese (II) content compared to atomic absorption spectroscopy, since EPR
does not require digestion of the plant leaves; the leaves are simply chopped in a blender, placed in a quartz EPR
tube, and studied at liquid nitrogen temperatures. On three separate occasions, we sampled yellow crowfoot
(Ranunculus flabellaris Raf.) at a multitude of locations in the Heim Wetland located in Warrick County, Indiana.
Yellow crowfoot is an obligate, found only in the wetland environment, either floating or submerged. The relative
manganese content in each plant sampling was determined by comparing the intensity of the manganese EPR
spectrum to that of a ruby crystal placed permanently within the EPR sample cavity. The manganese (II) content was
monitored with respect to position and depth of water in the wetlands. We also studied the presence of manganese
(II) in the leaves of a variety of tree species growing both in the wetland and in the adjacent upland. In most cases we
found that trees growing in the wetland contained more manganese than those in the upland.
SUCCESS OF LIMING A “SINKING STREAM” VERSUS A PERENNIAL SURFACE STREAM
Holly M. Tuck, Carla R. Landes(Dr. Daniel M. Downey), Department of Chemistry, James Madison University,
Harrisonburg, VA 22807
Mountain Run is located in the George Washington National Forest, part of the Massanutten Mountain Range of
Virginia. Brook trout and other species suffered severe losses in the 1980’s due to atmospheric acid injection (acid
rain). Mountain Run was treated with limestone to restore water quality initially at a location in a lower reach that
flows perennially and later in an upper reach that experiences several sinks into the soil. Water quality parameters
(WQP) were analyzed and fish inventories were taken from 1992 until present time. Key WQP values were as
follows for four locations: upstream of the perennial site, 1.25 km downstream of the perennial site, upstream of the
sinking site and 1.25 km downstream of the sinking site. Average values found were pH = 4.64, 5.73, 4.59, 5.05;
ANC = -22.2, 20.1, -28.2, -6.9 (&#956;eq/L); Ca:H = 1.3, 265.1, 0.87, 9.0; Al= 303, 108, 409, 184 (ppb). The only
location that accommodates trout is the area 1.25 km downstream of the lower reach with a perennial flow. This
study compares the effectiveness of liming Mountain Run in the two locations, and how the response from the upper
reach is less efficient due to the sinks in the stream and the acidic nature of the soil.
167
CHEMISTRY
The Use of Aptamers as a Novel Sensor of APC in Developmental Analysis of Drosophila
melanogaster
Edward Briercheck, National Science Foundation, Department of Biological Sciences, Carnegie Mellon University,,
4400 Fifth Avenue, Pittsburgh, PA 15213
Proteins have shown to be the key to unlocking the cellular mechanisms of organism development. Thus, the need for
efficient and accurate methods for proper identification of proteins and their interactions in vivo has arisen. Using the
fruit fly Drosophila melanogaster as a developmental model, our lab has determined that the Adenomatous polypolis
coli (APC) protein family plays a significant role in development. To determine mechanisms of APC function it is
crucial to be able to localize APC and other protein interactions within subcellular structures without disrupting the
functionality of the endogenous protein. Satisfaction of these requirements may be found in oligonucleotide aptamers.
Aptamers are short single-stranded DNA molecules which specifically bind to the target protein. These molecules
become biosensors through Watson and Crick base pairing interactions with fluorescently labeled peptide nucleic
acids (PNA). The extreme size difference between the 100 base DNA and present sensor methods such as green
fluorescent protein, suggest that aptamers may be less functionally disruptive to the target protein. To select for
aptamers that specifically bind to Drosophila APC2, we expressed and purified the C-terminal half of the protein as a
GST fusion, which was mixed with a synthetically produced randomized DNA pool generated by the systematic
evolution of ligands by exponential enrichment (SELEX) protocol. We then subjected this mixture of APC2 protein
with the DNA pool to nonequlibrium capillary electrophoresis of equilibrium mixtures (NECEEM), allowing us to
definitively separate free DNA, APC2 bound DNA, and free APC2. Using this separation procedure, we isolated
potential APC2 aptamers. We are currently amplifying these aptamers via PCR and will clone the optimum
sequences. One technical hurdle of aptamer biosensors is the control of fluorescence. The fluorescent difference
between bound and unbound can be determined in vitro using a fluorimeter. Previous aptamers for the protein
thrombin verified this technique. A PNA-linked Dapoxyl fluorophore, showed significantly more fluorescence when
bound to the protein than when free, making this a viable technique for future work on the aptamers for APC2.
PHASE CHANGES IN NEW LAMELLAR ZINC NITROBENZYL PHOSPHONATES
Brent L. Toland, Barbara Reisner, Department of Chemistry, James Madison University, Harrisonburg, VA 22807
Phosphonate ligands have been used to coordinate to metal ions in an effort to synthesize metal-organic frameworks.
These compounds can form microporous crystalline structures that contain large spaces within the crystalline lattice.
Two new zinc nitrobenzylphosponates were synthesized from Zn(NO3)3 &#8729; 6H20 and
diethyl(4-nitro-benzyl)phosphonate by changing the reaction temperature. The product is lamellar and the peak
corresponding to the interlayer spacing is split; reaction conditions affect the splitting. X-ray powder diffraction,
thermogravimetric analysis and differential scanning calorimetry have been used to characterize the materials.
Models explaining the splitting of the interlayer peak will be discussed.
SYNTHETIC PROGRESS TOWARDS A TEMPLATE ASSISTED SYNTHETIC PROTEIN
Karolina Z. Roszak, Ashleigh R. Borges, Christian D. Zeigler, Kevin P. C. Minbiole, and Kevin L. Caran. (Research
Corporation) Department of Chemistry, James Madison University, Harrisonburg VA 22807
Molecular scaffolds are molecules that provide functional groups as points of attachment to chemical groups or
biologically significant moieties. A rigid scaffold can serve as a molecular core that holds its substituents in defined
orientations in space. The relative positioning of these functional groups can have a profound effect on the
aggregation, self-assembly, binding ability of the molecules. Derivatives of these scaffolds bearing small peptides
may also be utilized as enzyme mimics where the enforced juxtaposition of amino acid chains provides a binding
pocket that imitates the active site of an enzyme. Currently, we are in the process of synthesizing two stereoisomeric
molecular scaffolds based on a C3-symmetrical, conformationally-fixed trans-trans-trans fused perhydrophenalene
system. The synthesis of the molecular scaffold will be accomplished by a triple ring-opening-ring-closing metathesis
starting with cyclobutanone. Presently, we are undertaking a model study using cyclohexanone. Progress on our
synthetic efforts will be reported.
168
CHEMISTRY
DETERMINATION OF THE COEFFICIENT OF NEGATIVE THERMAL EXPANSION OF ZrM2O8 (M = Mo,
W)
Nicole A. Brehm (Dr. Barbara A. Reisner), Department of Chemistry, James Madison University, Harrisonburg, VA
22807
An exercise for the physical and materials chemistry laboratory that will enable students to investigate how changing
the temperature can induce changes in cell constants and phase transitions has been developed. ZrM2O8 (M = Mo,
W) were synthesized by precipitating a precursor material from solution and heating a solid state mixture.
Measurements with a differential scanning calorimeter (DSC) and powder X-ray diffractometer were used to
determine the enthalphy change of the solid-solid phase transition and the negative thermal expansion of ZrM2O8.
Successful collection of cell constants and phase transition data was made and characteristics of negative thermal
expansion of the materials were observed.
Synthesis of 3-Methyl-Pentylcyclobutene--Initial Attempts to Capture a Fluorocyclopentadiene on
the Pathway to 1,3-Difluorobenzenes
Michelle B. Curtis (Scott B. Lewis), Department of , Chemistry, James Madison University, Harrisonburg, VA 22807
One of the key intermediates in the proposed mechanism to form 1,3-difluorobenzenes from substituted cyclobutenes
with difluorocarbene is believed to be a fluorocyclopentadiene. To date, this intermediate has proven to be too fast
kinetically to catch and characterize. Rather than try to catch the intermediate on its way to a 1,3-difluorobenzene,
controlling the reactivity of the intermediate is feasible. An appropriately substituted cyclobutene should allow the
fluorocyclopentadiene to be captured and characterized. This paper discusses the synthesis of
3-methyl-3-pentylcyclobutene, and its reactions with difluorocarbene.
EXPRESSION AND REGULATION OF CGRP AND ITS RECEPTORS IN HUMAN ENDOMETRIUM AND
ITS CELL LINE
Priscila Salgado, Senior, Department of Biology and Honors College
This study was part of a grant supported by the National Institute of Health in contribution to the study of
preeclampsia. Preeclampsia is a disorder that occurs only during pregnancy and the postpartum period. This condition
affects at least 5-8% of all pregnancies, and it is characterized by high blood pressure and the presence of protein in
the urine. This study was designed to provide evidence that Calcitonin Gene-Related Peptide (CGRP) serves as a
paracrine and autocrine modulator of human endometrial and
decidual cells. This will support the search of a treatment for the prevention of preeclampsia.CGRP is a 37-amino
acid neuropeptide produced by tissue-specific alternative splicing of primary RNA transcript of the gene encoding
calcitonin. CGRP
acts at the cellular level by binding to a seven-transmembrane domain G-protein coupled receptor. It is the most
potent endogenous vasodilator.
Immunohistochemical studies have shown the presence of CGRP in human decidua, indicating that it may be an
autocrine growth factor for human endometrial endothelial cells and is thus involved in endometrial angiogenesis.
However, the expression and regulation of CGRP and its receptor components, receptor activity modifying protein
(RAMP1) and calcitonin receptor-like receptor (CRLR) in human endometrium, remain
unknown. The present study investigated the hypothesis that CGRP and its receptors were expressed in human
endometrium and regulated by steroid hormones. Ishikawa cells, immortalized endometrial living cells derived from
adenocarcinoma were used as cell models for in vitro investigation.
Immunohistochemical studies revealed that CGRP and its receptor components
CRLR and RAMP1, were identified in glandular epithelial cells in human endometrium, and decidual cells in human
decidual tissues. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated that Ishikawa
cells expressed both CGRP and its receptor components, CRLR and RAMP1 and that mRNA expression for CGRP,
CRLR, and RAMP1 was upregulated by progesterone, but not by estrogen. Furthermore, proteins for vascular
endothelial growth factor (VEGF) and placenta growth factor (PlGF) were expressed by Ishikawa cells. This
expression was enhanced by CGRP treatment and blocked by CGRP antagonist, CGRP8-37.
169
CHEMISTRY
VARIATION OF THE POISONING CONDITIONS OF POLY(3-METHYLTHIOPHENE) COATED
ELECTRODES
Andrea Kelley (Dr. Isam Marwai), Department of Chemistry, Xavier University, 3800 Victory Parkway, Cincinnati, OH
45207
Poly(3-methylthiophene) (P3MT) as an electrode modification has been shown to improve the selectivity and
sensitivity of several important classes of biological molecules including catecholamines. Catecholamines are a class
of neurotransmitters involved in many important processes, and a method that improves their detection could
potentially elucidate more knowledge about their role in these processes. Previous experiments have shown that the
activity of this polymer was significantly hindered by certain substances, including the hexavalent molybdate ion and
anthraquinone-1,5-disulfonic acid, when the polymer was cycled in solutions of these substances. The poisoning
occurred even when the poisoning agent was present in very small amounts. This work further explores the poisoning
phenomenon by examining the interactions of the poisoning agents with the polymer under variety of experimental
conditions. It has been found that the inclusion of trace amounts of a substance such as catechol or ascorbic acid, for
which the polymer improves detection, in the poisoning agent solution during cycling causes the polymer to retain
much of its electrocatalytical activity. The results will include cyclic voltammograms of the cycles in various poisoning
agents under different conditions, as well as scans of catechol taken before and after the cycling of the polymer in the
poisoning agent mixtures.
2,2-DIMETHOXYPROPANE AS POTENTIAL DRYING AGENT FOR GREEN SOLVENTS
Michelle Sullivan (Markus Hoffmann), Department of Chemistry, SUNY College at Brockport, Brockport, NY 14420
The presence of water in organic solvents, specifically ionic liquids and surfactants, can interfere with
water-incompatible reagents in chemical synthesis. Ionic liquids and surfactants are notoriously hygroscopic, and
removal of water impurities can be difficult. In this investigation, 2,2-Dimethoxypropane (DMP) was used as a drying
agent for these green solvents. The drying reaction was studied in several different types of ionic liquids and
surfactants having a broad range of water content, 0.2-2 % H2O by mass. DMP reacts with water, in the presence of
toluenesulfonic acid as catalyst, cleaving the ketal and forming acetone and methanol as products. The volatile
products and remaining DMP reactant were then removed by stripping with nitrogen gas. Water content of most
samples was reduced to a range of 0.04-0.06% water by mass, signifying that DMP is an effective drying agent.
As an extension of our work we considered if our drying procedure could be used to quantitatively determine water
content. The Karl Fischer titration method which is commonly used to analyze water content involves carbon
tetrachloride and other toxic compounds, which are of environmental concern. Since in our method the reaction
products and reactants are stripped off using nitrogen gas, we attempted to analyze water content by mass loss. A
comparison of expected mass loss results as per Karl Fischer titration and actual mass loss were shown to closely
agree.
PROGRESS TOWARDS THE SYNTHESIS OF PYRIMIDODIAZEPINE-BASED FOLATES AS
POTENTIAL INHIBITORS OF GLYCINAMIDE RIBONUCLEOTIDE FORMYLTRANSFERASE
Danielle Shacklady, Morin Frick, Elizabeth Denton, Partha Ray*, Honors College and Chemistry Department of the
University of West Georgia, 1600 Maple St. Carrollton, GA 30118.
Glycinamide ribonucleotide formyltransferase (GARFT) catalyses the reaction between
10-formyl-5,6,7,8-tetrahydrofolic acid and glycinamide ribionucleotide (GAR) to give N-formylglycinamide
ribonucleotide (FGAR). The formyl carbon of FGAR is destined to become the C-8 carbon atom of inosine
monophosphate from which all purine nucleotides are derived. Consequently, GARFT is an attractive target site for
anticancer chemotherapy. We have designed a series of pyrimido[4,5-e][1,4]diazepine-based folates as potential
inhibitors of GARFT. We will report on our progress on developing synthetic methodology to access the
pyrimidodiazepine heterocyclic system and our targets.
170
CHEMISTRY
ENANTIOSELECTIVE OXIDATION OF RACEMIC PHOSPHINES
Jennifer Beierlein (Dr. Ray Baechler), Department of Chemistry, Russell Sage College, , Troy, NY 12180
Enantiomerically pure phospines and phosphine oxides can be used as ligands in many asymmetric syntheses. Due
to this use, the ability to easily isolate pure phospines and phosphine oxides would have a significant effect on the
ability to synthesize other enantiomerically pure products. The goal of this research is to find an oxidizing agent that
will react selectively with only one enantiomer of a chiral phosphine. Ideally, this would result in a 50% conversion of
the racemic phosphine to phosphine oxide and the products will be the enantiomerically pure (or enriched) phosphine
oxide and the enantiomerically pure (or enriched), unreacted phosphine.
A somewhat similar oxidation has been carried out on sulfur compounds through a modified Sharpless Oxidation
which uses titanium isopropoxide, diethyl L-tartrate, and cumene hydroperoxide to carry out the selective oxidation.
Since phosphines and sulfides are similar in some respects, it may be possible to apply oxidizing agents to
phosphines that have been effective on sulfides and obtain similar results. However an important distinction is that
chiral, organic phosphines possess just one reactive electron pair on the phosphorous atom, so selectivity requires
oxidation of just one enantiomer of the phosphine.
Another important distinction is that phosphines are more reactive towards oxidation than sulfides, potentially
making enantioselectivity more difficult to achieve. This fact was taken into consideration in selecting the phosphine
system which will be used in this attempt to explore selective oxidation. Diaryl methyl phosphines are less susceptible
to accidental air oxidation due to the presence of the stabilizing aryl groups, making oxidation easier to control.
Different functional groups can be incorporated onto one of the rings to explore effects on oxidation.Enantiopurity of
the products is being examined through the use the NMR chiral solvating agent, R-1-(9-Anthryl)-2,2,2-trifluoroethanol.
Using a methyl group provides a simplified NMR analysis of the products of the oxidation and the measurement of
enantiomeric composition.
THE SPECTRUM OF CHLOROPLAST DNA MUTATION CHANGES WHEN CELLS ARE GROWN IN
LOW LIGHT
Ngoc Nguyen (Dr. Barbara B. Sears), Department of Plant Biology, Michigan State University, East Lansing, Michigan
4882
This study tested the hypothesis that oxygen radicals produced by photosynthesis contribute significantly to mutations
that occur spontaneously in chloroplast DNA. The type and frequency of spectinomycin resistant mutants of
Chlamydomonas reinhardtii that arise in a low light regime were compared to previously characterized mutations
when cells were grown in constant high light. Amplification of the DNA region of interest by the polymerase chain
reaction (PCR) followed by DNA sequencing enabled the characterization of mutations that occur in the 16S rRNA
gene. A novel base substitution and a recurring 12-bp deletion were observed. The overall mutation frequencies were
similar under the two growth regimes, although the types of base substitutions were different. Under low light, fewer
transversion mutations arose, suggesting that the chloroplast repair system is able to eliminate most of the DNA
damage caused by reactive oxygen. Under high light and continuous photosynthetic activity, the oxygen radical
byproducts of photosynthesis may overwhelm the repair system.
SYNTHESIS AND CHARACTERIZATION OF TETRA [3-(3, 4-DICHLOROPHENOXY)] PORPHYRIN
Tiffany A. Rudeseal, Gretchen M. Rudeseal, Misty J. Bailey, John T. Hubbard (Cynthia P. Tidwell and Ken Belmore),
Department of Biology, Chemistry, and Mathematics, University of Montevallo, Montevallo, Alabama 35115
Tetra [3-(3,4-dichlorophenoxy)] porphyrin, TDClPP, was prepared using a condensation reaction of pyrrole and 3(3,4-dichlorophenoxy) benzaldehyde in an acid solution. The reaction mixture was refluxed for 30 minutes and
allowed to cool to room temperature. A sodium bicarbonate solution was used to neutralize the reaction mixture.
Vacuum filtration was used to collect the porphyrin which was then purified by column chromatography with silica gel
as the packing material and chloroform as the eluent. The purified porphyrin was characterized by 1H NMR, 13C
NMR, electronic absorption spectrophotometry, and fluorescence spectrophotometry. The absorption spectrum of
TDClPP showed a characteristic spectral pattern similar to those of tetraphenylporphyrin. TDClPP exhibited a Soret
band at 419 nm and four Q bands were observed at 515 nm, 549 nm, 589 nm and 646 nm. A strong fluorescence
was observed at 650 nm and a weak fluorescence at 703 nm. Other characterizations of the base porphyrin are
currently being carried out these include mass spectroscopy and CHN analysis. The zinc, copper, cobalt and nickel
complexes of this porphyrin have been isolated and characterized by electronic absorption spectroscopy. The
manganese complex is currently being synthesized.
171
CHEMISTRY
THERMODYNAMIC PROPERTIES OF NOVEL FLUORINATED COMPOUNDS AND THEIR MIXTURE AS
CHF2Cl (R-22) ALTERNATIVES FOR STRATOSPHERIC OZONE PROTECTION
Chris Blaszkowski, Fene Richemond and Ismail Kul, Provost Grant-Widener University, Department of Chemistry &
Biochemistry, Widener University, One University Place, Chester, PA 19013
Thermophysical properties including boiling point, critical tempreature, heat of vaporization at the normal boiling point
and ideal gas heat capacities of fluorinated ether, sulfur and hydrocarbon mixtures are presented. The coefficient of
performance (COP) for the mixtures is estimated and reported at two different application conditions for these
mixtures from thermodynamic literature data using methods of Morrison and McLinden (1986) along with a developed
mathematical model which requires only boiling point, critical temperature, heat capacity, heat of vaporization and
vapor pressure data. The relative COP values are reported as comparison to that of R-22. The relative COP values
of mixtures and pure compounds range from 0.511 to 1.132. The best COP values relative to those of R-22 are for
CH3CH2F with the values, 1.132 and 1.124 for both low temperature and high temperature applications respectively.
The alternatives identified in this work that have the highest potential as an R-22 replacement are CF3OCF2H, the
binary mixtures of CF3SCF3/CH3CF2F and the ternary mixtures of CF3OCF2H/CF2H2/CH3CH2F,
CF3OCF2H/CF2H2/CH3CF2H and CF3OCF2H/CF2H2/CF3CH2F.
FABRICATION OF GOLD COLLOID FILMS ON GLASS SUBSTRATES FOR RAMAN STUDIES OF
ALKANETHIOL SELF-ASSEMBLED MONOLAYERS
Leila J. Roberson,* (Brian W. Gregory),* Sarat Shanmukh,** and Richard A. Dluhy**, *Department of Chemistry,
Samford University, Birmingham, AL 35229-2236, **Department of Chemistry, University of Georgia, Athens, GA
30602-2556
In order to experimentally observe the vibrational spectra of self-assembled monolayers (SAMs) of alkanethiols
(CH3(CH2)nSH, where n = 1,2,3,etc.) by Raman spectroscopy, the use of microscopically rough gold or silver
surfaces are often necessary in order to provide sufficient enhancement of the Raman scattering rate. These types of
surfaces can often be produced by chemically attaching gold colloidal particles to glass slides (or related materials).
In this regard, gold colloid films were prepared on glass slides pretreated with the silane-coupling reagent
(3-mercaptopropyl)trimethoxysilane (MPS). MPS allows the attachment of small metal particles via the reactive
mercapto moiety at the propyl chain terminus, and hence can be used to irreversibly bind gold colloid from solution.
Overnight immersion of the MPS-coated slides in various concentrated gold colloid solutions (having average optical
densities of 1) resulted in adhesion of gold metal colloids to the MPS layer. Ultraviolet-visible (UV-vis) absorption
spectroscopy was implemented to demonstrate the attachment of gold colloid to the surface and the extent of colloid
aggregation. High density, highly aggregated gold colloidal films covered with alkanethiol SAMs were produced by
alternating exposures of the gold colloid slide to a solution containing 1.00 X 10-3 M decanethiol (CH3(CH2)9SH =
C10SH), followed by re-immersion in the original colloid solution. Following this, surface-enhanced Raman
spectroscopy (SERS) was used to obtain vibrational data of the C10SH SAM deposited on the gold colloidal films.
UV-vis absorption data of the aggregation state of the gold colloidal films, and SERS spectra of the C10SH SAMs, will
be presented.
(http://www.samford.edu/groups/honors/leila/NCUR_Abstract_Roberson.doc)
172
CHEMISTRY
THE EFFECTS OF MULTIVALENT CATIONS ON THE PHASE DIAGRAMS OF LYOTROPIC
CHROMONIC LIQUID CRYSTALS
Kelly S. Antion (Dr. Oleg D. Lavrentovich), Liquid Crystal Institute and Chemical Physics Interdisciplinary Program ,
Kent State University, Kent, Ohio 44242
Lyotropic chromonic liquid crystals (LCLCs) represent a distinct class of liquid crystals formed in aqueous solutions by
molecules with rigid polyaromatic cores and ionic groups at the periphery (usually deprived of aliphatic tails). The
LCLCs attract a considerable interest because of their potential applications as an amplifying medium in biological
sensors. The mechanisms responsible for their mesomorphic behavior are far from being understood well. It is
known that molecular self assembly with face-to-face stacking of the aromatic cores results in one-dimensional
aggregates with the ionic groups exposed at the aggregate/water interface. These aggregates serve as the building
units of the LCLC phases but it is not known whether the interactions among them are of a pure entropic nature or
involves electrostatic interactions of the ionic groups. If the latter is of any importance, then the phase diagrams of
LCLC should be altered by salts. In this work, we study how the phase stability of the LCLCs is influenced by salts
with multivalent cations, such as MgSO4, spermidine, a flexible polyamine molecule with charge 3+, and spermine
(4+). The multivalent cations are chosen because unlike their monovalent counterparts they might alter the sign of
Coulomb interactions of like-charged rods from repulsion to attraction thanks to their strong correlations. We
demonstrate that the phase diagrams of LCLCs are indeed changed dramatically upon addition of the multivalent
cations. Depending on the LCLC structure, they cause expansion of either the nematic or isotropic phase, often by
tens of degrees. Moreover, the multivalent cations also cause condensation of LCLC aggregates into domains of a
new phase with birefringence higher than that of the normal nematic phase. We explain these results qualitatively by
considering how the multivalent cations can alter the interactions within the aggregates and between the aggregates.
University of West Georgia, Carrollton, GA 30118
CHARACTERIZATION OF THIN-FILM GeC FABRICATED BY A HOLLOW-CATHODE PLASMA JET
DEPOSITION METHOD
Ruth N. Udey (Dr. Christopher L. Exstrom and Dr. Scott A. Darveau), Department of Chemistry, University of
Nebraska at Kearney, Kearney, Nebraska 68849, Dr. Rodney J. Soukup and Dr. Natale J. Ianno, Department of
Electrical Engineering, University of Nebra
Raman and infrared spectroscopy and spectroscopic ellipsometry were used to characterize a series of germanium
carbide (GeC) thin films fabricated by a hollow-cathode plasma jet deposition method. Raman spectroscopy
determined the crystallinity and carbon particle sizes present in the films. The crystalline Ge peak and the TO-like
amorphous Ge peak were fit with Gaussian curves. The relative degrees of crystallinity ranging from 3%-79% were
calculated from the Ge peaks. Fits of the carbon D and G peaks using a combination of Lorentzian and
Breit-Wigner-Fano (BWF) curves revealed carbon particle sizes of 8-300 A. The wide ranges were caused by the
non-uniformity of the films. The ellipsometric spectra were modeled to determine film thickness, composition, and
optical constants.
173
CHEMISTRY
TRACKED-IN ARSENIC FROM CCA-TREATED DECKING
Margaret C Ullman (Richard Maas and Steven Patch) Department of Environmental Studies and The Environmental
Quality Institute, The University of North Carolina at Asheville.
Through the course of this study the levels of arsenic tracked into homes via foot traffic from CCA pressure treated
decks will be determined. The wood preservative CCA (chromated copper arsenate) was used universally for deck
treatment until January 1, 2004 when the EPA banned its residential use due to health risks associated with arsenic
exposure. Arsenic exposure can result from ingestion, inhalation, or absorption and can lead to cardiovascular
disease, reproductive, developmental, immunological, and neurological diseases among others. In this study a total of
nine local decks treated with CCA will experience controlled foot traffic by student volunteers. Each of whom will
expose 4ft x 4ft carpet squares, one per site, to the arsenic tracked in on the soles of their shoes. Laboratory analysis
will be preformed on three different types of samples from this carpet to determine levels and rates of arsenic
exposure to the carpet from shoe bottoms. The three sample types are: a dust sample obtained from vacuuming 2ft x
4ft, wipe samples for dislodgeable arsenic over a 1ft x 1ft section, and carpet squares 1in x 1in. All of the samples will
go through extraction and digestion to then be analyzed using Graphite Furnace Atomic Absorption
Spectrophotometry. Results and conclusions will be made regarding the levels of arsenic tracked into homes and the
subsequent health risks associated.
DESIGNING AN INHIBITOR TO UNDERSTAND THE MECHANISM OF THE ENZYME TYROSINASE
Anna Verderame, mentors: Patrick Holland and Timothy Machonkin, NSF and University of Rochester Medical center,
, Department of Chemistry, University of Rochester, RC Box 270216, Rochester, NY 14627-0216
Tyrosinase is an important enzyme in the fields of medicine, food science, and bioremediation because it is efficient.
Unlike other enzymes which require organic cofactors to function tyrosinase requires only O2 and copper in the active
site. Tyrosinase’s binuclear copper active site activates O2 to form a peroxodicopper(II) complex. This complex
oxidizes phenols to catechols. Since there is no crystal structure of tyrosinase questions remain regarding its
mechanism, namely the structure of the transition state, and the chemical environment of the active site, remain. We
are attempting to learn about the active site and the mechanism of tyrosinase by designing inhibitors. The first set of
molecules we are synthesizing mimic the proposed transition state in the literature. These are tested as competitive
inhibitors of phenol oxidation. We are also examining some potential inhibitors that are commercially available and
some that we have synthesized to determine what aspects of a molecule make it a good inhibitor for the enzyme.
Recently molecules having the same geometry as that of the proposed transition have been proposed for synthesis.
There is precedent in the literature for inhibitors that have the same chemical properties as the transition state act to
as competitive inhibitors once the geometry has been corrected. These molecules will also be tested as competitive
inhibitors of phenol oxidation.
We have synthesized 2-hydroxymethyl-pyridone. This molecule was characterized and was tested for inhibition.
It was found not to be an inhibitor of Tyrosinase. We confirmed that the molecule did not decompose in water by
taking an NMR in D20. We have also synthesized 2-hydroxymethyl-5-methylpyridone and N-phenyl hydroxylamine.
We are in the process of characterizing and testing these molecules.
Upper Rim Functionalized Resorcarenes
Amanda L. Robbins (Dr. Frank L. Switzer) Department of Science and Mathematics, Coker College, 300 East College
Ave., Hartsville, SC 29550
Resorcarenes are truly marvelous nanomaterials with a variety of uses. They are readily prepared via a reaction that
regioselectively and stereoselectively forms eight new carbon-carbon bonds. Their synthesis and properties are
mediated by a network of hydrogen bonds that give them their basket shape and facilitate their ability to form dimeric
and hexameric capsules. Resorcarenes have also been elaborated into covalent cages and larger baskets and
capsules. More has not been done with these molecules largely because the reaction that forms resorcarenes is very
limited in scope. My research is focused on ways to form new carbon-carbon bonds at the resorcarene 2-position on
the basket’s upper rim. I am currently studying two approaches to this functionalization: direct electrophilic
substitution of resorcarenes and carbanion chemistry of protected resorcarenes. My main synthetic targets are the
2-formyl and 2-carboxy resorcarenes. They will be key intermediates toward new baskets, capsules, and cages.
174
CHEMISTRY
SITE SPECIFICITY AND EFFECT OF CATION-PI INTERACTIONS IN DNA CRYSTAL STRUCTURES
Mikaela D. Stewart and Tori O`Bannon (Lori Isom), Department of Chemistry, University of Central Arkansas,
Conway, AR 72035
Cation-pi interactions are a type of cation-induced DNA distortion that pulls bases out of the helical stack and into the
major groove where a bound ion or waters of its bound hydration shell interact with the exposed aromatic face of the
unstacked base. Many high resolution DNA structures have been deposited in the PDB since the initial publication
describing DNA cation-pi interactions in 2 DNA and 2 RNA structures (Isom, et al., Biochemistry, 1998). A survey of
the Nucleic Acid Database produced 8 structures with resolution 1.6 &#506; or higher that contain groove-bound
cations but no significant chemical modifications or bound ligands. Each structure’s coordinates were analyzed using
Visual Basic for Application programs written to calculate the distance (d) and angle (a) between the base centroids
and the bound cation. Interactions having distances less than or equal to 5.5 A and angles less than or equal to 55
degrees were considered cation-pi interactions. Using this procedure, cation-pi interactions were detected in 7 of the 8
high-resolution structures analyzed, including the surprising discovery of cation-pi interactions between DNA bases
and monovalent thallium ions (PDB 1JGR) and spermine (PDB 428D). Seventy-six percent of the interactions are
less than 5 &#506; distance with an angle less than 50°. They occur in both the major and minor groove, sequence
specifically, with all four DNA bases. These interactions were found to induce DNA distortion, including base
unstacking. Based on these data, we conclude that cation-pi interactions are common between cations and DNA
bases and that these interactions do, indeed, induce DNA distortion.
Synthesis and Reactions of 10-aza-Housanes
Wade M. Blackmon (Dr. Frank L. Switzer) Department of Science and Mathematics, Coker College, 300 East College
Ave., Hartsville, SC 29550
The double cycloaddition of münchnones with 1,5-cyclooctadiene has proven to be a versatile and efficient method
for the preparation of 10-aza-housanes. A similar reaction with 1,3,5,7-cyclooctatetratene provides a synthetic route
toward highly-strained pentaprismane ring systems which are of interest as fuel sources and explosives. The
10-aza-housanes may also be useful as antiviral agents. My research focuses on
9,11-diphenyl-N-piperonyl-10-aza-housane which has been prepared using methods analogous to those my advisor
has previously published. I plan that this housane will be an effective precursor to the free secondary amine ring
system. It may be further elaborated by nitrogen extrusion to a more highly-strained ring system. I have also
prepared a simple tertiary amine model system to study the removal of the piperonyl group from nitrogen via
acidolysis or hydrogenolysis. Work is now in progress to generate the free secondary amine
9,11-diphenyl-10-aza-housane.
Thiophene - Based Coordination Polymers
Abdullahi Ali (Dr. Christopher Ziegler), Department of Chemistry, The University of Akron, Akron Ohio
“Conducting plastics,” have recently been one of the most interesting subject to the research community because of
the multiplicity of their potential applications in modern electronic device. Due to their stability and structural polarity,
thiophene-based polymers are important among the candidates for a "conducting plastic;" furthermore, the thiophene
molecule is easy to functionalize and a wide variety of thiophene derivatives are commercially available, making the
synthesis of thiophene polymers straightforward and eventually, cost-effective. In this research, we have attempted to
synthesize thiophene- based metal network solids as a precursor to other advanced material. The coordination
polymers of [2, 3-(thiophenecarboxylic acid)] have been prepared with the metal ion Cu(II). Elemental analysis, IR
and x-ray crystallography will be used to characterize the solids. The metal content in the polymer was found to be 1:2
(metal: ligand) stoichiometry. A crystalline of the network polymer containing (Cu + 2-Thiophenecarboxylic acid) was
mounted on an x-ray diffraction and subsequent chemical elucidation flowed.
175
CHEMISTRY
Enthalpy Change of the Chemiluminescent Reaction of Luminol
Nicole Savino, Lisa Morkowchuk (Dr. Carl Salter) Chemistry, Moravian College 1200 Main Street Bethlehem PA
18018
The purpose of this experiment is to investigate the heat of reaction of luminol with hydrogen peroxide. This is a well
known chemiluminescent reaction that produces blue light. The preliminary data suggests that the heat of reaction is
-66kcal/mole. Normally the chemiluminescence escapes from the calorimeter and does not contribute to the
temperature rise. We are investigating whether we can experimentally determine the heat of reaction when all the
light is absorbed inside of the solution.
DETERMINATION OF HEAT CAPACITIES AND THERMAL CONDUCTIVITIES OF LIQUIDS WITH
THERMAL LENS CALORIMETRY
Kacey L. Meaker, Karishma R. Rajani and Ashleigh E. Baber (Kurt Seidman), Department of Chmeistry,
Randolph-Macon Woman’s College, Lynchburg, VA 24503
Thermal lens calorimetry can be used to determine the heat capacities of liquids. This method is much faster than
traditional calorimetry, but it originally produced results with very large errors. The large errors resulted from not
calibrating the procedure. We use the diameter of the thermal bloom as a method of calibration. With this method,
errors of less than 2% were obtained for heat capacities and thermal conductivities. Originally the absorbances of the
samples were kept constant for each calibration line, until it was noticed that the slopes and y-intercepts of the
calibration lines constructed at different absorbances were similar to each other. It was determined that the
calibration line is independent of the absorbance by producing one calibration line over a range of absorbances.
Using this calibration line, the heat capacities and thermal conductivities of methyl acetate, toluene, 1-propanol, THF,
cyclohexene, and acetone were determined. Errors ranging from 0.9% to 1.9% were obtained. Since the calibration
lines are independent of sample absorbance, the time it takes to make up the samples is greatly reduced. Originally,
the bloom diameter was measured visually, and the method required two people. This was very tedious and
somewhat subjective. A new method for measuring the bloom diameters has been developed which is rapid,
objective, and requires only one person to make the measurement. This method was used to produce a new
calibration line over a range of absorbances. Heat capacities of the same unknowns were determined with the new
line, and errors of 1.4%, 1.9%, 1.0%, 2.3%, 0.3%, and 0.5%, respectively, were obtained. The errors obtained from
the thermal conductivity determinations were similar. Using this modified procedure, heat capacities and thermal
conductivities of liquids can be measured more quickly and more objectively than with the original method, and the
accuracy is good.
An Investigation of DNA Damage Induced by the Chemotherapeutic Drug Cyclophosphamide
Akhil Mehta and Jake Smith (Dr. Nolan Carter), Department of Chemistry,University of Central Arkansas,201
Donaghey Ave.,Conway, AR 72035
The purpose of this project was to investigate the mechanism by which cyclophosphamide, a widely used
chemotherapeutic agent, damages DNA. Since cancer accounts for nearly 25% of all fatalities in the United States,
the development of efficacious new treatments for this disease is imperative. A more complete understanding of the
way in which currently used drugs kill tumor cells will facilitate the development of new therapeutic agents. Like many
anticancer drugs, cyclophosphamide results in the covalent linking of tumor DNA strands, a process which inhibits the
growth of tumor cells. This research project investigated the extent to which other chemicals present in cells affect
the efficiency of this process. Glutathione, a compound often present at unusually high levels in tumor cells, is known
to result in inactivation of cyclophosphamide. The extent to which this interferes with the reaction of
cyclophosphamide with DNA is not known. This study used a chemical model system to investigate the competition
between the beneficial DNA damaging reaction and the undesired deactivation of the drug by glutathione. The
reactive species formed from cyclophosphamide in vivo was obtained by chemical synthesis. Imidazole and
2’-deoxyguanosine were used to model the nucleophilic nitrogen atoms found in the DNA double helix. The results
indicate that at high glutathione concentrations, deactivation of the drug becomes significant. This indicates that this
is a significant issue which should guide the design of new treatments.
176
CHEMISTRY
Y-AROMATICITY: WHY THE TRIMETHYLENE METHANE DICATION IS MORE STABLE THAN THE
BUTADIENYL DICATION
Christopher D. Seigfred, (Joel M. Karty), Chemistry, Elon University,100 Campus Drive, Elon, NC 27244
Over the last few decades the trimethylenemethane dication, [(CH2)3C]2+, has received considerable attention due to
indirect evidence suggesting that it possesses a special resonance stabilization known as “Y-aromaticity.” Similar to
aromatic compounds such as benzene, the trimethylenemethane has a fully conjugated &#61552; system containing
Hückel’s 4n+2 electrons. Instead of conjugation about a ring, such as in benzene, the trimethylenemethane dication
is conjugated through a central carbon atom, giving it its Y-shape.
Recently, however, the notion of Y-aromaticity has become controversial. Our research therefore aims to determine
the resonance stabilization in the trimethylenemethane dication in a direct fashion and to compare it to that in its
linear counterpart, the butadienyl dication, [CH2CHCHCH2]2+. To do so, we employed a vinylogue methodology
similar to that which Holt and Karty used to determine the resonance contribution of formic acid’s enhanced acidity
over methanol (Holt, J.; Karty, J. M.; J. Am. Chem. Soc., 2003, 125, 2797). Specifically, we performed density
functional theory calculations to determine the enthalpy to remove two hydride (H&#8254;) ions from methylpropene,
as well as from 2-butene, for which the products are the trimethylenemethane dication and the butadienyl dication,
respectively. Such double hydride abstraction enthalpies were not only calculated for the parent molecules, but also
for their vinylogues, in which 1 to 4 vinyl units were inserted between the central carbon-carbon double bond and each
of the two reaction centers. Double hydride abstraction enthalpies were also calculated for reference compounds,
methylpropane and butane, as well as for their vinylogues. By extrapolation of the double hydride abstraction
enthalpies of the n=1 to 4 vinylogues to n=0 (i.e., the parent compounds), the resonance stabilization in the
trimethylene methane dication was found to be about three times greater than that of the butadienyl dication, thus
providing support for the notion of Y-aromaticity.
PROGRESS TOWARD TANDEM NITRO REDUCTION, IMINE FORMATION, NUCLEOPHILIC ADDITION
IN THE SYNTHESIS OF BETA-AMINO ESTERS
Jamie Garcia, Department of Chemistry, Seattle University, Seattle, Washington, 98122
The environmentally benign synthesis of b-amino carbonyl compounds via tandem nitro-reduction/Mannich reactions
is being investigated. b-Amino carbonyl compounds are biologically and synthetically important compounds, having
wide applications in the synthesis of numerous pharmaceuticals and natural products. A unique feature of the reaction
being investigated is the recycling of indium(III) byproducts of the nitro reduction step as catalysts for imine formation
and subsequent condensation reactions. Each step of this reaction has been explored individually; however,
combining nitro reduction with imine formation and reaction with the silylketene acetal in a one-pot synthesis has not
been reported. We will report our progress investigating this domino synthesis, along with reaction optimization
efforts.
TRANSPORTATION OF MAGNETOSOMES FROM MAGNETOSPIRILLUM MAGNETOTACTICUM (MS-1)
USING BDELLOVIBRIO BACTERIOVORUS (109J)
Anil Sindhurakar (Dr. Eileen Spain), Department of Chemistry, Occidental College, Los Angeles, California 90041
Predator-prey interactions between Bdellovibrio bacteriovorus 109J, Gram-negative bacteria, and another
Gram-negative bacteria Magnetotacticum magnetospirillum MS-1 are studied using Atomic Force Microscopy (AFM).
The main goal of this project is to find a novel method of transporting magnetosomes, biogenic magnetic
nanoparticles, found in the prey bacteria (MS-1) using a predator bacterium (109 J). Bdellovibrios are known to attack
and thrive on other Gram-negative bacteria and the strain 109 J in particular has been found to prey upon wide range
of such bacteria like Escherichia, Pseudomonas, Rhizobium, Chromatium, and Spirillum, to name a few. Thus, this
predator may be envisioned as a potential living cargo carrier. One of the main challenges of this project is that the
109 J and MS-1 strains thrive in completely different environmental and nutritional conditions. MS-1 prefers
microaerophillic conditions in its growth medium while 109 J are aerobic bacteria. We have circumvented this
problem with a unique method to prepare the bacteria for imaging by AFM. Preliminary data and results will be
presented, including nanometer scale magnetosomes of the MS-1, successfully imaged and resolved for the first time
using AFM. If this system is successful, it will provide further information on the nature of bacterial predation at
interfaces as well as a significantly different way to isolate and transport biogenic materials.
177
CHEMISTRY
Inhibition of Calcium Sulfate (gypsum) Crystals using Organic Molecules
Hollis T. Warren (Loyd D. Bastin), Department of Chemistry, Widener University, One University Place, Chester, PA
19013
Many salts, such as calcium sulfate (gypsum) and barium sulfate (barite), are found prevalently in oil well lines and
water treatment systems. These deposits are costly to oil refineries and water treatment plants, as they cause
operational issues and are difficult to remove. Organic compounds have been found to inhibit the crystal growth of
these salts. Previous studies by Roger Davey investigated the effect of organic molecules as inhibitors of barium
sulfate crystal growth. However, few research groups have studied the growth inhibition of calcium sulfate crystals.
In our study, we are synthesizing a group of potential crystal growth inhibitors through the Mannich Reaction from
simple diamines (varying only in length of the carbon chain). Once synthesized, each individual product was
analyzed through proton NMR and melting point determination. The molecules were tested for their ability to inhibit
the growth of calcium sulfate crystals in a supersaturated solution. Many factors, including solution pH, concentration,
and temperature, can be altered in an effort to increase the induction time once a potential inhibitor is introduced to
the system. Ideally, we hope to synthesize an inhibitor that will control growth of all the faces of calcium sulfate over
the longest induction time.
THEORETICAL STUDIES ON ADDUCT FORMATIONS OF PD(II) AND PD(IV) COMPLEXES WITH AN
ENGINEERED OLIGONUCLEOTIDE
Myrlene Jeudy (Beatriz H. Cardelino), Department of Chemistry, Spelman College, 350 Spelman Lane, Atlanta,
Georgia 30314
Metal-based anticancer drugs containing Pt (II) (e.g, cisplatin) are among the most effective drugs used in
chemotherapy. These agents interfere with DNA’s functions and inhibit its ability to divide, thus destroying cancer
cells. However, studies have shown that these types of drugs show limitations because of their lack of specificity in
destroying cells. Thus, it is of interest to investigate other candidates as possible anticancer drugs. In this
investigation the binding effects of metal complexes (analogue to cisplatin) with engineered oligonucleotide were
analyzed using theoretical approaches. The metal complexes considered contained the transition metals Pd(II) and
Pd (IV), with different halide ligands, as well as CO, CN, and OH substituents. The selected oligonucleotide was a
DNA octamer duplex, created by D. Yang et al. in 1995 (Biochemistry, pp.12912-12920), with its structure determined
by high-resolution NMR. The theoretical approach consisted of the hybrid computational procedure called ONIOM.
Within the ONIOM partition scheme, the “active” part of the systems (where chemical bonds break and form) were
treated at the quantum mechanical level using hybrid density exchange functionals (B3LYP and PW91). The “real”
system was treated using molecular mechanics with the UFF and AMBER force fields. Entropy, enthalpy and free
energy for the binding process of these adduct formations were calculated using statistical thermodynamics.
CYTOTOXICITY STUDIES OF 5, 10, 15, 20 TETRA [N-ETHYL-3-CARBAZOYL] PORPHYRIN AND ITS
METAL COMPLEXES FOR POSSIBLE USEFULNESS IN CANCER THERAPY
Leslie A. Alexander, Benjamin F. Cherry IV, (Cynthia P. Tidwell and Casey N. Bassett) Department of Biology,
Chemistry, and Mathematics, University of Montevallo, Montevallo, Alabama 35115
Porphyrin compounds are of interest in many areas of study, particularly in photodynamic cancer therapy. Porphyrins
are substituted aromatic macrocyclic rings that can be found in nature or synthesized in the laboratory. A novel
porphyrin, 5, 10, 15, 20 tetra [N-ethyl-3-carbazoyl] porphyrin, has been synthesized in our laboratory. Because of the
potential use of porphyrin chromophores in photodynamic cancer therapy determination of the porphyrin`s toxicity is
thus very important, both in determining its feasibility for use in living systems, as well as its efficacy in killing cancer
cells upon proper activation. The objective of this research project was to determine if 5, 10, 15, 20 tetra
[N-ethyl-3-carbazoyl] porphyrin and its zinc, copper, cobalt, manganese, and nickel complexes are toxic to cells grown
in culture. Chinese hamster ovary (CHO) cells, a common cell line used in cytotoxicity studies, have been treated with
varying concentrations of each porphyrin to determine its the lethal dosage, the dosage kills 50% of the cells (LD50).
Studies are currently ongoing for concentrations that fall below the LD50 to test the efficacy of the porphyrin as a
potential chemotherapeutic agent.
178
CHEMISTRY
THE EFFECT OF A BRITA® FILTER ON THE MINERAL CONTENT OF WATER SAMPLES
Traci L. Andersen, Stephanie D. Goshorn (Dr. Paul Hooker and Dr. Robyn Hyde) Department of Chemistry,
Westminster College, Salt Lake City, Utah 84105
The calcium, sodium, potassium, chloride, and fluoride concentration of municipal water samples was determined
before and after being passed through a Brita® water filtration system. The purpose of this research was to determine
the effect of a Brita® filter on the ion concentration of a typical water sample. The concentration of calcium, sodium,
and potassium cations was determined using atomic absorption spectrophotometry. Results showed that calcium
concentrations decreased, while sodium and potassium concentrations increased when the water samples were
passed through the filter. Chloride and fluoride anion concentrations were analyzed using specific ion electrodes and
ion chromatography. The results from these experiments showed that both the chloride and fluoride concentrations
remained essentially the same in the unfiltered and filtered water samples. Evaluation of the results indicates that the
filtration system contains a cation exchange resin. This research will be used to create an inquiry-based lab
experiment for an introductory chemistry course which will allow students to investigate the operation of a familiar
device by utilizing a variety of instrumental methodologies.
SPECTROSCOPIC INVESTIGATIONS OF NOVEL HOST-GUEST MATERIALS USING MEROCYANINE
DYES IN GAMMA-CYCLODEXTRIN CAVITIES
Jennifer L. Sidun, (Jennifer S. Holt), School of Science, Penn State Erie, The Behrend College, 5091 Station Rd. Erie,
PA 16563
Using host-guest chemistry, new composite materials can be created by inserting a guest molecule into a host
molecule. The new composite materials often have properties that are not seen in the guest or host molecule
individually. This project studied the behavior of host-guest interactions when the environment and concentrations of
the host and guest molecules were varied. The guest molecule used was Brooker’s Merocyanine (4-[(1-methyl-4(1H)
-pyridinylidene)ethylidene]-2,5-cyclohexadien-1-one). This molecule was chosen due to the unique way it responds to
different environments as observed in a UV/Visible spectrum. The host material used was a gamma-cyclodextrin
molecule. Using UV/Visible spectroscopy, the equilibrium constant of the system was measured. The effect of time
and other experimental conditions such as pH, temperature, and UV light exposure, were varied to determine the
effect of each on the complex formation. An equilibrium constant, K, was determined in two ways: using a linear
expression and using a nonlinear regression called the modified Benesi-Hildebrande equation. The calculated values
were 234 M^-1 and 195 M^-1, respectively. A temperature study on the guest-host complex determined that the
complex is not temperature dependent. The wavelength shifts in the complex as the temperature was increased were
consistent with the dye reference wavelength shifts of the respective temperatures. Time and UV light exposure
showed no spectral changes. The outcome of this research increased the understanding of the interactions of
guest-host systems based on their environmental conditions. Further research may develop a more effective way to
use &#947;-cyclodextrin molecules in guest-host systems.
179
CHEMISTRY
SYNTHESIS OF SUBSTITUTED 1-BENZOXEPIN DERIVATIVES FROM ISOXAZOLES
Matthew D. Jackel (Dr. Martin G. Kociolek), School of Science, Penn State Erie, The Behrend College, Erie,
Pennsylvania 16563
The 1-benzoxepin ring skeleton has been found in a variety of natural and unnatural biologically active molecules.
Some examples include pterulone, which shows antifungal activity, and heliannuol C, which has ubiquinone
oxidoreductase inhibiting properties. Currently, there are few methods known to synthesize the highly functionalized
benzoxepins from readily available starting materials. This work focuses on expanding the known methodology for
the synthesis of the benzoxepin ring system. A method to synthesize substituted 1-benzoxepin derivatives by a
tandem ring-opening/cyclocondensation sequence has been developed. The appropriately substituted
3-bromoisoxazoles have been readily opened by ferrous chloride tetrahydrate giving beta-cyanoketones. These
compounds have been found to spontaneously undergo intramolecular condensation followed by a dehydration to
give 1-benzoxepins in good yields. The application of this methodology to a series of 1-benzoxepins will be reported.
This methodology has the limitation of allowing only cyclocondensation with aldehydes giving benzoxepins
unsubstituted at the carbon adjacent to the aromatic ring. An analogous ring opening/cyclocondensation of
3-carboxyisoxazoles was also investigated. In the presences of base, the esters hydrolyze, with the resulting acids
undergoing decarboxylation followed by the opening of the isoxazole ring to give beta-ketonitriles. The resulting
enolate can spontaneously cyclize to give the benzoxepin. The ongoing progress of the application of this method to
the synthesis of a series of benzoxepins will be discussed.
STUDIES TOWARD THE SYNTHESIS OF ALPHA-HYDROXY-BETA-AMINO ACIDS FROM ISOXAZOLES
Robert C. Blanner III (Dr. Martin G. Kociolek), School of Science, Penn State Erie, The Behrend College, Erie, Pa
16563
Isoxazoles have proven to be useful intermediates in the synthesis of a number of biologically interesting molecules.
The focus of this research is the use of functionalized isoxazoles as precursors for the synthesis of biologically
important alpha-hydroxy-beta-amino acids. The importance of alpha-hydroxy-beta-amino acids can be attributed to
their well-documented biological activity. These amino acids have been utilized in the synthesis of a number of
biologically significant compounds including anti-HIV, anti-cancer, and anti-bacterial agents. A key step in the
synthesis of alpha-hydroxy-beta-amino acids involves the functionalization of the monosubstituted isoxazoles. A new
synthesis of isoxazoles substituted at the 3-position from silyl isoxazoles has been developed. The applications of
this to a series of isoxazoles will be reported. While there are numerous methods for the synthesis and opening of
isoxazoles, the chemistry of the carbon-carbon double bond has not been well documented. The desired chemical
reactions to be used to exploit the chemistry of this double bond include oxidations, with a variety of oxidants, as well
as additions of halogens using halogens or hydrogen halides. The results of these investigations will be discussed.
The ongoing progress of our synthetic studies towards the transformation of these functionalized isoxazoles into
alpha-hydroxy-beta-amino acids will also be reported.
MOLECULAR MODELING OF CAROTENES AND RELATED DERIVATIVES
Edward A. Motea, (Dr. Timothy Laher and Dr. Michael Bucholtz), Department of Chemistry, Gannon University, Erie,
PA 16541
The carotenes are a major class of pigments found in plants, microbes and animals and thus are among the most
widespread of natural pigments. They have a role as a secondary light har-vester in the photosynthetic apparatus of
plants, as a precursor to the formation of vitamin A, as an antioxidant and as a colorant for animals and flowers. The
carotenoids are a diverse family with C-20, C-30 and C-40 hydrocarbon molecules. In addition there are varying
degrees of oxy-genation and functional complexity distributed through nature. This study focuses on the mo-lecular
modeling of C-40 hydrocarbon polyenes and C-20 retinals. In this study several C-40 polyenes and C-20 retinals
were geometry optimized using the Spartan molecular modeling software; a subsequent computational analysis (using
the same software) was performed em-ploying the density functional B3LYP method to calculate the ultraviolet/visible
spectra maxi-mum (on the basis of HOMO-LUMO energy gaps obtained) for correlation with published data. This
analysis will provide useful information on determining the structure of new polyenes as well as a better understanding
of the role of carotenes as auxiliary pigments in photosynthesis. In addition, the correlation obtained can then be used
to examine the spectra shifts obtained in the cis-trans isomerization of the retinal system responsible for the
photo-response in vision.
180
CHEMISTRY
THE SYNTHESIS OF AMINO-SUBSTITUTED ALKYLARYLTELLURIDE PHOSPHONIUM SALTS
Jacqueline A. Simson (Margaret E. Logan and Rey A. Sia), Departments of Chemistry and Biological Sciences, SUNY
College at Brockport, Brockport, New York 14420
This presentation describes the successful synthesis and characterization of four electron-rich amino-substituted
alkylaryltelluride phosphonium salts. These compounds will be evaluated as probes of oxidative damage and repair
in yeast mitochondria. This is of interest because damage caused by oxidation in the mitochondria contributes
significantly to many human disorders, including neuromuscular degenerative diseases and cancer. Oxidative
damage is especially prevalent in the mitochondria because it is the site of cell respiration, and reactive oxygen
species are generated during this process. Alkylaryltellurides were chosen because they are known to act as
antioxidants by readily losing an electron under oxidizing conditions and by oxidizing to the telluroxide in the presence
of two-electron oxidants. More specifically, tellurides having amino-substituted aryl rings are most easily oxidized,
thus making them better antioxidants. The presence of a triphenylphosphonium group at the terminus of the alkyl
group of the alkylaryltellurides should result in improved transport into yeast mitochondria, as has been shown by
others for lipophilic cationic derivatives of the antioxidant vitamin E (a one-electron antioxidant). The four
amino-substituted alkylaryltelluride phosphonium salts were prepared in moderate yield in a three-step synthesis from
the corresponding anilines. They were then purified by chromatography and recrystallization, and characterized by
proton, carbon-13, and 2-dimensional NMR spectroscopy.
AMINE SUBSTITUTED MACROMOLECULAR ELECTROLYTES
Brad C. Miller (Dr. Michael B. Ramey), A.R. Smith Department of Chemistry, Appalachian State University, 525 Rivers
Street, Boone, North Carolina 28608
The long term objective of this research is to synthesize and develop a set of functionalized hexaphenylbenzene
polyelectrolytes with the ability to transport a variety of ionic species (H+, Li+, Cl-, etc.). These materials will be
candidates for use in fuel cell and rechargeable battery applications. The initial synthetic target is a
hexaphenylbenzene molecule in which each of the phenyl rings is substituted in the para position with
(N,N-diethylamine)-1-oxapropyl groups. The amine groups can be reacted with acid (HCl, HBr) or alkylhalides
(methyl or ethyl bromide) to provide a single molecule with six positively charged quaternized amine sites.
Production of this molecule hinges on the synthesis of 3 distinct components: a) Diethyl-[2-(4-iodo-phenoxy)
-ethyl]amine, Compound A; b) 4,4’-(3-[N,N-diethylamino]-1-oxapropyl)diphenylacetylene, Compound B; and c) cobalt
octacarbonyl promoted cyclization of the substituted diphenylacetylene into the hexaphenylbenzene molecule.
Synthesis of Compound A has been optimized using a variety of Williamson etherification techniques.
Para-iodophenol was reacted with 1.1 equivalents of 2-chloroethyldiethylamine hydrochloride and an excess of
potassium carbonate. Acetone and a mixed acetone/water solvent system were explored, along with the use of
standard reflux versus microwave promoted heating. The conditions leading to the highest percent yield (80%) and
simplest workup were sealed tube microwave reactions in acetone solvent at a temperature of 100 0C (~ 80 psi) for
30 minutes. Upon cooling, the reaction mixture was extracted using methylene chloride and 3M aqueous sodium
hydroxide, followed by evaporation of the organic solvent. Using this method, approximately 0.5 grams of product
could be isolated in 1 hour. Gas chromatography/mass spectroscopy indicated the iodoamine compound was 98%
pure. With a useable protocol developed for Compound A, attention is now turning towards the synthesis of
Compound B via Sonogashira coupling reactions.
181
CHEMISTRY
SYNTHESIS AND PROPERTIES OF POLY(4’-METHYL-2,5-BENZOPHENONE)-B-POLY(ARYLENE
ETHER SULFONE) MULTIBOLOCK COPOLYMERS
Domonique O. Downing, Charles A. Jones III, (Valerie S. Ashby)
The class of polymer known as poly(paraphenylene)s (PPP) and their derivatives have superior thermal and
mechanical properties, but are difficult to produce at high molecular weights due to oligomer precipitate during
polymerization. Higher molecular weight PPPs that are soluble in polar, aprotic solvents can be produced by adding
lateral substituents to the polymer backbone. Specifically, poly(2,5-benzophenone)s (PBP) are a promising class of
PPP derivatives with high thermal stability and better mechanical properties than other higher performance
thermoplastics such as poly(arylene ether sulfone) (PES). Unfortunately, these materials are unable to produce
tough, flexible films which are often necessary for many applications. The main objective of this project was to
synthesize new materials that possess the excellent thermal and mechanical properties of PBP with the excellent film
forming capabilities of poly(arylene ether sulfone)s. A macromonomer approach was chosen in which two short-chain
polymers are reacted together via nucleophilic aromatic substitution chemistry to form multiblock copolymers. The
two macromonomers used were fluorine-endcapped poly(4’-methyl-2,5-benzophenone)s and hydroxyl-terminated
poly(arylene ether sulfone)s of various molecular weights. The end-functionalized poly(4’-methyl 2,5-benzophenone)s
were synthesized by a catalytic nickel-(0) coupling reaction while the various poly(arylene ether sulfone) were
prepared via nucleophilic aromatic substitution. Subsequent reaction of these macromonomers with each other in a
nucleophilic aromatic substitution reaction yielded multiblock copolymers. The chemical structures, thermals, and
mechanical properties of these new multi-block copolymers were determined by proton, carbon, and fluorine nuclear
magnetic resonance spectroscopy, differential scanning calorimetry and thermogravimetric analysis, and dynamic
mechanical analysis, respectively.
Funding: Research Education Support Program, National Science Foundation: #HRD-0450099, UNC-CH Chemistry
Dept. Start-Up Funds
TOWARD THE SYNTHESIS OF (-)-INCARVILLINE and (-)-INCARVILLATEINE
Jessica R. Chavey (Dr. Karl Peterson), Department of Chemistry, University of Wisconsin-River Falls, 410 South
Third Street River Falls, WI 54022-5001
Recent investigations into the therapeutic properties of the plant Incarvillea sinensis have revealed the presence of a
novel monoterpene alkaloid that was named (-)-incarvillateine. This compound has been found to have
antinociceptive activity comparable to that of morphine, but at slightly lower dosage levels. Additionally, (-)
-incarvillateine is reported to operate through the adenosyl rather than the opiate receptor mechanism. The first total
synthesis of (-)-incarvillateine was reported in 2004. The overall goal of this project was to complete the
enantioselective total synthesis of (-)-incarvilline and (-)-incarvillateine by following a significantly shorter synthetic
scheme. The synthesis of (-)-incarvillateine was broken down into the separate syntheses of the
2,4-diaryl-1,3-dicarboxylic acid core and the bicyclic aminoalcohol, (-)-incarvilline. The 2,4-diaryl-1,3-dicarboxylic
acid core has been successfully synthesized from vanillin in four steps. The key reaction involved a [2+2]
photochemical dimerization of the 4-O-tosyl derivative of ferulic acid. A nine step synthesis of (-)-incarvilline has
been designed starting with (-)-carvone and including a notable Favorskii rearrangement of the O-protected
chlorohydrin derivative of (-)-carvone. The final step of the synthesis assembles (-)-incarvillateine through
esterification of (-)-incarvilline with the 2,4-diaryl-1,3-dicarboxylic acid core.
http://www.uwrf.edu/~kp0b/NCUR2006-JC.doc
182
CHEMISTRY
BIOINFORMATIC AND ENZYMATIC STUDIES ASSOCIATED WITH D-LACTATE AND GLYCOLATE
METABOLISM IN CHLAMYDOMONAS REINHARDTII
Yi Li1 and 2Elizabeth Reese (Dr. Diane Husic1) 1Department of Biology, Moravian College and 2Northampton
Community College, Bethlehem, PA 18018
In unicellular eukaryotic green algae like Chlamydomonas reinhardtii, glycolate is produced as an intermediate of the
oxidative photosynthetic carbon cycle (C2 cycle) or photorespiration. In contrast, the D-lactate accumulates
significantly only under anaerobic conditions in Chlamydomonas in a reaction presumably catalyzed by a soluble
NAD+-linked D-lactate dehydrogenase (D-LDH). However, a number of studies suggest that a single enzyme may be
responsible for the oxidation of both glycolate and D-lactate in some green algae and that the enzyme is linked to
mitochondrial electron transport. Because D-LDHs have not been well characterized in eukaryotes (other than from
simple yeast), the objective of this project was to gain more information about the genes and enzymes associated
with D-lactate metabolism in C. reinhardtii. Preliminary studies utilized a variety of bioinformatics tools to identify
genes related to D-lactate and glycolate metabolism as well as to predict structural information about the putative
soluble D-LDH protein (molecular weight, pI, amino acid composition, etc.). The gene for the NAD+-linked D-LDH
appears to contain 11 coding regions and the predicted protein sequence has significant homology (46 to 51%) with
D-LDHs and 2-hydroxy acid dehydrogenases from cyanobacteria, H. influenzae and E. coli. The putative
GDH/D-LDH gene has 23 exons and encodes for a much larger protein that has homology with FAD binding proteins
and Fe-S proteins. The subunit molecular weight for the NAD+-linked D-LDH is predicted to be 45.6 kDa; however,
data from nondenaturing polyacrylamide gels and activity staining suggests that the enzyme exists as a tetramer in
vivo. This is unusual since many D-specific LDHs exist either as monomers or dimers. Results from these studies
provide direction for designing protein purification strategies as well as insights into the phylogenetic relationships
between C. reinhardtii (and other Chlorophycean algae) to both prokaryotes and higher plants.
MOLECULAR MECHANICS STUDY OF CHEMISORBED HYDROGEN ON FULL MODELS OF CARBON
NANOTUBES AS A FUNCTION OF RADII, GEOMETRY, AND HYDROGEN COVERAGE
Nicole L. Vaccariello (Dr. Ron Brown), Department of Chemistry, Mercyhurst College, Erie, Pennsylvania 16546
Recent studies have shown that carbon nanotubes adsorb significant amounts of hydrogen. High adsorption levels
could lead to the use of hydrogen in vehicular fuel cells. Some models have suggested that hydrogen chemisorption,
the direct formation of C-H bonds, is necessary to explain the high adsorption levels. We have performed a series of
molecular mechanics calculations in order to investigate the dependence of hydrogen chemisorption upon tube
geometry and adsorption pattern. Full circumference nanotube models with capped ends were constructed with zigzag
and armchair geometries. The radii of the tubes ranged from 2.37 to 4.10 Ångstroms. Hydrogen coverage, using a
variety of coverage patterns, was investigated on (6,0) zigzag nanotubes of varying lengths. The MacroModel
software package was used to perform MM3 energy minimizations. Results include the effects of tube radius,
hydrogen coverage amounts, and coverage pattern on the relative strengths of the C-H bond. In general, it was found
that tubes with more than 50% hydrogen coverage were unstable and that 100% chemisorbed hydrogen coverage is
improbable. An exceptionally stable hydrogen coverage pattern was found when hydrogen was bonded to every other
carbon on the tube. Ab initio calculations were performed on models with this pattern to further investigate the
strength of the C-H bonds.
183
CHEMISTRY
MATRIX ISOLATION INFRARED SPECTROSCOPY OBSERVATION OF THE AZIDE RADICAL (N3)
FROM A NITROGEN MICROWAVE DISCHARGE PLASMA SOURCE
Christopher T. Dine, Jessica R. Collier, Nancy L. Saxton, and Roy M. Schleicher (Jay C. Amicangelo), School of
Science, Penn State Erie, The Behrend College, Erie, PA 16563-0203
The asymmetric stretching band of the azide radical (N3) at 1657.5cm-1 has been observed by directing the output of
a nitrogen microwave discharge plasma source onto a CsI window at 12 K using matrix isolation infrared spectroscopy
experiments. The identification and assignment of this band to the N3 radical has been accomplished by performing
isotopic experiments with 15N2, matrix annealing experiments in which the CsI window is warmed to 30 K and back to
12 K, photolysis experiments in which the light from a mercury-xenon lamp is directed at the 12 K sample, and
theoretical calculations (ab initio and density functional theory methods) using the Gaussian 98 program. As well, the
observed N3 isotopic band positions are compared to literature frequencies observed by other experimental methods.
Temperature dependent deposition spectra (10 – 20 K) were also performed in an effort to determine if the N3 radical
is being formed in the gas phase or by matrix surface reactions. The intensity of the N3 band was observed to
decrease with increasing deposition temperature, which is consistent with a gas phase formation mechanism.
DEVELOPMENT OF METHODS FOR ASYMMETRIC INSTALLATION OF A HYDROXYMETHYL GROUP
Amanda Ramey and Kyle Chormanski (Dr. Richard J. Mullins), Department of Chemistry, Xavier Univeristy,
Cincinnati, Ohio 45207
Many endeavors in natural product synthesis require the stereoselective addition of a hydroxymethyl group. While
seemingly straightforward, this has proven to be a very difficult process because of conditions necessary to attain
stereoselectivity. The development of two general methods for achieving this goal is currently underway in these
laboratories. Primarily, chiral, nonracemic acetals derived from formaldehyde are being investigated as electrophilic
sources of a protected hydroxymethyl equivalent. Upon reaction with a Lewis acid, the intermediate chiral oxonium
ion should react in a stereoselective manner with stabilized crotyl nucleophiles. The second method makes use of
enantiopure N-enoyloxazolidinones for directing the conjugate addition of the organocopper reagent derived from
commercially available chloromethyldimethyl-isopropoxysilane. Oxidative cleavage of the alkylsilane under
Tamao-Kumada conditions would reveal the hydroxymethyl nucleus. Our efforts for the development of these
complimentary hydroxymethylation methods will be presented.
METALLATETRABORANES: PHOTOCHEMISTRY AND LIGAND SUBSTITUTION
George A. Hamilton (Dr. Craig M. Davis), Department of Chemistry, Xavier University, Cincinnati, OH 45207-4221
The arachno-octahydrotriborate anion ([B3H8]-) typically acts as a bidentate ligand towards metal atoms. Only one
reported compound shows the anion acting as a tridentate ligand: Mn(CO)3B3H8. This compound was formed from
Mn(CO)4B3H8 by heating or photolysis. We attempted to synthesize new tridentate complexes by a new approach.
Three of the carbonyl ligands in Mo(CO)6 can be displaced by three acetonitrile ligands to form
fac-Mo(CO)3(CH3CN)3. Unfortunately, upon addition of [B3H8]- only two acetonitrile ligands were displaced.
Nonetheless, we obtained a new compound, [Mo(CO)3(CH3CN)(B3H8)], which was characterized by 11B-NMR
spectroscopy. Attempts were made to substitute the last acetonitrile with phosphanes; however, the [B3H8]- was
displaced instead.
We also employed photolysis in our pursuit of new tridentate complexes. Photolysis of a solution of Mo(CO)6 and
[B3H8]- produced the known bidentate complex [Mo(CO)4B3H8]-, with nearly 100% yield. A polynuclear metal
carbonyl, Fe3(CO)12, was photolyzed with [B3H8]-. The 11B-NMR spectrum showed peaks which have also been
observed by our group in the “dark” reaction between [B3H8]- and various metal halides. The chemical shifts are
identical, regardless of metal (transition metal, main-group, or lanthanide) or halide (chlorine or bromine). Our
hypothesis is that these peaks belong to a previously unreported boron hydride. The addition of
bis-diphenylphosphinoethane (“dppe”), a bidentate phosphane, to the above mixtures gave two new peaks, again,
regardless of metal or halide. The observation of B-P coupling suggests that the metal halides mediated the binding
of the phosphane to the [B3H8]-.
Finally, we have studied ligand exchange on copper complexes. Earlier work in our group suggested that both
mono- and bidentate phosphane ligands can displace the halide from [XCuB3H8]- (X = Cl, I). We employed
31P-NMR spectroscopy in attempt to validate these results, and extended this work by using sulfur-containing ligands
(thiocyanate and the pentasulfide dianion).
184
CHEMISTRY
REGULATION OF MAMMALIAN LARGE-CONDUCTANCE POTASSIUM CHANNELS BY HEME
Jacqueline T. Wesolow (Dr. Mark Reynolds), Department of Chemistry, 5600 City Avenue, Saint Joseph’s University,
Philadelphia, Pennsylvania, 19131
The Slo1 BK channels (large conductance, Ca2+-activated K+ channels) restrict Ca2+ influx and excitability, and they
are involved in many biologically important processes, such as muscle relaxation, blood pressure regulation, and
cytoprotection from ischemia where heme stress may be particularly crucial. Our collaborative project with the lab of
Dr. Toshinori Hoshi at the University of Pennsylvania, recently discovered that the mammalian Slo1 BK channels
have a CXXCH motif similar to cytochrome c which can bind heme and regulate the ion channel. Our studies of the
Slo1 BK channel use spectroscopic techniques such as UV-vis, CD spectroscopy, and EPR to determine how the
heme cofactor binds to the intracellular, regulatory domain of the BK channels and inhibits the channel. The inhibition
of the BK channels by heme may be important after cellular injury where levels of heme are high. We are currently
studying how heme interacts with a 27 kD segment of the intracellular, regulatory domain of the channel. More
specifically, we have created three specific mutants A115C, H119A and the double mutant, to test the viability of the
CXXCH motif as the heme binding site
DETERMINATION OF ELECTRONIC STRUCTURE OF PHOTOCHROMIC RUTHENIUM(II) COMPLEXES
Beth Anne McClure, (Dr. Jeffrey J. Rack), Department of Chemistry and Biochemistry, Ohio University, Athens, OH
44118.
Photochromic molecules change color when irradiated with light. This property is caused by a mechanism that is
driven by the absorption of light in which the electronic structure of the molecule is altered by changes in its physical
structure. Due to this mechanism, these molecules may be developed for optical memory storage devices.
Temperature dependent emission lifetime measurements of the photochromic complex ruthenium(II)
2,2:6`,2"-terpyridine 2,2`-bipyridine dimethylsulfoxide, [Ru(tpy)(bpy)(dmso)]^2+, were collected over the range of 24 300 K. Analysis of the kinetic data yielded the activation barriers between the triplet metal-to-ligand charge-transfer
(3MLCT) state and two thermally accessible higher excited states as well as the corresponding pre-exponential
factors. The radiative rate constant was also determined. The lower excited state was found to have an activation
barrier of 8.7 ± 1.6 cm^-1, which is much less than kT (17 - 209 cm^-1). The pre-exponential factor was only 7.53 ±
0.28 x 10^6 s^-1, which indicates that the state is not strongly coupled with the 3MLCT state, but because the
activation barrier is so low, these states are likely to be in thermal equilibrium. The higher excited state was found to
have an activation barrier of 952 ± 53 cm^-1, which could correspond to either a ligand field state or a higher excited
3MLCT state. However, the pre-exponential factor of 1.77 ± 0.47 x 10^9 s^-1 is about three orders of magnitude lower
than the expected value for a ligand field state, indicating that it is more likely a strongly coupled 3MLCT state. This
information is valuable in the determination of the electronic structure of this complex. By determining the electronic
structure of this complex it is hoped that the photochromic mechanism can be better understood. Research was
funded by Clare Boothe Luce Foundation and Condensed Matter and Surface Science Program. Ms. McClure is from
John Carroll University in Cleveland, OH 44118.
MAKING SOAP FROM READILY AVAILABLE AGRICULTURAL WASTE MATERIALS CAN ICREASE
CLEANLINESS IN RURAL AREA
Eugenia Lucas, Thomas Ciaglo (Kenneth Frost), Department of Natural Sciences and Mathematics, Dominican
University of California, 50 Acacia Avenue, San Rafael, CA 94901
In some areas of the world, soap is currently not readily available, and it may be very expensive. Therefore, among
the benefits of developing a simple recipe for soap, which can be adopted by thousands of rural people is to help
prevent the spread of disease associated with lack of overall cleanliness. In addition, we expect that the overall
utilization of family resources will be improved. For example fat that is now thrown away as garbage, can be used,
and unused vegetable waste, can be dried and burned to provide valuable base for the saponification. This will
improve the overall utilization of family resources and improve overall health. The purpose of this project is to
develop a process for making soap that can be used in rural households to make soap from readily available
agricultural waste materials and other inexpensive chemicals. Soap is made from animal fats or vegetable oils by
saponification using strong base. The simple soaps can be isolated as cakes or bars, or they can be used as water
solutions. Many reaction conditions were studied to develop a recommended process that can be done using simple
equipment and reaction conditions that can be achieved in a kitchen or a fireplace. The soaps were characterized
using infrared spectroscopy and various other analytical techniques as well as tests to show their effectiveness.
185
CHEMISTRY
Investigation of the Active Site of 5,10-methenyltetrahydrofolate synthetase
Amber Hancock, Mark Bouley (Dr. Timothy Johann, Dr. Catherine Sarisky) Department of Chemistry and Physics,
Radford University, Radford, VA
The enzyme 5,10-methenyltetrahydrofolate synthetase (MTHFS) from Mycoplasma pneumoniae has been studied to
further the understanding of the roles of individual amino acids in its proposed active site. MTHFS is the only enzyme
known to catalyze the irreversible conversion of 5-formyltetrahydrofolate to 5,10-methenyltetrahydrofolate. This
reaction is coupled to the hydrolysis of ATP to ADP. This process is important because
5,10-methenyltetrahydrofolate is a precursor for cofactors essential to the synthesis and repair of DNA and the
synthesis of proteins. The roles of individual amino acids are being investigated through site directed mutagenesis
followed by kinetic and structural characterization. Preliminary kinetic studies on the purified wild type enzyme show
folinic acid and ATP Km values of 2.7 micromolar and 190 micromolar respectively. Reported values for the wild
type MTHFS enzyme purified from other organisms are consistent with our determined values. Several alanine
substituted mutations to residues in the most conserved region among species, from the 100th to the 127th amino
acids, have been successfully produced and purified. Kinetic and structural analysis of the mutant R115A has been
performed. Kinetic data collected for R115A displays no activity for the mutant enzyme. Circular dichroism spectra
are consistent with a correctly folded enzyme. Based on a recent crystal structure, the arginine present at the 115
position in the wild type is thought to participate in ionic interactions essential to substrate binding. The inactivation of
a mutant where this arginine was changed to alanine is consistent with this hypothesis.
Investigation of Photoinitiators in Ionic Liquids
Joanna Rush (Dr. Steven W. Anderson), Department of Chemistry, University of Wisconsin- Whitewater, Whitewater,
WI 53190
Photoinduced cationic polymerization has been employed in a wide variety of industrial applications, including
adhesives and non-stick release coatings. The key to the development of this technology is the availability of highly
photosensitive and efficient cationic photoinitiators, which can be designed to be responsive to various UV
wavelengths. Our research has been directed toward the design and synthesis of two sulfonium cation initiators, which
are photoactive at short wavelengths of irradiation. We have synthesized
2-dimethyl(2-phenyl-1,3-indandionesulfonium)triflate (DPIST) and
2-dimethyl(2-phenyl-1,3-indanedionesulfonium)tetraphenylborate (DPIP), in conventional solvents like benzene and
toluene, which now pose health hazards (carcinogenicity and toxicity). Some syntheses have been accomplished in
somewhat safer solvents, such as 3-pentanone and acetone; however, the synthesis of DPIP in these solvents was
not entirely successful (yield <30%) because we need a higher temperature for a complete reaction. Therefore,
because of health concerns and the need for a higher temperature, we looked into using ionic liquids. Ionic liquids are
nearly considered green solvents because they appear to be environmentally benign, have virtually no vapor
pressure, can be used over a wide temperature range, and work as well as many conventional solvents. We have
done some preliminary work with 1-butyl-3-methylimidazolium tetrafluoroborate and are still investigating the outcome
(non-isolated yield ~80%) of that synthesis. We are testing other ionic liquids in search of the best solvents for our
reaction. Once the salts have been synthesized, they are evaluated for their efficiency as photoinitiators. This is done
through kinetic and electrochemical studies. While we have data for studies in conventional solvents, to our
knowledge, no studies have been done using ionic liquids. Therefore we are exploring the use of ionic liquids to not
only carry out the synthesis of photoinitiators but to use this medium for testing their efficiencies as well as conducting
polymerization reactions. These will include Stern-Volmer quenching and quantum yield studies. Our results, to date,
will be reported.
186
CHEMISTRY
Electrochemical Studies of Ketones in Ionic Liquids
Midn 1/C Dana S. Canby (Prof. Graham T. Cheek), Chemistry Department, United States Naval Academy, 572
Holloway Road Stop 9b, Annapolis MD 21402
The purpose of this research is to examine Lewis acid-base reactions in ionic liquids via electrochemical methods.
Unlike conventional solvents, ionic liquids consist entirely of ions. Their very low vapor pressures make them
environmentally friendly and appealing for future industrial use. This research specifically involves the ionic liquid
1-ethyl-3-methyl imidazolium tetrafluoroborate (EMI+BF4-) in which the electrochemical behavior of the ketones
benzophenone and 9-fluorenone have been studied. Cyclic voltammetry of fluorenone shows that reduction occurs in
two processes, separated by 0.40 volts. Comparative electrochemical studies of fluorenone have been carried out in
the molecular solvent system composed of propylene carbonate with tetraethylammonium tetrafluoroborate as
supporting electrolyte. In the latter system, the distance between the reduction peaks increased significantly to 0.7V,
with the second reduction process shifting to more negative potentials. This behavior indicates that the EMI cation
interacts specifically with the fluorenone dianion. In separate experiments, addition of boron trifluoride etherate to
fluorenone in EMIBF4 shifted the reduction process to more positive values, consistent with the complexation of the
ketone carbonyl group with BF3. Similar studies with benzophenone exhibited analogous behavior. Further work will
involve examining the products of the complexed ketones with various substrates.
Investigation of the Mechanism of Formation of Periodic Precipitation Reactions
Collin J Bright, Austin Toman, Joel Karty, Department of Chemistry, Elon University, 6605 Campus Box Drive, Elon,
NC 27244
Atomic absorption spectroscopy, solid state probe pH measurements and acid/base titrations were employed to gain
insight into the mechanism of pattern formation in the Mg(OH)2 periodic precipitation reaction. Periodic precipitation,
also know as the Liesegang Phenomenon, occurs when one reactant salt of a precipitation reaction diffuses into the
other. Typically, this is accomplished by dissolving one reactant salt in a gel in a test tube and placing an aqueous
solution of the second reactant salt on top of it. When the reactants meet in the gel, horizontal bands of precipitate
form, and on either side is a “band free” region where no precipitate is visible. Although this phenomenon has been
studied for over a century, having both geological and biological implications, the mechanism by which the reaction
occurs remains poorly understood. In this study, a band forming region and an adjacent band free region of the gel
were sliced away and removed for analysis at various points in time during the course of the reaction. Using the
techniques mentioned above, each slice of gel was analyzed for both aqueous and nonaqueous forms of the reactant
ions, Mg2+ and HO&#8254;. Profiles of concentration vs. time were constructed for the two different regions in the
gel. Our results are discussed in terms of the two leading theories of pattern formation: the autocatalytic growth
theory, and the coagulation theory.
REACTIONS OF 2-HALOGENO-3-METHYL-1,4-NAPHTHOQUINONE WITH PRIMARY AMINES
Nathan o. Adkins and Cameron P. Iverson (Dr. Tetsuo Otuski), Department of Chemistry, Occidental College, 1600
Campus Road, Los Angeles, CA 90041
The derivatives of 1,4-naphthoquinone are known to possess a wide variety of biological activity. One important
reaction of 1,4-naphthoquinones responsible for their biological activity is the one toward nucleophiles. The focus of
this study is the reactivity of 2-halogeno-3-methyl-1,4-naphthoquinone with primary alkylamines in methanol at room
temperature. The chosen primary alkylamines that we examined here were methylamine, ethylamine, propylamine,
isopropylamine, cyclopropylamine, butylamine, and tert-butylamine. In the reaction with an alkylamine, we found that
2-bromo-3-methyl-1,4-naphthoquinone exclusively yielded 2- alkylamino-3-bromo-1,4-naphthoquinone through an
unusual demethylation process. Similarly, 2-chloro-3-methyl-1,4-naphthoquinone yielded
2-alkylamino-3-chloro-1,4-naphthoquinone through this same demetylation as the major product. Different from the
reactions of 2-bromo-3-methyl-1,4-naphthoquinone, however, 2-chloro-3-methyl-1,4-naphthoquinone produces
another product, 2-alkylamino-3-methyl-1,4-naphthoquinone, in which the a substitution between the primary amine
and the chlorine atom occurs. Since halogens such as chlorine and bromine are known to be good leaving groups,
the substitution at the halogen site was originally expected for both 2-bromoe-3-methyl-1,4-naphthoquinone and
2-chloro-3-methyl-1,4-naphthqouinone with reactions with primary alkylamines. We believe that the size of the
primary alkylamine affects the yield of the products due to steric hindrance. We also believe the unusual
demethylation reaction found is caused by the acidic nature of the &#945;-proton of
2-halogeno-3-methyl-1,4-naphthoquinone.
187
CHEMISTRY
ATROPISOMERISM IN SUBSTITUTED ACRIDINES
Sarah Marshall (Dr. Michael Mosher), Department of Chemistry, Univeristy of Nebraska at Kearney, Kearney,
Nebraska 68849
The acridine nucleus can intercalate double-stranded DNA. This property is most likely implemented by m-amsacrine
(m-AMSA), a common pharmaceutical agent used in the fight against childhood leukemia. In an effort to explore the
possibility of a chiral acridine interacting with chiral DNA, we have examined the stereochemistry of substituted
9-phenylacridines. Depending on the substitution, this class of molecules is predicted to exist as atropisomers. Herein,
we report the preparation of 9-o-toluyl-3-methylacridine and a potential route to the isolation of its atropisomers.
(The complete abstract and with figures can be found at
http://acs.confex.com/acs/mwrm05/techprogram/P22033.HTM)
EFFECT OF PORPHYRIN STRUCTURE ON BINDING TO HUMAN SERUM ALBUMIN (HSA)
Jennifer A. Miller Meyer and Amanda Hannemann (Dr. Olga Rinco), Department of Chemistry, Luther College,
Decorah, Iowa 52101
Supramolecular systems are structures held together by intermolecular interactions and not chemical covalent bonds.
Many biological systems are actually supramolecular architectures. These systems are capable of performing
specific functions and/or altering the reactivity of their constituents. Understanding how probe molecules (guests) can
interact with biologically relevant supramolecular structures (hosts) was the focus of the research. This study looked
at the photophysical interaction of porphyrin probes with a protein solution. The data shows that the structure of the
prophyrin probe affects the ability of the probe to interact with the host protein. Various spectral and kinetic data
along with binding experiments and fluorescence quenching experiments were used to help characterize the
host-guest interaction between porphyrins and the human serum albumin (HSA) protein system. Data has been
collected for protoporphyrin IX (PPIX) and Hematoporphyrin (Hme) (as well as their diester counterparts: HmeDE and
PPIXDE) in homogeneous solvents, buffer, and HSA protein solutions. The data for fluorescence emission spectra is
promising as all porphyrins studied had blue shifts of at least 14 nm in the emission maximum between polar and
non-polar solvents. In the presence of protein binding sites all of the probes except HmeDE showed blue shifts, which
suggest incorporation in a less polar environment. The discrepancy for HmeDE is the first indication that not all
porphyrins behave in the same manner. In the steady-state quenching experiments, Stern-Volmer rate constants
were found to vary for all four probes. The rate constants were decreased by an order of magnitude for Hme, PPIX
and PPIXDE, but the decrease for HmeDE was only 2 fold. Again, the decrease in Stern-Volmer constants indicates
inclusion within a binding region for the porphyrin probes.
Ligand binding studies of PDZ domains
Kyriacos M. Koupparis (Dr. Mark Spaller), Chemistry, Wayne State University, Detroit, Michigan 48202
PDZ domains are sequence repeats apparent in diverse signaling proteins. They are predominantly found in
cytoplasmic proteins, in which they occur in one or multiple copies. The focus of this research is the three PDZ
domains of postsynaptic density 95 protein (PSD-95), also known as synapse associated protein 90 (SAP-90). The
first two domains are studied separately and also as a PDZ1-2 dual domain. PDZ1, PDZ2 and coupled PDZ1-2
domains were expressed by transforming E. coli competent cells with DNA plasmid encoding the amino acid
sequence of these domains. The protein domains were subsequently isolated using affinity chromatography and ion
exchange chromatography. This protocol was repeated for PDZ3, the third domain of PSD-95. Once purified, the
binding properties of these domains with peptides and organic molecules were studied using Isothermal Titration
Calorimetry (ITC) and thermodynamic variables were obtained for each ligand tested. The ligands tested are derived
from known PDZ binding partners such NMDA receptors, Kv 1.4 protein, and CRIPT protein. The goal is to
understand the interactions between these domains and their binding partners, as well as to identify essential residues
within the binding pocket. Initial binding studies have been performed involving Kv 1.4 derived peptides, AVETDV
and its acetylated analog Ac-AVETDV. These peptides were titrated against PDZ 1-2 using ITC, resulting in
dissociation constants of 2.9 &#956;M and 2.7 &#956;M, respectively.
188
CHEMISTRY
SEMICONDUCTOR PHOTOCATALYSIS OF FERROUS IRON
Jessie M. Penich and Samir Raut (Dr. Steven J. Gravelle and Dr. Caryl Fish), Department of Chemistry, St. Vincent
College, 300 Fraser Purchase Road, Latrobe, Pennsylvania 15650
The purpose of this research was to determine whether or not the oxidation rate of ferrous iron [Fe(II)] to solid iron
[Fe(III)] can be increased using semiconductor photocatalysis. Many streams in western Pennsylvania are
contaminated by iron due to a long history of coal mining in the area. In wetland treatment systems, a high rate of
Fe(II) oxidation would allow more water to be treated and released back into the stream over a shorter period of time.
The method for increasing the Fe(II) oxidation rate proposed in this research was semiconductor photocatalysis.
Powdered (solid) titanium dioxide was used as the semiconductor photocatalyst for each experimental trial and the
light source used was a 400 watt Xenon arc lamp. Each experimental trial was carried out in a rotating 25 mL
reaction cell and small samples of solution were drawn out after regular time intervals of light exposure. The data for
each experimental trial was obtained by using a UV/VIS spectrometer to generate a spectrum for each sample at
each time interval. For comparison purposes, a control was performed for each experimental trial in which no
photocatalyst was used. The data for each trial was analyzed by comparing the rate of decrease in the concentration
peak for Fe(II) in the control spectrum to the rate of decrease of the Fe(II) concentration peak in the spectrum from
the actual experiment. Several different experiments were performed to observe the behavior of Fe(II) in
photocatalysis reactions. The two experiments that provided the most definitive conclusions were the photocatalyis of
Fe(II) alone and the photocatalysis of Fe(II) with hydrogen peroxide. The results from these two experiments (and
several others that were performed in this research) suggest that Fe(II) will not oxidize to Fe(III) during semiconductor
photocatalysis. In summary, the data from this research indicates that semiconductor photocatalysis would not be an
effective method for increasing the oxidation rate of Fe(II).
ANALYSIS OF AIR SAMPLES FOR ROXARSONE BY A GAS CHROMATOGRAPHY-MASS
SPECTROMETRY METHOD
Michelle D. Dickinson (Dr. Barbara M. Hopkins), Chemistry Department, Xavier University, 3800 Victory Parkway,
Cincinnati, OH 45207
The compound roxarsone (4-Hydroxy-3-nitrobenzenearsonic acid) has been used in the agricultural sector as a
growth promotor in chicken feed. While research has concluded that consumers are not exposed to toxic levels of
arsenic in poultry that is purchased, feed-plant workers can be exposed to unknown levels of roxarsone in the air.
The purpose of this work is to develop a gas chromatography-mass spectrometry (GC/MS) method for the qualitative
and quantitative analysis of roxarsone using a modification of the work of Roderdink and Aldstadt (J. Chromatogr. A
1057 (2004) 177.), and to couple this method with an air sampling method developed by the National Institute for
Occupational Safety and Heath (NIOSH). Because roxarsone is not volatile, a derivative was generated by reacting
acidified methanol solutions of roxarsone with 1,3-propanedithiol. Following extraction with toluene, a 1 &#956;L
sample of the volatile product was introduced into the GC on a Factor Four Varian Column. The resulting calibration
curve showed linearity over a concentration range appropriate for the determination of roxarsone that could be
present in samples of air. An instrumental limit of detection (LOD) of 5.6 &#956;g/mL as roxarsone was determined.
To validate the method, results will be presented that test the method against the established criteria of NIOSH for air
sampling methods.
189
CHEMISTRY
SYNTHESIS AND CHARACTERIZATION OF LAYERED TRANSITION METAL PHOSPHATE
NANOPARTICLES—PRECURSOR MATERIALS TO PHOSPHIDE NANOPARTICLES
Christopher J. Young (Mr. Palaniappan Arumugam and Dr. Stephanie Brock), Department of Chemistry, Wayne State
University, Detroit, Michigan 48202
Nanoparticles have been of recent interest to researchers because of the myriad practical applications to which they
may be applied, as well as their interesting chemistry. Prime candidates for studies of physical properties as a
function of size are transition metal (TM) pnictides (pnicogen = Group 15 element) because of their interesting
magnetic and catalytic properties in the bulk phase. The Brock group has recently employed hydrogen annealing for
the reduction of TM phosphates to produce corresponding TM phosphides. In addition to hydrogen annealing, our
group has synthesized TM arsenide nanoparticles by reduction of lamellar TM arsenates on a carbon substrate. This
carbothermal reduction method results in an ordered array of nanoparticles.
Currently our studies are focused on perfecting a solvothermal assisted microemulsion synthesis of lamellar TM
pnictates, specifically iron and nickel phosphates. In addition to the solvothermal synthesis route, we have had some
success in producing lamellar phosphates by exfoliation of bulk phosphates. The resulting phosphates will be used to
extend the aforementioned reduction techniques to produce ordered arrays of FeP, Fe2P, and NiP nanoparticles.
The Role of Estrogen on Retina Regeneration in the Zebrafish: Testing Estrogen and Aromatase
Levels post trauma via ELIZA
Jessica Lynn Fulton (Dr. James Turner, Dr. Dan McCain), Swope Summer Research Program, Jackson Hope Fund,
Biology Department and Chemistry Department, Virginia Military Institute, Lexington, VA 24450
Although estrogen has been primarily regarded only as a female sex hormone, recent studies show that estrogen and
its receptors are found in the brain and act as neuro-protectors, slowing the progress of such diseases as Alzheimer’s
disease. Telost fish, such as the zebra fish, possess the capacity to regenerate damaged parts of their central nervous
system, such as the eye. It has been hypothesized that this regenerative capacity is due to the fact that zebrafish
have 1000 fold more estrogen present in their central nervous tissue than do humans. This study attempts to test
estrogen’s effect on neural regeneration by inducing damage to the retina of the zebra fish via an optic nerve crush
and then analyzing the concentration of estrogen and aromatase post crush as opposed to a control via an ELIZA
test. Higher post crush levels of estrogen and aromatase would suggest a vital role of estrogen’s role in posttraumatic regeneration. The significance of this work revolves around the fact that if estrogen does play a major role
in post traumatic regeneration, clinical and pharomlogical applications could result in cures for many neurological
diseases.
190
CHEMISTRY
Overexpression of Glutathione Peroxidase (GPx) in Islets of C57 Black/6 Mice
Sin Man Mak (Dr. R. Paul Robertson), Department of Chemistry, Seattle University, 900 Broadway, Seattle, WA,
98122
Diabetes is a non-curable, common disease in which people suffer greatly from having high glucose concentration in
their blood. It is segregated into two categories, type 1, an autoimmune disease, and type 2, a polygenic disorder
rather than a single specific entity [1]. Many studies are under investigation to control type 2 diabetes using
approaches other than medications, which become ineffective over time. Chronic hyperglycemia leads to a
phenomenon termed glucose toxicity—when glucose remains at supraphysiological levels for protracted periods of
time, the homeostatic relationship of insulin and blood glucose level is disrupted and leads to cellular dysfunction that
becomes irreversible. One mechanism of glucose toxicity is chronic oxidative stress due to an overproduction of
reactive oxygen species (ROS), which are toxic agents that can cause defective insulin gene expression and insulin
secretion as well as increased apoptosis [2]. One way to dispose of ROS is by antioxidant enzymes, which are
normally found at low levels in the beta cells of the pancreas, the only cells of the body that make insulin. Our
hypothesis is that an increase in beta cell antioxidant enzyme activities would provide better protection against
oxidative stress and glucose toxicity. To study this hypothesis in vivo, we chose to create a beta cell specific
transgenic mouse model with an overexpression of glutathione peroxidase (GPx), a major antioxidant enzyme that
metabolizes ROS and is normally undetectable in pancreatic islets, to determine whether this will prevent the adverse
effects of glucose toxicity. Western analysis of our transgenic (Tg) mice demonstrated an increase level of GPx
protein in islets compared to wild type (WT) mice, and as much as the level observed normally in the livers of WT
mice. No overexpression was seen in other tissues from Tg mice. We are now challenging Tg and WT mice with
high fat diet (which causes hyperglycemia) and streptozotocin (a beta cell toxin) in vivo and hydrogen peroxide (a
type of ROS) in vitro to ascertain whether the increase in GPx expression will protect beta cells from ROS and
consequent functional damage.
References
1. Robertson, R. P.; Harmon, J.; Tran, P. O.; Tanaka, Y.; Takahashi, H. Diabetes. 2003, 52 (3), 581-587.
2. Robertson, R. P. JBC. 2004, 279, 42351-42354.
Revealing Correlated Motions of Biomacromolecules via Wavelet Transforms
Elijah J. Gregory, (sponsor) Thomas Cheatham III, Dept. of Medicinal Chemistry and Pharmaceutics and
Pharmaceutical Chemistry, 30 South 2000 East, Rm 307 SLC, UT 84112-5820, (sponsor) Julio C. Facelli, Center for
High Performance Computing and Department o
One of the tenets of classical science is that, given an understanding of the underlying physics and molecular forces,
all of the motions and dynamics of an interacting system may be predicted exactly. Atomistic simulations of
bio-molecular systems in their native solvent environment have progressed so that it is now possible to fold a protein
and to understand the atomic details of a drug binding to its target. These simulations are computationally
demanding, resulting in massive amounts of data, Gigabytes or even Terabytes in size.
In this work, we have explored new computationally efficient versions of the wavelet transform known as lifting
schemes. We have created a lifting scheme analog to the non-subsampled wavelet transform in order to temporally
localize frequencies while using computational resources most efficiently. Performing the wavelet transform splits the
data into two time series. One of these is a projection of the data onto a smooth subspace, while the other is onto a
high-frequency subspace. By repeatedly applying the transform one is able to obtain an approximation of the data on
a subspace containing large-scale correlated motions of macromolecules.
These methods can be applied to aid in the design of new drugs and to develop novel drug delivery systems and
biomaterials, because they can readily expose critical structure/function relationships. We are using them to analyze
large-scale simulations of the dynamics of nucleic acids and complex multi-unit proteins, with the hope of being able
to expose hidden correlations in the biomolecular simulation data that may provide insight on how to design the
medicines of the future.
191
CHEMISTRY
SYNTHESIS AND CHARACTERIZATION OF SUBSTITUTED DITHIENYLBENZO [G] QUINOXALINES
Stefanie A. Cantalupo (Dr. Guy Crundwell), Department of Chemistry, Central Connecticut State University, 1615
Stanley St., New Britain, CT 06050
Whereas quinoxalines have been the subject of many studies in organic, bioorganic, and materials chemistry, little is
known about the electronic and metal-binding behaviors of benzo[g]quinoxalines. The significance of studying
dithienylbenzo [g]quinoxalines can aid in further understanding electronic interactions in thiophene based polymer
networks or fabricating methods for preparation of thiophene containing compounds. Our lab is also interested in
dithienylbenzo[g]quinoxalines as potential bidentate [N,S] ligands— an approach that might be useful in metal-based,
crude oil refinement processes. The investigation of these compounds required dithienylbenzo[g] quinoxalines with
different constituents and thienyl linkages. A variety of substituted dithienylbenzo[g]quinoxalines were synthesized
via the direct condensation of diaminonaphthalein and the corresponding dithienylethanedione in order to fully explore
these properties. All compounds underwent extensive purification during an REU experience at the University of
Connecticut under the guidance of Dr. Amy Howell. Various purification techniques were carried out such as
recrystallization and flash column chromatography in order to purify samples for X-ray quality crystal growth. X-ray
quality crystals were grown by various techniques, including vapor diffusion. Diffraction data for all samples was
collected on an Xcalibur kappa geometry single crystal X-ray diffractometer which is equipped with a CCD detector
(Sapphire3) which was recently acquired by CCSU through an NSF MRI grant. Crystal structures for three
dithienylbenzo[g]quinoxalines are refined and solved. A full characterization of these compounds has been
completed and will be presented. Spectra that have been collected include 13C and 1H NMR (300MHz), mass
spectrometry, UV/Visible, and Infrared Spectroscopy. Aside from dithienylbenzo[g]quinoxalines, asymmetric
ethanediones (such as 1-phenyl-2-thien-2’-yl-1,2-ethanedione) are also being synthesized so they can be reacted to
make asymmetric substituted 2,3-diarylquinoxalines and 2,3-diarylbenzo[g]quinoxalines.
ROTAXANES AS PRECURSORS FOR A MOLECULAR MUSCLE
Tiffany M. Mingin, Christian P. Jensen, and Mark Clark,(Dr. William R. Kwochka), Department of Chemistry and
Physics, Western Carolina University, Cullowhee, NC 28723
Rotaxanes are large, mechanically-interlocked molecular systems consisting of a ring component threaded by a string
component. Stoppers placed at the ends of the string prevent the ring from falling off. Rotaxanes have attracted
quite a bit of attention lately because of their potential to act as switching devices at the molecular level. A molecular
switch is a nanoscale device which switches reversibly between two or more positions. Groups located within the
framework of the ring can then selectively interact with stations on the string of the rotaxane. If the position of the
ring on the string can be controlled by some external stimuli such as light or electrical current, the rotaxane becomes
a system that can communicate in binary logic – the basis of digital computing. Thus, rotaxanes, when organized into
polymeric arrays, could act as molecular switching devices capable of performing tasks such as information
processing and storage.
A common approach for preparing rotaxanes makes use of self-assembly, a process that is ubiquitous in nature. An
elegant example of both the simplicity and complexity of a self-assembled system is found in the superstructure of
DNA. Essentially, this process uses weak, non-covalent interactions (hydrogen bonding) to assemble small molecules
(the nucleotides) into larger supramolecular structures (the double helix). Our work is focused on preparing rotaxanes
that have functional groups incorporated in the ring component; we will report on our progress in making these
functionalized rings and our plans for the preparation of a molecular muscle based on our rotaxane work.
192
CHEMISTRY
POLYMERIZATION OF D,L-LACTIDE WITH TITANIUM (IV)-BINOLATES
Elizabeth Matzen, Suzanne Crumley, Amanda Fenner, and Heidi Schwanz (Dr. Bradley M. Chamberlain), Department
of Chemistry, Luther College, Decorah, Iowa 52101
Biodegradable polymers derived from renewable resources (e.g., starch, cellulose, lactic acid) provide a foundation
for sustainable commercial technologies; these technologies, in turn, alleviate environmental problems associated
with the increasing demands of industrialized society for carbon-based chemical processes. Recently, rapid advances
have been made in the development of biodegradable polymers, most notably the commercialization of polylactide.
Polylactide (PLA) is a polyester with considerable potential in fiber, film and packaging application; it is prepared via
the ring-opening polymerization of lactide, a cyclic ester ultimately derived from the fermentation of corn. PLA’s
physical properties, along with its rate of chemical and biological degradation, are determined by the
three-dimensional orientation of substituents along the polymer chain (i.e., its microstructure). Considerable effort
has been devoted to controlling PLA microstructure, and a number of tin, aluminum, zinc, yttrium and iron catalysts
have been developed. Herein we report the preparation of two titanium(IV) alkoxides bearing optically active BINOL
ligands. Both complexes (2a-b) are active catalysts for the ring-opening polymerization of D,L-lactide. The effect of
ligand architecture upon polymerization rates and polymer microstructure will be presented; the results of mechanistic
analyses will also be discussed.
http://admin1.luther.edu/~brandtja/Abstract - Matzen.doc
COMPUTATIONAL STUDY OF HYDROGEN CHEMISORPTION TO CARBON NANOTUBES
Sara A. Houston (Dr. Ron Brown) Department of Chemistry, Mercyhurst College, Erie, PA 16546
The potential for hydrogen fuel in transportation applications has been the focus of much recent research. According
to the DOE, in order to achieve the required efficiency for widespread use, the fuel cells must hold a 6.5 weight
percent of hydrogen. To achieve these levels of hydrogen adsorption, chemisorption may be a significant factor. We
have performed a systematic computational study of hydrogen chemisorption to carbon nanotubes. The methods
used include density functional theory (DFT) and molecular mechanics. For the DFT calculations, small clusters
containing 9 to 45 carbon atoms were designed that model zigzag and armchair carbon nanotubes. The radii of the
tubes was varied from 1.97Å-3.54 Å to model (5,0)-(9,0) zigzag tubes and 2.05Å -4.10Å to model (3,3)-(6,6) armchair
tubes. The relationship between geometry and chemisorption energetics was investigated at the B3LYP/6-31G** level
of theory. Full optimizations of models were performed that allowed for the study of carbon deformation effects. We
found that, due to a higher degree of curvature, chemisorption was more favorable on tubes with smaller radii.
Furthermore, chemisorption was somewhat more favorable to armchair tubes. The effects can be explained by
examination of the local geometry of the adsorption site. Molecular mechanics calculations were run on full-length
tubes and clusters in order to correct for the truncated size of the models in the ab initio calculations. The results of
this study are significant towards understanding the mechanism of hydrogen chemisorption to carbon nanotubes.
The Study of the Nucleophilic Aromatic Substitution Reactions of 1,3,5-Trichlorobenzene and
1-Bromo-3,5-dichlorobenzene with Cyclic Amines
Jason Crumpton (Dr.Claudia P.Cartaya-Marin and Dr.Steve D. Willimas), A.R. Smith Department of Chemistry,
Appalachian State University,, Boone, NC 28608
1,3,5-Tribromobenzene and 1-bromo-3,5-dichlorobenzene undergo nucleophilic aromatic substitution when reacted
with cyclic amines such as pyrrolidine, piperidine and morpholine. Each reaction of 1,3,5-Tribromobenzene and
1-bromo-3,5-dichlorobenzene with morpholine, piperidine or pyrrolidine was conducted under three different reaction
conditions: reflux, microwave and sealed tube (200C). Ratios and percent yield of products were found using gas
chromatography and mass spectrometry. Products were separated using column chromatography. Ab initio
calculations were used to model these reactions. Structures of the products were determined using NMR
spectroscopy and mass spectrometry. The results from this study will be presented.
193
CHEMISTRY
THE STUDY OF COMPLEXATION OF BROOKER`S MEROCYANINE IN ALPHA- AND
BETA-CYCLODEXTRIN
Anthony Campitella, (Jennifer Holt), School of Science, Penn State Erie, The Behrend College, 5091 Station Rd, Erie,
PA 16563
Guest host complexes are used in a variety of fields of study such as chemistry, material science, plastics and
biology. A guest host complex is a host system that has a cavity into which another molecule can enter. Guest host
complexes are important and useful because they can vary the physical and chemical properties of a material by the
way the molecules bind to each other. The binding constant for both alpha- and beta-cyclodextrin with Brooker’s
Merocyanine was determined using UV-Vis spectroscopy. Brooker’s Merocyanine was chosen as the guest due to its
simple conjugated structure that can be detected using UV/Vis spectroscopy. The spectrum of Brooker’s Merocyanine
is especially sensitive to the environment of molecule, which was used to probe the formation of the complex. It was
determined that the binding constant for the beta-cyclodextrin was 413 M^-1 which is good agreement with the
literature value of 480 M^-1 [Hamasaki, K., Nakamura, A., Uneo, A., Toda, F., J. Inclus. Phenem. And Mol. Rec.
Chem., 1992, 13(4),349.]. The binding constant was also determined for alpha-cyclodextrin with Brooker’s
Merocyanine to be 72 M^-1 indicating less preference for binding of Brooker’s Merocyanine to alpha-cyclodextrin than
to beta-cyclodextrin. The binding constant of the data was fit with linear and nonlinear models using a modified
Benesi-Hildebrande equation. These results were confirmed by measuring the binding constant using fluorescence
spectroscopy.
SYNTHESIS OF SUBSTITUTED ORTHO-PHTHALADELHYDES FOR THE DERIVATIZATION OF AMINO
ACIDS
Andrew J. Mueting; David C. Hawkinson, PhD; Miles D. Koppang, PhD, Department of Chemistry, University of South
Dakota, Vermillion, South Dakota 57069
Since amino acids are generally undetectable using standard spectroscopic (absorbance or fluorescence) or
electrochemical methods, analysis of these compounds requires that they be derivatized with an appropriate
chromophore or electrochemically active group before or after chromatographic separation. One standard method for
low-level analysis of amino acids is derivatization with ortho-phthalaldehyde (OPA) in the presence of an alkylthiol to
yield N-substituted 1-alkylthioisoindoles. Despite the widespread use of OPA in amino acid analysis, this reagent has
some shortcomings, in particular, instability of the isoindole derivatives. In the hope of improving the stability of these
derivatives, we are preparing substituted OPA derivatives containing internal nucleophiles for potential use in amino
acid analysis. While thiols have traditionally been used as the nucleophile in the derivatization reaction, examination
of the proposed reaction mechanism suggests a secondary amino group may be more desirable as the internal
nucleophile. Key steps in the synthesis of these compounds include the preparation of functionalized 2,4-hexadienes
from sorbic acid, cycloaddition of dimethyl butynedioate to the 2,4-hexadienes, aromatization of the Diels-Alder
adduct, and subsequent functional group interconversions to form the desired product. Progress on the synthesis of
these compounds to date will be presented.
DISSOLUTION OF THE IONIC LIQUID TRIHEXYLTETRADECYLPHOSPHONIUM
BIS(TRIFLUOROMETHYLSULFONYL)AMIDE IN SUPERCRITICAL CO2
Peter Apps, (Dr. Mark Heitz), Department of Chemistry, State University of New York College at Brockport, Brockport,
New York 14420.
Recently much attention has been given to the study of room temperature ionic liquids. Ionic liquids are salts of an
organic cation and a wide variety of inorganic anions. The interest in these liquids lies in their potential towards
replacing traditional solvents used in various organic, separation, and electrochemical reactions. Previous research
has mainly focused on the imidizolium-based ionic liquids, which are insoluble in supercritical carbon dioxide
(scCO2). However, ionic liquids with a phosphonium cation have shown solubility in CO2. Our research focused on
the solubility of the room temperature ionic liquid trihexyltetradecylphosphonium bis(trifluoromethylsulfonyl)amide in
scCO2. Fluorescence from coumarin 153 (C153) was used to characterize the solvation of this ionic liquid. Steady
state emission spectra were recorded at CO2 pressures ranging from 1040-3200 psi, at 323K. To provide a basis for
comparison, emission spectra of C153 were also collected for the ionic liquid in a variety of organic solvents. The
spectra show that a partition exists between a CO2 rich phase and an ionic liquid phase. This partition, along with
shifts of the emission spectra, provides evidence of the dissolution of the ionic liquid in scCO2. This presentation will
focus on the preliminary results of our research as well as detail upcoming areas of investigation.
194
CHEMISTRY
REFECTION ABSORPTION INFRARED SPECTROSCOPY (RAIRS) – FIBER OPTIC AS A SURFACE
ANALYZER OF ORGANIC RESIDUES ON SURFACES
Yadira Soto-Feliciano, Oliva M. Pirmera-Pedrozo, Pedro Fierro-Mercado, Leonardo C Pacheco-Londoño and Samuel
P. Hernandez-Rivera, Chemical Imaging Center / Center for Sensors Development, Department of Chemistry,
University of Puerto Rico-Mayagüez, PO BOX
Refection Absorption Infrared Spectroscopy - Fiber Optic Coupled has been investigated as a potential technique to
develop a new methodology for detection of explosive and pharmaceutical residues on surfaces. The methodology is
remote sensed, in situ and can detect nanograms of the compounds. It is solvent free and requires no sample
preparation. Smearing technique was used for transferring the target analyte on the substrates to be used as
standards. The sample transfer methods gave good sample distribution, reduced sample loss on transfer and were
easy to manipulate giving good reproducible distributions. Samples with surface concentrations ranging from
micrograms/cm2 to nanograms/cm2 of explosives: 2,4,6-trinitrotoluene (TNT), pentaerythritol tetranitrate (PETN) and
of the active pharmaceutical ingredient (API) ibuprofen were deposited on stainless steel metal surface. Methanol,
Acetone, dichloromethane, tetrahydrofuran and acetonitrile were used as transfer solvents for smearing. Mixtures of
Acetonitrile, methanol and 2-propanol were used for mass transfers in some cases. The data were analyzed using
Chemometrics routines. In particular, Partial Least Squares (PLS) multivariate analysis was used. The results of the
experiments gave intense absorption bands in the fingerprint region of the infrared spectrum that were used to
quantify and calculate the detection limit for each compound. This study has demonstrated that Fiber Optic Coupled
FT-IR is capable of detecting low loading concentrations of 0.1 &#956;g/cm2 on metallic surfaces.
Link: http://grad.uprm.edu/tesis/primerapedrozo.pdf
SORPTION STUDIES OF Cu2+ ION ON VARIOUS SOLID MATRICES SUCH AS GEL AND
MEMBRANES
Hong N. Tran, Fallon L. Lowery and Siva Mandjiny, Department of Chemistry and Physics, University of North
Carolina at Pembroke, Pembroke, NC 28372
In this study it is intended to decontaminate Cu2+ ion from drinking water. Simple chromatographic experiments were
conducted with various solid matrices such as iminodiacetic acid (IDA) gel and various IDA membranes such as
Sartobind and Convective Interactive Media (CIM) disk. Elution conditions and flow rate were optimized for better
performance of the separation process. It is clear from these studies that the membranes do have larger binding
capacity of Cu2+ ion over the gel under similar conditions. Also, membranes are easier to operate than the gel.
Finally, affinity constant, KD, between the Cu2+ ion and the IDA gel was determined using Langmuir adsorption
isotherm. This study proved that the drinking water containing Cu2+ ion even at lower concentration such as 10 ppm
could be decontaminated by these solid matrices.
Nuclear Charge Stabilization in Characterization of Metastable-State Energies of Fluorocarbon
Anions in Dissociative Electron Attachment
Emily B. Moore (Prof. Jack Simons ), Department of Chemistry, University of Utah, Salt Lake City, Utah 84112
Finding ways to functionalize the C-F bond in fluorocarbons, one of the strongest bonds known, is currently an active
area of research with industrial as well as environmental significance. The focus of this project involved
characterizing the energy for C-F bond cleavage through dissociative electron attachment (DEA). DEA is the process
in which a chemical bond of a molecule is broken by being struck by an electron having an energy appropriate to
attach to the &#963;* molecular orbital of the C-F bond causing bond cleavage. The theoretical simulation of such a
DEA event involves characterizing both the energy profile of the neutral molecule and the energy profile of the
molecule’s anionic species. By comparison of the two energy profiles, the electron energy necessary to cause bond
cleavage by attaching to the &#963;* molecular orbital is determined. Because the anion is an electronically unstable
species, characterization of its energy profile involves using the so-called nuclear charge stabilization device, a
process that involves systematically increasing (artificially) the nuclear charges on the C and F nuclei. After
successful testing of the nuclear charge stabilization device on H2 and N2 molecules, this process was completed for
primary, secondary and tertiary fluorocarbons, and will be explained and results summarized.
195
CHEMISTRY
PREPARATION OF LITHIUM-INTERCALATED COBALT VANADATES.
Patrick H. Redwood (Keenan Dungey), Department of Chemistry, University of Illinois at Springfield, Springfield,
Illinois 62703
We are interested in improving the lithium ion batteries used in portable devices by preparing a bimetallic oxide
cathode material which allows more efficient charge transfer than current cathode materials. We have prepared
samples of cobalt vanadates through coprecipitation under basic conditions and ultrasound under acidic conditions.
Two new compounds were produced: a cobalt hydroxy pyrovanadate and a layered polyoxyvanadate intercalated with
cobalt ions. The samples were characterized through elemental analysis, infrared spectroscopy, and powder X-ray
diffraction. We have also investigated the thermal decomposition patterns of the samples. We have explored
several routes (LiI, nBuLi, LiOH, various pH and thermal programs) for intercalating lithium ions into these materials.
The resulting lithium-inclusion compounds were structurally compared with commercial and prepared lithium
cobaltate. We are currently working to increase the lithium ion storage capacity of the cobalt vanadates.
CIRCADIAN GENES ARE EXPRESSED DURING EARLY DEVELOPMENT IN XENOPUS LAEVIS
Brittany L. Bronson (Dr. Kristen L. Curran), Biological Sciences Department, University of Wisconsin-Whitewater, 800
W. Main Street, Whitewater, Wisconsin 53190
Almost all organisms exhibit circadian rhythm; a 24 hour cycling of physiological processes and behaviors. In these
organisms there exists a central oscillator that operates as a time-keeping mechanism in the absence of external
stimuli. A negative feedback loop involving several different genes is implicated in the timing of circadian rhythm. In
situ hybridization was performed to elucidate the developmental expression pattern of these particular genes in
Xenopus laevis. It was hypothesized that although each gene studied shared similar expression patterns (i.e., the
nervous system), each gene would also possess its own unique expression pattern. Data has been analyzed for
Xbmal-1, Per1, Per2, and Nocturnin , and expression was observed in the developing central nervous system and
sensory tissues. Unexpected mRNA expression was observed in the pronephros, and pronephritic tubules for
Nocturnin, and Xbmal-1 respectively. The reason that these genes are present in the developing kidney and the duct
is unknown, and is being tested further. We intend to next analyze three other circadian genes (Cry1, Cry2b, and
Clock). This data combined with our previous results will act as a foundation for future studies concerning the
ontogeny of the circadian rhythm clock and will help understand the importance of circadian genes in tissues that are
not assumed to be circadian in nature.
A Chemical Assessment of Heavy Metals in Peak Creek
Diane G. Youker, Joseph I. Wirgau, Department of Chemistry and Physics, Radford University, Radford,VA 24142
Peak Creek is a historically contaminated stream which runs through downtown Pulaski, Virginia and ultimately flows
into Claytor Lake and the New River. Peak Creek has multiple sources of heavy metal contamination and has
previously exhibited ecological impairment. The primary source of inorganic pollutants is the site of the former Allied
Chemical and Dye Company, which produced sulfuric acid, vanadium pentoxide, and ferrous sulfide from 1904 to
1968. The stream was surveyed in 1989 and found to be in the ninety-ninth percentile for copper, lead, and zinc, as
well as being void of aquatic life near the Allied Chemical site. A study from the late 1990’s observed only a partial
recovery of the stream both chemically and biologically. This past summer we reinvestigated the levels of inorganic
compounds in Peak Creek. Overall the chemical levels in the stream have continued to improve, although toxic
levels of copper and iron were observed. The highest concentration of copper, iron, and zinc observed were 1780,
40,000, and 210 ppm respectively. Lead concentrations were below our detection limits. We also observed
increased conductivity, hardness, and pH upon entering the town of Pulaski toward the Allied Chemical site, which we
attribute to urban runoff and the canalization of the river.
196
CHEMISTRY
MIMICING THE CD4 BINDING SITE ON GP120 USING NGAL.
Camden M. Driggers, (Dr. Martin Teintze), Chemistry & Biochemistry Department, Montana State University,
Bozeman, Montana 59717
The entry of HIV into cells requires interaction between the viral surface protein gp120 and the human CD4 receptor.
Antibodies to the CD4 binding site (CD4bs) on gp120 are rarely produced but are particularly efficient at neutralizing
the HIV virus. These antibodies contain a finger-like projection of peptides which fit into the CD4bs. We are trying to
find molecules that mimic the CD4bs to use as vaccine antigens. The purpose of this project is to mimic the CD4bs
using human neutrophil gelatinase-associated lipocalin (NGAL), a cup-shaped protein which will be able to elicit
antibodies with a finger-like projection. NGAL has been expressed on the pIII protein of bacteriophage M13. After it
is determined that a functional NGAL is expressed, regions in the gene which encode for the internal concave
structure of the protein, where the siderophore ligand normally binds will be randomized. Then this library of phage
with NGAL variants will be screened against antibodies specific for the CD4bs. This will select for the random peptide
regions that mimic the CD4bs. This mutant NGAL could then be developed as a vaccine which would elicit antibodies
that inhibit multiple variants of HIV from entering the cell.
VARIABLE TEMPERATURE AFM STUDY OF COATED PET FILMS
Kenny Johnsen (Laura Wright, Rodney Carter, and Robert Posey), Department of Chemistry, Furman University,
3300 Poinsett Highway Greenville, SC 29613
Polyester films have been characterized at temperatures ranging from room temperature to 160 degrees Celsius
using a MultiMode AFM equipped with a hot stage. The films being studied have coatings that allow them to be used
in heat seal applications. These applications include lidding materials for microwave food packaging and for polymer
based serving dishes that go directly from the freezer to the oven. The behavior of these materials upon heating is
therefore of interest. Upon controlled heating specific changes are easily observed in the phase image when tapping
mode AFM is employed. In some cases the coating flows at relatively low temperatures. One example of this
behavior was illustrated using a DuPont product which contains erucamide as an additive. In other films higher
temperatures are required and a growth of spherical crystallites is detected as material migrates away from some
sites and builds up in neighboring sites. Each of these processes have been monitored in real time. The results of
this study and the implications of the film behavior at various temperatures will be presented in this poster.
PREPARATION OF NOVEL DIPHOSPHINE-BRIDGED MOLYBDENUM-DIIMINE COMPLEXS
Preston T. Larson, (Christopher L. Exstrom), Department of Chemistry, University of Nebraska at Kearney, Kearney,
NE 68849
We report two novel binuclear [Mo(phen)(CO)3]2(&#956;-PP) (phen = 1,10-phenanthroline; PP =
1,2-bis(diphenylphosphino)ethane (dppe) or 1,4-bis(diphenylphosphino)butane (dppb) complexes that may serve as
precursors to future solvatochromic clathrate materials. The above complexes are prepared from Mo(phen)(CO)4
and a diphosphine in refluxing toluene. The structures of the isolated complexes have been characterized by 1H and
31P NMR, FTIR, and UV-vis spectroscopy. Solid-state structure analyses and solvatochromic properties will also be
discussed. Regardless of the Mo(CO)4(phen)/diphosphine ratio used, bridged structures of the complexes resulted.
This is surprising given the reported examples Mo(CO)4(PP) and [Mo(CO)5]2(&#956;-PP) in which chelating and
bridging coordination modes of diphosphines are sensitive to reaction stoichiometry. One possibility is that the
reactions carried out do not provide enough energy for a fourth carbonyl to be lost. However, it is likely basicity and
&#960;-acceptor abilities of diphosphines relative to monophosphines may influence their reactivity with Mo(phen)
(CO)4. Further investigations are underway.
197
CHEMISTRY
GEOMETRY AND SEQUENCE DEPENDENCE OF H2O INTERACTIONS WITH THE FACES OF DNA
BASES
Tori B. O`Bannon Garen Holman Mikaela Stewart, (Dr. Lori Isom), Department of Chemistry, University of Central
Arkansas, Conway, Arkansas 72035
This study investigates water interactions with the conjugated pi system of the rings of DNA bases, called water-pi
interactions. Because helical formation is driven by the favorable pi-pi stacking between the hydrophobic bases of
DNA, the interaction between groove-bound water molecules and the pi systems of DNA bases, and the effect of
these interactions on DNA structure has never been investigated. However, cation-pi interactions have been detected
between DNA bases and bound cations in DNA. Due to water’s high dipole moment, we suspected that water will
orient such that the H atoms are interacting with the DNA pi systems. To investigate this hypothesis, the Protein Data
Bank (PDB) was screened for B-form DNA structures with a resolution of 1.6 Å or higher that were free of significant
chemical modifications and bound ligands. These structures were then analyzed using Visual Basic programs to
determine the distance and angle between each water molecule and the centroid of every DNA base ring. The waters
with an angle less than 55° and a distance less than 5.0Å from a centroid were isolated. These interactions were then
analyzed for sequence specificity, major/minor groove patterns, and average distance and angle of interaction.
According to these criteria, water-pi interactions were found in all structures analyzed. The water-pi interactions
occurred most often with adenine in the major groove and thymine in the minor groove. Water interacts in the minor
groove with bases on both chains at AT steps. Water-pi interactions only occurred with the 3’side of the bases in the
major groove and the 5’ side in the minor groove. The interacting waters tended to form a double-layered ribbon in
the major groove and a spine in the minor groove. It is concluded that water-pi interactions are common in DNA.
These interactions tend to be sequence dependent and potentially unstack DNA bases contributing to DNA distortion /
flexibility important in many cellular processes such as transcription regulation and cancer formation.
ANALYSIS OF FOOD PRODUCTS FOR IRON, COPPER, AND MANGANESE BY FLAME ABSORPTION
AND FLAME EMISSION SPECTROSCOPY
Marguerite C. Ortman (Dr. Barbara Hopkins, Dr. Isam Marawi), Department of Chemistry, Xavier Univesity, Cincinnati,
Ohio 45207
Spectroscopy has been used as an analytical tool for a couple hundred years; however, the modern Atomic
Absorption Spectrophotometer (AAS) has only been in existence for several decades. The Varian 240 Fast
Sequential (FS) AAS, features the ability of simultaneously analysing multi-elements by either flame absorption
and/or flame emission. In this research project the analytical performance of both modes of this instrument will be
tested. Linear dynamic range, detection limits (DL), and relative standard deviations (RSD) will be assessed for each
element under study. Dietary iron, copper, and manganese in wheat, rice, bran and possibly other natural organic
products will be determined. An acid digestion method followed by a charring step has been used to bring the analyte
into solution for the determination by the instrument. The iron and copper will be simultaneously analysed by flame
absorption while manganese is measured by flame emission. Thus far, standards have been analysed for the
establishment of the analytical figures of merit. A calibration curve for copper had an regression coefficient equal to
0.9984 and for iron the regression coefficient was equal to 0.9975. The DL for copper is 0.054 ppm and for iron 0.01
ppm. In preliminarily testing for wheat the results for RSD, iron was 0.3%, copper was 0.24%, and manganese was
0.26%.
198
CHEMISTRY
THE SYNTHESIS OF AN ANALOGUE OF BLATTELLAQUINONE, THE COCKROACH PHEROMONE
Justin T. Fey, Colleen E. Moriarty (Daniel J. McLoughlin) Department of Chemistry, Xavier University, Cincinnati, OH
45207-4221
The natural sex attractant pheromone for the German cockroach Blattella germanica was recently isolated, its
structure determined, and synthesized. We have two goals to our research design: First the re-synthesis of the
pheromone (3,6-dioxocyclohexa-1,4-dien-1-yl)methyl 3-methylbutanoate (common name blattellaquinone). The
purpose of this re-synthesis is to examine the possible use of the reaction sequences as a method of teaching an
oxidation reaction involving Cerium Ammonium Nitrate. The synthesis has been performed in both the presence and
the absence of DMAP as a catalyst and proton-NMR has been utilized to follow the progress of the reaction and to
identify the products. Attempts to following the progress of the reaction with GC techniques did not prove to be an
advantage over use of proton-NMR techniques. In addition to repeating the literature synthesis of blattellaquinone,
our second goal is to make a new analogue of the Blattella germanica pheromone by utilizing a Friedel-Crafts
reaction type in the synthetic sequence. The reaction steps necessary to produce 2-(3-oxohexyl)benzo-1,4-quinone
using 2,5-dimethoxybenzyl alcohol and 1-chloro-3-pentanone as starting reactants will be presented.
IMPROVED SYNTHESIS AND CHARACTERIZATION OF 5, 10, 15, 20-TETRA(9-ETHYL-3-CARBAZOYL)
PORPHYRIN
Misty J. Bailey, Leslie A. Alexander, and Katherine L. Lanier, (Cynthia P. Tidwell) Department of Biology, Chemistry,
and Mathematics, The University of Montevallo, Montevallo, AL 35115. USA
There is considerable interest in electron transfer involving porphyrins, driven by the need to better understand
photosynthesis and potential applications such as solar energy and photodynamic cancer therapy. Substituted with
strongly electron withdrawing carbazoyl groups 5, 10, 15, 20 tetra (9-ethyl-3-carbazoyl) porphyrin is expected to
undergo ready photo reduction with suitable donors. The objective of this work is to improve the yield of this
porphyrin so that it can more easily be studied for these many applications. The synthesis previously reported with a
10% yield was the condensation of pyrrole with 9-ethyl-3-carbazoyl benzaldehyde. The reaction was refluxed in a
50:50 mixture of propionic and octanoic acids. The resulting reaction mixture was filtered and purified with column
chromatography using silica gel as the packing material and chloroform as the eluent. The yield has been improved
by adding a strong oxidizing agent 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, DDQ, at the completion of the
reaction. The reactions mixture was then purified using column chromatography and characterized by 1H NMR, 13C
NMR, mass spectroscopy, electronic absorption spectroscopy and fluorescence spectroscopy. Other modifications of
the procedure are currently being investigated which include varying the concentration of the reactants in the reaction
mixture as well as employing other techniques in the purification of the product such as HPLC and crystallization.
Wide-ranging bioassay for the presence and relative toxicity of genotoxic agents
Vanessa R. Palmer (Emily H. Turner, Haley Pugsley, and Dr. Norman J. Dovichi), Department of Chemistry,
University of Washington, Seattle, WA 98195
A handful of species of bacteria and yeast have been successfully engineered as biosensors for the presence of
genotoxic agents. While the methods of detection may differ, all of these assays share a common drawback: low
thresholds for DNA damage. Our group has created a Deinococcus radiodurans construct containing a translational
fusion of the DNA repair protein RecA and green fluorescent protein (GFP). When the organism produces RecA to
initiate the SOS pathway in response to DNA damage, GFP fluorescence is detectable at proportional intensities.
The key advantage of our biosensor is the extremely wide scope of tolerance of D. radiodurans to genetic damage via
alkylation, double strand breaks, and oxidative stress; it is, in fact, the most radioresistant organism known. Several
genotoxic compounds, including formaldehyde and mitomycin C, were tested. Survival curves and plates were
generated after exposure to determine maximum and optimal dosages to induce RecA production. Fluorimetry was
used to determine the linear dynamic range of response; the kinetics of the RecA response were also analyzed. To
determine stochastic variability in RecA expression between individual cells, fluorescence microscopy was performed.
Using these methods, DNA damage in D. radiodurans relative to control cells with baseline RecA levels is
quantifiable, making the RecA:GFP fusion strain potentially viable as a sensitive detector for dangerous contaminants
in environments too hostile for other organisms.
199
CHEMISTRY
ENANTIOSELECTIVE SYNTHESIS OF (-)-CONIINE
Jeffrey S. Cannon, Jason Papazian, Michael W. Heffner (Dr. Donald R. Deardorff), Department of Chemistry,
Occidental College, 1600 Campus Rd., Los Angeles, California 90041
(-)-Coniine is a toxic six-membered ring alkaloid that is isolated from spotted hemlock. The molecule has been a
popular synthetic target for showcasing new methods of enantioselective synthesis. Our own strategy for the synthesis
of (-)-coniine blends the enantioselectivity of an enzyme with the rich organometallic chemistry of pi-allyl palladium
and ring-closing metathesis.
The enzyme, oxynitrilase, is an especially robust enzyme found in raw almonds. This enzyme is able to produce the
corresponding unsaturated cyanohydrin of trans-2-hexenal in greater than 99% enantiomeric excess. The cyano
moiety is then converted to the ethyl ester and the hydroxy function transformed to an acetate leaving group in
preparation for a subsequent palladium—catalyzed allylic substitution step. Importantly, the stereochemistry is
conserved throughout this process. Exposure to palladium(0) initiates a novel substitutive 1,3-chiral transfer where
the crucial 3-butenylamino fragment is incorporated at the gamma-position of the alpha,beta-unsaturated ester with
full enantiofidelity. The unsaturated amine then undergoes an intramolecular Grubbs-catalyzed metathesis to form an
unsaturated piperidine ring system. Catalytic hydrogenation over Pd/C affords the desired natural product in high
enantiomeric excess and yield.
The successful synthesis of (-)-coniine provides proof-of-concept for our sequential enzymatic-organometallic
strategy. This unique methodology has widespread applications in the synthesis of other biologically important
piperidines.
AN ACTIVE BIOMIMETIC CATALYST FOR H+ REDUCTION USING THE FERRITIN PROTEIN CAGE
ARCHITECTURE
Chad Shoopman (Zachary Varpness, John W. Peters, Mark Young, Trevor Douglas), Lancy Scholars Program,
Department of Chemistry and Biochemistry, Montana State University - Bozeman, Bozeman, MT 59717
A biomimetic approach has been used to create a synthetic hydrogenase mimic capable of H+ reduction to form H2.
Catalytic Pt metal nanoparticles were synthesized within the protein cage architecture of ferritin to generate a
protein-based catalyst with high activity. Ferritin is a naturally occurring protein that functions to sequester Fe as a
nanoparticle of Fe2O3 within the protein cage. The protein is composed of 24 subunits that self-assemble into a 12
nm diameter cage-like architecture with a 8 nm interior cavity. In the ferritin-based, biomimetic enzymes, the Pt
clusters act as active sites analogous to metal clusters in hydrogenase enzymes. The protein cage architecture
provides a template for the formation of small highly active Pt nanoparticles, and acts to prevent Pt particle
association and aggregation. The H+ reduction reaction is driven by a coupled photochemical reaction with
Ru((bpy)3)2+ present as a photocatalyst, methyl viologen as an electron transfer mediator, and EDTA as a sacrificial
electron donor. The measured hydrogen production rates for these catalysts were found to be comparable, or better
than those of, hydrogenase on a per protein basis and exceed reported per Pt production rates of other Pt-based
catalysts. These results demonstrate the utility of a biomimetic approach towards the challenges of hydrogen
production.
200
CHEMISTRY
DETERMINATION OF PHYSICAL AND CHEMICAL PROPERTIES OF FLUORINATED CARBOXYLIC
ACIDS
Kelley A. O’Hara, Ray J. Clifford, Nisreen M. Madhoun, Justin A. Miscavige (Dr. Shirley G. Fischer-Drowos and Dr.
Linda R. Betz ), Department of Chemistry, Widener University, Chester, Pennsylvania 19013
Fluorinated processing aids (FPAs) are used in the industrial production of fluoropolymers. The demand of these
materials in the consumer market is great due to their desirable properties. Non-stick frying pans, stain-resistant
fabrics and carpeting, cleaning and waxing products are just a sample of the products in which fluoropolymers are
used. Their thermal and stain-resistant characteristics make this family of polymers of particular interest.
Unfortunately, these very same properties raise concerns about their environmental impact. In order to ascertain the
persistence of these processing aids, their properties need to be determined. In addition, the process of degradation
and the byproducts of that process need to be characterized as well. This will allow us to better understand the
mechanism of environmental transport. Two byproducts of fluoropolymer processing aids were examined, specifically
2H-Hexadecafluoro-2-decenoic acid, an unsaturated fluorinated carboxylic acid and 2-Perfluorooctylethanoic acid, a
saturated fluorinated carboxylic acid. Characteristics determined include melting point, solubility and acid dissociation
constants. Instrumentation that was utilized to obtain these data include, differential scanning calorimeter, titrimetry
and HPLC/MS/MS. Preliminary data indicates that these acids have a low solubility in water at the level of 60 ng/uL.
The melting point of the unsaturated system was determined to be 105&#61616; C and the saturated system was
determined to be 85&#61616; C. Studies were conducted on hydrocarbon and perfluorinated analog systems for
comparison.
DEVELOPMENT OF A FIBER OPTIC SPECTROELECTROCHEMICAL ABSORBANCE SENSOR FOR
IN-SITU METAL ION DETERMINATIONS
Marquite N. Lilly (Paul A. Flowers), Chemistry & Physics Department, University of North Carolina at Pembroke,
Pembroke, NC 28372
Progress on the development of a fiber optic spectroelectrochemical (FOSEC) absorbance sensor for in-situ
determination of transition metal ions is reported. The sensor is constructed by incorporating small working, reference
and auxiliary electrodes into the inlet and outlet ports of a commercially available fiber optic flow cell, permitting
absorbance measurements on flowing samples subjected to controlled potential electrolysis. The electrochemical
characteristics of the sensor were evaluated via voltammetric measurements using aqueous ferricyanide as a test
system and determined to be acceptable. Aqueous copper(II) was chosen as a model system to investigate the
analytical merit of an analysis scheme involving (a) preconcentration via cathodic electrodeposition, followed by (b)
anodic stripping in an aqueous medium containing ethylenediamminetetraacetic acid (EDTA) and (c) subsequent
measurement of the absorbance of the copper/EDTA complex at 730 nm. Results of calibration studies employing a
10 minute electrodeposition step indicate a linear response (r2 = 0.97) up to approximately 6 mM Cu(II) with a
sensitivity of ~0.08 L/mmol and an estimated detection limit on the order of 0.1 mM. This work was supported by the
National Science Foundation`s Research Experiences for Undergraduates program (CHE-0353724).
201
CHEMISTRY
SPECTROSCOPIC INVESTIGATIONS OF THE BENZENE-HYDROGEN PEROXIDE COMPLEX IN
LOW-TEMPERATURE MATRICES
Khanh H. Do (Dr. Thomas M. Halasinski) , Department of Chemistry, Saint Joseph’s University, Philadelphia, PA
19131
Conventional hydrogen bonds involve the attraction of a hydrogen atom to the lone-pair electrons of fluorine, oxygen,
or nitrogen atoms. Nonconventional hydrogen bonds are formed when the lone-pair electrons are replaced by
another acceptor group such as an aromatic ring. Although these relatively weak nonconventional hydrogen bonds
may play an important role in biochemistry by influencing interactions between molecules, they have not been well
studied. Several investigations have been done on the simplest OH-aromatic hydrogen bond, the benzene-water
complex. We have extended these studies by investigating the benzene-hydrogen peroxide complex.
We have utilized the matrix-isolation method to study the benzene-hydrogen peroxide complex spectroscopically.
The spectral absorbances in our initial experiments indicate that more than one type of the benzene-hydrogen
peroxide complex may have formed in the argon matrices. We have explored several possibilities for the origin(s) of
these spectral absorbances. The first possibility that may account for these absorbances is the variation of the matrix
environment (matrix site effects). The second involves different conformations of the dimer complex. In particular,
we have studied the benzene-hydrogen peroxide complex involving the enantiomer of hydrogen peroxide. The third
possibility involves different polymers of benzene and hydrogen peroxide. To investigate all of these possibilities, we
have performed experiments using krypton matrices, annealing experiments, as well as theoretical calculations
utilizing density function theory (DFT). We will present the results of our investigations and discuss the possibilities
for the origin(s) of the spectral absorbances.
CONSTRUCTION AND CHARACTERIZATION OF A FIBER OPTIC SPECTROELECTROCHEMICAL
FLUORESCENCE SENSOR FOR IN-SITU METAL ION DETERMINATIONS
Kristen A. Arnett (Paul A. Flowers), Chemistry & Physics Department, University of North Carolina at Pembroke,
Pembroke, NC 28372
This poster describes the construction and initial characterization of a fiber optic spectroelectrochemical (FOSEC)
fluorescence sensor for the in-situ determination of transition metal ions. The sensor was fabricated by incorporating
small working, reference and auxiliary electrodes into the inlet and outlet ports of a commercially available fiber optic
flow cell, thereby permitting fluorescence measurements on flowing samples subjected to controlled potential
electrolysis. The electrochemical performance of the sensor was confirmed to be adequate based on voltammetric
measurements using aqueous ferricyanide. Aqueous copper(II) was chosen as a model system in order to investigate
the analytical merit of an analysis scheme involving (a) preconcentration of the analyte via cathodic electrodeposition,
followed by (b) anodic stripping in an aqueous medium containing calcein and (c) subsequent measurement of calcein
fluorescence quenching at 517 nm induced by its complexation with the analyte. Preliminary results obtained
employing a 5-minute electrodeposition step indicate the method’s detection limit to be on the order of 0.05 mM. A
summary of the data gathered thus far and plans for future work on this project will be presented. This work was
supported by the National Science Foundation`s Research Experiences for Undergraduates program (CHE0353724).
202
CHEMISTRY
PHEROMONES AS PESTICIDE ALTERNATIVES: METHODS FOR INCREASED RETENTION AND
EFFICIENT APPLICATION
Brent L. Lance, (Dr. Cynthia Atterholt), Chemistry and Physics, Western Carolina University, Hwy. 107 , 231 Natural
Sciences , Cullowhee, NC 28723
In an effort to produce a safer alternative to pesticides, the use of pheromones to disrupt insect mating cycles is both
intriguing and beneficial. These safe alternatives are designed to control specific pests and to leave other organisms
unharmed. This also reduces harmful pollutants that may enter the environment and affect the organisms that inhabit
it for many years. Laboratory data and field tests indicate that this method is highly effective for controlling pest
populations and is safe for the environment. The agricultural industry has a commercial interest in the successful
development of such products.
The purpose of this research is to test formulations of wax emulsions containing specific pheromones such that the
retention times of these pheromones will increase and allow for a longer lasting, more effective product. In addition to
increasing retention times, it is also ideal for the formulation to have a viscosity low enough to allow it to be applied to
the designated areas in an efficient manner.
Each formulation is tested in flow cells stored in a temperature controlled environment with a designated air flow rate.
Columns packed with a “Super Q” stationary phase are attached to the flow cells to trap the released pheromone.
The columns are eluted with solvent periodically. The eluted solutions are analyzed using Gas Chromatography to
determine concentrations of pheromone released at each interval. This data is then compared with previous data to
further improve the formulations, thus improving the effectiveness of environmentally-safe pest control methods.
DNA BENDING OBSERVED IN DNA/PROTEIN COMPLEXES IS CORRELATED WITH PHOSPHATE
COLLAPSE IN THE VICINITY OF CATIONIC PROTEIN RESIDUES
K. Courtney Huff, James W. S. Lewis (Dr. Lori Isom), Department of Chemistry, University of Central Arkansas,
Conway, Arkansas, 72035
DNA deformation is crucial in many fundamental biological processes such as DNA packing, proper functioning of the
immune system, gene activation, and DNA mutation repair. In this project we tested the theory that cation
interactions with DNA phosphate groups results in partial charge neutralization inducing DNA bending. We used
X-ray crystal structures of protein/DNA complexes to investigate the influence of cations on DNA structure. Unlike
monovalent ions, the position of cationic side chains, such as lysine and arginine are quantifiable and so their location
with respect to DNA phosphate oxygens can be calculated. We selected DNA/protein crystal structures with a
resolution of 2.7 &#506; or higher and screened for phosphate crowding around high densities of cationic protein
residues. The structures obtained were then used to investigate the relationship between phosphate crowding, cation
density, and the direction of DNA bending. Analysis of 121 structures led to results suggesting that phosphate
crowding is related to cation density. A positive correlation value (0.321 ± 0.070) was calculated for DNA/protein
complexes containing DNA that bends toward the bound protein, while a negative value (-0.215 ± 0.139) was
observed for complexes with DNA bending away from the protein. Complexes containing unbent DNA have a
correlation close to zero (0.018 ± 0.076). The observed correlations support our hypothesis that a relationship exists
between phosphate crowding, cation density, and DNA bending.
203
CHEMISTRY
Studies of Olefin Metathesis Initiators
Christian Coe (Dr. Robert K. Szilagyi), Department of Chemistry, Montana State University, Bozeman, 108 Gaines
Hall, Bozeman, MT 59715
The purpose of this research project was to explore the ligand effects on olefin metathesis activity for a series of
Mo(II) and (II) carbonyl-halide-phosphine complexes. These air and light tolerant complexes are active precursors in
the ring opening metathesis polymerization (ROMP) of norbornene (NBE). Both the mechanism of this reaction and
the variations in ROMP activity resulting from changes in the ligand environment are yet to be understood. These
initiators, which were synthesized in the Szilagyi lab at MSU, were subjected to a rigorous in-silico (computational)
investigation to elucidate the mechanism of the reaction and to identify the initiating carbene species. The
computational investigations, combined with X-ray absorption spectroscopic (XAS) measurements, were utilized to
define the electronic and geometric structures of these inorganic initiators. These insights will be used to provide
quantitative structure and activity relationships, which will aid in the design of novel catalyts for future olefin
metathesis reactions.
The Kinetics and Thermodynamics of Ternary Complex Formation Between 2,2’-Bipyridine and
Ferrioxamine B
Amy E. Ratliff, Joseph I. Wirgau, Department of Chemistry and Physics, Radford University, Radford, VA 24142.
The treatment for &#61538;-thalassemia involves repeated blood transfusions, which results in an iron-overloaded
patient, and eventually death. The most common method for iron removal from these patients involves chelation
therapy, most often utilizing desferal. Desferal is the trade name for the mesylate salt of desferrioxamine B, a
naturally occurring siderophore. Iron-overload is often associated with vitamin C (ascorbic acid) deficiency, which
results from an increase in the conversion of ascorbate to oxalate. Ascorbate increases the levels of chelatable iron
by delaying the transfer of iron from ferritin to insoluble hemosiderin. Administration of both desferal and ascorbate
for iron-overload treatment results in an increase in urinary iron excretion. Unfortunately, large doses of ascorbate in
these cases leads to a variety of toxic effects. This could be due to the presence of ascorbate increasing the amount
of iron available for Fenton chemistry and free radical generation, whereas a large amount of ascorbate may create a
reductive environment, which prevents iron recycling. Our current research is interested in the redox chemistry
associated with the reaction of ferrioxamine B with ascorbate. This reaction is found to be exceedingly slow, unless
an Fe(II) specific chelator is present. We report that the reduction of ferrioxamine B by ascorbate is facilitated by the
presence of 2,2’-bipyridine. The kinetics and thermodynamics of ternary complex formation between 2,2’-bipyridine
and ferrioxamine B will be discussed and compared to sulfonated-bathophenanthroline and ferrioxaimne B.
CONTINUED PROGRESS TOWARD THE SYNTHESIS OF (-)-TETRANORLABDANE OXIDE FROM
ABIETIC ACID
Gary L. Willman (Robert D. Rapp, Christian S. Hamann) Department of Chemistry and Biochemistry, Albright College,
Reading, Pennsylvania 19612
(-)-Tetranorlabdane oxide is an extremely valuable fixative used by the perfume industry that was formally produced
from a natural product of the sperm whale (Physeter macrocephalus) called Ambergris. Due to environmental
concerns, Ambergris is now banned for use in many industrial countries, including the United States. The perfume
industry had to devise a synthetic way of making this valuable natural product. Today, (-)-tetranorlabdane oxide is
currently synthesized by the perfume industry by one synthetic pathway with (-)-scarelol as the starting material. This
production cost of (-)-tetranorlabdane oxide from (-)-scarelol is very high, which is passed on to the consumer. The
high demand of (-)-tetranorlabdane oxide on the international market also contributes to the high cost of perfumes
containing (-)-tetranorlabdane oxide.
We are reporting on the progress of synthesizing (-)-tetranorlabdane oxide from abietic acid, which is a naturally
occurring product found in pine rosin. We have successfully isolated gram quantities of abietic acid from pine rosin.
The first six steps of the proposed synthetic pathway been successfully completed and their products analyzed. We
also propose future steps in the synthetic pathway that will eventually lead to the product, (-)-tetranorlabdane oxide.
204
CHEMISTRY
Enzymatic Labeling of Phosphoproteins
Mary S. Jacob (Mr. Keith Green and Dr. Mary Kay Pflum), Department of Chemistry, Wayne State University, Detroit,
Michigan 48202
The phosphorylation of serine, threonine and tyrosine amino acid residues by protein kinases is an important
transformation in the cell. The formation of cancer is associated with these protein kinases and phosphorylation
events. Despite the importance of phosphoproteins and phosphorylation, in vitro methods of determining which
proteins are phosphorylated and which kinase catalyzed the reaction are challenging or ambiguous. These methods
often involve harsh reaction conditions or unnatural proteins and substrates. Therefore, techniques that allow rapid
identification of kinase protein substrates have the potential to reveal new protein targets for the development of
future cancer therapies. We describe a purely chemical method that has the potential to unequivocally determine
phosphoproteins and kinase-substrate partners. From the analysis of multiple kinase crystal structures(1), a
noteworthy observation is the presence of an area of empty space near the phosphate end, suggesting solvent
exposure. We hypothesize that the solvent exposed area is likely to accommodate the addition of a small group
added to the phosphate terminus of ATP and not hinder catalysis. Consistent with our hypothesis, phosphate-terminus
modified ATP analogues were previously synthesized and acted as competitive inhibitors of ATP binding.(2)
Specifically, we propose the development of ATP derivatives that have the capability to label phosphorylated
peptides for unambiguous detection. Upon phosphorylation by a kinase, the labeled phosphate end will be transferred
to the peptide/protein and the specific phosphorylation site can be identified by using standard spectroscopic methods
and immunoassays. This proposed strategy describes a novel method to selectively label and purify phosphoproteins.
Furthermore, the natural substrate and kinase are used, making the use of ATP derivatives a useful strategy in signal
transduction and proteomics research.
(1)Hubbard, S.R. Crystal Structure of the Activated Insulin Receptor Kinase Complex with Peptide Substrate and
ATP Analog. EMBO J., 1997, 16(18), 5573-5581
(2) Samukov, V.V.; Knorre, D.G.; Kurbatov, V.A. General Method for the Synthesis of ATP Gamma-Derivatives.
FEBS Lett. 1976, 70(1), 105-108
Identification of Essential Histidines in Arsenite Oxidase
Shannon F. Renfrow, Keri L. Hershberger, Mark A. Smith, Kari A. Kubalanza, (Dr. Gretchen Anderson), Department
of Chemistry, Indiana University South Bend, 1700 Mishawaka Avenue, South Bend, IN 4663-7111
The presence of arsenic as an environmental contaminant is a recognized concern. Due to the relative abundance of
arsenic as a naturally occurring element, it is of special interest that some bacteria are able to survive in the presence
of significant levels of arsenic. The bacterium Alcaligenes faecalis survives in such an environment because it
produces the enzyme arsenite oxidase. Arsenite oxidase allows the bacterium to convert arsenite, a highly toxic form
of arsenic, to arsenate, a significantly less toxic form of arsenic, by a redox reaction involving the exchange of two
electrons. Intramolecular electron transfer occurs via a molybdopterin cofactor and two unique iron sulfur clusters.
Other enzymes which also use a molybdopterin cofactor for redox reactions are strongly inhibited by arsenite, which
binds irreversibly within the coordination sphere of the molybdenum atom. In contrast, arsenite oxidase remains
reactive in the presence of arsenite. Arsenite may therefore bind to amino acids in the active site rather than directly
to the molybdopterin cofactor. Earlier data suggest that histidines are essential for the enzyme’s activity, and the
X-ray crystallographic structure indicates the presence of two histidines in the putative active site. To test whether
histidines may serve as functional binding sites for arsenite, the active site of the enzyme was protected with arsenite
while the remaining histidines were modified with DEPC. The histidines in the arsenite binding site were exposed by
removal of arsenite, and modified with radiolabeled DEPC. The active site histidines were identified following tryptic
digestion of the protein, and purification and sequencing of the radiolabeled peptides.
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CHEMISTRY
MUSHROOMS AROUND THE WORLD: IS D. CONFRAGOSA FOUND IN BERKSHIRE COUNTY, MA,
USA, THE SAME SUBSPECIES AS D. CONFRAGOSA VAR. TRICOLOR OF KOREA?
Nida Intarapanich (Dr. David Myers), Department of Chemistry, Simon’s Rock College of Bard, Great Barrington,
Massachusetts 01230
The purpose of this project was to determine if the white-rot fungus Daedaleopsis confragosa exhibits different
properties varying with locale. We examined whether samples from two places located roughly halfway around the
world from each other possessed different properties and may in fact be distinct subspecies. Dried fruit-bodies of
Daedaleopsis confragosa were gathered in Berkshire County, MA, USA. Preliminary bactericidal screenings of the
crude petroleum ether extract were performed against Escherichia coli, Staphylococcus aureus, and Pseudomonas
aeruginosa. The results showed little to no antibacterial activity against these microorganisms, which was in direct
contrast to the results published by Kim, Jung, and Min (2001) about Daedaleopsis confragosa var. tricolor picked in
Korea. These results caused us to further examine the local Berkshire mushrooms with the purpose of determining if
they were in fact a different subspecies than those researched in Korea. In addition, other solvent systems as well as
the antioxidant activity of the extract were investigated, and will be reported upon.
Kim, E.-M.; Jung, H.-R.; Min, T.-J. Bull. Korean Chem. Soc. 2001, 22(1), 59.
HIGH CURVATURE MOLECULAR STRUCTURE
Per D. Gunst (Dr. Joshua Smith), Department of Chemistry, Humboldt State University, Arcata, California
95521-8299, , (Charlie Robertson and Dr. Benjamin T. King), Department of Chemistry, The University of Nevada at
Reno, Reno, Nevada 89557-0042
Introducing curvature into molecular structures has been of considerable interest in recent years, e.g. fullerenes,
carbon nanotubes, and coranulene. Coranulene, the first of these compounds that was studied with respect to its
curvature, is a bowl shaped molecule composed of five benzene rings ortho-fused to form a five membered central
ring. The goal of this project is to synthesize a bowl shaped molecule like coranulene, but with four benzene rings
ortho-fused to form a four membered central ring. This arrangement is expected to introduce curvature far greater
than that found in coranulene. The central cyclobutadiene ring acts as the template for this compound as a complex
to cobalt cyclopentadiene. Once made, this compound’s characteristics vs. corannulene and other curved polyarenes
will be studied. For this synthesis, a five step reaction pathway is proposed. Coupling of
1-bromo-3,5-dimethoxybenzene with trimethylsilylacetylene is the first step of the pathway, followed by removal of the
trimethylsilyl protecting group giving 1-ethynyl-3,5-dimethoxybenzene. Both of these steps have been carried out with
good yields (>80%). 1-Ethynyl-3,5-dimethoxybenzene is then coupled with 1-iodo-3,5-dimethoxybenzene, generated
via a Sandmeyer reaction from 3,5-dimethoxyaniline, yielding the tolane 3,3’,5,5’-tetramethoxydiphenylacetylene.
Yields for this step were very poor, and current work is now centered on optimization of reaction. The tolane will be
used to form [tetrakis(3,5-dimethoxyphenyl)cyclobutadiene]cyclopentadienylcobalt(I). Oxidation of this compound
should produce the target molecule.
EXTRACTION AND IDENTIFICATION OF ORGANIC MATERIAL ASSOCIATED WITH ROCKS AT
POSSIBLE MAMMOTH SITES
Jeffrey C. Shearer (Dr. Karen L. Steelman), Department of Chemistry, University of Central Arkansas, Conway, AR
72035
Plasma chemistry was used to extract organic carbon from rocks on the Californian coast purported to have been
sites of mammoth rubs. A high vacuum apparatus was built to conduct plasma oxidation. Low-temperature plasmas
are ideally suited for these samples instead of high-temperature combustion because it is below the decomposition
temperature of inorganic minerals. Milligram-size samples were treated with base to remove potential humic acid
contamination before exposure to the plasma. In contrast, traditional radiocarbon sample preparation involves an
acid-base-acid treatment, which often dissolves trace organic compounds along with mineral and soil contamination.
Organic carbon was extracted for stable-isotope and radiocarbon measurements. Infrared spectroscopy was used to
characterize material associated with the rock surface. Fatty acids that might be present in the rocks were derivatized
to fatty acid methyl esters for identification by gas chromatography/mass spectrometry.
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CHEMISTRY
A Novel Pathway for Polysaccharide Precursor Synthesis
Author: Liron Bar-Peled, Research Sponsor: Maor Bar-Peled, Department: Plant Biology, Institution: Complex
Carbohydrate Research Center at the University of Georgia, 315 Riverbend Rd. , Athens, GA 30602
Polysaccharides are the most abundant class of molecules that exist in nature. They have numerous functions
such as structural support (cellulose in plant cell walls), storage (starch), cell-to-cell communication (glycoproteins and
glycolipids), protein trafficking and protein folding. Polysaccharides are synthesisized from activated sugar-moieties
known as nucleotide sugars. A question which we are trying to address is the different pathways by which these
precursors are synthesized. One route by which nucleotide sugars are made involves a family of enzymes known as
nucleotide sugar pyrophosphoyrlases (PPase) which convert a nucleotide (NTP) and sugar-1-P to NDP-Sugar and
inorganic pyrophosphate PPi. PPases are very specific enzymes and convert only one type of sugar-1-P and
nucleotide to respective nucleotide sugar. We have identified a gene in Arabidopsis thaliana that shares some amino
acid sequence similarity to the well-characterized UDP-Glucose PPase (BLAST analysis shows 29% sequence
identity and 39 % similarity). The recombinant gene was over-expressed in Escherichia Coli and the protein was
purified via affinity chromatography. A series of assays were conducted varying the type of nucleotide and sugar-1-P
used and it was discovered that this enzyme converts 5 different sugar-1-P’s (Glucose-1-P, Galactose-1-P, Xylose-1
-P, Glucuronic acid-1-P and Galacturonic acid-1-P) and UTP to respective nucleotide sugars. This enzyme has been
termed UDP-Sugar PPase due to its broad sugar-1-P specificity. Initial biochemical characterization shows
differences in kinetics between UDP-Sugar PPase and other substrate specific PPases. Additionally, this enzyme
differs in its substrate specificity when compared with UDP-Sugar PPase from Rice. Since there are other enzymatic
pathways by which nucleotide sugars are made, it remains unclear the biological role that UDP-Sugar PPase has in
vivo. Our objective is full biochemical characterization of this enzyme.
INVESTIGATION OF BIFUNCTIONAL CROSS-LINKING REAGENTS AS PROBES FOR STUDYING
PROTEIN-NUCLEIC ACID INTERACTIONS BY MASS SPECTROMETRY
Elizabeth A. Crosland (Dr. Daniele Fabris), Department of Chemistry & Biochemistry, University of Maryland,
Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250
The bifunctional cross-linking reagents sym-triazine trichloride (s-TT) and 1,4-phenyl-diglyoxal (PDG) were evaluated
as potential bridging probes for the investigation of the 3-dimensional structure of large macromolecular complexes
using high-resolution mass spectrometry (MS3D). The s-TT adduct with the dipeptide glycine-lysine (GK) and the
PDG alkylation of the dinucleotide dideoxyguanylate (GG) were characterized by electrospray ionization (ESI)
Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry. Using this technique, the reaction products
were rapidly identified from their characteristic mass signatures, associated with the covalent bonding of the reagent
with the substrate. It was found that the reaction proceeds through the attack of the nucleophilic amine functional
groups of the substrates by the electrophilic chlorine atoms on s-TT and the 1,2-dicarbonyl groups of PDG. The
reaction process was optimized in near-physiological conditions to facilitate the probing of folded complexes in their
native state.
Author`s Home Institution: Villa Julie College
1525 Greenspring Valley Road
Stevenson, MD 21153
Investigation of Ammonia Inversion via Numerical Integration Techniques
Jack DaSilva, Clyde Stanton and Rodney Dunning, Chemistry and Physics, Birmingham-Southern College, 900
Arkadelphia Road, Birmingham, AL 35254
The goal of this work was to develop a computationally efficient method of directly applying Schroedinger’s equation
to systems which have traditionally been treated with computationally intensive methods such as Hartree-Fock. Our
method does not rely on solving for the electronic structure explicitly. We have investigated the tunneling artifact
associated with the inversion of ammonia. We developed software to solve Schroedinger’s Equation for a
double-well potential which approximates the ammonia potential energy surface. Our calculations showed qualitative
agreement with experimental results and Hartree-Fock calculations, ie., we find the energy level splittings of the
system to increase as the energy levels themselves increase.
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CHEMISTRY
The Synthesis of small molecules for use as Chemopreventives
Courtney Henry and Timika Williams, (Dr. Marion Franks), Chemistry Department, North Carolina A&T State
University, 1601 E Market St.
Carcinogenesis is a chronic and multi-step process, resulting from mutagenic damage to growth regulating genes and
their products, which ultimately leads to development of invasive or metastatic cancers’. During the change from
normal to metastatic cells, carcinogenic steps can be arrested or reversed through treatments known as cancer
chemoprevention. In 2001, there were 553,768 cancer deaths in the US.” African-American men and women have the
highest rates of cancer mortality among all the races.” The term “Chemoprevention” is coined for cancer prevention
and cancer control by use of naturally occurring and/or synthetic compounds. The occurrence of the disease may be
completely prevented, blocked, or reversed. This approach is promising because chemotherapy and surgery have not
been fully effective against the high incidence or low survival rate of most of the cancers. The long term goal of this
proposal is to understand the detailed structure-activity relationships for possible cancer chemopreventive agents.
The goal of our research:
1. To prepare and completely characterize the synthesis of compounds that can be used as chemopreventive agents.”
We have facilities whereby we can make compounds and characterize them using NMR, IR, and GO/MS.
2. To test our compounds potency using biochemical and cellular biology techniques.
CORRELATION OF KETO-ENOL EQUILIBRIUM AND ACID DISSOCIATION CONSTANTS
Sherri Young (Dr. Christian Hamann), Department of Chemistry and Biochemistry, Albright College, Reading, PA
19612-5234
The primary objective of this project was to investigate the correlation between acid dissociation constants and
keto-enol equilibrium constants of a group of keto-enol tautomers with varying steric bulk. We also planned to
analyze these compounds under a variety of solvent and temperature conditions and to investigate if the same trends
could be applied to their nitrogen analogues.
The compounds that we investigated thus far include: 2,4-pentanedione, dimethyl malonate, 3,5-heptanedione, diethyl
malonate, 2,6-dimethyl-3,5-heptanedione, diisopropyl malonate, 2,2,6,6-tetramethyl-3,5-heptanedione, and
di-tert-butyl malonate. This series of compounds is listed in order of increasing steric bulk, which forces nonplanarity
in a molecule.
We used proton NMR spectroscopy to determine keto-enol equilibrium constant values for these compounds using
the solvent deuterated chloroform. We calculated the keto-enol equilibrium constant values by integrating the NMR
peaks for the keto and enol hydrogens for each compound and dividing the amount of enol hydrogens by the amount
of keto hydrogens.
At the conclusion of our summer research, we found that for the investigated diketones the acid dissociation constant
values increase with increasing steric bulk. In other words, forcing nonplanarity causes the keto-enol equilibrium to
favor the enol form and thus increases the keto-enol equilibrium constant. Literature data indicate that acidity
decreases with increasing steric bulk for these compounds, suggesting that there is some inverse correlation between
keto-enol equilibrium and acidity. The data we collected were less clear for the diesters and further investigation
using more sensitive NMR techniques may be necessary. Also, the literature dataset is incomplete concerning acid
dissociation constants so our future work must include the determination of these values. This provides us with the
opportunity to determine acid dissociation constants for all of the compounds using one technique.
We plan to use this preliminary dataset to propose a model for the stabilization of enols and enolates due to structural
constraints that force planarity or nonplanarity, and determine if a similar model can be applied to the nitrogen
analogues of these compounds.
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CHEMISTRY
Essential Histidines in the Active Site of Arsenite Oxidase
Keri Hershberger (Dr. Gretchen Anderson), Department of Chemistry, 1700 Mishawaka Ave South Bend, IN 46634
The purpose of this project was to explore the active site arsenite oxidase. Arsenite oxidase is an enzyme that
oxidizes arsenite into arsenate. Arsenate, a less toxic form of arsenic, is charged and therefore more easily removed
from tap water. Arsenic contamination of ground water is a major environmental problem in Bangladesh, but also here
to a lesser extent in the United States. Arsenic poisoning has been linked to lung, uterus, bladder, and skin cancer.
The information that can be learned from studying how the oxidation occurs via arsenite oxidase could provide a way
to remove arsenic from tap water on a large scale. The goal of the project was to determine if histidines are essential
to arsenite, substrate, binding. By the addition of arsenite to purified enzyme, only histidines not in the active site will
be modified upon addition of diethyl pryocarbonate. Excess substrate and modifier are removed, and radioactive
diethyl pryocarbonate is used to modify active site histidines. Arsenite oxidase is then cleaved in to small peptides
and separated by size with High Pressure Liquid Chromatography. The fractions of peptides containing radioactivity
are sequenced. The identification of histidines in the arsenite binding site is accomplished by comparison of the
amino acid sequence in the radiolabelled peptides to the sequence observed in the active site by X-ray
crystallography.
RADIOCARBON MEASUREMENTS OF BLACK CARBON IN MEXICO CITY AEROSOL
Amanda MacMillan (Dr. Karen Steelman), University of Central Arkansas, Conway, Arkansas 72035
We report measurements of radioactivity in fine aerosol in one of the largest megacities in the world, Mexico City.
Naturally occurring radionuclides enable us to study the transport of ozone and aerosol in the troposphere.
Radiocarbon measurements for carbonaceous aerosol can indicate sources of carbon as either from fossil or biomass
derived carbon sources. Samples were collected on quartz fiber filters by using cascade impactors and high-volume
air samplers on the rooftop of the main laboratory of El Centro Nacional de Investigación y Capacitación Ambiental
(CENICA) during the month of April 2003. Radiocarbon levels were determined by accelerator mass spectrometry
and demonstrated a 70% modern biogenic-to-fossil ratio indicating that carbonaceous aerosol was from three
sources: fires in the Yucatan; oxidation of monoterpenes and sesquiterpenes emitted from a nearby fruit-drying
facility; and trash burning within the city.
QSAR Modelling of Agonism at the LPA3 Receptor
Titilola Afolabi (Dr. Abby Parrill), The Department of Chemistry, The University of Memphis, Memphis TN 38111
QSAR MODELING OF LPA3 ACTIVITY
Abstract:
Lysophosphatidic Acid (LPA) is a phospholipid growth factor that activates the cell through a family of G protein
coupled receptors which are LPA1, LPA2, LPA3 and LPA4. Due to the growing understanding of LPA initiating a
myriad of responses which includes cell apoptosis, migration, cytoprotection, chemotaxis and oxygen radical
production, the study of the specific receptor activities is vital to the understanding of the pathological and
physiological role of LPA. Agonism at LPA3 and LPA1 receptors activate platelet aggregation leading to
atherosclerotic plaque rapture, a precursor to stroke and heart attack. In this research, agonism at LPA3 is studied
using Quantitative Structure Analysis Relationship (QSAR) to relate biological activity to a mathematical model. This
computational project involves the modeling of LPA analogs on MOE (Molecular Operating Environment) and the
superimposition to a fixed structure of LPA 18:1. After superimposition, selected conformation of each LPA analog is
described based on the topology and the geometry of their atoms and molecules. On another computational program,
Cerius2 uses algorithm of Genetic Function Approximation (GFA) to calculate the regression by optimizing 3
descriptors to generate a QSAR model. The result of this research will be used to predict the activity of compounds of
unknown activity at the LPA3 receptor and improve the process of drug synthesis.
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CHEMISTRY
PROTEIN FINGERPRINT FOR BREAST CANCER CELLS USING GEL AND CAPILLARY
ELECTROPHORESIS
Joan Bleecker (Melissa Harwood and Norman Dovichi PhD.) Department of Chemistry, University of Washington,
Seattle, Washington 98105
I am studying protein expression “fingerprints” of adenocarcinoma breast cancer cells. Protein expression differs from
case to case, and cancer cells undergo major changes through different cancer stages. I have been working on
finding more specific protein fingerprints using gel electrophoresis and capillary electrophoresis (CE). The ultimate
goal of this research is to improve cancer prognoses and to develop better patient-specific treatment regimens.
First off the samples we use into CE and gels are from lysed cancer cells grown in culture. I have been studying the
effects of ethanol fixation, which stops protein degradation during lysis, on the solubility of proteins in lysis buffer and
resolution of samples in gels. I have made steps in creating a standard protocol to give the highest resolution of
protein separation.
For gels, I used 1D and 2D SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) to separate
proteins based on different characteristics. 1-D gels separate proteins based on molecular mass. 2D gels separate
proteins first by their isoelectric point on an isoelectric focusing strip (IEF) and then by molecular weight on a gel
(SDS-PAGE).
In capillary electrophoresis, protein sample, labeled with the fluorescent dye, is suspended in a buffer and passed
through a capillary. The proteins are separated in the capillary by two methods, one based on size of proteins and the
other the proteins’ hydrophobicities. The buffer and sample exit the capillary and focus into a narrow zone, which is
then illuminated by a laser. The laser beam excites the labeled proteins to fluoresce and that fluorescence passes into
a detector. The detector counts fluorescent photons and a computer records that signal as peaks over time. These
peaks represent separated proteins that can be mapped to protein expression in breast cancer cells.
ACCELERATOR MASS SPECTROMETRY MEASUREMENTS OF CALCIUM OXALATE ACCRETIONS
ASSOCIATED WITH SPAINISH ROCK PAINTINGS
Jacque DuPriest (Dr.Karen Steelman), Department of Chemistry, University of Central Arkansas, Conway, Arkansas
72035
At the Cueva del Tío Modesto in Spain, rock paintings had become coated with a mineral accretion over time. Using
X-ray diffraction and Fourier transform infrared spectroscopy, we determined that the accretion contains whewellite,
the monohydrate of calcium oxalate. Oxalate accretions on rock surfaces are usually attributed to metabolic activity
of fungi and lichens that live or have lived on these surfaces. Accelerator mass spectrometry radiocarbon dating of
oxalates may be used to establish minimum ages of pictographs painted over these accretions or maximum ages for
paintings that are covered with oxalate. Samples were acid treated to remove any carbonate contamination, followed
by oxidation of organics using an oxygen plasma. Remaining solid oxalate mineral was combusted to carbon dioxide,
which was then reduced to graphite over a metal catalyst. Accelerator mass spectrometry measurements of
radiocarbon place the older paintings at this site to at least the 5th and 6th millennium BC (cal BC 5230-5010 and cal
BC 4800-4610), demonstrating that the art is older than expected according to stylistic parallels.
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CHEMISTRY
SYNTHESES OF IMINES AND A Ni(II) COMPLEX FROM TETRAKETONE BIS-1,4(1,3-DKETOBUTYL)BENZENE AND HEXAKETONE BIS-1,4-(1,3,5-TRIKETOHEXYL)BENZENE
Robin L. Mizzell (Dr. Sandra Boatman), Department of Chemistry, Hollins University, 8003 Fishburn Drive Dana 220,
Hollins University, Roanoke, VA 24020
Tetraketone bis-1,4-(1,3-diketobutyl)benzene (I) and hexaketone bis-1,4-(1,3,5-triketohexyl)benzene (II) were
prepared by condensing acetone or acetylacetone with dimethyl terephthalate using sodium hydride according to the
method of Von Schriltz et al (J. Org. Chem. 32 1774 (1967)). Reactions of I and II with aniline and diamines p-, m-,
and o-phenylenediamines gave products in which both terminal carbonyls of the respective polyketone reacted. With
aniline, I and II gave a diiminodiketone and a diiminotetraketone, respectively. Under the conditions reported here,
only one amino group of each diamine reacted, so the products obtained from I and II were diaminodiiminodiketones
and diaminodiiminotetraketones, respectively.
For imine preparation, in a typical reaction, 0.001 mole of I or II was refluxed with 0.002 mole of anililne or diamine in
5 mL of absolute ethanol for two hours. Products were recrystallized from methanol or 95% ethanol. Structures of the
imines were confirmed by C,H analysis and infrared (IR) spectra, which contained appropriate carbonyl absorbances.
IR spectra of the diamine products contained doublets at 3200-3400 cm-1, indicating the presence of a primary amino
group.
In earlier research Boatman and Clark (unpublished results) combined benzoylacetone (BA) with nickel(II) chloride
and diethylenetriamine (dien) followed by NaPF6 to form the orange complex BA(dien)Ni(II) PF6 (III). Its structure
was determined by crystallography. In the current work, I was combined with nickel chloride and dien in a 1:2:2 ratio
to afford an orange complex that had almost exactly the same IR spectrum as III. The complex`s structure is
proposed to be the bis-di-(Ni(dien)) complex analogous to that of BA. Attempts to prepare the mono-Ni(dien) complex
have been unsuccessful.
ESTABLISHING AN IN VIVO MODEL TO STUDY THE EFFECT OF INTERLEUKIN-17
Asieh Zarghami, (Reen Wu), Internal Medicine, University of California at Davis, CCRBM & Internal Medicine, 451
East Health Science Dr.,Davis, CA, 95616
Nowadays, microorganism infections and allergies are among the most common diseases initiated in the respiratory
tract system. Interleukin-17 (IL-17), a cytokine that is frequently found elevated in various lung diseases and
infections, has been shown to play a critical role in the host defense against Gram-negative bacterial infections and to
serve as a regulator that modulates the mucosal immunity in allergies. Due to manipulation difficulties however, most
of the recent studies regarding the role of IL-17 have been done in vitro, or in an over-simplified condition. Our lab is
devoted to the task of developing an in vivo model wherein the diverse functions of IL-17 can be reasonably
demonstrated. Our approach is thus to engineer an inducible construct in which the expression of mouse IL-17 is
controlled by the tetracycline regulator responsive element (TRE), pTRE2-mIL-17. Using this construct, together with
two additional airway epithelial cell-specific constructs, CC10-rTA and CC10-sTA that target the expression of both
tetracycline regulated activator and suppressor, we generated several transgenic mice carrying these triple genes.
Using these mice, airway epithelial cell-specific expression of IL-17 can be induced with the administration of
doxyclycline (a tetracycline derivative) in the mice drinking water. Using these mice, we plan to examine if induced
IL-17 plays an essential role against the bacterial infection and the alteration of the mucosal immunity in
allergen-induced allergic airway disease. This triple-transgenic mouse model will significantly facilitate the study of
IL-17 in various lung diseases.
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CHEMISTRY
AN ANIONIC, WATER-SOLUBLE POLYMER AS A FLUORESCENT SENSOR
Joseph V. Goodwin and Michelle Wood (Dr. Robin Lammi), Department of Chemistry, Physics and Geology, Winthrop
University, Rock Hill, South Carolina 29733
Anionic conjugated polymers are promising cation sensors, offering bright fluorescence and superior
fluorescence-quenching response upon analyte binding, due to highly efficient intramolecular energy transfer along
the polymer chain. We have investigated the sensing capability of an anionic polythiophene, poly[2-(3-thienyl)ethoxy-4-butylsulfonate] (Mw = 1,000,000 g/mol), in aqueous solution, using methyl viologen dication as a quencher.
A detection limit of 5.0 E-8 M methyl viologen was determined for polymer solutions 1 E-4 M in monomers. In studies
with added SSC buffer, increasing buffer-ion concentration resulted in decreased quenching efficiency, suggesting a
static quenching mechanism involving ground-state polymer-ion complexation. Results of quenching measurements
in ethylene glycol support this assertion. The viscous solvent had no deleterious effect on measured polymer
sensitivity for cation, as would be expected if quenching occurred via a collisional mechanism. Stern-Volmer kinetic
analysis of data in both solvents yielded Ksv values (a measure of sensitivity) of 7 E5 1/M, similar to the published
value for a related polythiophene. Unfortunately, poor polymer solubility limited sensitivity. Three cationic
surfactants, OTAB, DTAB and CTAB, were tested for their ability to break up polymer aggregates. Of the three,
CTAB (cetyltrimethylammonium bromide) showed the most promise, giving an 11-fold increase in polythiophene
fluorescence intensity and a narrowed, blue-shifted polymer absorption spectrum, consistent with decreased polymer
aggregation. The polythiophene-CTAB solution (1 E-4 M CTAB) showed a strong quenching response to methyl
viologen, with a similar Ksv value (3 E5 1/M), indicating that polymer-surfactant complexation only slightly decreases
polymer availability to quencher. Future efforts will focus on determination of metal-ion concentrations and
examination of related polymers with side chains tailored to improve solubility.
CATALYTIC MECHANISM OF CARBONIC ANHYDRASE
Suzanne English (Dr. Robert Zurales), Division of Natural Sciences, Mathematics and Business, Middle Georgia
College, Cochran, Georgia 31014
Carbonic anhydrase is the enzyme that catalyzes the reaction of carbon dioxide and water to form carbonic acid. This
enzyme is important in maintaining the body`s acid-base balance. The active site of the enzyme is believed to
contain a zinc(II) ion bound to three neutral histidine amino acid side chains and one hydroxide ion. The first step in
currently proposed mechanisms is the addition of the zinc-bound hydroxide to the carbon atom of carbon dioxide.
This leads to the formation of the bicarbonate ion. We have proposed a novel mechanism that assumes that the
coordination sphere for the zinc(II) ion expands to include a water molecule. In our proposed mechanism, as the
hydroxide ion attaches to carbon, a proton is also transferred from the zinc-bound water to directly form carbonic acid.
We tested this novel mechanism using computational chemistry. Our simplest model of the enzyme-substrate
complex consisted of a zinc(II) ion coordinated to a hydroxide ion and a water molecule, with the water molecule
being hydrogen bonded to carbon dioxide. Calculations were performed using the Gaussian98 software package at
the B3LYP level using a 6-31G(d) basis set. The transition state geometry and activation energy of our proposed
mechanism will be compared and contrasted to other proposed mechanisms.
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CHEMISTRY
Determining Proton-Transfer Reaction Rate Constants of Volatile Organic Compounds Using
Quadrupole Ion Trap Mass Spectrometry
Jeremy Felton, (Dr. Gary Glish), Chemistry Department, University of North Carolina at Chapel Hill, Chapel Hill, NC
27599-3290
Proton-transfer reaction rate constants of hydronium reagent reacting with volatile organic compounds (VOC’s) have
been studied previously using proton-transfer mass spectrometry (PTRMS); however, reaction times in PTRMS
experiments cannot be controlled as they can using Quadrupole Ion Trap Mass Spectrometry (QITMS). The two
goals of this work are to compare the reaction rates determined from QITMS with the rates previously published using
PTRMS and to compare the experimentally-determined rate constants with their theoretical values.
Hydrodium ions were used as the reagent to ionize the VOC’s via proton transfer. Reagent ions were generated using
atmospheric sampling glow discharge ionization (ASGDI) of water vapor in the source region of the quadrupole ion
trap. In contrast, the headspace above the neutral VOC analyte was sampled directly into the trapping volume of the
QITMS. Freeze-pump-thaw of the analyte liquid was used to ensure a pure vapor from each VOC. The VOC
concentration was in excess, so pseudo-first-order reaction kinetics apply. The concentration of VOC was determined
from the pressure of the analyte in the trapping volume of the quadrupole ion trap. After the pressure of the VOC was
recorded, hydronium ions were introduced into the ion trap from the ASGDI source, where they reacted for a specified
time period. The decrease in the intensity of the hydronium ion signal during successively longer reaction times was
used to determine the proton-transfer rate constant.
Preliminary results with the VOC isoprene gave a rate constant of 3.0E-10 + 5E-11 cm3/molecules-sec. Future work
will focus on studying how other VOC’s react with the hydronium reagent as well as determining proton-transfer rate
constants using different reagents, such as protonated acetone.
CATALYTIC MECHANISM OF CARBOXYPEPTIDASE A
Elizabeth Dietz (Dr. Robert Zurales), Division of Natural Sciences, Mathematics and Business, Middle Georgia
College, Cochran, Georgia 31014
We used molecular modeling to investigate the catalytic activity of the enzyme carboxypeptidase A.
Carboxypeptidase A is present in the small intestine of mammals and breaks down proteins to provide essential
amino acids. The active site of the enzyme is believed to contain a zinc(II) ion bound to two histidine and one
glutamate amino acid side chains, as well as one hydroxide ion. In a currently proposed mechanism, a protein
oxygen attaches to the zinc, then the hydroxide attaches to the carbonyl carbon. This produces a tetrahedral carbon
with an oxyanion stabilized by the zinc ion. We have proposed a novel mechanism that assumes that the
coordination sphere for the zinc(II) ion expands to include a water molecule, not the protein. In our proposed
mechanism, the oxygen of the protein is hydrogen bonded to the zinc-bound water. As the hydroxide ion attaches to
carbon, a proton is also transferred from the zinc-bound water to form a neutral intermediate. In a second step, the
proton is transferred to nitrogen resulting in ammonia as a leaving group. To test our mechanism, we used a simple
model of carboxypeptidase A that consisted of zinc hydroxide and determined the structure and energy of the
transition state. Calculations were performed using the Gaussian98 software package at the B3LYP level with a
6-31G(d) basis set. The activation energy of our proposed mechanism will be compared to the activation energy of
the currently accepted mechanism.
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CHEMISTRY
DEVELOPMENT OF AN IN-SITU METHOD TO MONITOR BIODIESEL PRODUCTION USING
ATTENUATED TOTAL REFLECTION INFRARED SPECTROSCOPY (ATR-FTIR)
Robert Moore and Kyle Wilmsmeyer (Dr. H. Francis Webster), Department of Chemistry and Physics, Radford
University, Radford, VA 24142
The interest in alternative fuels has increased dramatically in recent years due to rising cost and environmental
concerns related to fossil fuels use. Biodiesel represents an environmentally friendly and cost-effective fuel,
consisting of methyl esters produced from transesterification of various alcohols with triglycerides found in commonly
available vegetable oils. While the kinetics and methyl ester composition of biodiesel are typically monitored using
gas or liquid chromatography, these prove to be time consuming, making real time analysis difficult. The purpose of
this work was to develop a novel in-situ technique to rapidly monitor the kinetics of alcohol / triglyceride
transesterification using soybean oil to produce biodiesel under a variety of experimental conditions. A custom stirred
reactor was used to produce methyl esters using both chemical and enzyme catalysts. Reaction kinetics were
monitored by attenuated total reflectance infrared spectroscopy (ATR-FTIR) in real time using a variable angle ATR
infrared accessory fitted with a ZnSe hemispherical crystal and flow cell. Enzyme catalysis was evaluated using lipase
from both Candida Antarctica immobilized on a commercial acrylic resin and Pseudomonas Cepacia immobilized in a
sol-gel matrix. Transesterification using both methanol and ethanol were evaluated, and the inhibition of enzyme
activity in excess methanol was studied. The reaction progress was followed by monitoring changes in the ATR
infrared spectrum including increases in the intensity of a distinctive infrared band at 1034 cm-1 associated with
glycerol production. While the infrared spectra of the methyl esters produced and the triglyceride starting material
were almost the same, small but significant shifts in the peak frequencies and intensities near 1375cm-1 and
1150cm-1 were also used to monitor the reaction kinetics. Results using this in-situ technique compared favorably
with those found for samples analyzed using gas chromatography.
FREE RADICAL CREATION BY OZONE
Christina Chu (Dr. Robert Zurales), Division of Natural Sciences, Mathematics and Business, Middle Georgia College,
Cochran, Georgia 31014
Although important in the upper atmosphere, ozone in the lower atmosphere is one of the principal components of
smog. Ozone is known to react with unsaturated fatty acids in the lungs. The fist step is believed to be the addition
of ozone to a double bond. In the Criegee ozonation mechanism, ozone adds to a double bond to produce a primary
ozonide. The primary ozonide then rearranges to form a more stable secondary ozonide or reacts with water to form
a ketone and hydrogen peroxide. When ozone reacts with certain polyunsaturated fats, however, radicals are formed.
William A. Pryor of Louisiana State University has proposed that the primary ozonide can split into radicals before
rearranging or reacting with water. To test this mechanism, we calculated the activation energy for this step using the
Gaussian98 software package at the B3LYP level with a 6-31G(d) basis set. We suggest that this mechanism is not
competitive with the Criegee mechanism, and propose instead that radical formation may result from hydrogen
transfer. Our model of the polyunsaturated fat, 2.5-heptadiene, could lose a hydrogen atom from carbon four to form
a resonance-stabilized radical. We will present the calculated activation energies for our proposed mechanism and
the previously proposed mechanism.
214
CHEMISTRY
BETA-LACTAMASES, THE ANTI ANTIBIOTIC
Kevin Waits (Dr. Robert Zurales), Division of Natural Sciences, Mathematics and Business, Middle Georgia College,
Cochran, Georgia 31014
The purpose of this project was to understand the enzymatic activity of a beta-lactamase. This enzyme provides
bacterial resistance to drugs such as penicillin by adding a water molecule, converting the drug into an inactive form.
The active site of the enzyme is believed to contain a zinc(II) ion bound to three neutral histidine amino acid side
chains and one hydroxide ion. We assume that the zinc(II) ion binds an additional water molecule that forms a
hydrogen bond to the oxygen of the four-membered lactam ring. In our proposed mechanism, as the hydroxide ion
attaches to carbon, a proton is also transferred from the zinc-bound water to this oxygen. This mechanism differs
from a recent mechanism that has the water move toward the zinc as the reaction progresses. We tested this novel
mechanism using computational chemistry. Our simplest model of the beta-lactamase consisted of a zinc(II) ion
coordinated to a hydroxide ion. We used only ring of four atoms to model penicillin. Calculations were performed
using the Gaussian98 software package at the B3LYP level using a 6-31G(d) basis set. The transition state geometry
and activation energy of our proposed mechanism will be compared and contrasted to the other proposed
mechanism.
INDIUM-MEDIATED PYRIDINONE SYNTHESIS
Sarah R. Slauson (Research Corporation), Department of Chemistry, Seattle University, Seattle, Washington 98122
Pyridinones are a notable class of N-heterocyclic compounds due to their high level of biological activity and potential
pharmacological use. Pyridinone scaffold synthesis is reported successful through an aza Diels-Alder type reaction
between an imine and Danishefsky’s diene catalyzed by indium salt. The coupling of imine formation and heterocycle
synthesis has been reported, as well as reduction of nitro-containing compounds to amines, but literature report of a
combined nitro reduction, imine formation, and pyridinone synthesis. Here a range of indium catalyst and solvent
conditions have been studied to determine the optimal environment for successful one-pot synthesis of the pyridinone
scaffold from nitro-containing starting reagents.
The reduction of nitro-containing compounds to amines is successfully achieved in an acidic In(0) environment,
producing oxidized In3+ as a result. Amines and aldehydes in an In3+ environment readily react to form imines,
which may then perform an aza Diels-Alder reaction with a very reactive diene, such as Danishefsky’s diene, to form
the pyridinone scaffold.
Initial studies of the tandem reaction with nitrotoluene, ammonium chloride, and benzaldehyde in ethanol showed
successful reduction and imine formation, but heterocycle formation was not observed. Further studies involved
varied nitro compounds, including nitrobenzene and p-chloro-nitrotoluene, as well as other solvents such as
acetonitrile and methanol. Reduction and imine formation were observed in each reaction, but again no heterocycle
formation was detected. GC/MS analysis of each reaction’s products detected degradation products of Danishefsky’s
diene, suggesting that the reactive diene is sensitive to one of the reaction conditions held constant.
We have found that the current acidic conditions used for reduction of the nitro group to the amine may prematurely
induce the diene to decompose, preventing the formation of the N-heterocycle. An investigation of the tolerance of
Danishefsky’s diene to Lewis acid, as well as the acid requirements of the reduction step are in progress to determine
a low, but effective acid level that will allow the successful one-pot synthesis of pyridinones from nitro-containing
compounds.
215
CHEMISTRY
Using Gold Nanoparticles to Study Protein Conformational Structures
Logan Leslie (Drs. Richard Zare, Soonwoo Chah, and Andrew Leavitt), Department of Chemistry, Stanford University,
Stanford, CA 94305-5080
The conformational structure of a protein determines its function, so being able to determine the structure of a given
protein is an important component of biochemical research. In previous work done by the Zare research group (Chah
et al., 2005), it was demonstrated that gold nanoparticles could be used to determine protein conformational structure
by looking at the maximum absorption wavelengths of UV-vis spectra. In the present study, we built upon those
findings, working with a mixture of proteins rather than individual ones. We used common proteins to demonstrate
the feasibility of developing a simple technique using gold nanoparticles to compare mixtures of proteins and quickly
determine their ratios.
To prepare the gold nanoparticles, we boiled HAuCl4 in water, then then stopped heating the solution and added
sodium citrate. Changing the amount of sodium citrate used influences the size of the gold nanoparticles produced.
The size of the nanoparticles was checked using a transmitting electron microscope. Because different proteins
change structure at different pH levels, with a resultant color change, we then combined the nanoparticles with
different percentages of cytochrome C (CYTC) and horseradish peroxidase (HRP), and observed how the color of the
solution changed at different pHs. Using UV-vis spectroscopy to determine the amount of color change at different
pH levels enabled us to determine the amounts of the different proteins present.
This research was done during the summer at Stanford University. The University of West Georgia is my home
institution.
THERMODYNAMICS
OF THE EXTRACTION OF THE RARE EARTH ELEMENTS WITH
P,P`-DI(2-ETHYLHEXYL)ETHYLENE-DIPHOSPHONIC ACID
Jason E. Spencer (Emmanuel O. Otu), Department of Chemistry, School of Natural Sciences, Indiana University
Southeast, New Albany, IN 47150
Competitive extraction of the rare earth elements from aqueous perchloric acid medium by
p,p`-di(2-ethylhexyl)ethylenediphosphonic acid dissolved in o-xylene was studied and will be presented. The
extractions were investigated as a function of aqueous perchloric acid concentration and extractant organic phase
concentration in the 25 – 60oC temperature range. The extraction stoichiometry did not change with temperature.
The extractant aggregation has been shown to remain constant in the temperature range of this work. The
thermodynamics of the extraction process was studied by measuring extraction coefficients in this temperature range.
The thermodynamic parameters give insight into the mechanism associated with extraction.
THE ROLE OF WATER IN HUMAN QC
Ashland Doomes (Dr. Robert Zurales), Division of Natural Sciences, Mathematics and Business, Middle Georgia
College, Cochran, Georgia 31014
In 2005, the x-ray crystal structure of human glutaminyl cyclase was reported. This enzyme catalyzes the conversion
of N-terminal gultaminyl residues into pyroglutamic acid, a posttranslational event important to activate many
enzymes. The enzyme active site was found to contain a zinc(II) ion bound to a histidine, a glutamate and an
aspartate amino acid side chain, as well as one water molecule. This is very interesting as water is not a reactant in
this reaction. But other zinc enzymes, with very similar structures, do catalyze the reaction of water and a protein. In
the currently proposed reaction mechanism, this water molecule is assumed to be displaced by the incoming protein.
Although this may also occur in the other zinc enzymes, we wondered if the water molecule may be part of the
enzyme`s catalytic action. This would suggest a similar role of water in all zinc enzymes. To test this hypothesis, we
performed calculations using the Gaussian98 software package. We used density functional theory at the B3LYP
level, and our basis set was 6-31G(d). We replaced the protein with the small molecule 4-aminobutyramide and the
enzyme by zinc hydroxide. We will compare our calculated activation energy to that of the currently proposed
mechanism.
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CHEMISTRY
PROGRESS TOWARD THE SYNTHESIS OF TRIALKYLSILOXY PROTECTED
AMINODIHYDROPYRIDINONES FROM 1-AMINO-3-TRIALKYLSILOXY-1,3-BUTADIENE AND IMINE
Robert O’Brien, Jamie Garcia, Sarah Slauson, Adam Johnson, Lorien Wallace, Peter Alaimo.
Dihydropyridinones have a range of biological activity, and are therefore of potential therapeutic value. 1-amino-3(tert-butyldimethylsiloxy)-1,3-butadienes are useful substrates for synthesis of dihydropyridinones due to the ease of
their synthesis as well as their high reactivity.
The reaction of 1-pyrollidinyl-3-(tert-butyldimethylsiloxy)-1,3-butadiene with N-benzylideneaniline using a variety of
solvent/catalyst combinations was studied, and it was determined by GC/MS that the product of this reaction was the
TBDMS protected pyrollidinyldihydropyridinone cycloadduct, with yields ranging from 40-80% (as determined by
GC/FID). This product has two important features: the amino group was not eliminated after cyclization and the
product contains a TBDMS protected enol. These features may allow for further synthetic steps such as selective
deprotection of the enol, followed by subsequent addition at the resulting keto group while preserving the amino
group. The isolation and characterization of the product is in progress as well as the study of diastereoselectivity of
the reaction.
BINDING MODE OF INHIBITORS OF HUMAN QC
Bianca Lester (Dr. Robert Zurales), Division of Natural Sciences, Mathematics and Business, Middle Georgia College,
Cochran, Georgia 31014
This enzyme glutaminyl cyclase catalyzes the conversion of N-terminal gultaminyl residues into pyroglutamic acid.
Osteoporosis may result from the malfunctioning of this enzyme. One way of learning about the enzyme is to find
chemicals that inhibit its activity. In 2005, the x-ray crystal structure of human glutaminyl cyclase was reported, with
and without inhibitors. The enzyme active site was found to contain a zinc(II) ion bound to a histidine, a glutamate
and an aspartate amino acid side chain, as well as one water molecule. To test our model of the enzyme, we
calculated the equilibrium constant for binding of the enzyme substrate and the inhibitor imidazole. We performed
calculations using the Gaussian98 software package. We used density functional theory at the B3LYP level, and our
basis set was 6-31G(d). We used a model of the enzyme that include the first level of ligands. Equilibrium constants
were determined in air and in various dielectric media. We will compare our calculated constants to the known
values.
AMMONIA SENSORS, HYDROGEN STORAGE, METHANE ACTIVATION AND TRIPODAL
NICKEL-NITROGEN COMPLEXES
Christopher A. Sutton (Patrick J. Desrochers) Department of Chemistry, University of Central Arkansas, Conway,
72035
Hydrogen storage and methane activation represent significant challenges for the increasing use of these simple
gases as commercial energy sources and industrial raw materials. Research in this laboratory on tripodal (C3v)
N3NiX complexes has produced results suggesting potential solutions to both of these challenges. To date, X in the
above complexes includes Cl-, Br-, I-, BH4-, NH3, and -SPh. The N3 nitrogen donor atoms are provided by the facial
tridentate chelate, Tp* (hydrotris(3,5-dimethyl-1-pyrazolyl)borate). An example of the interaction of Tp*NiX with
ammonia involves its rapid binding of the gas, forming [Tp*Ni(NH3)3+][X-]. This ammonia gas uptake is reversible;
ammonia is released with gentle heating. These solid [Tp*Ni(NH3)3+][X-] compounds are interesting both as
precursors to the elusive fluoride derivative (Tp*NiF) and as the basis for efficient ammonia sensor/storage materials.
The electronic structure of the halide and borohydride derivatives has been studied using high frequency/field EPR
spectroscopy and angular overlap theory. Low level DFT calculations of fac-(H3N)3NiBH4+ reproduced structural
and spectroscopic features of fac-Tp*NiBH4. Because BH4- is isostructural with methane, results for Tp*NiBH4
suggested the possibility for facile methane activation at a tripodal N3Ni center like fac-Tp*Ni or fac-(H3N)3Ni. The
nickel-nitrogen tripod is proving to be a durable and controllable center for the interaction of nickel with hydrogen-rich
substrates.
217
CHEMISTRY
THE FIRST STEP IN A FUNCTIONAL HYDROGENASE MODEL INVOLVING NICKEL AND CYSTEINE
Ariel S. Marshall, Davis Duong, (Patrick J. Desrochers) Department of Chemistry, University of Central Arkansas,
Conway, AR 72035
Cysteine coordination is a recurring feature of the biochemistry of nickel. One significant association of this amino
acid with nickel is in bacterial hydrogenase enzymes where protonation of a nickel-cysteine active site is believed to
precede the catalytic formation of energy-dense hydrogen gas. To model some of this chemistry, the pH-dependent
stability and redox activity of phosphine-nickel-cysteine systems (dppeNiCys) has been investigated. Reversible
protonation of the nickel-cysteine group of dppeNiCys in phosphate-buffered aqueous solutions was documented by
NMR. A promising result of this chemistry is the moderately acidic aqueous solutions in which it is observed.
Aqueous media will almost certainly be the hydrogen source for future cost-effective large scale hydrogen production.
Experimental progress to achieve the second step of catalytic hydrogen generation, accumulation of electrons at the
nickel center, will also be discussed. This has involved electrochemical measurements using cyclic voltammetry and
modifications of the ancillary ligands to tridentate phosphines. Phosphorus-rich environments are expected to favor
reduced nickel. Nickel reduction and cysteine protonation are two key steps in functional hydrogenase activity.
SYNTHESIS OF NEW THIOPHENE-CONTAINING ALUMINUM COMPLEXES RELATING TO OLED
APPLICATIONS
Kevin T. Mellem (Andrew G. Sykes), Department of Chemistry, University of South Dakota, Vermillion, SD 57069
Previous research has shown that many inorganic aluminum complexes have excellent electroluminescent properties,
leading to their use in organic light emitting diode (OLED) devices. The purpose of our project is to combine the
electropolymeric properties of novel organic ligands with the electrolumiscence properties of aluminum, ultimately
forming a PLED (polymeric light emitting diode) type device. Thiophenes are known to electropolymerize and form
conducting polythiophene. We provide both the synthesis and characterization of new thiophene-containing organic
ligands and their resulting tris-substituted aluminum complexes. The advantages of growing electropolymerized films
include the fine control of electrochemical film growth (mainly film thickness), the control of anion dopant (electrolyte
counterion), and the codeposition of a hole injection layer (polythiophene itself). All of these factors are important in
manipulating and optimizing the electroluminescence of the OLED devices. Using common amine coupling
chemistry, we have synthesized several modified hydroxyquinoline ligands that contain terminal, pendent thiophene
functional groups, and we have subsequently bound these ligands to aluminum (III) cations. The synthesis,
characterization, and electrochemical results for these complexes are presented in detail by the authors.
Experimental Unimolecular Rate Constants for 2,3-HF and 2,3-HCl Elimination from CF3CFClCH2Cl.
Oksana Zaluzhna (Bert E. Holmes), Department of Chemistry, University of North Carolina at Asheville, One
University Heights, Asheville, NC 28804, USA.
Chlorofluorocarbons are one class of chemical substance that depletes the ozone layer. Because of their deleterious
effect on stratospheric ozone, the production of CFCs is being phased out and other organic molecules are being
introduced as substituents. Hydrochlorofluorocarbons (HCFCs) are one class of compounds under consideration as a
replacement because they are similar to CFCs but are not as destructive to ozone.
The present work studied chemically activated HCFC, CF3CFClCH2Cl, which was formed in the gas phase from the
combination of CF3CFCl and CH2Cl radicals. The radicals were generated from the ultraviolet photolysis of the
corresponding iodides using a high pressure Hg lamp. The decomposition products were E- and Z-CF3CCl=CHCl
from a 2,3-FH elimination and E- and Z-CF3CF=CHCl from a 2,3-ClH elimination reaction. Unimolecular constants for
CF3CFClCH2Cl were 1.7 x 10-4 s-1 and 0.3 x 10-4 s-1 for 2,3-HF elimination forming E- and Z- products,
respectively, and 7.5 x 10-4 s-1 and 2.2 x 10-4 s-1 for 2,3-HCl elimination to form the E- and Z- products,
respectively. No evidence was found for a 1,2-FCl interchange reaction.
218
CHEMISTRY
AB INITIO AND DENSITY FUNCTIONAL THEORY CALCULATIONS OF Co, Cr, AND Fe CARBONYL
COORDINATIONS WITH C6H6 AND C5H5 AS PROTOTYPES FOR BINDING SITES IN CAPPED
NANOTUBES
Jason Crawford (George Heard), Department of Chemistry, The University of North Carolina at Asheville, Asheville,
NC 28804
Nanotechnology is an exciting field with potentially far-reaching applications, among which are those involving carbon
nanotubes. These allotropes of carbon are repeating arrays of hexagons similar to graphite rolled into a tube.
Experiments synthesizing single-walled carbon armchair nanotubes using transition-metal catalysts observed that
nanotubes can be capped off at both ends with (conceptually) half a molecule of buckminsterfullerene (C60); this cap
is a hemispherical arrangement of pentagons and hexagons. A model to describe a capped nanotube would consider
introduction of pentagons into a nanotube as “defects” that would cause the extending nanotube to cap off.
Understanding how transition-metal complexes coordinate with C6H6 and C5H5 (as the two possible binding sites for
metals to a capped nanotube) could shed light on formation of “defects” in extending nanotubes. With the exception
of carbonyl coordinations of chromium, few energetics calculations of transition-metal complexes with C6H6 and
C5H5 have been carried out. This project used ab initio and density functional theory techniques to find the
lowest-energy geometry of Co, Cr, and Fe carbonyl complexes (h-6)- and (h-2)-coordinated to C6H6 and (h-5)- and
(h-2)-coordinated to C5H5. Several possible symmetries of coordinated molecules were examined to determine
which, if any, symmetry constraints have lower energies.
A COMPUTATIONAL STUDY OF THE BARRIER HEIGHT OF THE 1,2-FCl EXCHANGE REACTION OF
SUBSTITUTED ETHANES
William C. Everett (Dr. George Heard), Department of Chemistry, TheUniversity of North Carolina at Asheville,
Asheville, NC 28804
A 1,2-FCl interchange mechanism has been shown by our research group to be an important intermediate step in
many reactions of chlorofluorocarbons(CFCs) and hydrochlorofluorocarbons(HCFCs) in the atmosphere. This
mechanism may produce hitherto unexpected products which may contribute to ozone depletion. This investigation
examines the 1,2-FCl interchange reaction for all possible two-carbon CFCs and HCFCs and determines the effects
of fluorine and chlorine substitution on the barrier height and transition geometry for this mechanism. For the
molecules CH2FCH2Cl, CHF2CH2Cl and CH2FCHCl2, we examine the reaction at B3PW91, B3LYP, MPWB1K,
MPWB95, MP2, CCSD, QCISD, G2MP2 and G2MP2B3 levels of theory. Basis sets 6-31g(d',p'), 6-311+g(2d,p),
6-311+g(2df,2p), 6-311++g(2d,p), cc-pvtz, aug-cc-pvtz and cc-pvqz under B3PW91 are considered in order to
determine a consistent and representative level of theory. For all other exchanges, comparisons are made at the
B3PW91/6-311+G(2d,p) level of theory. The relative energies of the transision geometries of isomeric species
correlate well with the relative energies of isomeric substituted alkenes that would result from the elimination of FCl
and this can be used to rationalize the trend in barrier heights.
219
CHEMISTRY
INVESTIGATION OF THE PGC-1-ALPHA CO-ACTIVATOR INTERACTING WITH THE HUMAN
ESTROGEN RECEPTOR ALPHA
Rachael R. Beach (Noah W. Allen), Department of Chemistry, The University of North Carolina at Asheville, Asheville,
North Carolina 28804
The development of cancer in breast, thyroid, and uterine tissue is largely due to the uncontrolled expression of the
estrogen receptor (ER). A well-founded understanding of the various factors and their interplay in the regulation of ER
activity is imperative for the development of treatment and prevention strategies for these cancers. This research
proposes to focus on the interactions between ERalpha and the co-activator PGC-1-alpha. The PGC-1-alpha actions
are less sensitive towards the nature of the bound ligand in the ER, whereas classical co-activators interact with the
ER in a highly ligand dependent fashion. Additionally cases have been found where PGC-1-alpha interacts with ER in
the absence of a ligand suggesting a binding mode, diverging from that of classical co-activators. Potential
co-activator binding interfaces on the ligand binding domain (LBD), DNA binding domain (DBD), and hinge region of
the ERalpha are of particular interest here. Single point mutations and changes in rigidity of these interfaces as well
as of the complementary interfaces on the co-activator are performed systematically. Interactions are simulated and
analyzed, utilizing computational methods such as molecular dynamics, geometry optimization, and visualization.
The resulting data should then provide insight into the possible binding modes of the PGC-1-alpha co-activator to the
ERalpha. Furthermore, a basis will be established for future in vitro studies.
CREATION OF A MUTANT HUMAN ESTROGEN RECEPTOR GUIDED BY IN SILICO
METHODOLOGIES FOR USE IN EXPLORING NATIVE LIGAND, CO-ACTIVATOR AND DNA BINDING
AFFINITY
Mitch Lazaro (Noah Allen), Department of Chemistry/Biochemistry, The University of North Carolina at Asheville,
Asheville, NC 28804
for human estrogen receptor alpha (hER alpha) has been completed. In addition, sf9 cells have been grown for the
specific purpose of producing hER alpha after being infected with this particular baculovirus. Following the production
of the native hER alpha a mutant hER has been created using recombinant DNA technology, rendering a new
baculovirus for subsequent sf9 cell infection. In preparing for the creation of the new baculovirus, in silico studies
generating electrostatic potential maps were used to design novel constructs. These novel constructs will then be
used to explore in vitro binding of numerous oligonucleotide sequences, native co-regulators and ligands to the
mutant receptor. The purpose of this research is to compare the binding affinities of native hER alpha with that of the
mutant hER alpha. In this specific case, we have modified a sequence of ten amino acids in the DNA binding domain
of the hER alpha to study novel oligonucleotide binding in the presence of the native ligand, 17 beta-estradiol and the
native co-activator, SRC-1(2). These comparisons have been made by using radiolabeled oligonucleotides to detect
binding affinity and comparing these to the native receptor construct. This technique has allowed for a uniquely
guided experiment based on in silico data, and also provided a method to generate mutant hER alphaconstructs for
further study.
220
CHEMISTRY
STUDIES TOWARDS IMPROVING THE SYNTHESIS OF THE MITOMYCIN FAMILY OF ANTITUMOR
AGENTS
David R. Mabe (Herman L. Holt, Jr.) Department of Chemistry, The University of North Carolina, Asheville, One
University Heights, Asheville, NC 28804
The mitomycins are a family of anticancer compounds that are currently in widespread clinical use because they are
effective in halting growth and reducing the size of malignant tumors. Mitomycin C is the most common drug in the
mitomycin family used. It binds to a cell's DNA (guanine specifically), rendering the cell incapable of replication and
of protein synthesis. The synthesis of mitomycin compounds is typically costly and involves many intermediate
compounds that are produced in moderate yields. A novel synthesis of the four fused ring system that is the
backbone of all the mitomycins has been developed. Our research utilizes a novel A-B-C-D approach to the
synthesis of the ring system, beginning with a vinyl azide rearrangement to the indole backbone that proceeds in
close to quantitative yield. In order to determine the best method of initiating formation of the C-ring, several different
types of reactions were investigated. Intramolecular Claisien-Schmitt condensations utilizing different combinations
of reagents were examined, as well as a Grubbs catalyzed sequence. Subsequent steps towards the formation of the
D-ring of the mitomycin skeleton involve aziridination of the resulting alkene by a nitrene source or by utilizing a
triazole intermediate. Progress towards the mitomycin skeleton and other model systems will be presented.
SYNTHESIS AND BIOACTIVITY OF ANTI-TUMOR COMBRETASTATIN ANALOGUES
David R. Mabe (Herman L. Holt, Jr. and Ted Meigs), Departments of Chemistry and Biology, The University of North
Carolina Asheville, Asheville, NC 28804 USA
Since the isolation and characterization of the anti-tumor agent combretastatin A-4 from the African bushwillow tree
Combretum caffrum (Combretaceae), a large family of synthetic combretastatin analogues has been produced that
show great potential for use as chemotherapy drugs. The combretastatins slow tumor growth and reduce cellular
function by proceeding through the same mode of action as the natural product colchicine. Colchicine and the
combretastatins (natural and synthetic) are excellent disruptors of the cellular cytoskeleton, which is composed of
microtubules. Microtubules are polymers of the proteins α and β tubulin, and the combretastatins inhibit their
polymerization. The cytoskeleton controls the arrangement and movement of organelles in a cell, as well as many of
the processes involved in mitosis (cellular replication). Since the combretastatins inhibit the formation of the
cytoskeleton, many cellular functions are halted by the combretastatins, effectively reducing tumor growth and size.
This study aims to produce novel combretastatin analogues that contain functional groups new to the combretastatin
family of compounds. The ethene group that bridges the two phenyl rings of the combretastatins is aimed to be
replaced with an aziridine group as well as an azirine group. The aziridine and azirine groups are hypothesized to
increase the activity of the compounds by increasing water solubility as well as general reactivity. Bioassay studies
will determine how effective these new compounds are versus other combretastatins and compounds currently used
in chemotherapy. Specifically, K562 cells (human myelogenous leukemia), MDA-MB-435 cells (human breast
cancer) and SW480 cells (human colon adenocarcinoma) are to be tested with the compounds produced.
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CHEMISTRY
INVESTIGATION OF THE HUMAN ESTROGEN RECEPTOR ALPHA DNA BINDING DOMAIN BOUND
WITH ESTROGEN RESPONSE ELEMENT
Cassaundra Papaj (Noah Allen) Chemistry Department, The University of North Carolina at Asheville, Asheville, NC
28804
Studies of the estrogen receptor (ER) have implicated it in the role of cancer. ER, a nuclear receptor, binds to DNA
and then in combination with another part of the entire protein, acts as a transcription activator for the production of
increased proliferation of estrogen receptors, which may lead to full-blown metastatic cancer if left deregulated. It is
the DNA binding domain (DBD) of the estrogen receptor that binds DNA at a palindromic sequence site, called an
estrogen response element (ERE). Homology searches for the ERE were run using the software BLAST, available
from the NCBI website. The homology search found DNA sequences that had shared sequence similarity with the
ERE. The search for an ERE provided an ERE in close proximity to the nucleotide sequence for estrogen receptor.
Computational studies using molecular dynamics simulations examined interactions of the DNA binding domain and
the ERE bound in silico. The simulations provide a model in vitro structure of the complex. Further computations
examined the DBD in complex with multiple variations of an ERE with single point mutations. The purpose of
studying the interaction between the estrogen receptor DBD and the mutated ERE is to distinguish the interactions of
the DBD/ERE interface. The findings will be used to create an algorithm that predicts DNA properties (such as
electrostatic interactions) that are necessary to bind to the estrogen receptor. The results will also be used to provide
data for future in vitro studies.
EXPERIMENTAL INVESTIGATION OF THE UNIMOLECULAR REACTION PATHWAYS AND RATE
CONSTANTS FOR ENERGIZED CF2CLCH2CL.
Sarah Boshamer (Bert Holmes), Department of Chemistry, The University of North Carolina at Asheville, Asheville,
NC 28804
Hydrochlorofluorocarbons (HCFCs) are relatively stable, non-reactive, non-poisonous, and odor-free gases. These
properties are why they are often used as refrigerants, aerosols, and as cleaning agents. However, HCFCs are
dangerous in another way: they attack ozone molecules in the stratosphere. Therefore, it is important to study the
decomposition reactions of HCFCs in the gas-phase to determine whether the products formed are safe for the
environment. This research investigates the reaction pathways and reaction rate constants for a specific HCFC,
1,2-dichloro-1,1-difluoroethane (CF2ClCH2Cl). The CF2ClCH2Cl was prepared with about 95 kcal/mol of vibrational
energy by the combination of CF2Cl and CH2Cl radicals. These radicals were generated by the UV photolysis of
CF2ClCOCF2Cl and CH2ClI in clean quartz vessels. The reaction mixture was analyzed using a gas
chromatograph/mass spectrometer (GC/MS) in order to separate and identify the reactant and products. The
decomposition products observed by the GC/MS indicate that CF2ClCH2Cl undergoes 2,1-HCl elimination forming
CF2=CHCl, 2,1-HF eliminations forming E- and Z-CFCl=CHCl, and a novel 1,2-FCl interchange reaction forming
CFCl2CH2F. In addition, collisions may remove the internal energy forming a stabilized CF2ClCH2Cl. The product
ratios of these molecules give rate constant ratios of 20:1:1:0.2 for the 2,1-HCl elimination reaction, E- and Z-2,1-HF
elimination reactions, and the 1,2-FCl interchange reaction, respectively.
COMPUTATIONAL MODELING STUDIES OF ESTRADIOL STEREOISOMERS AND
RECEPTOR-LIGAND DOCKING INTERACTIONS
Catherine Reilly (Noah Allen), Chemistry Department, The University of North Carolina at Asheville, Asheville, NC
28804
Estradiol (E2) has five stereocenters and can, therefore, exhibit 32 (25) isomers, each optically distinguishable from
the others. Two of these isomers are the naturally occurring 17α-estradiol (E2α) and 17β-estradiol (E2β). Recent
literature has revealed, however, that the energetically most stable isomers are not those that are naturally occurring.
This find is interesting in that it suggests that the energetic characteristic of the ligand is not sufficient for the
explanation/prediction for its binding to estrogen receptor. We, therefore, explored the conformational space of each
isomer, in silico, utilizing Molecular Dynamics (MD) simulations. Isomers used in simulations were first geometrically
optimized at Hartree-Fock level of theory. MD simulations resulted in up to 20 additional conformations for each
isomer, each of which was geometrically optimized on the Density Functional Theory level. These will be
automatically docked in the binding pocket of the estrogen receptor. The interactions between each structure and the
receptor will be calculated, scored and ranked against each other. Analysis of the resulting ligand-receptor
interactions will offer a possible explanation to questions on isomer stability and binding.
222
CHEMISTRY
UNIMOLECULAR RATE CONSTANTS FOR THE 1,2- AND 1,1-HF ELIMINATION FROM CHEMICALLY
ACTIVATED 1,1,1,2,2-PENTAFLUOROETHANE
Hayley Schiebel (Bert E. Holmes), Chemistry Department, The University of North Carolina at Asheville, Asheville, NC
28804.USA
Hydrofluorocarbons (HFCs), the class to which CF3CF2H belongs, have been replacing chlorofluorocarbon
compounds (CFCs) that have been implicated in the accelerated depletion of ozone in the earth's stratosphere. When
chlorine is regenerated in CFCs, it is free to continue to break down other ozone molecules. This process continues
for the atmospheric lifetime of the chlorine atom (one to two years), during which it destroys an average of 100,000
ozone molecules. Chlorine radicals are removed from the stratosphere after forming two compounds that are
relatively resistant to dissociation by ultraviolet light: hydrogen chloride (HCl) and chlorine nitrate (ClONO2).
Dissociation is slow enough so that these compounds can diffuse down to the troposphere, where they react with
water vapor and are removed in rain. Because HFCs do not contain chlorine, they are better suitable for the
atmosphere. CFC’s were developed in the early 1930s and are used in a variety of industrial, commercial, and
household applications. CFC’s are non-toxic, non-flammable, and non-reactive with other chemical compounds. They
are highly effective refrigerants that were developed in response to the pressing need to eliminate toxic and
flammable substances, such as sulfur dioxide and ammonia, in refrigeration units and air conditioners. The most
common commercial CFCs, marketed under the trade name Freon, are trichlorofluoromethane (CFC-11) and
dichlorodifluoromethane (CFC-12). This work will report the reactions of a chemically activated HFC, CF3CF2H,
which was prepared with approximately 95 kcal/mol of internal energy by the combination of CF3 and CHF2 radicals.
The total rate constant was measured as 1.3 x 105 s-1 using CF2HCF2H as an internal standard and the rate constant
for the 1,2-elimination of HF was measured from the pressure dependence of the [CF2=CF2/[CF3CF2H] ratio as 0.2 x
105 s-1. If the only decomposition channels are the 1,2- and 1,1-HF elimination, then the rate constant for the
1,1-channel is 1.1 x 105 s-1, a factor of 5 faster than the more common 1,2-HF elimination pathway. It also appears
that the carbene formed by 1,1-HF elimination lacks sufficient energy to cause a 1,2-F migration to form CF2=CF2.
223
CHEMISTRY (NO CATEGORY)
MÖSSBAUER STUDIES OF IRON-DOPED ZINC OXIDE
Jeremy Fitzpatrick (Amar Nath), Department of Chemistry, The University of North Carolina at Asheville, Asheville NC
28804
It is well known that ZnO is a non-magnetic substance. However, recent evidence indicates that adding a small
amount of iron (only about 1 to 3 percent by mole) to the structure may cause it to have ferromagnetic properties.
Much effort is being made to understand this intriguing effect. To create the samples, metal acetates were mixed with
oxalic acid and freeze dried. The precipitate that forms allows for evenly distributed oxalates. Some samples were
made containing .5 to 3 percent cobalt and/or copper in addition to the iron. After drying, they were heated to 800
degrees Celsius to form an oxide. In addition, samples created by laser evaporation were also analyzed. Mössbauer
spectroscopy has been used to look for magnetic properties in the iron doped zinc-oxide. X-ray diffraction determined
structure of the samples and EDX gave elemental analysis of the samples. X-ray diffraction has shown that the
samples created have a zinc oxide structure. Elemental analysis confirms that the zinc and oxygen have a proper
stoichiometric ratio, showing a pure compound. Cobalt and copper were in the structure in appropriate amounts.
Mössbauer spectroscopy shows that the iron was in a Fe+3 high spin state.
224
CLASSICS
A DIACHRONIC STUDY OF SOME "SACRED" TERMS IN LATIN
Leah Suzanne van Ameyden van Duym (Lora Holland), Department of Classics, The University of North Carolina at
Asheville, Asheville, NC 28804
This research focuses on the etymological and semantic fields of the related Latin words sanctus and sacer. While
some research in this area has been done by others in the past, it has been primarily in languages other than English.
The research includes a diachronic etymology of the words through present-day usage in English and Italian, going as
far back in time as possible with available resources. It also involves a limited study within the semantic field of these
words, focusing on semantic change, particularly as it relates to the introduction of Christianity. The primary
methodology of these investigations involved examining and collecting occurrences of the words, their precursors,
and all members of their lexemes in Latin, Oscan, Umbrian, Etruscan, and any other available Italic or other relevant
dialects. These have been gathered from inscriptions and texts over a period of time extending from as far back as
possible to approximately 500 CE. Sources include books and articles on the fields of historical linguistics and
semantics, and in particular their methodology; a wide variety of Latin texts, and translations and annotations of those
texts; concordances; and inscription sources, including books and databases. Specific sources selected thus far are
listed in the reference section below. The goal of this research was to find a verifiable common root for sanctus and
sacer, and to determine how it evolved in pre-Latin languages as well as in Latin and contemporary dialects. It also
aimed to determine the precise linguistic forces behind the nasalization that seems to have precipitated the
separation of these into two distinct words, and thus to learn whether they are really as distinct as they appear to be.
Finally, through the semantic study, evidence has been found of a fundamental shift in the meanings of these words
after the introduction of Christianity into the Latin-speaking world.
CONNECTIONS BETWEEN POETRY AND MAGIC IN HOMER`S ODYSSEY
Alexander Hall, Edmund Cueva, Classics, Xavier University, 3800 Victory Pkwy, Cincinnati, OH, 45207
To the modern mind, poetry and magic are distinct phenomena. While it is true that both are complex and
multifaceted, and that to define either in a succinct fashion is nearly impossible, a modern thinker would still be
unlikely to see them as intimately connected, much less as springing from a common source. Ancient thinkers and
writers, on the other hand, believed exactly this: that poetry and magic were inextricably intertwined, sharing many of
the same powers, and even the same names and terminology. My analysis focuses on a single work, the Odyssey of
Homer. The Odyssey is one of the oldest pieces of literature in the Western Canon, and thus reflects western thought
at its most ancient. Furthermore, the Odyssey deals directly with both magic and poetry. On his journey, Odysseus
encounters many gods, monsters, and witches, whose powers can only be described as magical. Similarly, he meets
the court bards and singers, the first pictures captured in literature of poets at work. These two groups, the poets and
the magic-users, are linked in every case through language and the powers which are attributed to them. The word
used for “song” is identical to that used for “spell.” Similarly, the “enchantment” that a poet works over his audience is
exactly the same “enchantment” worked by a witch or a siren, trying to control the minds and feelings of her victims.
Not only in language, but even in power, we see the connections between poetry and magic. As was mentioned
above, both poetry and magic could affect the mind, changing what people think and feel. They could also affect the
physical world, healing or harming a person directly. Given these strong overlaps, in language and in power, I
conclude that poetry and magic were not originally distinct entities, but two aspects of a single phenomenon.
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CLASSICS
SLITHERY AGENTS OF FATE: SNAKE IMAGERY IN VIRGIL`S AENEID
Steven T. Wilco (Prof. Angela Taraskiewicz) Department of Foreign Languages (Classics), Valparaiso University,
Valparaiso, Indiana 46383.
Snakes are a frequently recurring symbol in Virgil’s Aeneid, but their varied use makes them at times an ambiguous
symbol. In the time of the early Roman Empire, snakes had already been established as a symbol of evil and
violence, of passionate love, and of boundary crossing (male/female, life/death). This diverse background makes it
difficult to determine the nature of the many references in the Aeneid. The first and dominating reference to snakes in
Book Two, tells of the twin serpents of Tenedos who kill Laocoon, symbolizing the fate of Troy. Snake references
occur in seven of the ten remaining books, almost always at key events that move the plot forward. In the original
Latin, an analysis of textual evidence surrounding the snake references strongly suggests that they are used with
specific intent. While they occur in varied ways, their appearance tends to drive the plot forward, and drive Aeneas to
his destiny. They operate independently of both gods and men, indicating their supreme power and underscoring their
ambiguity. This parallels the action of fate in the poem – the reader questions how and never if Aeneas will found
Rome. It is beyond his power and understanding and the path to his destiny is winding and serpentine, driven by fate.
The snake imagery serves to emphasize the role of fate in the poem and vividly portray the forces at work behind
Aeneas’ journey.
AN INVENTORY OF GHOSTS: CULTURAL ATTITUDES TOWARD HELEN OF TROY AS OBSERVED
BY APHRODITE
Allyson F. Blomeley (Lora Holland), Department of Classics, The University of North Carolina at Asheville, Asheville,
North Carolina 28804
According to Homer’s Iliad, Aphrodite is very much the young, spoiled puppet-master, while her puppet, Helen, suffers
the effects of Aphrodite’s wants and decisions. This creative writing project explores the possibility of an Aphrodite
who has lived long enough to see the downfall of the Greek pantheon due to other religions, and has subsequently
found herself eclipsed by the memory of Helen. Research for this project include both ancient sources as well as
modern: the more widely read classical Greek dramas and classical compendiums and accounts of history and myth,
as well as works such as Isocrates’s Encomium of Helen, the Dyctis Cretensis, and several Greek epic fragments
concerning the Trojan War, such as the Cypria and The Sack of Ilion. Modern sources include both strictly-Helen
related material as well as works discussing the relationship between Aphrodite and Helen. The process of
implementing the research into the writing of a piece of fiction involved selecting certain aspects of the stories found
in various sources and choosing between varying interpretations or versions of the many different Helen and
Aphrodite myths. These were reinterpreted and woven into a new story that reflects the depth of research this project
entailed.
226
CLASSICS
THE PROCESS OF EDITING HENRY OF GHENT’S ARTICLE 53, QUESTION 1 OF HIS QUAESTIONES
ORDINARIAE
Kristina P. Robinson (Lora Holland), Department of Classics, The University of North Carolina at Asheville, Asheville,
North Carolina 28804
This research is the editing of one question from Henry of Ghent’s Quaestiones Ordinariae Article 53, Question 1.
Henry of Ghent’s Quaestiones Ordinariae is a collection of his quaestio from the Paris University. The medieval
quaestio, or inquiry, are the arguments for and against a weekly debate topic a master teacher would provide for his
students. During the mid 13th century the quaestio method became the standard form for university lectures and
textbooks. The quaestio provided detailed expositions of, and the problems raised by a text, section by section.
Henry of Ghent has been described as the “most illustrious thinker in the last quarter of the 13th century.”[1] Henry of
Ghent taught at the Faculty of Theology at the University in Paris from 1276 until his death in the 1293, and this
makes him the important link between Thomas Aquinas, who died in 1274, and John Duns Scotus, who was just
finishing his studies in the 1290’s. Article 53, Question 1 is important for several reasons. First, it has not been edited
by a modern scholar. Second, one of the manuscripts for this question may be the exemplar, the official ‘bookstore’
copy that would have been approved by the professor and then rented out piecemeal for students to have their own
copies made. The editing of this article will add to the debate of whether or not that particular manuscript is the
exemplar. That question was the central point of this research. The third reason to edit this article is because it
contains a lengthy discussion of the Greek word prosopon. The examination of this discussion imparts a sense of
Henry of Ghent’s understanding of the Greek language. The process of creating an edited edition is one that has
fascinated scholars for centuries. This research will also contribute to a series of editions which will replace the 1508
Renaissance edition, which has been the standard text for nearly 500 years. This edition will then be the basis for the
subsequent vernacular translations.
227
COMMUNICATIONS
PIZZA, PEPSI, AND MULTICULTURALISM - A CONTENT ANALYSIS OF ETHNICITIES IN SUPER
BOWL COMMERCIALS FROM 1996-2005
Kathryn L. Bodenhamer (Dr. Ruth Brown), Department of Communication, The University of Nebraska at Kearney,
905 West 25th Street, Kearney, Nebraska 68849
This paper is an analysis of the underlying messages drawn from factual evidence presented within a decade of
strictly Super Bowl commercials. The focus was centered on ethnic minorities as commercial actors/actresses, and
data was collected from different perspectives such as how much representation they received, the type of role each
played, what general product category each was associated with, and how their character was depicted. Our central
hypothesis was an expectation to see a steady increase in representation and more positive depictions as years
passed. Results showed that (1) an inconsistency of increases and decreases occurred throughout the decade
possibly due to corresponding events in the United States as well as in the advertising industry, (2) an abundance of
ethnic minorities were seen in supporting roles, though, once again, no definite pattern could be determined, (3)
ethnic minorities can be said on average to show representation in half of the commercials within each general
product category, and (4) types of depictions varied throughout the decade depending on the ethnicity used and the
year, which had several influences as to what was occurring at that time.
An Analysis of the Code of Athens as a Model for a Global Ethics Code
Justine Weber, Department of Communication Arts, Xavier University , 3800 Victory Parkway, Cincinnati, OH 45207
Globalization requires public relations practitioners to understand and work in international contexts; however, the
variety of economic, political and religious norms makes ethical standard-setting difficult. This paper examines the
International Public Relations Association’s (IPRA’s) 1965 Code of Athens as one model for a universal code of
ethics. A well-developed universal code contributes to the distinction of public relations as a profession rather than an
occupation, the recognition of public relations in management functions and the identification of a scholarly body of
research. The Code of Athens follows from the United Nations Universal Declaration on Human Rights and
establishes a set of values to guide ethical decision-making around the world.
The inherent power of public relations in corporate, government and activist structures requires responsibility on the
part of individual practitioners and the organizations they represent. The call becomes more urgent in a shrinking
world where individuals not necessarily targeted by a public relations campaign can recept its messages. This
requires an obligation to accuracy and truthfulness in explicit and implicit messages dispersed to explicit and implicit
publics. Practitioners must be involved in the ongoing ethical discussions within the industry.
Two philosophical perspectives shape the theories used to evaluate this universal code. This paper suggests that by
following Aristotle`s Doctrine of the Mean practitioners avoid the extremity of absolutism and the deficiency of
relativism. Public relations scholars frequently criticize absolutist and relativist theories for their tendencies to relieve
the practitioner of accountability in his or her work. Furthermore, Kant`s deontological theories support the systems
model and subsequent normative model for decision-making. These models enable the development of ethics codes
containing general principles with specific applications (J. Grunig, 2001).
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COMMUNICATIONS
Are all Arthur Page Society Members Ethically Practicing Public Relations?
Adrienne Blumthal (Jeong-Nam Kim)Department of Communication Arts, Xavier University, 3800 Victory Parkway,
Cincinnati, OH 45207
Today in the United States there are several organizations that praise members for ethical practices. Among these is
the Arthur Page Society; an elite organization consisting of senior public relations counselors and corporate
communication executives who inquire to enhance their profession. Members are chosen for membership based upon
their practices and advancements in the organization in which they belong. The society prides itself on practicing
public relations by the standards of Arthur Page. After thorough evaluation of current Arthur Page Society members, it
has become evident that not all of the representative members belong to organizations with ethically clean track
records, including companies such as IBM Global, Weber-Shandwick, Fleishman-Hillard and Ketchum.
The Arthur Page Society views public relations as “the art of increasing, understanding and communicating corporate
and individual character” (Page Principles). This was formulated based on Page’s strong beliefs in humanism and
freedom as America’s guiding distinctiveness and qualifications for a free enterprise. The dynamic principles and
ethical guidelines on which the Arthur Page Society was founded are shown through its members keeping the public
informed through good character and action serving practices. Page believed organizations must operate based on
public interest and manage on long term scales to make customer satisfaction the primary goal. With this in mind, it is
vital for the society to assure that proper ethical decisions are being made on this basis, in representing members.
By reevaluating the acts of IBM global and their cooperation with Nazi Germany, Weber-Shandwick breaching the
PRINZ code of ethics while working with Timberlands West Coast Pty Ltd, Fleishmen-Hillard Ex-SVP pleading guilty
to the act of over billing clients, and Ketchum’s participation in the case involving Armstrong Williams failing to
disclose he was paid by a Ketchum client for lobbying efforts, the society can assure that the situations have been
liquidated and better practices have been established for the future.
This article suggests that by employing a systems theory and model for ethical issues and ethical decision making
management based on excellence in practicing public relations, the society can judge how the situations were
handled and therefore determine the ethical state of the organizations with members represented within the society.
FOR THE GOOD OF THE FAMILY: ALCOHOL ADVERTISING AFTER PROHIBITION
Emily H. Heiple (Dr. Julie Williams), Department of Journalism/Mass Communication, Samford University,Birmingham,
AL 35229
Prohibition came to America in 1920, causing liquor companies and breweries to stop the manufacture and sale of
their product. The manufacture, sale, and consumption of alcohol were outlawed by the 18th Amendment to the
Constitution that year. In 1933, Congress passed the 21st Amendment which repealed the 18th Amendment. Once
again, people were allowed to purchase and consume alcohol, and alcohol manufacturers were allowed to advertise
and sell.
It took alcohol companies a while to take advantage of the right to advertise. Because many people had agreed
with Prohibition, alcohol manufacturers faced the same dilemma that drug pushers might face today if they were
suddenly granted the right to advertise marijuana. They had to proceed delicately to sell the once-forbidden product.
This paper examined alcohol ads in Harper’s Magazine published in 1938 and 1939. Harper’s was selected due to
the fact that it was the one of the first general interest magazines to advertise alcohol after Prohibition, even though
those ads didn’t appear for several years. Every alcohol advertisement in Harper’s for the two years under study was
examined for its apparent target audience, theme, and style of advertising. The analysis determined the approach
alcohol producers took to reintroduce a product that had recently been illegal.
The research showed surprising themes in alcohol ads after Prohibition. For instance, Pabst beer ads in 1938 and
1939 had a family theme and targeted the “family man” as a drinker. Alcohol consumption, no longer a secret or
illegal affair, was depicted as happening in family settings. It was implied that generations of a family would enjoy this
particular brand of beer, from grandfather, to father, to son. Ads in Harper’s did not, however, target female drinkers.
While alcohol companies worked hard to develop a family theme in the post-Prohibition era, they usually avoided
suggesting that women should drink.
229
COMMUNICATIONS
Creating an Endless Oil Fantasy: America`s Media-Perpetuated Denial of Global Peak Oil
Ryan O`Connor, Jenna Robbennolt, Kimberly Smith, Lindsay Den Herder (Dr. Terry Robertson), Speech
Communications Department, The University of South Dakota, Vermillion, South Dakota 57069
Scientists, corporations, and the United States Government all agree that America’s current level of oil consumption
is unsustainable. Furthermore, all three groups recognize that peak oil—the point at which more conventional oil has
been extracted than remains in the Earth—will occur on a global scale within the next thirty years. The effects of
global peak oil will be significant; indeed, when domestic oil extraction peaked in the United States in 1971,
Americans were forced to re-examine their consumption practices and governmental policies. Yet as global peak oil
nears, the media has done little to inform American citizens about the potential effects of a substantial energy
shortage. The purpose of this project was to analyze five major U.S. newspapers—the New York Times, Chicago
Tribune, Washington Post, LA Times, and USA Today—along with one international news service—the BBC—to
rhetorically critique the reasons behind the media’s decision to treat global peak oil as a second-rate news story,
specifically between the dates of September 14, 2005 and November 1, 2005. By applying Ernest Bormann’s fantasy
theme principles to the media’s coverage of global peak oil, it is clear that three distinct paradigms are responsible for
the lack of candid reporting and information sharing. First, while Americans understand that oil is a finite resource,
few citizens believe that an energy shortage is imminent, and the media has therefore chosen to produce stories that
cater to these naïve expectations. Second, Americans believe that technology has the ability to solve all societal
problems, and the media is currently unwilling to explain to citizens that an unequivocal trust in technology may in
fact prove detrimental. Finally, Americans have been conditioned to believe that economic growth is intrinsically
connected to a successful society, and this socially constructed belief prevents the media from educating citizens
about sustainable energy alternatives. As long as these three paradigms remain unchanged in the hearts and minds
of the American public, meaningful media coverage will remain dormant, and the possible effects of global peak oil
will become increasingly negative.
EYES WIDE OPEN: NEWS MEMORIES IN CHILDREN
Kimberly M. Holland (Julie Williams) Journalism/Mass Communication Department, Samford University, Birmingham,
Alabama 35229
News events such as the September 11 attacks make their way to the minds of young children. These children, in
turn, are affected in various ways by the news. Sometimes they become fearful and seek out care and explanation
from parents or caregivers. Sometimes the news amuses them. Other news influences how they perceive politics or
other key institutions. Discovering news memories, how children were affected by the news events as well as if or
how parents comforted or explained the news to children will help to show the impact news events have on children.
The purpose of this project was to determine the earliest memories of news events that college students had and the
effects those news event memories had on the respondents.
The researcher surveyed 116 respondents enrolled in eight freshman-level introductory courses at Samford
University. Questions sought to discover each respondent’s earliest news memory by asking the respondent to
remember and identify the earliest event or events he or she could bring to memory and how he/she became aware
of that news. The respondents were also asked how this event and the memories of this event made him or her feel.
If a parent or caregiver comforted or explained the news event to the child, the respondent was asked to explain that.
Responses from the surveyed students found that approximately half remembered the election of Bill Clinton over
George H.W. Bush and the criminal proceedings against O.J. Simpson. The remaining half of responses recalled
many different events. The outlets from which respondents learned this information varied, with some citing
television as a source and many citing interpersonal communication as a source. Indeed, a large number of the
respondents learned their first news memory not by the mass media but through interpersonal communication. The
respondents reported various feelings about the event, but most reported either shock about a parent’s reaction to the
news or confusion about the event.
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COMMUNICATIONS
FULFILLING RELATIONAL COMMUNICATION DIMENSIONS THROUGH CELL PHONE INTERACTION
Stephanie K. Van Stee (Dr. Deirdre Johnston), Department of Communication, Hope College, Holland, Michigan
49423
Although prior research focusing on relational communication is plentiful, most of it uses face-to-face interactions as a
basis for study. Constantly evolving technology warrants a closer look at relational communication taking place in
new contexts. The purpose of this study was to investigate the role of the cell phone in relational communication;
specifically, the study investigated cell phone versus face-to-face interaction preferences along relational
communication dimensions. The relational dimensions used for the study were modified from the Relational
Communication Scale developed by Burgoon & Hale (1987). The modifications specifically addressed preferences
for cell phone versus face-to-face interaction in order to accomplish certain relational communication dimensions. An
additional series of items was used to assess the use of the cell phone for self-comforting behaviors. Surveys were
conducted with a sample of 247 Hope College students. The relational dimensions of relaxation, control and
self-comforting were found to be significantly related to preference for cell phone over face-to-face interaction.
Relaxation measures the ease with which a person interacts. The control variable measures the degree to which a
person feels personal power during a conversation. Comfort measures the likelihood that a person would use the cell
phone as a tool for self-comforting. Self-comforting is a reflection of people’s use of the cell phone in order to
communicate emotional states. An ANOVA revealed that females are more likely than males to prefer cell phone
interaction for fulfilling the relational dimensions of comfort, trust, and connection. There is a high correlation
between the connection and trust, suggesting that those who feel a greater connection on the cell phone than in
face-to-face interaction will also feel a greater level of trust on the cell phone relative to face-to-face interaction. The
implications of these results will be discussed at length, concentrating on whether cell phone interaction is a substitute
or a means for augmenting face-to-face interaction.
THE EFFECTS OF SOCIAL ANXIETY ON CELL PHONE BEHAVIOR
Patrick McConahy
Cell phones have broken down the time and space barriers to communication. There is the possibility for any cell
owner being contacted by or contacting almost any one, at any time, anywhere in the world. It is surprising that more
research has not been conducted on cells considering the impact they have had on communication and social
behavior. The purpose of this study was to identify how social anxiety affects cell behavior. The Liebowitz Social
Anxiety Scale (1987) and other cell behavior items were incorporated on a survey administered to 247 college
students. This study showed significant relationships between individuals with higher social anxiety and cell use.
Individuals who scored higher on the anxiety scale preferred cell communication over face-to-face communication in
a public setting (r =.19, p<.01). There was also a significant relationship between social anxiety and using cells to
escape one-to-one communication situations (r =.16, p<.05). These findings raise questions of whether cells may be
beneficial or detrimental to the development of communication competence. “Always speaking over the mobile phone
with one’s own intimate circle of persons, instead of conversing with people met anywhere, outside home, implies
remaining closed inside a rigid and inert kind of discourse.” (Fortunati, 2002, p.516) Thus, the cell can become a
barrier to the development of communication skills and relationships for those with social anxiety; instead of being
forced to cope with their social anxiety individuals can use their cell to escape the social setting they are in.
Alternatively, cells may help people with social anxiety to engage in more relational communication than they might
otherwise. Future research is needed to explore whether cells may be detrimental to communication in public
settings, but beneficial to interpersonal communication.
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Television Violence and the Cultivation Effect on Viewers
Lucinda Parker (Dr. Betty Attaway-Fink), Communication, Southeastern Louisiana University, Hammond, LA 70402
This study researched the effect of violent television on viewers’ perceptions of actual violence. This topic has been
researched extensively by the developer of the cultivation theory, George Gerbner. Past research by Gerbner and
other researchers has found a correlation between the amount of television viewed and elevated perceptions of
violence. The purpose of this study was to test the following hypothesis: Violent television, specifically crime drama,
affects the way viewers perceive real violence. This study was developed by survey research and included 113
participants. The study found a connection between types of television viewed and elevated perceptions of violence.
The overestimation of the total crimes committed in the United States was 2.4% higher by regular viewers of crime
drama than non-viewers and 1.5% higher by regular viewers of news than non-viewers. The overestimation of total
crime in the United States was 5.9% higher among the heaviest viewers than the lightest viewers. The effect of
viewers’ perceptions of crime in particular cities, however, was found to have little or no correlation with particular
crime dramas set in those cities. Only one city, New York City, had a higher overestimation of crime among regular
viewers of crime dramas set in the city than non-viewers of the programs. This was not found with crime dramas set
in other cities. The results of this study support the findings of past research. There was a correlation between the
amount and types of television viewed and elevated perceptions of violence. The expected correlation between
particular cities where crime dramas are set and viewers’ perceptions of crime in those cities, however, was not
discovered.
ADVERTISEMENTS DURING THE DEPRESSION: WHAT DID THEY REALLY SAY?
Ann Claire Vaughn (Julie Williams), Journalism/Mass Communication Department, Samford University, Birmingham,
Alabama 35229
This study will explore the directions advertisers chose in order to attempt to convince consumers to spend money
during the Great Depression of the 1930s. Whereas most studies of Depression advertising involve examining ads,
this study will ask old-timers to recall the Depression-era ads that impressed them the most. The researcher will
interview octogenarians and nonagenarians from the Somerby Retirement Community in Birmingham, Alabama. The
researcher will, of course, follow the lines of conversation that the subjects bring up. However, in general, the
researcher will be asking such questions as “What brands do you remember being advertised during the 1930s?”
“How did your family make decisions about product purchase when you were growing up?” and “Did advertisers of the
1930s discuss the Depression, or did they continue advertising as though nothing was wrong?” The researcher will
look for patterns in the type of ads the ladies are able to remember. Using the women’s oral history interviews as a
basis, the researcher will then look at ads of the era to see how the memorable themes articulated by the
octogenarians compared to actual ads that were published in the era. The advertisements the researcher will study
appear in 1930, 35, and 39 issues of Good Housekeeping.
Primary Media Research Concerning the Release of Birth of a Nation (1915)
Kristin A. Rose (Dr. David B. Schock), Department of Communications, Hope College, Holland, Michigan 49423
I have recently been collaborating with Hope College Professor of Communications Dr. David B. Schock to gather
primary research for the next segment of a four-part documentary series he is producing concerning the Ku Klux Klan
in Michigan. The segment I am in which I am assisting concerns the emergence of the first feature-length film, Birth
of a Nation, a highly controversial, racially-charged film based on Thomas Dixon’s novel, The Clansman, and its
implications on the African American community, particularly the its effort to ban the film from theaters across the
country. My research has involved gathering articles from newspapers, magazines, and journals from the time when
the film was first released, primarily the year 1915. These publications have included such titles as the New Republic,
The Crisis, New York Times, and Detroit News. The arguments against Birth of a Nation I have read thus far all
portrayed the deep ethnic divisions ever-present in our society that only grew deeper with what some would describe
as slanderous films such as this one that distort and demean the average African American citizen. In a time when
racism flourished and the KKK grew daily, Birth of a Nation may have, in fact, caused the birth of a whole new
generation of members more devoted to the ideals of the KKK than ever before. The African American community,
however, was also more devoted to stopping these endorsements of racism and tirelessly striving to attain the civil
rights they deserved.
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COMMUNICATIONS
TRUE LIFE...COLLEGE...IS IT REALLY LIKE THE MOVIES???
Aimee M. Holcomb and Heather M. Bush (Dr. Betty Attaway-Fink), Department of Communications, Southeastern
Louisiana University, Hammond, Louisiana 70402, , , Southeastern Louisiana Univeristy, P.O. Box 10451, Hammond,
La 70402
Popular culture in the form of movies and television portray college life as fun, worriless, and trivial. Furthermore,
some movies such as Van Wilder, Old School and Animal House portray college students as party animals
consuming large amounts of alcohol, participating in illegal activity such as drug use and engaging in numerous
casual sexual activities.
The researchers of this paper conducted a narrative analysis of Van Wilder, Old School and Animal House and
utilized the content of the movies to examine how it portrays college life. Not only do these movies exhibit a large
amount of binge drinking and illegal behaviors, but also gives a negative look at the Greek letter wearing community
of universities.
The fantasy theme analysis and symbolic convergence theory was used as the basis of the studies of the
researchers. High school students and incoming freshmen may develop their own fantasies about what college is
really about based on the movies that they have seen.
The researchers of this paper obtained salient research data from 350 surveys completed by college students.
The survey consisted of questions that inquired whether or not the student had watched any of these three movies,
and if they did, did they really believe college was like that which was portrayed in the movies. The respondents also
answered questions pertaining to if they were part of the Greek letter wearing community, how much alcohol they
consume a week, if they participated in drinking games and if they are satisfied with their college experience.
The researches then compiled the results of the survey into four different categories. The categories consisted of
male fraternity students, female sorority students, male non-fraternity students, and female non-sorority students. It
was divided to compare if being male or female or part of a fraternity or sorority will affect the results of the survey.
Between surveys and narrative analysis, enough quantitative and qualitative data was obtained to suggest the
hypothesis to be true and support the fantasy analysis theme.
Get Real?! The Reception and Response Of College Women To Dove’s Campaign For Real Beauty
Cristina Mayer (Dr. Jessica Gisclair), Communications, Elon University, 100 Campus Drive Elon, NC 27244
The Campaign For Real Beauty is an ongoing, integrated marketing campaign for the Dove branch of the Unilever
Corporation. The television, print, and online advertisements have created a buzz with the public because of the
usage of average, normal-looking women. This paper examines the positive and negative effects The Campaign for
Real Beauty has on college women. The focus group results indicate that college women have their own definitions of
beauty. The respondents clearly defined beauty as trait separate from physical attractiveness, and their definitions of
beauty included traits such as confidence, spirituality, character and independence. Furthermore, the results show
that although the respondents had a positive reaction to the usage of normal people in the advertisements, they did
not feel The Campaign For Real Beauty would be successful in the long term.
HOMELESSNESS: A STUDY OF THE HOMELESS AS A MUTED GROUP
Tatiana Kireiev (Dr. Sook-Young Lee), Department of Communication Studies, Luther College, Decorah, Iowa 52101
The purpose of this study was to understand the homeless culture, to see how homeless individuals view themselves
in their own words. The culture of the homeless was investigated by conducting qualitative interviews with ten
different homeless individuals in Salt Lake City, Utah. All of the interviews were handwritten due to the nature of the
research. All of the research was conducted from May through August of 2005. The researcher spent over 90 hours
volunteering at the Road Home in Salt Lake City, Utah. There were three interview phases involved in the research:
the pre-interview phase, the informal interview phase, and the formal interview phase. Each of the phases led up to
an established trust with members of the homeless community. Muted group theory and cultural studies theories
were used as a theoretical framework. The results showed that the homeless as a culture are a muted group with
rules, beliefs and values of their own. In addition there were many interesting commonalities and differences found
among the members of the homeless culture having to do with language, friendship and social norms. Despite the
fact that the homeless make up a very diverse community, their experiences have been named for them. Many of
the participants had internalized the dominant ideological view of homelessness. This has serious theoretical
implications including problems with society’s view of homelessness and the way that homelessness is dealt with in
our society.
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COMMUNICATIONS
Race, Law, and Order in Small Town Appalachia
Adrienne Avery, Sponsor: Will Dulaney, Communication, Theatre, and Dance, Western Carolina University,
Cullowhee, NC, 28723
In the fall of 2005, a young black man, Dewann McCollum, was shot four times when a team of white Jackson County
Deputy Sheriffs attempted to question him about the recent robbery and shooting at a local Chinese restaurant,
adjacent to Western Carolina University, in Cullowhee, North Carolina. A potential suspect and convicted drug felon,
McCollum had moved to this largely white community early this year to be close to a family support system and to
distance himself from previous legal problems in Fayetteville, North Carolina. It can be seen that in the aftermath of
this robbery and murder, regional news media reported contradictory reports and stories that reflected biased
perspectives. Some local and state papers presented the police shooting as a simple matter of law and order.
Numerous articles in the Asheville Citizen Times raised questions about police procedure, evidence, and interviewed
family members of McCollum. Across the nearby campus, white and black faculty, staff, and students responded to
the event with different fears and apprehensions that provided a window into the persistent divide between the
perceptions of the white majority and the black minority This paper combines George Gerbner’s Cultivation Theory
and Cheris Kramarae’s Muted Group Theory in order to analyze university, local, and regional reactions and media
coverage of the Cullowhee, North Carolina incident. Visual images, textual analyses, and oral interview expose how
the following variables:1) racial bias, 2) stereotyping, and 3) majority-minority reactions, interpretations, and
responses, affected the local community. With a keen awareness of historical forces and the relationship between
race, poverty, environment, and the justice system, this paper argues that incidents like the one in Cullowhee reveal
that even in the post-civil rights era, communication still falls short in arresting the legacies of racial discrimination.
Saving Face: Presidential Image Repair Discourse
Jamie Chappell (Dr. Cheryl Jorgensen-Earp), Communication Studies, Lynchburg College, Lynchburg, VA 24501,
1501 Lakeside Dr.
Maintaining a positive public image is essential to any public figure, particularly world leaders such as presidents.
Much of their legitimacy and ability to govern effectively rest in the way in which the public perceives them, and it is
important for them to be able to address the nation after a damaging event so that their image can be repaired. This
study looks at the speeches of four U.S. presidents who had to address the public after they were accused of some
offensive event to try to restore their image. I analyzed the presidential addresses using William Benoit’s typology of
image repair strategies, and I assessed the effectiveness of the strategies in each of the speeches.
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COMMUNICATIONS
Advertising America: European Settlement Literature as the Originator of the American Dream
Hannah Atchley (Dr. Julie Williams), Journalism/Mass Communication, Samford University, 800 Lakeshore Drive
Birmingham, Al. 35229-7020
Until Columbus stumbled upon the Americas, Europe, Africa, and Asia were all Europeans ever knew. The newly
found land intrigued many; however, the unknown and uncertainty of settling new colonies frightened them.
Settlement leaders had to find ways to compel people to make a journey to the new world and leave everything they
had ever known. This study shows how early European advertisements, in the form of pamphlets and letters written
to persuade settlers, were actually the first incarnation of the American Dream. European settlement literature
intended to promote American settlement by telling of the great possibility of wealth that a person could gain in
America. Although there have been many definitions of the American Dream in many ages, certainly the possibility of
growing wealthy by one’s own hard work is part of every version of the American Dream.
To support the correlation between settlement literature and the American Dream, this paper examined letters written
in the 17th century about America, trying to encourage readers to settle there. It also examined pamphlets from the
17th century. These, too, were targeted at European readers and encouraged them to settle in the growing nation.
The research showed that early settlement literature offered all sorts of hyperbole and outright falsehoods about
America as it tried to advertise the America’s opportunities. In publishing such misleading jargon, these early mass
media not only encouraged settlement but also gave birth to the American Dream; and thus, the settlers that arrived
clutching pamphlets had actually brought the notion of the American Dream with them as a European import.
"You`ve crossed the line": A study of relational boundaries within recalled conflict episodes.
Brittany Barbera, Colleen Hughes, Michelle Krug (Dr. Lynn S. Cockett), Department of English, Communication and
Theatre Arts, Juniata College, 1700 Moore Street, Huntingdon, Pennsylvania 16652
This study examines how undergraduate students perceive conflict. Specifically this research describes individuals’
conflict behavior as oriented towards relational boundaries. By taking a symbolic interactionist approach, we seek to
understand the meaning people ascribe to their world and how those meanings are negotiated through discourse. In
this study, we see conflict as the site of interaction through which people assign meaning to their communicative
behaviors. Undergraduate students were solicited through flyers and electronic announcements on a small college
campus. We conducted eleven interviews with participants who self-selected using a hybrid of semi-structured and
narrative methods. Participants responded regarding serious conflict in their lives. Data were transcribed and
analyzed using a variety of coding techniques. Preliminary findings suggest that relationship type, while unique to
each participant, influences a person’s conflict behaviors and their reflections upon conflict outcomes. We argue that
these perceptions mirror Mead’s notion of the “I” and the “Me” as components of the self, and that conflict is a
particularly salient relational context in which to examine symbolic interactionism.
THE DEVELOPMENT OF A CULTURALLY SENSITIVE HEALTH INFORMATION BROCHURE FOR
YOUNG FATHERS ON THE ISLAND OF ST. KITTS
Andrew J. King (Dr. Ray Baus), Department of Communication, University of Wisconsin at Whitewater, Whitewater,
Wisconsin 53190
Research has established that teen fathers’ involvement in the upbringing of their children benefits both fathers and
children. This project seeks to encourage the involvement of teenage fathers on the Caribbean island of St.
Christopher (St. Kitts), where teenage paternity is widespread. The goal is to create a brochure that will help teenage
fathers improve their parenting skills, specifically targeting methods of effectively dealing with infant crying, an issue
which has been identified in the literature as a serious precipitator of child abuse. The design of effective, culturally
sensitive patient education materials such as brochures has become an important area of health communication
research. The brochure being designed for this project will apply the research of Hofstede, Leininger, and others on
cultural differences and health education to the problem of informing teen fathers about effective responses to infant
crying. The completed brochure will be distributed in the island’s community health clinics and through the St. Kitts
Youth Senate. In addition to presenting the brochure itself, my poster will explicitly outline the theoretical rationale
supporting the brochure, specifically, the social and interpersonal problems of infant crying, the issues of teen
paternity in the context of St. Kitts, and current research on the design of culturally sensitive health education.
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COMMUNICATIONS
THE TRICKY TRANSITION: COMMUNICATION IN THE MOTHER AND ADOLESCENT-DAUGHTER
RELATIONSHIP
Ashley A. Kraemer, (Dr. Barbara Penington), Department of Communications, The University of
Whitewater-Wisconsin, Whitewater, WI 53190
The purpose of this project was to investigate characteristics of communication in the mother-adolescent daughter
relationship. Adolescence is a time of physical, social, and emotional changes which can positively or negatively
impact the quality of the relationship the adolescent has with her mother. Since a key to understanding a relationship
is observing the communication within that relationship, this study’s goal was to obtain detailed descriptions of how
mothers and their adolescent daughters talked about their communication. A focus group methodology was used.
The four focus groups consisted of 1) seventh grade daughters 2) the seventh graders’ mothers, 3) high school junior
girls, and 4) high school junior girls’ mothers. Each group was interviewed individually, but asked similar questions.
A thematic analysis was conducted to identify the most important themes relating to mother adolescent-daughter
communication. Results demonstrated that older adolescents had slightly different relationships with their mothers
than did younger adolescents and were more open in communication with mothers. The tension between the need for
freedom and the need to maintain connection was observed between the majority of mothers and their daughters
regardless of age group, supporting dialectical tensions theory (Baxter, 1988, 1990). The theme of “disagreements”
between mothers and daughters was common and focused on chores, listening behavior, and the daughters’ freedom
to engage in desired activities. Future research should focus on mothers and daughters of different regions of the
country, cultures, or social classes as this study was conducted in the Midwest and used adolescents from middle
class European-American families.
A Quantitative Analysis of the Golden Era of Children’s Television
Apperson, Melissa (Dr. Constance L. Book), School of Communications, Elon University, Elon, NC 27244
In his anthology, Hi There Boys and Girls: America’s Local Children’s TV Programs, researcher Tim Hollis (2001)
details a three year national effort collecting information about children’s television programming. This study builds on
that catalog by quantifying the wealth of information collected by Hollis to create the first national database of local
children’s television programming and to use that database to offer quantitative descriptions of local children’s
television in the United States from 1940 to 2000. Trends in the 976 local children’s television programs researched
and analyzed, illustrate that local children’s television programs were embraced nationwide during the 1950s and
1960s, and that the decline of local children’s television shows, while frequently cited as caused by the 1973 NAB
voluntary policy of not allowing children’s programs to include within the program sponsorship or advertising, actually
was more likely due to the availability of syndicated and procured programming options. The thematic orientations of
local children’s television programs illustrate cultural trends in the southern states where local children’s television
was more dominant and that no market size was
too small to have local talent. The database also illustrates the popular themes of children’s television over time, the
domination of males as local children’s host and reveals significant insights into a key attribute of local television
during its initial presence in the United States.
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COMMUNICATIONS
FAMILY ENVIRONMENT VARIABLES AND THEIR EFFECT ON THE ADOLESCENT SELF-CONCEPT
Rachael M. Vance (Dr. Betty Attaway-Fink), Department of Communications, Southeastern Louisiana University,
Hammond, Louisiana 70402
Adolescence is a time of self-discovery and self-evaluation. During this time, the self-concept is shaped and molded
based on feedback from significant others, such as family members and friends. The family has the largest impact on
the shaping of the adolescent self-concept due to the longevity of the family relationship as well as the frequency and
number of interactions. Cooley’s theory of “the looking glass self”, symbolic interactionist theory, and Mead’s
self-fulfilling prophecy site the perceptions that significant others have of a particular person as the main factors that
shape the self-concept.
In a 1981 study, Forman and Forman found parental effects on adolescent self concept can be categorized into
relationship, personal growth, and systems maintenance variables. A reproduction of this study using a sample of
Southeastern Louisiana University sociology students aimed to test the validity of the Forman and Forman study to
Southeastern students. Eighty-eight students answered questionnaires on the perception of their own personality
traits, as well as the perception of their family environment. After analysis, a number of traits were found to have a
direct correlation with each family environment variable. Students that scored high in the family relationships area
tended to be more self-assured, respectful of rules and emotionally stable. Students that scored high in the personal
growth area viewed themselves as independent, adventurous, and warmhearted. High scores in systems
maintenance correlated with respect for rules, obedience, and excitableness.
"GOD HAS A HARD-ON FOR MARINES": THE PORTRAYAL OF MASCULINITY AND MALE SEXUALITY
IN THE AMERICAN WAR FILM
Lindsey A. Sherrill (Mrs. Sally B. Bell and Dr. David Callaghan), Department of Communication Arts, University of
Montevallo, Montevallo, AL 35115
The purpose of this project is to critically examine the rhetoric of the American war film genre in relation to
masculinity and male sexuality. The link between warfare and sexuality has been noted by scholars in many
disciplines and is the keystone of this project. This link is vital to the American war film and places an emphasis on
the concepts of masculinity and male sexuality in the genre. Masculinity/sexuality is enforced in the war film by
linkage to father-son relationships, emphasis of the male body`s prowess or superiority, the use of culturally symbolic
characters such as John Wayne or Sylvester Stallone, and denial or destruction of anything regarded as "feminine" or
"weak." This emphasis on masculinity and male sexuality and denial of the feminine creates the paradigm of the
"Three Evils" of the American war film: the infantile, the female, and the homoerotic. In each of the films critiqued
(including, but not limited to, Platoon, The Deer Hunter, Midnight Clear, Born on the Fourth of July, Apocalypse Now,
Guadalcanal Diary, The Green Berets, The Sands of Iwo Jima, Courage Under Fire, G.I. Jane, and Rambo) these
"evils" are introduced and must be expunged in some way. The emphasis on annihilation of these "evils" along with
glorification of masculinity and machismo is paramount to the rhetoric of the films and is a reflection, even today, of
American gender consciousness in entertainment and society.
WIZARDING WOMEN: FEMINISM IN THE WORLD OF J.K. ROWLING’S HARRY POTTER (A LITERARY
CRITIQUE)
Kristina Kuzma (Ms. Sally Bell), Department of Communication Studies, The University of Montevallo, Montevallo,
Alabama 35115
The purpose of this literary critique was to determine the significance and influence of third wave feminist ideologies
in the works of British author J. K. Rowling, focusing specifically on her Harry Potter novels. The criticism employs the
method of feminist literary criticism put forth in Peter Barry’s Beginning Theory and primarily utilizes the definition of
‘third wave feminism’ used in the text Gendered Lives, a gender studies book by Julia T. Wood. The primary foci of
the rhetorical analysis are the titular male lead Harry Potter and all female characters who have a direct impact on
Harry’s behavior or who appear consistently throughout the six artifacts, thus proving themselves to be relevant and
influential characters. Examples of characters discussed include Hermione Granger, Molly and Ginny Weasley, and
Harry’s foster mother Petunia Dursley. This essay uses a combination of biographical information about the author
and evidence from a rhetorical analysis of the six novel Harry Potter series to argue that the work can be interpreted
as an example of third wave feminism.
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COMMUNICATIONS
ANTI-CONSUMERISM, ANARCHY, AND FEMINISM: THE UNIQUE PATH OF FIGHT CLUB’S PURSUIT
FOR GENDER EQUALITY
Kyle V. Morris (Dr. Rebecca Bell-Metereau), Mitte Honors Program, Texas State University, San Marcos, TX 78666
As the dominance of American culture continues to transcend borders and oceans, its very core is being questioned
at home. Many concerns have been raised as to what the social implications may be for a nation that has placed so
much value on consumption and created a need to purchase. One of the most provocative sources for social dialogue
on the critical implications of consumerism has been the 1999 film Fight Club. While the critics of this film may label it
as an overly masculine blood fest, this paper analyzes the pursuit of feminism that occurs within the plot of the film.
Ultimately, Fight Club can be viewed as an advocacy film for the end of conventional consumerism with an attempt to
create a heightened sense of gender equality through a masculine means of anarchist deconstruction. By analyzing
the unique relationships and actions of the key characters in this movie, it is clear that three basic arguments have
been made by the film itself. First, consumerism must be abolished in the name of preserving traditional masculinity;
specifically the masculinity of middle class males. Second, adopting a system of anarchy is the appropriate method
for achieving the preservation of masculinity. This is especially true since every other model for a society relies on
consumption at some level; even if that dependence is minimized in comparison to capitalism. Finally, the film makes
an argument that true progress will only occur in our society if we become more accepting of the feminist paradigm.
Ultimately, the juxtaposition of the first two arguments against the final argument leads to the climax of the film.
When the dust settles, both literally and figuratively, feminism stands and brute masculinity is defeated. The
questions that Fight Club raises will be important for our national and global leaders to consider in a shrinking world;
which makes the social dialogue of the film significant to every member of the global village.
ACADEMIC AND SOCIAL GROUP COMMUNICATION DYNAMICS FOR COLLEGE STUDENTS
Bessie Dietrich (Dr. James Manning) Communication, Theatre & Dance Dept., Western Carolina University,
Cullowhee, NC 28723
In addition to adapting to new environs and situations, college freshman develop many new relationships as they
enter college. Because of this, the academic and social small group dynamics that new students were previously
familiar with change in drastic and sometimes frightening ways. This paper will examine the dynamic of healthy small
group communication as they apply to college freshmen. Specifically, it will focus on how these communication
dynamics work both in and out of the classroom and how new college students often yield to “groupthink” when in
these surroundings. Recent research has noted the importance of healthy small group dynamics as they relate to
college freshmen. University of Central Florida professor and researcher Rufus L. Barfield stated in 2003 that
“[individuals] in student work groups in the college classroom often struggle to maintain their individuality within the
peer-based groups.” These dynamics are also important socially according to researchers Rempel and Fisher in their
1997 study who studied 31 groups of college males and “examined the impact of perceived threat and cohesion on
the ability of groups to solve problems in a situation of social conflict.” By investigating these small group
communication dynamics and how they apply to college freshmen, the information in this paper will be of great value
to professionals within the communication field, college administrators and faculty, and others interested in student
life and student affairs.
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COMMUNICATIONS
THE PORTRAYAL OF ISLAM AND MUSLIMS IN THE AMERICAN TV NEWS.
Fouad Touzani (Dr. Belle Tuten), Department of History, Juniata College, Huntingdon, PA 16652.
This study examines how Islam and Muslims are portrayed in the American media in general, with a focus on TV
news as the main medium. Moreover, it shall demonstrate how this portrayal has evolved in wars, especially the war
in Afghanistan and Iraq. The CNN and the Fox News will be taken as case studies to show the picture that those
channels are offering to the American audience about Islam and Muslims. This audience can be either active or
passive; the passive audience can be easily influenced by the media, whereas the active audience is more likely to
make more active decisions about how to use the media.
The findings of this study shall be analyzed in the light of the Theories of Message Reception and Processing. In other
words, we shall describe how American TV news receives, organizes and presents information, and how that
information affects the American perception of Islam and Muslims. In addition, we shall discuss the importance of TV
in the judgment processes and message interpretation with reference to Islam and Muslims. Additionally, we will also
examine this study using the Frankfurt School which deems the media, especially TV, as a means of constructing
culture.
Promises Made: A documentary exploring commitments and the fight to honor one promise in rural
South Carolina
Lauren Waring, (Susan Hogue), Department of Art, University South Carolina, Department of Art McMaster College
Columbia, SC 29208
Does protecting a man made lake that displaced 177,000 acres of trees, people, churches and untold and mostly
unrecorded wildlife trump eliminating poverty of people in a small rural Southern town?
Seventy years ago, a dam and lake were created in rural SC to facilitate electric power in the area.
Two communities separated by the lake were promised reconnection by a bridge, and a law was passed in 1968 for
that purpose, but it was un-funded legislation. Four decades later there is still no bridge, and decay continues.
A black congressman in SC has taken up the issue and begun the process of realizing the promise made decades
ago.
Environmentalists, landowners, sportsmen and taxpayers go on record opposing the bridge.
Residents of the communities represented by the congressman tell a short but old story of living out lives in hope and
waiting for real and imagined progress.
Amidst all the practicalities of bridge building, funding and environmental changes, this documentary explores the
many elements of keeping promises and maintaining hope in the face of disappointment, the change of commitments
over time, and the classical American drama: conflict between whites and blacks, the wealthy and the impoverished,
those with political power and those without, progress and tradition.
239
COMPUTER SCIENCE
STABILIZATION OF A SINGLE-PHOTON INTERFEROMETER FOR ONE-WAY QUANTUM KEY
DISTRIBUTION SYSTEMS
Vasil S. Denchev (Dr. Alexei Trifonov and Dr. Anton Zavriyev), MagiQ Technologies, Inc., 11 Ward Street, Somerville,
MA 02143
The two major implementation schemes for Quantum Cryptography are Plug-and-Play (or Autocompensating) and
One-Way. Plug-and-Play systems use Weak Coherent Pulses (WCP) over optical fiber to implement the BB84
Quantum Key Distribution (QKD) protocol, while One-Way systems can utilize both WCP and single photon sources.
Even though Plug-and-Play systems have received considerable attention in a variety of theoretical research efforts
as well as practical realizations in commercial Quantum Information Processing (QIP) products, some firmly
established limitations of Plug-and-Play and the advent of high-quality single-photon sources and detectors give a
competitive edge to One-Way systems. The main weaknesses of the Plug-and-Play approach are relatively low
key-generation rates due to Rayleigh scattering and increased vulnerability to Quantum attacks due to the geometry
of the transmitting station. Both of these drawbacks can be eliminated in One-Way systems, which can also take full
advantage of the newly available single-photon sources. One of the most practical implementations of the One-Way
method when used with optical fiber is the so-called Mach-Zehnder interferometer scheme. The most important
difficulty related to a robust implementation of such a system is the necessity of a continuous active control of the
interferometer. In order for the system to work reliably, the phase of the interference curve must be stabilized with
respect to various mechanical and thermal disturbances. Without stabilization, random phase drifts cause detections
to lose any correlation with encodings. That is why a system for real-time phase-tracking active stabilization was
designed. The system was developed using LabView programming environment and National Instruments Data
Acquisition interface. Presented here are the methodology of the stabilization, results from testing, and a preliminary
simulation of QKD on the stabilized interferometer.
Department of Computer Science, SUNY College at Brockport, 350 New Campus Drive, Brockport, NY 14420
The Measure of Foot Pronation by Stress Gauge Load Cells Utilizing Wireless Data Communication
Joshua Gauthier, Ryan Mclean, Isaac Ostlund, (Dr. Mac Baker), Department of Computer Science, Virginia Military
Institute, Lexington, Va 24450
When fitting a consumer for athletic shoes, most retail stores only measure the length and possibly the width of the
foot. However, to ensure a proper fit, foot pronation and supination should also be taken into account. Foot pronation
and supination is the body`s natural reaction to shock associated with the striking of the foot. Pronation causes the
arch to flatten out and the ankle to roll inward during toe off. Supination, conversely, makes the ankle roll outward.
Excessive ankle rotation caused by improper foot wear is quite prevalent in society and has been determined to be
the cause of some long-term musculoskeletal injuries. If detected, however, one could be fitted with a shoe that
would reduce shock to the skeletal system, especially the feet, thereby reducing their risk of long term injuries.
The purpose of this project is to develop a system that can be used to measure excessive ankle rolling as a
customer is running. The system will include a set of stress gauge load cells embedded in the sole of a shoe that
measure the forces acting on specific areas of the shoe. Using a wireless device mounted on the shoe, the force
information will be transmitted in real-time to a computer, which will analyze the data using a unique algorithm and
determine the proper shoe for the customer based on the calculated pronation or supination of their foot.
The project will involve the development of a prototype system, including shoes with embedded stress gauges and
wireless transmitters, and development of the necessary software to receive and analyze the data from the shoes.
The project will involve the development of a prototype system, including shoes with embedded stress gauges and
wireless transmitters, and development of the necessary software to receive and analyze the data from the shoes.
The microcontroller housing the wireless device will be programmed exclusively in C, while the software for analysis
will be in Java.
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COMPUTER SCIENCE
Social Spiders for Color Image Segmentation
Adam Murphree, J. Thomas Allen, Furman Advantage, Dept. of Computer Science, Furman University, 3300 Poinsett
Hwy, Greenville, SC 29613
This paper deals with evaluating and extending a recent multi-agent model for region detection proposed by Christine
Bourjot, Vincent Chevrier, and Vincent Thomas. This model, inspired by the collected web-weaving of social spider
colonies, involves agents laying virtual silk lines on pixels in the image. This silk weaving, according to the model’s
creators, allows integration of “non-local information in local processing.”
In the paper, we investigate some of the important parameters in the model. We attempt to verify the parameter
values set by the model’s creators by testing the model on a different set of images. We also extend the model to
color, and examine the effects that different color models have on the results produced by the system. Finally, we
address the overall effectiveness of the current spider agent model in mimicking an appearance of global
coordination
POOH: A Theoretic Approach to Building a Robust Hybrid Agent Architecture
Michael Pellon, Heather Richardson, Brent Peckham and Phil Coleman (Dr. Yu Zhang), Department of Computer
Science, , Trinity University, , San Antonio, Texas 78212
Hybrid agent systems attempt to combine two or more agent philosophies into a single agent architecture. One
variant of hybrid agents aims to achieve a balance between the quick responsive behavior of reactive agents with the
proactive behavior of goal-directed agents. We present a novel approach, POOH (Percept Object Oriented
Hierarchy), to constructing this variant of hybrid agents and we discuss the preliminary results that we obtained from
our initial implementation of this approach. Our approach is novel in the following two aspects. First, our approach
allows an agent to pursue multiple goals autonomously. Second, our decision-making process is based on utility
functions, resulting in behavior that is characterized as both efficient and responsive to dynamic and changing
environments. The utility functions express in the form of tuples both the agent`s cost and expected return after
carrying out a potential action. A single execution of the decision-making process returns the action that when
executed by the agent will yield the highest expected value of utility. The results of our experimental analysis show
that the combination of these two aspects results in an agent architecture that is capable of autonomous behavior but
yet robust enough to handle the unforeseen circumstances that may arise in dynamic environments.
Analysis of Usage Statistics of Library E-Resources
Cadet Catrina D. Helsel (Dr. VK Walsh, LTCOL J. Holly), Department of Mathematics and Computer Science, Virginia
Military Institute, Lexington, VA 24450
This research analyzed the usage statistics of nineteen electronic databases owned by Virginia Military Institute’s
Preston Library. Preston Library’s collection contains electronic resources, which are full-text and citation databases.
The database vendors provide information on searches and sessions that occur within each. The databases from
each vendor were reviewed to compare the searches and sessions that were used. The data was compiled by time
periods to make comparisons within and between databases. This analysis covers the usage data from the nineteen
databases during the time period of July 2000 to June 2005, both monthly and annually. The analysis of the VMI
owned databases has assisted the Preston Library staff in their overall analysis of the resources. Using these statistics
the Preston Library staff is able to determine the need for a subscription to a particular online resource. Additionally,
the library staff is able to compare the usage of full-text resources to citation resources. This comparison allows the
staff to select future subscriptions as well as examine the retention for current subscriptions. With new statistics every
year, the process of analyzing the usage of online databases at Preston Library is a continuous task. Future analysis
can be accomplished by creating a Microsoft Access program for Preston Library’s online databases. This program
will allow the library staff to easily maintain the usage statistics of all the online databases.
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COMPUTER SCIENCE
X-SEED SUPER COMPUTER CLUSTER: APPLE G5 X-SERVE HIGH PERFORMANCE COMPUTING
FACILITY AT BOWIE STATE UNIVERSITY
Daniel Green (Dr. Mark Matties), Department of Computer Science, Bowie State University, Bowie, Maryland 20715
Apple built the PowerPC G5, the world’s first 64-bit desktop processor at the time to solve the world’s immense
computing needs. This allowed Bowie State University to create a Super Computer Cluster to handle projects that
were too great for the average computer. The release of the X-Serve PowerPC G5 has allowed scientists to
accomplish even more rigorous calculations, in Bowie State University’s X-Seed High Performance Networking and
Computing facility. The PowerPC G5 is the result of a fruitful collaboration between Apple and IBM, two companies
known as rivals that have solved the problems posed to them from their customers for many years. IBM and Apple
co-created a PowerPC architecture that could run 32-bit and 64-bit programs. When a computer has 32-bit or 64-bit
processors the bit size describes the width of the microprocessor’s data stream. The data stream is a process of the
sizes of its registers and the internal data paths that run the registers. A 64-bit processor compiles data and
instructions across 64-bit-wide data paths, in comparison with the 32-bit-wide paths on 32-bit processors. Also 64-bit
processors have wide registers that hold very precise 64-bit numbers. Now available at Bowie State University’s
X-Seed High Processing Networking and Computing, the PowerPC G5 has next-generation architecture built for
speed and parallel operations. This great computing power is available on Apple systems today at Bowie State
University. In addition students built and manage the X –Seed High Performance Networking and Computing facility.
CIBAC IMAGE PROCESSING USING LIDAR
Navraj Ahuja, (Dr. Paul Chi), Computer Science Department, Bowie State University, Bowie, Maryland 20715
CIBAC is the Chesapeake Information Based Aeronautics Consortium, a consortium backed by NASA to promote
research in engineering and aeronautics. Among the many tasks of CIBAC, my task involves LIDAR data processing.
LIDAR stands for LIght Detection And Ranging, and essentially can be thought if as a radar. In this case, LIDAR will
be used for detecting runways in complex airport image scenes. The LIDAR data, which will consist of connected
component labeling graphs, edge filters, and edge points of airport runway scenes, will be inputted into a c++ program
that must compile and process the data to reproduce the runway image. This program will be loaded and executed
onto a dedicated CIBAC node that is connected to the XSeed super computer located at Bowie State University. This
program must be able to make sense of the LIDAR data to be able to piece together the runway image. This must be
done by making use of auto correlation functions, vector functions, and/or intensity gradient vectors. The algorithms
have yet to be developed, but the program to interpret the data is near completion and will fully utilize the processing
power of the XSeed super computer.
Wireless Remote Access Point (AP) Host Exploitation and Detection
CDT Tyler Steffenson, CDT Joseph Weston, CDT Andrew Wilcox, MAJ Damon Becknel (Dr. Aaron J. Ferguson),
Department of Electrical Engineering and Computer Science, United States Military Academy, 601 Thayer Road,
Room 1128, West Point, NY 10996
WRAP HExD is a tool which allows network administrators to monitor the security posture of their network by
detecting and locating unauthorized wireless access points connected to their network. The system consists of three
subsystems: (1) a “locate and map” subsystem which allows a user to locate and map a wireless AP; (2) an access
control subsystem that exposes unauthorized AP’s. This is done by checking against values given in a user defined
profile and allows the user to view collected information about the AP; and (3) an alert subsystem that is used to
contact the system administrator and inform her of potential network vulnerabilities, and attempt to deny access to the
unsecured APs. All subsystems will be integrated into one graphical user interface.
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COMPUTER SCIENCE
Secure Web-based Distributed Authoring and Versioning (WebDAV)
CDT Ryan Collins, CDT Daniel Stewart, MAJ Rick Brown, Department of Electrical Engineering and Computer
Science, c/o Dr. Aaron J. Ferguson, United States Military Academy, 601 Thayer Road, Room 1128, West Point, NY
10996
A relatively new protocol known as Web-based Distributed Authoring and Versioning (WebDAV) is becoming more
prolific on the Internet. This protocol, which sits on top of the more traditional HTTP protocol, allows multiple
distributed clients to author a document as if they were on the same network and without fear of over-writing one
another’s work. WebDAV, while functional and fast, lacks significant security safeguards and creates another vector
of attack for malicious insiders and hackers alike. Leveraging prior WebDAV security research conducted at the
National Security Agency (NSA). We will develop a new WebDAV server that implements and enforces strong
security policies, making WebDAV safe for public use and in military-based single and multiple-security domain
environments.
NetFires
CDT Brian Boyles, CDT Humberto Nieves, LTC John Hill, 2LT Dee Dee Rich, Department of Electrical Engineering
and Computer Science, (Dr. Aaron J. Ferguson) United States Military Academy, 601 Thayer Road, Room 1128,
West Point, NY 10996
What if an Army unit could rehearse a multi-million dollar mission for free? The solution to this is Netfires. Netfires is
a three dimensional environment that can reliably simulate Precision Attack Missiles (PAM) and Loitering Attack
Missiles (LAM) during launch, flight, and target acquisition. This project provides enhancements to previous versions
of NetFires--a camera view from the missile’s perspective, advanced physics to its flight path, and waypoints to steer
it along its route. In addition to simulating realistic missile traits in a 3D world, an enhanced environment is provided
to support graphical overlays that include military unit designators, phase lines, and other useful 2D visual aides used
in military mission planning. The ability to create attribute-based scenarios makes Netfires a cost-effective alternative
for PAM and LAM mission rehearsal. This paper outlines the design, test, and build methodologies used to solve this
research problem and discusses lessons learned along the way.
Multi-sensory Autonomous Ground Vehicle Inter-Collegiate Competition (MAGIC)
CDT Brian Bifulco, CDT Matthew Hicks, CDT Rodney Johnson, CDT Matthew Smalley, CDT Austin Story (Dr. Pete
Hanlon, Dr. Tommy Wagner) Department of Electrical Engineering and Computer Science, c/o Dr. Aaron J. Ferguson,
United States Military Academy, 601 T
Autonomous vehicles are still in the early stages of development and practical functionality. Their application has
been limited to industrial construction, error detection, and data gathering in remote locations. The goal of MAGIC is
to advance capabilities of autonomous ground vehicles. The roles these vehicles play may one day play include
clearing mine fields, improvised explosive device detection, scientific research in dangerous or remote environments,
and autonomous commercial payload transportation. Our research vehicle is designed to operate within the
environment defined by the Intelligent Ground Vehicle Competition. It will be capable of following two white boundary
lines through an obstacle filled course. The vehicle will operate autonomously to navigate the course, basing its
decisions upon sensory data received from both a laser range finder and an optical system. This paper will analyze
the results garnered from building and testing an intelligent ground vehicle within a path following and obstacle
avoidance environment.
243
COMPUTER SCIENCE
TRUEL – “3 PLAYER DUEL”
Bhaarat Sharma,(Dr.Clyde Kruskal and Dr.William Gasarch), Department of Computer Science,University of Maryland
at College Park,College Park, Maryland 20742
Truel is a duel among three people. In this game three people try to eliminate each other in series of one-to-one
competitions, following the rules of the game. There are many possible variants of the game. The variant we are
considering is sequential (fixed order), the players fire one at a time in order: A, B, C, A, B, C.... Each player has a
Hit Rate, which depicts how accurate he is at shooting the other player. Moreover, they also shoot at the player who
has a higher hit ratio, other than themselves. Given the Hit Rates, we compute the Survival Rates of players using
hand derived formulas involving probability. Furthermore, we have programmed a simulation of the game, solely
based on different hit rates of players, in java. We show that the results derived from the simulation coincide with the
results derived from the formulas. Our aim is to find equal survival rates for players (1/3), given different Hit Rates.
We also try to find patterns between the different hit rates, which give us equal survival rates. Moreover, our future
plans include: extending the game to more than 3 players, finding hit rates that give us equal survival rates for more
than three players, generalizing the formulas, considering different scenarios and discover strategies for players to
maximize their survival rates.
Bowie State University
Perimeter Defense
CDT Anthony Andrews, CDT Brendon Baker, LTC John Hill, 2LT Dee Dee Rich (Dr. Aaron J. Ferguson), Department
of Electrical Engineering and Computer Science, United States Military Academy, 601 Thayer Road, Room 1128,
West Point, NY 10996
The purpose of the Perimeter Defense project is to create a system to respond quickly to an intruder without alerting
the enemy. This will enable friendly troops to gain an offensive edge on the opponent. The Perimeter Defense
System will aid in achieving this goal by allowing a perimeter to be passively monitored by sensors, keeping the
enemy unaware of his surveillance. Due to the system’s ability to determine when an intrusion occurs, our troops will
be able to respond accordingly, thereby attacking with devastating accuracy. The Perimeter Defense System, then,
will play its part in the digitization of our Armed Forces. This is the third year for the Perimeter Defense project. For
its first build, a design team of electrical engineering and computer science cadets designed a system made up of a
sensor box, a control box, and one hand held device. The system could monitor a static array of sensors on a single
hand held device. The second build team comprised of just computer science cadets attempted to expand on the
previous build. They attempted to design a system of variable sensors for a flexible perimeter and a small network of
hand held devices that would share this information. While they made some important strides in achieving their goal,
the compilation of their project proved that the different systems they had designed did not work together. Given last
year’s unsuccessful attempt at designing a realistic and flexible perimeter defense system, this year’s design team
hopes to reanalyze, design, and build such a system. The end state is a system that can be used by the Armed
Forces, as well as in the civilian sector, to provide close reliable security in a variety of environments.
244
COMPUTER SCIENCE
Cost Effective Home Robotics Technology
Jonathan H. Schwartz, Mathematics and Computer Science Department, Bridgewater State College, 131 Summer St.,
Bridgewater, MA 02325
Home Robotics is a technology that has not yet developed a broad market largely because of prohibitive cost. The
purpose of this project was to develop new home robotics technology that makes an advance in terms of existing
software and maintains affordability. To do this, I constructed a robot designed to serve drinks. The hardware includes
a pre-assembled electronics board, hobby servos, various structural elements, and three infrared sensors. An analog
to digital converter uses the infrared sensors to compute a distance from objects. The software was written in Visual
Basic and includes code that uses the computed distance from objects to avoid those objects. Existing technology
performs this same function and utilizes camera vision systems and on-board computers to distinguish between
obstacles and objects that require a response. However, this technology is prohibitively expensive for the home
environment. The robot constructed for this project represents an affordable advance from existing technology by
including new code that determines the shape of an object and distinguishes between obstacles and people by only
relying on the infrared sensor. This new avoid and response technology could potentially have much wider application
in the Home Robotics market.
http://webhost.bridgew.edu/jschwartz/gir.htm <-- Online Lab Journal
PARALLEL COMPUTING USING MPI AND TASK ALLOCATION ALGORITHMS
Jon Quarfoth, Andrew Korth, (Dian Lopez), Division of Science & Mathematics, University of Minnesota, Morris, 600 E
4th St, Morris, MN 56267
The goal of our research is to implement solutions to complex problems in a parallel computing environment, using
task scheduling algorithms to distribute the workload efficiently across many computers. Many large jobs which run on
single computers can be split into a series of smaller tasks, each of which can be run on a different computer.
Algorithms we designed over the past few years schedule these smaller tasks onto a network of computers in order to
minimize the computation time for the job as a whole. This prior research, however, was purely theoretical. Jobs were
modeled as precedence graphs. Each node of the graph represented a different task as a part of the larger job. Edges
on the graph represented latency and communication time between associated tasks. We simulated the length of time
it would take a job to run. Our algorithms produced very promising results on these theoretical problems, so we
decided to implement them in a real system. We are using the Message Passing Interface (MPI) libraries to
coordinate jobs on a group of networked computers.
The problem of parallel matrix multiplication was examined. The problem can be easily scaled to different sizes,
which allows us to create a wide variety of test cases. The size of each individual matrix can be scaled, as well as the
number of matrices being multiplied. By altering the number and order of matrices multiplied, we can easily create
multi-leveled dependencies for our precedence graph models. This problem will serve as an excellent base for testing
the performance of our scheduling algorithms. If these tests demonstrate that our scheduling algorithms are viable,
this research could easily be applied to other parallel computing problems, such as data compression, machine
learning, video analysis, and data mining.
A Computational Model of Mars Craters-Size Frequency Distribution
Axel Rivera, Department of Mathematics, University of Puerto Rico at Humacao, , Elio Ramos, Department of
Mathematics, University of Puerto Rico at Humacao, , William Bruckman, Department of Physics, University of Puerto
Rico at Humacao, , Abraham Ruiz, Departm
We present a computational model that reproduces essential features of the frequency distribution vs. diameter, of
the 42,000 craters contained in the Barlow`s Mars Catalog. The model, based in the Monte Carlo method, assumes
that the martian surface is a two dimensional grid and simulates the crater formation process due to the meteor`s
impacts, of different diameters, falling uniformly at random locations. The diameters of the simulated meteors were
randomly generated from a power law distribution (slope=-4) were smaller meteors are more frequent than larger
meteors. The simulation takes into consideration the reduction of craters numbers as a function of time caused by the
impact of large meteors falling over small craters already formed (obliteration). The model provides a simple and
natural explanation for the presence of different slopes in the observed log-log plot of numbers (N) vs. diameter (D).
245
COMPUTER SCIENCE
IMPACT OF DEPARTMENT CLIMATE AND GENDER IN PROFESSOR PRODUCTIVITY
Margi Dubal, Amanda Bakian (Dr. Kim Sullivan, Dr. Ronda Callister, Christina Brower), (National Research
Foundation), Department of Biology, Advance, Utah State University , 0187 Old Main Hill, Logan, UT 84322-0187
Under the ADVANCE grant, we investigated productivity of science & engineering tenured and tenure-track
professors as a measure of external grant funding awarded and department climate. Teaching hours, previously
analyzed, was also considered in the research analysis. Data on external grant funding was collected over 8 years. 4
years of completed and cleaned data was used in our initial analysis. Results show the median amount of external
grant funding awarded to female and male professors to be equivalent. Medians over 4 years show an increase of
total external grants awarded to women while total awards to men have declined. This directly matches recent hiring
increases in female tenure-track professors and the retiring of male professors. Regression on the College of
Engineering data shows a slightly higher relevance between females and external grant funding even though the n for
females was significantly low. As far as climate, departments who have less than 50% of their faculty bringing in
external grant funding scored higher in their dept. climate satisfaction but also had higher teaching loads.
Departments who have 80% or more of their faculty bringing in external funding reported the largest dissatisfaction in
their department climate. Does teaching less or more have a larger impact on dept. climate than working on grant
funding activities? We will discuss our research results and how department climate and gender plays a part in
faculty productivity with regard to teaching, and obtaining external grant funding and how institutions may want to
change to increase faculty productivity.
DISTRIBUTED COMPUTATION IN AN INTERACTIVE ENTERTAINMENT ENVIRONMENT
Robert J. Zinchak (Dr. Gerald Pitts), Department of Computer Science, Trinity University, San Antonio, Texas 78212
The proliferation of multiplayer games has led to an increase in the total network capacity for processing in games,
but this capacity is rarely fully utilized or balanced. One prominent problem of distributed processing in a gaming
environment is increased latency time, which causes player disinterest in the game potentially causing poor sales and
the termination of future commercial development of this technology. Existing distributed techniques such as
OpenMP, MPI or VMPI are not well suited to gaming applications and may introduce additional overhead.
It is the purpose of this paper to describe a simple, yet effective technique (based on existing ideas in both multiplayer
network games and parallel processing) that can be implemented into existing networked games to allow for
distributed computation, which in turn will balance the work load across all connected machines while simultaneously
decreasing visible latency to players. Most multiplayer games use a client-server architecture, where the server acts
as a referee between clients to manage the game state. A game world with many A.I. agents (simple non-playing
characters) overloads the server and causes higher latency levels. Load may be balanced by allowing trusted clients
to compute these actions. Development and maintenance costs can be kept low while not diminishing production
quality, leading to a successful game. In addition to gaming applications, this technique can be incorporated in any
existing client-server network application, including: communication, transportation, and process control.
This paper seeks to discover whether parallel processing offers a viable option for reducing latency and enabling
larger, more realistic virtual worlds in multiplayer games. A comparison between the traditional client-server method
and the distributed processing approach will be provided. An example is created that simulates many A.I. agents in a
larger N.I. (narrative intelligence) environment. Empirical tables and graphs will show the relationship between
overhead and optimization this technique creates, illustrating whether parallel processing in networked multiplayer
games deserves implantation.
246
COMPUTER SCIENCE
THE RISE OF A NEW OBJECT GENERATION (BREAKING THE TRADITIONAL RELATIONAL
DATABASE)
Scott I Wakefield,(Floyd Wilkes), Information Systems and Technology, Utah Valley State College, 800 W University
Pkwy, Orem, UT 84058
The purpose of this research is to compare certain aspects of an Object-Oriented Database Management System
(ODBMS) with a Relational Database Management System (RDBMS.) Many organizations are finding that their data
requirements are becoming more complex than what can be delivered by a RDBMS. Since the early seventies,
relational databases have become the standard for most database development. Companies adopted this model
because it is flexible than its predecessor systems making it possible to obtain many answers to the question that a
business asks of its data. By using a RDBMS, a business has access to a large constituency of third party tools and
support for relational databases. If a company were to switch to an ODBMS, they would lose this support, and find it
necessary to rely on themselves or a small number of vendors for support. It would also require the retraining of many
employees currently familiar with RDBMS methods and technologies. However, from a developers’ standpoint,
ODBMS requires less code, and eliminates the need for connecting drivers such as ADO.net to access information
from the database. This research will investigate three areas where differences seem to exist: The number of lines in
code required, the amount of time required to produce the transactions, and the relative performance of each system
to handling different data types. SQL Server 2000 and db40 will be the RDBMS and the ODBMS respectively, and
code will be written in Java 1.5. The lines of code required will be measured by a program named CCCC available
from www.sourceforge.net. The required time to process the transactions will be measured by methods already
available in Java. The null hypothesis to be tested is that there will be no difference between the systems in the
areas studied.
EYE FEATURE DETECTION TOWARDS AUTOMATIC STRABISMUS SCREENING
Kenneth Allen, Khanh Nguyen (alphabetical order), (Dr. Sunghee Kim -- sponsor) , Department of Computer Science,
Gettysburg College, 300 North Washington St, Gettysburg, PA 17325
Strabismus is an eye defect occurring in infants which, if left untreated, can result in inability to attain binocular vision
or blindness in the affected eye. Early detection is critical for successful treatment but current methods involve going
to an eye
specialist or going to a lab with specialized hardware and lighting equipment.
The purpose of this project was to determine the feasibility of a software-only solution that could be deployed to photo
studios as a low-cost early screening mechanism. The eventual goal is to be able to detect signs of strabismus from a
single high resolution portrait photograph of an infant.
The strabismus screening algorithm consists of two phases. In the first phase the locations of certain eye features
such as pupil center, eye corners, specular highlights, etc are determined. In the second phase the locations of the
highlights in both eyes are examined in relation to the pupil centers. The success of the project depends heavily on
the accuracy of eye feature detection and is the focus of our work.
We investigated several existing computer vision algorithms for detecting various eye features and chose the ones
that would handle the requirements best for implementation. The results so far are very promising and we are
currently working on fine-tuning our program to improve the accuracy.
247
COMPUTER SCIENCE
Implementation of a Binary Tree Driver in Cactus
Jeff DeReus, Yaakoub El-Khamra, Frameworks Group, Center for Computation and Technology, Johnston Hall,
Lousiana State University, Baton Rouge, LA 70803
“Implementation of a Binary Tree Driver in Cactus”
Cactus is an open source environment designed primarily for scientists and engineers. Cactus is based on a
framework design in which the core or “flesh” acts as a central unit to which external modules or “thorns” can
interface. Cactus is used on many different architectures and can implement different codes from various disciplines.
While initially developed for solving problems in numerical relativity, it has since expanded to include thorns for
computational fluid dynamics, climate modeling and bioinformatics.
Binary trees are fast insert and lookup recursive data structures with at most two children at each node.
Considering the wealth of applications that require binary trees it is only natural to implement a binary tree module in
Cactus. Some examples of current applications include: high performance databases, visualization hierarchies,
discrete mathematics, Monte Carlo simulations, logic programming and computational econometrics. The module we
are developing is special in that it can allocate binary tree nodes for other thorns to use, in effect realizing the role of
a Cactus driver thorn. Due to the high computational loads involved in most real world problems, one basic
requirement of this binary tree driver is parallelism to make use of high performance computing environments. In this
paper we discuss the design and implementation of a complete binary tree driver in Cactus along with other
supporting thorns including a web-interface thorn.
CONSTRUCTION OF LISTS OF NEIGHBORS OF POINTS BY MEANS OF DELAUNAY
TRIANGULATIONS
Francisco J. Izquierdo, (José O. Sotero-Esteva), Department of Mathematics, University of Puerto Rico at Humacao,
Humacao, Puerto Rico 00791
Given a set of points in the R³ space we want to construct efficiently, for each point of the set, the list of all other
points that are whithin a prescribed distance. This problem is related with the computation of short range interactions
in molecular dynamics simulations. These neighbors lists could be constructed by the inspection of all possible pairs
of points. This naive algorithm takes time O(n²). On the other hand, we could assume that we have a structure
already defined using those points that could help us construct such lists. The data structure tested here was the
Delaunay Triangulation (DT). A DT is a triangulation that satisfies the empty circle property, that is, the
circumference defined by the vertices of each triangle cannot contain another point of the triangulation. In practice,
DTs connect each vertex with their nearest neighbors. Depending on which data structure is used to represent DT,
the teorethical construction times vary from O(n²) for a flat structure, to O(n log n) for a hierarchical structure. Once
the DT is constructed, we construct the lists of neighbors of one point by doing a depth first search starting from that
point until the prescribed distance is reached. In order to measure actual running times for data sets of the order of
magnitude and distributions commonly found in molecular dynamics simulations, we compared empirically the three
types of constructions mentioned above: naive, flat Delaunay, and hierarchical Delaunay. We used the CGAL
package which supply easy access to geometric algorithms and DTs. We observed empirically that the hierarchical
structure is better than the others for problem sizes ranging from 10³ to 10^5 points. Different distributions of points
do not seem to affect this performance.
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COMPUTER SCIENCE
Remote Real-Time Visualization in High Performance Computing Environments
Ian Wesley-Smith and Yaakoub El-Khamra, Frameworks Group,, Center for Computation and Technology,, Johnston
Hall,, Louisiana State University,, Baton Rouge, LA, 70803
Three dimensional visualization is arguably one of the most valuable tools supporting high performance computing. It
allows researchers to fully understand what is occurring inside a simulation, helping them gain insight into the problem
they are investigating. Additionally, it allows them to monitor the status of a simulation, ensuring that expensive
computational time is not wasted on a simulation that has gone awry. Three dimensional visualization, however, is by
no means a trivial task. Many simulations have very large data sets which need to be rendered in real-time, a task too
computationally intensive for all but the most powerful of workstations. To further complicate matters, users often
need to run their code in heterogeneous environments, e.g. moving from highly parallel clustered systems to
high-end, multi-processor shared memory systems.
Despite these difficulties, several open-source systems provide easy methods to generate three dimensional
visualizations in various environments. Cactus is an open source high performance computing framework that allows
researchers to use a variety of modules in their simulations. One such module, IsoSurfacer, provides a parallel
rendering engine that runs with the simulation and can output a variety of common visualization data formats. IsoView
is a remote real-time visualization client that can be used in conjunction with Cactus and IsoSurfacer. Using Cactus,
IsoSurfacer, and IsoView, users can run their simulation on any number of nodes, and then remotely visualize their
data, in real-time, on the machine of their choosing. Several techniques utilized in IsoView allow for data to be viewed
on a wide variety of systems, from a laptop with integrated graphics, to a high-end visualization workstation. This
paper compares IsoView to other popular remote visualization tools (such as Amira and OpenDX), analyzes specific
methods used in IsoView (and their effect on performance), and finishes by highlighting possible areas for
improvement.
A Parallel Artificial Neural Network Implementation
Ian Wesley-Smith and Yaakoub El-Khamra, Frameworks Group,, Center for Computation and Technology,, Johnston
Hall,, Louisiana State University,, Baton Rouge, LA, 70803
Traditional computational methods are highly structured and linear, properties which they derive from the digital
nature of computers. These methods are highly effective at solving a certain class of problems, for example physics
simulations, mathematical models, or the analysis of proteins. Classical computational methods are not effective at
solving other problems, such as pattern recognition, adaptive learning, and spam filtering. Some biological systems,
however, excel at the latter class of problems. For example, the human mind can quickly identify a face, even if it has
changed heavily from the last time it was seen, while traditional computational systems are unable to accomplish
facial recognition efficiently and accurately even if minor facial or environmental alterations occur. Attempts to create
facsimiles of these biological systems electronically have resulted in the creation of artificial neural networks. Similar
to their biological counterparts, artificial neural networks are massively parallel systems capable of learning and
making generalizations. The inherent parallelism in the network allows for a distributed software implementation of
the artificial neural network, causing the network to learn and operate in parallel, resulting in a significant performance
improvement. This paper will address a parallel neural network implementation in Cactus, a high performance
computing framework, the network`s relative strengths and weaknesses, and conclude by considering future
improvements to the system.
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COMPUTER SCIENCE
Using a Graphics Processor Unit (GPU) for feature extraction from turbulent flow datasets
Andrew Davidson, Yaakoub El-Khamra (Dr. Mayank Tyagi), Frameworks Group, Center for Computation and
Technology, Johnston Hall, Baton Rouge, LA 70803
Recent advances in programmable commodity graphics processing unit (GPU) cards allow many visualization and
numerical algorithms to be mapped efficiently onto hardware. Much literature shows that for compute bound
applications as well as for memory bandwidth bound applications, GPU has the potential to outperform CPU-based
solutions. In addition, due to its high cost effectiveness (Megaflops/dollar), the GPU has become a tempting
co-processing unit. However, the programmability on a GPU has remained a challenge. There is little flexibility with in
the current languages along with many hardware constraints, and general CPU codes cannot be converted directly
into corresponding GPU codes. Much progress is being made towards effective GPU co-processing. New graphics
languages such as Cg, Brook, GLSL, and HLSL have allowed programmers more control and options on the GPU.
Using these languages, and compatible GPU algorithms, a significant increase in speed (up to five fold) has been
observed when compared to a stand-alone CPU calculation.
In this research work, use of GPU as a co-processor is demonstrated by computing pressure Laplacian and
velocity gradient tensors from a turbulent flow field dataset generated by a computation fluid dynamics (CFD)
software. Using textures as data storage, and fragment programs for computation, we have been able to upload data
to a GPU, process data on a GPU, and return data from a GPU. Extraction of iso-surfaces from these relevant fields
(e.g. pressure Laplacian) reveal coherent structures (i.e., vortices) in these CFD datasets. All the tasks including
calculation of diagnostic fields, extraction of iso-surfaces, and rendering of features has been performed entirely on a
GPU.
A STUDY OF SECURE CLIENT/SERVER COMMUNICATION
Joshua N. Louden, (Dr. Brian King),Indiana University-Purdue University Indianapolis, School of Engineering and
Technology,, 723 W. Michigan Street., SL 160 Indianapolis, IN 46202-5160
Internet communication revolves around the client/server model. Information not encrypted when transmitted can be
seen by anyone. A method of encryption typically used requires the same secret key on both client and server. Often,
the client and server have never met, and a key exchange must take place. Recently an attack was discovered on
one of the key exchanges, requiring a modification of the cryptographic standard. Our goal is to develop a
client/server program, using the programming language Java, which simulates how a client and server securely
communicate. Then implement this attack demonstrating how an attacker can decrypt the communication.
250
COMPUTER SCIENCE
Integration of Trilinos Into The Cactus Framework
Josh Abadie and Yaakoub El-Khamra, Frameworks Group, Center for Computation and Technology, Johnston Hall,,
Baton Rouge, Louisiana 70803
Many of todays pressing problems in science and engineering involve the solution of systems of partial differential
equations on large scale computing systems. A number of frameworks and libraries have been designed and
implemented to expedite such research requirements in these fields.
The Cactus Code is an extensible, collaborative, parallel, computational application framework for large high
performance computing. Through its generalized and elegant design, Cactus capabilities are provided by a plethora
of plug-and-play modules, or `thorns`. Although Cactus was written in ANSI C for maximum portability, thorns can be
written in C, C++, Fortran 77, and Fortran 90. Currently, Cactus is being used in numerical relativity, bio-informatics,
climate modeling, astrophysics, computational fluid dynamics, and grid computing, among other fields.
Trilinos is a collection of self-contained interoperable `packages`, developed at Sandia National Laboratories.
The primary purpose of Trilinos is to ease integration of different mathematical software libraries. Some of the
packages in Trilinos implement distributed matrix and vector data types, general solvers, algebraic preconditioners,
linear solvers and non-linear solvers. Interoperability between packages is achieved through a number of
mechanisms designed to minimize inter-package dependence. Though mostly written in C++ with heavy use of
objects, packages can also contain Fortran source.
While there are a great number of different libraries which Cactus supports, several of the libraries that have
been assimilated into Trilinos, such as the solvers Aztec, Anasazi and Amesos, the preconditioner ML and many
others are not accessible in Cactus. To provide a broader range of solvers to the Cactus user community, we have
integrated Trilinos with Cactus.
We will briefly discuss the design of the Cactus Framework and then describe the design and implementation of
Trilinos integration into Cactus.
THE ROLE OF ITERATION IN THE EARLY PHASES OF A TRACEABILITY-ORIENTED SOFTWARE
DEVELOPMENT PROCESS.
Borislava I. Simidchieva, Stefan C. Christov (Dr. Sandeep Mitra), Department of Computer Science, State University
of New York, College at Brockport, Brockport, NY 14420.
According to the recent Standish Group’s CHAOS reports, the “software crisis” persists. Royce’s Waterfall Model
correctly identifies the phases of software development (requirements capture, problem analysis, design,
implementation, and testing), but does not facilitate iteration between phases. A recent IEEE Computer (September
2005) article presents a comprehensive, multi-faceted overview of iteration. It illustrates how iteration is an inevitable
aspect of software development. Many later methodologies attempt to facilitate iteration (e.g., the Spiral Model, RUP,
etc.), including Agile methodologies like eXtreme Programming (XP), which claim to “embrace” iteration. XP
discusses “refactoring”, whose primary emphasis is improving implementation code quality. However, techniques for
embracing iteration during earlier phases, focusing on artifacts created during these phases, remain relatively
untouched. Few methodologies explicitly demonstrate how iteration may actually be achieved in specific types of
applications.
This paper reviews existing iteration strategies, noting their focus on the design/implementation level – principally,
“code refactoring”. It then explores the Software Engineering Effectiveness Model, (SEEM™) and its unique focus on
ensuring accurate and obvious traceability (presented at NCUR 2005). SEEM™ advocates using certain principles to
create a precise, unambiguous analysis model for applications that have a GUI front-end and a database back-end. It
then uses standard design patterns to map from the technology-independent analysis model to the implementation
technology-specific design/implementation models. The paper demonstrates this process using a detailed case study
of a “Journal Maintenance System”. It then focuses on techniques for evaluating the quality of the analysis and design
models. A high-quality analysis model should enable the programmer to systematically map it to later phase models.
If this mapping is not easily achievable, the paper shows how a developer can reexamine the analysis model,
attempting to improve it. Thus, the developer effectively iterates at the analysis/design phases, improving quality and
correctness early. Our results show that this approach yields high-quality code, with drastically reduced refactoring
needs.
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COMPUTER SCIENCE
THE DEVELOPMENT OF AN INTEGRATED MODULAR EXTENSIBLE ROBOTIC PLATFORM
Julus S Schlosburg, Goran Trajkovski - research sponsor, Computer and Information Sciences Department, Towson
University, 8000 York Road, Towson, MD 21252-0001
During the development of POPSICLE (Pattern in Orientation: Pattern-Aided Simulated Interaction Context
Experiment) and through various other research projects at CARoL (Cognitive Agency and Robotics Lab), various
problems arose from the need to build a custom robot for every task. POPSICLE involves using robotic agents to
simulate learning in a physical environment. The experiment is currently being carried out by robots equipped with a
color sensor to ascertain their location on a color-coded floor. The robots are also equipped with infrared and sonar
proximity detectors to avoid collisions with objects in the environment and are controlled via a microcontroller called
the Acroname Brainstem. The platform that I have been using, however, uses a type of motion known as holonomic
motion, meaning the robot can rotate along a center axis without changing the direction it is moving in. This is
accomplished using a three-wheeled, hexagonal design utilizing specialized wheels, and, although useful in some
situations, its limited accuracy of movement is prohibitive for use with POPSICLE. I have research the possibilities
and methods involved in developing a module-based robotic platform for the lab that can be modified easily to be
used in any circumstances, mainly for POPSICLE, but also for general future research and education. The Lego
Mindstorms which are currently used in the lab for education, while versatile and modular, do not offer the type of
higher-level functionality that CARoL hopes to offer in the future. The platform built is a small base of rectangular
shape which includes servos and a treaded drive system, a universal header bus for peripherals, a universal adapter
for our most common microcontrollers, a universal power adapter for use with multiple types of power sources and
batteries, and an extensible body with removable placement points for sensors. Within this project, the prospect of
modularization of most of the peripherals in the lab to make them compatible with the new platform has also been
investigated.
Building a Neural Network CPU
Brian Whitman, Goran Trajkovski, research Sponsor, Computer and Information Sciences Department, 8000 York
Road, Towson, MD 21252-0001
It should be apparent to anyone in a technology industry that embedded computers are starting to saturate the
consumer space. With this increase in computing power available, literally, at everyone’s finger tips, the expectations
of typical users is increasing dramatically. Science fiction of yesterday and science fact of today is frequently the
same thing. Computer based voice and video recognition are two applications that are in the forefront of user
expectations, voice automated help systems and dialing directories are just a couple examples that are already
available. There a numerous ways to solve this type of classification problem, but Neural Networks provide an elegant
solution that inherently accommodates parallelism. Unfortunately software implementations of Neural Networks can
not leverage this parallelism as effectively as hardware. Our idea is to implement a basic Neural Network CPU using
a FPGA (Field Programmable Gate Array). Current FPGA hardware can accommodate very complex designs, and
should be a good match for prototyping our CPU.
To make this a useful exercise, there are three main problems we will need to address. The first problem is that of
network storage and instantiation. To be useful a CPU will need to handle more than one network, not necessarily at
one time, but certainly many with-in a short time frame, maybe hundreds or thousands per second. A Neuron is a
simple construct, and a Neural Network is not much more complex, but the number of connection possible between
Neurons can make the process building a network and its connections time consuming; a good analogy might an
Operating System’s job of task scheduling. The goal in our system is to minimize the context switch so the network
can respond to input in a timely fashion. Context switches must be managed by balancing internal versus external
storage memory usage. Internal memory is fast but limited while external memory is slow but abundant, and
determining how much of network must reside in internal memory is not a trivial problem. This internal versus
external problem is also related to the second problem. Neural Networks use weighted inputs to store what they learn,
and again to perform operations with a fast response time some balance between internal versus external storage of
these weights must be devised. The final problem to overcome occurs at the borders of our design: input and output.
Many devices use serial interfaces and many use analog interfaces, developing a flexible means of input and output
is essential for our CPU to be useful.
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COMPUTER SCIENCE
ADJUSTABLE ORGANIZATION OF DATA USING CHI-SQUARE SIMILARITY
Maeghan G. Gault (Ms. Aiyesha Ma and Dr. Ishwar K. Sethi), Department of Computer Science & Engineering,
Oakland University, Rochester, MI 48309
Clustering is a means of assembling objects into groups of similar items. Clustering data for exploration is very
important to many domains; therefore, many techniques exist for clustering data. The significance of clustering is
evident for high-dimensional data. It is difficult for humans to distinguish similarity in items when more than two
descriptive components are present. One problem with existing clustering methods, however, is the fixed
determination of whether data elements are similar enough to form a group. The chi-square test is a confidence
measure with intuitive thresholds for grouping data together, which allows for further data exploration by the user.
This research examines the ability of the chi-square similarity measure to cluster high-dimensional data using
standard data sets. The chi-square method is successful in combining data elements by allowing for decreased
similarity as the confidence level changes from 1 to 0. Initially small, highly similar clusters are generated, and then
larger clusters are generated as the confidence decreases. This confidence level may be specified with ease based
on the user’s needs and intentions in exploring the data. Due to the ever-increasing nature of multimedia information,
the chi-square similarity clustering approach is then applied to organize color images in an adjustable manner to
distinguish image similarity. The HSV color space is utilized for comparing global color histograms of the images that
are refined through window-based smoothing. Thus, the chi-square method establishes an effective organization of
color images.
MIGRATION FROM MICROSOFT WINDOWS TO LINUX BASED SOFTWARE IN A GERMAN
ENTERPRISE ENVIRONMENT
Jordan M. Koch, Dr. Russ Christensen, Department of Computer Science and Modern Languages, Hamline
University, 1536 Hewitt Avenue, Saint Paul, MN 55104
The option of using open source software that is suitable for enterprise conditions has become more readily available
since the introduction of Linux in 1991. Linux is open source, which means all of its underlying source code is free to
read, edit and redistribute, as opposed to Microsoft Windows’ proprietary code that is locked up and available only to
Microsoft. This research investigates the open source practices of Linux and Microsoft in German enterprise venues.
Aspects taken into consideration are the contribution of Linux’s prompt upgrades and patches, greater machine
control, reduced total cost of ownership and free software download with automatic installation. Microsoft will also be
looked at regarding these concepts, as well as any adaptations they would be required to contend with. As a growing
concern, issues of security are also considered: of the last forty security flaws found in both operating systems, thirty
eight percent of them were critically flawed in Windows, while only ten percent of the Linux flaws were critical
(Security Report: Windows vs. Linux, The Register online journal). Moreover, there are only five known viruses
capable of infecting Linux, whereas Windows is susceptible to 60,000.
German enterprise environments provide this study with venues which are particularly revealing of the advantages
and disadvantages of an open source migration. Munich, the third largest technology hub in Europe, has recently
switched its city administration over to Linux, much to the bristling response of Microsoft. The research concludes by
examining in detail the potential benefits as well as the current and possible consequences for Germany of this
migration.
253
COMPUTER SCIENCE
Ontology Based Search Engine for a Decision Support System
Brian C. Becker, Christina M. Vargas, and Avelino J. Gonzalez, National Science Foundation, University of Central
Florida, 4000 Central Florida Blvd., Orlando, Florida, 32816
As the amount of digital information increases, finding a specific piece of information becomes increasingly difficult.
Historically, search engines have endeavored to use a search term or question to sift through large amounts of
information to find relevant data. Traditional search engines usually have difficulties finding relevant information
unless the search term occurs explicitly within the information. While this approach oftentimes yields good search
results, it fails to account for variations or related words not specified in the search term, forcing users to rephrase
their question and perform another search. In a decision support system, searching becomes more critical as users
will expect an intelligent response on the first try. Utilizing a tree of word relationships such as an ontology
strengthens linkage between words and phrases. An ontology contains commonly used words within a domain of
knowledge and links them with other words in the ontology through a combination of 5 relationships: hypernym,
hyponym, holonym, meronym, and synonym. An ontology is best used in a localized domain, such as a decision
support system (DSS), since it allows a search engine to perform more in-depth matching between the user’s question
and the knowledge database that contains the answer. This paper investigates the application and effectiveness of a
search engine designed for a real-world domain-specific DSS called AlexDSS. The knowledge within AlexDSS is
categorized into contextual graphs (CxG) with each graph describing a specific practice, process, or fact. When
selected through the ontology-aided search engine, a contextual graph dialogues with the user to tailor the information
to the user’s unique situation. Comparisons between a traditional search engine and an ontology based search engine
in AlexDSS will determine the increased effectiveness of utilizing an ontology.
LIST MANAGER: SOFTWARE DEVELOPMENT LIFE CYCLE
Erica L. Lesyshyn, Department of Computer Science, Salem State College, Salem, Massachusetts 01970
Today`s developers and project managers face many pressures to meet expectations for faster, cheaper and more
efficient software development. Resources are wasted each year on projects that either fail or exceed their budget. To
avoid these pitfalls, programmers must utilize software development methodologies designed to maximize software
integration, code reusability, communication and resource estimation.
To accomplish these goals, the user requirements for the software underwent a structuring process. Multiple meetings
with the end users determined the storage, management and reporting requirements. These meetings painted a
picture of a software program designed to manage marketing list data. The end users needed a way to store, manage
and share information about marketing lists in a single, central repository.
After the requirements for the application were established, the design phase began. This stage was broken down into
three distinct tiers. Each tier allowed for careful consideration to ensure that it would seamlessly integrate with other
software and enable future growth for both the application and the data it would govern.
The first tier consisted of data normalization and database design. This provided a robust foundation for storing and
managing the data in the underlying repository. The second tier consisted of the core entity modules that
encapsulated the business logic. This tier also focused on integration with pre-existent software. The final tier
illustrated the user interface and program flow of data from interface to storage.
With the design phase complete, construction of the user interface began. When the interface became interactive, the
end users were brought back into the process to provide feedback. This process continued iteratively until all end
users were satisfied and the software requirements were met.
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COMPUTER SCIENCE
POCKETLENS: A GENERALIZED IMPLEMENTATION
Jeffery A. Beck, Adam M. Burk, Kristopher J. Williams (Dr. Bradley N. Miller), Department of Computer Science,
Luther College, Decorah, Iowa 52101
As available content continues to increase at an alarming pace, new ways to find relevant content are necessary.
Personal recommender systems can help individuals find quality content. Our research is the implementation of the
PocketLens algorithm, which is an important first step in personal recommenders. Scalability, privacy cost, and user
integration are important issues we consider. The PocketLens algorithm addresses problems with cost and privacy.
Because it is a distributed system, users retain complete control over their own data while eliminating the need for a
costly central server. To provide better scalability we changed the PocketLens algorithm so it allowed users to get
recommendations even when no other users were currently using the client. This change allowed for testing on a
much smaller scale. We generalized this method in order to explore the PocketLens algorithm and to test the
practicality of making cross-type recommendations using MultiLens, for instance using one’s taste in music to
recommend web pages. User integration is an important issue because the better the system fits into preexisting user
habits, the more likely users are to rate and use recommendations. Allowing users to import ratings easily from
preexisting sources such as iTunes and their favorite bookmarks is an example of this kind of integration. We test the
quality of our implementation by comparing it to standard published metrics and by surveying users about how
relevant the recommendations are and how easy the system is to use. If users feel that the recommendations are
relevant, then we will know that we can trust the cross-type recommendations generated through MultiLens. With
ease of use, we cannot only judge our system but also further improve it by finding out how they expected it to work
or how they would like it to work. We hope that our research will show that a large-scale distributed personal
recommendation system based on PocketLens is viable.
CORE EXTRACTION SOFTWARE SUBROUTINE
Tyler Huntley (Vito Gervasi), Rapid Prototyping Center, Milwaukee School of Engineering, 1025 North Broadway,
Milwaukee, WI 53202
This research was aimed at preparing a portion of a larger software package, which employs the “Shell-Slice”
technique, reducing complex three-dimensional objects into manufacturable elements. Currently, there are a number
of software packages available that are designed to slice three-dimensional objects in preparation for building parts
via solid freeform fabrication (SFF). There are various methods that are utilized, most exercising a layer-based raster
procedure. Using SFF, each of the cross-sectional layers is transformed into the larger object by curing, sintering, or
bonding. Current, layer-based methods lead to stair stepping defects. Thinner layers reduce stair stepping but lead
to undesirable lengthy build times. This research is attempted in order to create complex lattice structures more
efficiently and with reduced error. The main focus of this core extraction software is to identify and extract the
internal geometry from a computer aided design (CAD) object through the use of C++. CAD files are in an .stl format,
and the C++ program manipulates the CAD in order to extract the core(s) of CAD objects. This work presents 38
cases of possible extruded triangle to triangle interactions. The methodology employed here lays out the entire
process of being able to determine whether or not a line segment intersects a given triangle which is the critical
function of the extraction software.
Bethel College
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COMPUTER SCIENCE
CREATING A DECISION SUPPORT SYSTEM FOR PHENOLOGY ANALYSIS
Jose Martinez (Dr. Sherri Harms), Department of Computer Science & Information Systems, University of Nebraska at
Kearney, Kearney, Nebraska 68849
Phenology studies the seasonal timing of different developmental stages and the life cycles of plants and animals.
Understanding how biological phenomena such as rainfall and temperature impact the developmental stages for
crops are important to growers, food processors, crop insurance and seed companies, and the commodities markets.
This research consists of the development of a decision support system that allows for the analysis of
environmental impacts on important crop plants. This process will require computer science research to enable: 1)
the design and development of web-based tools to be able to display graphics and data in unique combinations; 2)
pre-processing and integrating existing data from several disjoint data sets; and 3) the creation of an web-based
interface for user accessibility. The long-term objectives of this project are to: 1) accept data sets from users, thus
enriching the quality and diversity of the data available within the infrastructure; 2) allow users to analyze the new
data sets; 3) make data sets readily available to users; and 4) develop analytical tools that will allow for the study of
complex environmental impacts such as global warming on the phenology of plants and animals.
NS-2 IMPLEMENTATION OF A SAMPLE NETWORK MODEL
Laidlaw Michael, Kyle Loomis (Dr. Chunyan Bai), School of Engineering, Computing and Construction Management,
Roger Williams University, One Old Ferry Road, Bristol, RI 02809
This project aims at enhancing our ability of Linux administration and familiarizing us with the usage of NS-2 – REAL
Network Simulator. We will design and implement a sample network model which will be used for teaching
communication and network security courses in the future. Also student oriented research in network security area will
benefit from the built network model.
NS-2 is a discrete event simulator targeted at networking research and teaching. It provides substantial support for
simulation of TCP, routing, and multicast protocols over wired and wireless (local and satellite) networks. In this
project, we will start with verifying the availability of NS-2 package in Linux system. Linux was chosen as the platform
for this project because of its growing success and open development environment. We will then act as a Linux
administrator and install the newest version NS-2.28 to the Linux lab server, which makes NS-2 simulator available to
all the Linux boxes in the lab. Validating the installation will be done by running some small bundled Tcl/Tk programs.
We need to learn the Tcl/Tk language in advance to read, execute and verify those programs. Finally, we will be
given a blueprint of a sample network model with specific parameter requirements such as network size (number of
nodes in the network), communication protocols (TCP/IP), connection mode (full-duplex or half-duplex), transmission
rate (bps or mbps), and overflow handling techniques for each node (drop or save). Following the specification of the
blueprint, we will write Tcl/Tk program to implement the model, test the implementation by running the program in
NS-2 with pre-defined data and verify the implementation by collecting data from the nam-1 file and browsing the
network layout and traffic flow from the XGraph.
OPERATING SYSTEMS SECURITY SURVEY AND SIMULATION:DOS,WINDOWS,UNIX/LINUX
Chris Koukol, James Kendera, Machael Laidlaw and Scott Kennedy (Dr. Chunyan Bai), School of Engineering,
Computing and Construction Management, Roger Williams University, One Old Ferry Rd, Bristol, RI 02809
Recognizing the critical role of the security mechanisms in supporting a reliable and trustable operating system
design, we aim at doing a case study on security threats for modern operating systems with emphasis on recently
happened attacks in this project.
Fundamental background knowledge such as security threats, protection mechanisms, type of malicious software and
intruder techniques will be introduced to us first. Then each of us will be given a modern operating system (MS-DOS,
Windows, Unix/Linux) to do the case study. The primary goal is to find out all recent attacks happened to each
operating system following the guidance of the faculty advisor. Causes of the attacks will be analyzed and tentative
mechanisms to handle those attacks will be proposed. Further more, each of us should try to propose our own view of
possible vulnerabilities of the system and present corresponding solution to prevent the vulnerability from being
discovered or utilized by intruders. Denial of Service attacks will be investigated with details. Simulation will be
utilized for constructing a sample network, repeating the attacks and im