tamhsc summer research program

Transcription

2014
Summer Research Program
Poster Session and Reception
August 5, 2014
9:00 AM - 4:00 PM
Health Professions Education Building
Bryan, TX
College of Medicine
SUMMER RESEARCH PROGRAM
Schedule of Events
August 05, 2014
8:00am
Registration Table Opens
HPEB LL Lobby
9:00am-4:00pm
Poster Viewing
HPEB LL43 A&B
11:00am
Luncheon
HPEB LL11 B
12:00-1:00pm
CST*R Grand Rounds – SRP Featured Speaker
Timothy D. Phillips, MS, PhD, ATS, Senior Faculty
Fellow (Texas AgriLife)
College of Veterinary Medicine & Biomedical Sciences
Texas A&M University
HPEB LL11 A
1:30-3:00pm
Poster Presentations – Students Presenting
HPEB LL43 A&B
3:00-4:00pm
Closing Remarks and Presentation of Certificates
Dr. Warren Zimmer, Summer Research Program Director
HPEB LL38
Presentation of Dean’s Recognition Awards
HPEB LL38
4:00pm
Adjourn
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Speaker’s Biography
Timothy D. Phillips, MS, PhD, ATS, Faculty Fellow
Distinguished Professor
Reed Endowed Chair in Toxicology
Veterinary Integrated Biosciences
College of Veterinary Medicine, Texas A&M University
Dr. Timothy (Tim) D. Phillips is a professor of veterinary integrative biosciences in the College of Veterinary
Medicine and Biomedical Sciences and a Texas AgriLife Senior Faculty Fellow, who holds the Chester Reed
Endowed Chair in Toxicology. He is a Professor of the Faculty of Toxicology and the Faculty of Food Science,
Texas A&M University and holds a joint appointment with the Texas A&M Health Science Center School of Public
Health. Dr. Phillips earned a B.S. from Mississippi State, and M.S. and Ph.D. from the University of Southern
Mississippi. He joined the faculty of Texas A&M University in 1979 and over his 34 year career with A&M has
been recognized with many honors including Senior Faculty Fellows Distinction (Texas AgriLife), a 2007
Innovation Award for Research, the 2006 Association of Former Students' Faculty Distinguished Achievement
Award for Research and a 2005 Bush Award for Excellence in International Research. He has also been
recognized nationally and internationally for his research in food safety and toxicology most recently being
awarded the 2014 Society of Toxicology Translational Impact Award, which distinguishes scientists whose
research leads to improving human health.
Dr. Phillip’s research is focused on developing strategies to alleviate risk factors of disease found in the
environment. His recent research centers on aflatoxins, which are naturally occurring mycotoxins and are
among the most toxic substances known. A major success for his research is the development of a novel,
inexpensive method to resolve toxicity in food staples thus preventing aflatoxin diseases of the liver,
compromised immunity and malnutrition. These studies helped form the basis of Dr. Phillip’s talk entitled:
“Ancient Medicine for the Mitigation of Aflatoxicosis in Animals and Humans”.
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Acknowledgements
This was a banner year for the COM Summer Research Program as we had the largest student population
(51) in my 7 years as director. The increased numbers is due, in part, to our taking and mentoring
volunteer medical students in addition to the 20 stipend positions we normally provide. In addition we
have maintained program partners that sponsored undergraduate students for the Temple Health and
Bioscience District and the Prairie View Undergraduate Medical Academy and we have new affiliations with
UT Pan American as partners through an NIH RISE grant. The strength and quality of our program is also
reflected in the record number of applications (>125) we received this year for the summer positions. I am
most thankful for the faculty that give graciously of their time to help the program through serving on the
committee that reads and evaluates each application to find the best students; presenting stimulating and
informative lectures; and allowing students to work in their labs. These latter interactions have also
provided the strong mentoring that has allowed the program to operate at the highest levels. It is the COM
faculty that should get the loudest “round of applause” for their continued strong commitment and
involvement in the program; the program would not be able to sustain its quality and existence without
them.
Even though we serve two distinct student populations, medical students between their first and second
years and undergraduates, all participate equally in the program. This enriches the experience as each
group can learn from the other and share their ups and downs of laboratory experience. Each has worked
extremely hard this summer and the posters displayed at today’s reception are the products of this hard
work. Please take the time to visit the posters and ask the students what they did during their summer
vacation; be prepared to be amazed by their work and their abilities!
We have obtained funding from a number of sources and are especially thankful for sustained funding from the
COM Deans office, Baylor/Scott and White Research, Temple Health and Bioscience District Board, and the HSC
VRP Office. I think we will continue to grow our funding as we begin to reap benefits of submitted grant
proposals in addition to cultivating strong partners such as the UT Pan American RISE and Texas A&M System
BUILD grants. In addition, we could not have had a successful year without the generous support of many
department chairs, both clinical and basic science. It is difficult, but not impossible, to provide content
simultaneously to three locations. The work of Drs. Mitchell (Temple) and Huston (Houston) as site
coordinators keeping things running efficiently is greatly appreciated. Finally, I would like to thank Dr. Van
Wilson and his staff in College Station, Mary Ann Wolff, Peggy Brigman; Dr. Huston’s staff in Houston, Anna
Wirt, Tammy Kocurek; and the Dean’s staff in Temple, Cari Cummings, Loria Lynce for making certain that
the entire program got off the ground and kept running effortlessly.
Warren Zimmer, PhD
Director, SRP
3|Page
Abbas, Darren, Xin Wu, Bryan Clossen, and D. Samba Reddy*
LC-MS/MS Analysis of Brain Neurosteroid Levels in a Mouse Epilepsy Model
Page 9
Babalola, Ebunoluwa, Cody Dornhecker and Johanna Villaseñor*, Teminioluwa Ajayi, Jennifer Ross, Dr. PH,
Robin Fuchs-Young, Ph.D.
Page 10
Mission BREATHE: Better Recognition of Exacerbating Asthma Triggers in the Home and Environment
Bosquez, Kimberly, Marcella D. Cervantes, Geoffrey Kapler
The Phenotype of Reducing RAD51 mRNA in Tetrahymena
Page 11
Bowman, Jason, MS, EMT-P1,6; Matthew McClure MS, EMT-P2; David E. Dostal PhD3,4,5
Page 12
Sodium Nitroprusside, Adenosine and Levosimendan Improve Cell Survival Over Epinephrine and
Amiodarone in a Rat Model of Cardiac Arrest
Capetillo, Mario, Andrés Santos, Robin Fuchs-Young
Page 13
Assessing the DNA Damage Response in p53 Polymorphic Variants Present in Different Ethnic Groups
Chen, Chris, Pablo Ormachea, J.D. , Gabe Haarsma, Ph.D., Sasha Davenport, David M. Eagleman, Ph.D. Page14
Exploring the Relationship Between Ambient Temperature and Aggression or Impulsivity as Measured by
Criminal Behaviore
Chi, Alvin, Duren M. Ready
An Exploratory Study of Medications Prescribed to Individuals Diagnosed with Headache
Page 15
Cochran, David, Darren, and Emily Wilson
Page 16
The Role of Alpha Smooth Muscle Actin in Regulation Of Nuclear Translocation of Myocardin and
Serum Response Factor
Coffman, Jason, Jason Coffman, Sam-Moon Kim, Nichole Neuendorff, Amutha Selvamani, Farida Sohrabji, and
David Earnest
Page 17
Impact of Shift Work Schedules and Circadian Clock Disruption on Estrus Cycles and Neuroprotection in
Response to Stroke in Adult Female Rats
Colman, Ellen, D. and D. Samba Reddy*
A Comparative Analysis of Human Organophosphate Poisonings Using Social Media
Page 18
Daniels, Michlene, Joseph Masdeu, MD, PhD, Belen Pascual, PhD
Neuropsychological Evaluation in Spastic Paraparesis with SPG 11 Mutations
Page 19
Dawson1, Ashley, Sanjib Mukherjee MD, PhD1, Sören Singel MD2, Karen Newell-Rogers PhD1,2
Aquaporin-4 and ICAM-1 Expression after Traumatic Brain Injury
Page 20
4|Page
Desai, Kurren, Bharathi Hattiangady, Geetha Shetty, Ashok K. Shetty
Page 21
Gulf War Illness Related Chemicals Adversely Affect the Proliferation of Human Neural Stem Cells
Dhanani, Naila, Joseph Fernandez-Moure, M.D., M.S.
Evaluating Efficacy of Nitric Oxide Releasing Nanoparticles
Page 22
Hamilton1, Alina, Vinod Srivastava2, Jill Hiney2, Robert K. Dearth1, W. Les Dees2
Page 23
Manganese Increases Proliferation within the Prepubertal Mammary Gland through Estrogen-Mediated
Pathways
Hayworth, Carla, Rachel Petrofes Chapa, Warren Zimmer, Ph.D.
Identification of the E2/E3 Ligase Activity Within the Spll Spectrin Progein
Page 24
Herrera de Lechuga1, Camille Duran1, L. Gerard Toussaint2, Raquel Sitcheran1
Differential miRNA Expression in Highly Invasive Glioblastoma Cell Lines
Page 25
Howell, John, B Sanchez, A Coletta, E Galvan, P Jung, R Dalton, K Levers, M Koozehchian, S Simbo, A
O’Connor, A Reyes, S Springer, C Goodenough, M Cho, C Rasmussen, RB Kreider
Page 26
Influence of ADRB2-79 and ADRB3 Metabolic Gene Single Nucleotide Polymorphisms on Body
Composition and Fitness
Huynh, Victoria, Laura Hargrove, Sharon DeMorrow, Allyson Graf, Lindsey Kennedy, Quy Nguyen,Brittany
Ladd, Maryah Baker, Christopher Johnson and Heather Francis
Page 27
Inhibitionof Histamine Synthesis Decreases Cholangiocracinoma Progression, EMT and Metastatic
Potential
Kearney, Kate, Xu Peng, Shenyuan L. Zhang
Page 28
Characterization of the Molecular Identity and Functional Roles of Store-Operated Calcium Channels in
Human Lymphatic Endothelial Cells
Lane, Mar’Kiffany, Yifan Zhang, Danielle M. McGrath, Magnus Hook, and Dekai Zhang
Modeling TLR Agonist Protective Effects in Vitro
Page 29
Luna, Sarah, Aris J. Maguddayao, Ryan T. Wade, and William C. Culp Jr., M.D.
Page 30
Assessing the Flammability Characteristics of Alcohol Skin Prep Solution to Reduce Operating Room Fire
Risk
Maguddayao, Aris, William C. Culp Jr., MD; Bradly A. Kimbrough; Sarah Luna
Prevention Operating Room Fires: Development of a Carbon Dioxide Suppression Device
Page 31
Mahmood1,2, Tasneem, Mohaiza Dashwood2, Praveen Rajendran2, and Roderick H. Dashwood2
Page 32
Does Tannase from Intestinal Bacteria Modify the Anticancer Effects of Tea Catechin in Colon Cancer
Cells?
5|Page
Maloney, Taylor, Ronak Tilvawala, Preeti Sule, Suat L.G. Cirillo, Jeffrey D. Cirillo
Quorum Sensing Molecule Identification in Mycobacterium Tuberculosis
Page 33
McGhie, Brandon, Georgina Kolcun, Warren Zimmer
Smooth Muscle Gamma Actin Expression in Prostate Cancer Cells
Page 34
Megahed, Tarick, B. Hattiangady, B. Shuai, and A. K. Shetty
Page 35
Differential Vulnerability of Hippocampal GABA-ergic Interneuron Subpopulations in an Animal Model of
Gulf War Illness
Milad, Moheb, Valorie Chiasson, Piyali Chatterjee, Olga Gasheva,Kelsey Foster, Marcos Hernandez, Brett
Mitchell
Page 36
Depletion of CLIP Decreases Toll-Like Receptor-Induced Preeclampsia in Mice
Mohiuiddin, Maha, D. Oweis, G. Tavana, P. Moore, DP Huston
Human Basophil Degranulation Mediated by the Anaphylatoxin C5a is IL3-dependent
Page 37
Munir, Yunib, Sanjib Mukherjee; Sören Singel, M.D.; M. Karen Newell-Rogers, Ph.D.
3-Bromopyruvate Stresses Mouse GBM Cells to Express MHC Class-II in vitro
Page 38
Naumann1, Heather, Patrick McCulloch, MD2 and Patrick Massey, MD2
Sensitivity and Specificity of a New Clinical Test for Anterior Cruciate Ligament Tearse
Page 39
Nguyen, Quoc-Bao, Jonathan A. Friedman, MD
Page 40
Evaluation of Two Neurosurgical Approaches to Cervical Spondylotic Myelopathy: Posterior and
Combined Posterior-Anterior
Niakan, Lillian, Ricky Savjani, David Eagleman, Ph.D.
Pupillary Reactivity to Cocaine Cues in Cocaine-Dependent Patients
Page 41
Ochoa, Brennan, , Hao Feng, and David E. Dostal
H2O2 Induces HEK Cell Apoptosis By Disrupting the Interaction Between p38 and MKP-1.
Page 42
Ogden, John, Qian Chen Yong, Kenneth Baker, Rajesh Kumar
Regulation of the Renin-Angiotensin System in the Diabetic Heart
Page 43
Oweis, Divina, M. Mohiuddin, G. Tavana, P. Moore, D.P. Huston
Human Basophil Degranulation byBacterial-Derived N-Formylated Peptides is IL-3 Dependent
Page 44
Phillips, Deanne, Candice M. Thomas, Amanda L. Roth, Kenneth M. Baker, Rajesh Kumar
The Role of the Intracellular Renin Angiotensin System in Diabetic Cardiomyopathy
Page 45
Robinsona, Lance, Megha Bijalwanb, Andrew J. Steelmanb, Luke Baconb, Caleb Gottlichb, Colin R. Youngb, Richard
P. Tobind, M. Karen Newell-Rogersd, C. Jane R. Welshb,c
Page 46
Administration of CAP Peptide Ameliorates Theiler’s Virus-Induced Seizures
6|Page
Shin1, Hope, Matthew McMillin1, Cheryl Galindo1, Gabriel Frampton1, Sharon DeMorrow1,2,3
Page 47
Specific Serotonin Reuptake Inhibitors Promote Cholangiocarcinoma Growth and Serotonin Production
Sohner, Mark, , A. Martínez, H. Francis, J. Venter, H. Standeford, M. Guerrier, J. Greene, G. Alpini, and S.
Glaser
Page 48
Complete Inhibition of the SEC/SR Axis Triggers Increased Biliary Damage
Stone, Ryan M., Sanjib Mukherjee, Richard Tobin, Susannah Rogers, Stephanie Henderson, Heather Motal,
Jessica Kain, Karan Newell-Rogers, Lee Shapiro
Page 49
The Role of the Vagus Nerve in the Peripheral Immune Response Following TBI
Thomas, Rose, Isabel Lambertz & Robin Fuchs-Young
Page 50
Analyzing a Racially Disparate Polymorphism and its Role in Early Onset Breast Cancer in AfricanAmerican Women
Tran1,2, Dickson, Ana Cohen1, Vannakambadi K. Ganesh1, Magnus Höök1
Bioinformatic Analysis of MSCRAMMs in S. Aureus
Tsai, Yuwei, Gyhye Yoo, and Clinton D. Allred
Estradiol Suppresses Cell Growth of Non-Malignant Colonocytes Induced by NF-κB signaling
Page 51
Page 52
Wiewiorowski, Elizabeth, Seyed Hossein Mousavi-Fard, and Steve Maxwell
Page 53
Upregulation of Superoxide Dismutase in Ovarian Carcinoma Cells Increases Their Resistance to
Doxorubicin
Young, Corinne, Jun-Yuan Ji
Obesity Increases Cancer Risk by Dysregulation of CDK8-Cyclin C
Page 54
Younus, Iyan, Jenessa Short, Victoria Dunlap, R. Kuruba, Xin Wu, and D. Samba Reddy*
Page 55
Quantification of Neuroinflammation by Area Fractionation Method in Epilepsy Brain Injury Models
Zaayman, Marcus, Ramkumar Kuruba, Ellen Colman and D. Samba Reddy*
The Time-Course of Acetylcholinesterase Inhibition and Recovery Following Organophosphate
Intoxication in Rats
Page 56
Zingg, Leonardo, A. Selvamani, S. Bake, M.J. Park, and F. Sohrabji
Developing a Model to Study MicroRNA-20a Function in Post-Stroke Astrocytes
Page 57
7|Page
Participants
Babu1Christina A., Thomas A. Jeffreys2, and Ashton J. Richardson3
Page 58
Texas A&M One Health Initiative: Investigating the Health Disparities of Humans and Animals on Isla de
Ometepe, Nicaragua
Bounds, Rachel, Daria Muller, Michael Golding
The Effects of the Organochlorine Methoxychlor on the Transcriptional Activation of Sox2
Page 59
Cobb1,,W. Jacob and Sarah Genzer2
Page 60
Texas A&M University One Health Initiative: FDA Center for Veterinary Medicine Internship on
Antimicrobial Resistance
Hammond, Jacob B. B. Gadad, W.Li, U. Yazdani, T. Johnson, L. Hewitson, D.C. German
Effect of Pediatric Vaccines on the Hippocampus: Relevance to Autism
Page 61
Nguyen, Jeannie M., Angela M. Harrington, Louise C. Abbott
Effect of Zinc Exposure Before and During Methylmercury Toxicity in Zebrafish Embryos
Page 62
Reddy, Sandesh, Iyan Younus, and D. Samba Reddy*
Neurosteroid-Independent Synaptic Antiseizure Activity of Midazolam
Page 63
Short, Jenessa, Jonathan Brewer, Victoria Dunlap, Megan Swonke, Iyan Younus, Divya Raju, Ramkumar Kuruba,
Xin Wu and D. Samba Reddy*
Page 64
An Optimized Brain Stereology Protocol for Neuron Counting
Shutze, Sr., William P., Katherine Kane, Taylor Hicks, John Kedora, Steve Hohmann, Toby Dunn, Brad Grimsley,
Dennis Gable, Greg Pearl, Bertram Smith, Richard Lueking, Elizabeth Nguyen, Grace Lassiter
Page 65
The Incidence and Outcome of DVT after Endovenous Laser Ablation
Thomas, Clarence, Jadye Kee, M.S.1, Quoc-Bao D. Nguyen1, Jason M. Hoover, M.D.2, Daniel J. Lamont, PA-C2
Page 66
Intraoperative Ultrasound and Isolated Lumbar Schwannomas
Walker, Kindra M., Syeda H. Afroze, Damir Nizamutdinov, Micheleine Guerrier, Allyson Martinez, David E. Dostal,
Shannon Glaser
Page 67
Effect of Secretin in Cardiac Contractile Regulation
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TAMHSC SUMMER RESEARCH PROGRAM
August 5, 2014: 9:00 AM - 2:00 PM
Bryan, Texas
LC-MS/MS Analysis of Brain Neurosteroid Levels in a Mouse Epilepsy Model
Darren Abbas, Xin Wu, Bryan Clossen, and D. Samba Reddy*
Department of Neuroscience and Experimental Therapeutics
Texas A&M University Health Science Center College of Medicine, Bryan, TX 77807
Introduction: Catamenial epilepsy is caused by fluctuations in endogenous neurosteroids, which modulate
GABAergic phasic and tonic inhibition and play a key role in the pathophysiology of epilepsy. The precise
molecular pathophysiology of this gender-specific seizure condition remains poorly understood. Recently, we
developed a mouse model of catamenial epilepsy based on the clinical neuroendocrine milieu. The model is
based on the premise that seizure susceptibility decreases when neurosteroid levels are high (luteal phase)
and increases during their withdrawal (perimenstrual periods) in association with GABA-A receptor subunit
plasticity and function. The objective of this study was to determine the gonadotropin-induced elevations and
withdrawal in neurosteroid levels in the brain and plasma using highly specific liquid chromatography/tandem
mass spectrometry (LC-MS/MS) assay.
Hypothesis & Objectives: It is hypothesized that seizure susceptibility decreases when neurosteroid levels
are high in the hippocampus and increases during their withdrawal such as that occurs around perimenstrual
periods in close correlation with their circulating levels. Here, we seek to test this premise by determining the
extent of correlation between the neurosteroid allopregnanolone levels in the brain and in the plasma to the
seizure activity in a mouse epilepsy model.
Methods: Female WT and DKO (-subunit knockout) mice were utilized for this study. Catamenial-like
neurosteroid milieu was created by gonadotropin protocol. Elevated neurosteroid levels were induced by
sequential gonadotropin treatment, and withdrawal was induced by the 5a-reductase and neurosteroid
synthesis inhibitor finasteride. To mimic the catamenial-like condition, mice were treated with PMSG (day -2)
followed by HCG 44 hours later (day 0). On day 9, mice were treated with finasteride. Plasma samples and
brain tissues were collected on each day of the treatment timeline (-2 through 10). Plasma and brain levels of
allopregnanolone were estimated by a quantitative LC/MS-MS method using 3-AP as internal standard. The
steroid and IS were extracted with 0.9 ml of hexane. Each sample was analyzed by using the APCI technique
under acidic conditions in a triple Quad system (Applied Biosystems) under the multiple reaction monitoring
(MRM) detection modes.
Results: The mass conditions used for the analytes AP and IS were m/z 301.2→283.1 and 315.3→297.3,
respectively, with collision induced fragmentation. A standard plot was created using 50 and 1000 nM AP
levels. Standard AP added to mouse plasma has been successfully analyzed with excellent linearity,
specificity, and reproducibility. The sensitivity of the method was < 20 ng/ml with a detection limit of 10 ng/ml
and a linear range of 10–10,000 ng/ml. The method was used for the analysis of gonadotropin-induced
increase in plasma and brains AP levels in mice. The Gn protocol produced a steady increase and sustained
elevation in brain AP from day 1 to day 9, which was extensively reduced by treatment with finasteride,
indicating the withdrawal phenomenon that occurs during perimenstrual period.
Conclusions: The gonadotropin regimen produced a state of sustained elevations in neurosteroids in the
hippocampus and cortex in close parallel relationship with plasma levels. Thus, this analytical study confirms
that this LC–MS/MS method allows accurate, high-throughput analysis of neurosteroids in plasma and brain
tissue samples.
**Supported by NIH Grant R01 NS051398 & WHIN project**
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PROGRAM TAMHSC SUMMER RESEARCH
August 5, 2014: 9:00 AM - 2:00 PM
College Station, Texas
Mission BREATHE: Better Recognition of Exacerbating Asthma Triggers in the Home and
Environment
Ebunoluwa Babalola, Cody Dornhecker and Johanna Villaseñor*, Teminioluwa Ajayi, Jennifer Ross, Dr. PH,
Robin Fuchs-Young, Ph.D. (*these two authors contributed equally to this work)
Department of Molecular and Cellular Medicine
Texas A & M University Health Science Center College of Medicine, College Station, Texas
Introduction: McAllen, the largest city in Hidalgo County, is located in the southern tip of Texas in the Rio
Grande Valley, where some of the poorest cities in the state are located. According to the McAllen asthma
coalition, individuals who reside in border cities are especially vulnerable to asthma due to the poor air
quality caused by pollutants, and the use of pesticides.9 McAllen continues to be a vulnerable population, as
childhood asthma rates are highest among minorities, families with low educational levels, and those who
reside in low-income communities.3 Previous studies have demonstrated the effectiveness of asthma
education in improving asthma knowledge and management in parents of children with asthma. The purpose
of this study is to assess the effect of a 30-minute asthma informational presentation in educating parents
about their child’s asthma and the identification and removal of asthma triggers in the household. We will
also examine the effect of the educational session on the level of control parents feel they have over their
child’s asthma in order to help improve the quality of life for their child with asthma.
Hypothesis: We hypothesized that a short 30-minute asthma education session geared towards parents will be
more appealing to participate in, improve parental knowledge of asthma and its management, and help
improve the quality of life for a child with asthma.
Methods: A community based participatory research approach was used in this study. Participant recruitment
was conducted in partnership with local physicians that referred patients with moderate to severe asthma to us
as possible candidates for our Asthma Educational Program. After obtaining consent and meeting all
inclusion/exclusion criteria, participants completed an asthma knowledge assessment pre-test, and an asthma
control survey. A 30-minute asthma education was then provided, followed by a post-test, and an evaluation
of the session. Program efficacy was evaluated at two to four weeks and included a control survey measuring
parental control and management of their child’s asthma as well as quality of life. We worked with a
Promotora, a certified community healthcare worker, and the Rio Grande Regional Hospital in order to
address any issues related to cultural differences and language barriers, and to ensure the availability of
language and culturally appropriate translation of all materials used.
Results: Twelve participants have successfully enrolled in the study and completed the educational portion of
the intervention, and seven participants completed the follow-up control survey. Participants demonstrated
improved knowledge of asthma and its management immediately following the education session. Improved
parental control of their child’s asthma was also seen two to four weeks later. In addition, 92% of the
participants reported that some of the information being presented was new to them, and agreed that the
presented information was useful. The recruitment and intervention process are still underway, and data
collection and analysis will continue until the 30 participants sample size has been reached.
10 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
The phenotype of reducing RAD51 mRNA in Tetrahymena
Kimberly Bosquez, Marcella D. Cervantes, Geoffrey Kapler
Molecular and Cellular Medicine Department
Texas A & M University Health Science Center College of Medicine- College Station
Tetrahymena thermophila is a single-celled eukaryote with two nuclei. The germline nucleus
contributes genetic information to offspring. The somatic nucleus is the source of all gene
expression. The Rad51 protein is a key component of homologous recombination and DNA
repair machinery, and plays a crucial part in sexual and asexual reproduction in Tetrahymena.
Deletion of Rad51 can lead to a loss of viability. This study will verify transformation of cells
with an inducible Rad51 hairpin construct that will be used to rapidly deplete cells of the Rad51
mRNA and protein, to observe the phenotype of Tetrahymena progeny with reduced levels of
RAD51 under specific experimental conditions. DNA was prepared from single cell isolates of
presumed hairpin construct transformants. From polymerase chain reactions we were able to
verify the mating type of germline transformants and the insertion of the Rad51 hairpin within
the genome. To further my investigation, a pair of single cell isolates must be found that are of
different mating types. Once identified, the transformants must be starved and mated. During
mating, the phenotype of the Rad51 hairpin knockdown in late development will be determined
by checking for normal endoreplication of the DNA and genomic rearrangement events. This
will confirm an arrest in late development due to reduction of Rad51p and reveal whether
Rad51 is required for programmed DNA rearrangement and/or DNA replication.
11 | P a g e
Sodium Nitroprusside, Adenosine and Levosimendan Improve Cell Survival
Over Epinephrine and Amiodarone in a Rat Model of Cardiac Arrest
1,6
2
Jason Bowman MS, EMT-P ; Matthew McClure MS, EMT-P ; David E. Dostal PhD
3,4,5
1
Texas A&M Health Science Center College of Medicine, Temple Campus; 2UNT Health Science Center Texas College of Osteopathic
Medicine; 3Central Texas Veterans Health Care System; 4Division of Molecular Cardiology, Cardiovascular Research Institute, The Texas
A&M Health Science Center; 5Division of Cardiology, Scott and White Memorial Hospital; 6Emergency Medical Services, Scott and White
Memorial Hospital;
Introduction: During the restoration of circulation, the standard of care resuscitation drug combination
epinephrine and amiodarone, used in advanced cardiac life support, may induce ischemic cell death. Two
recent proposals; a novel drug combination with a unique hemodynamic profile consisting of Sodium
Nitroprusside, Adenosine and Levosimendan (SNPAL); or controlled interruptions at the start of resuscitation
called “ischemic post-conditioning” may prove advantageous in preventing ischemic cell death. We assess
the impact these interventions have on the survivability of cardiomyocytes exposed to an ischemic challenge
and resuscitation.
Hypothesis: Sodium Nitroprusside, Adenosine, and Levosimendan given during resuscitation from cardiac
arrest will improve cell survival over ACLS and will induce the effects of ischemic post-conditioning.
Methods: We developed a novel study design consisting of langendorff perfusion of 12 Sprague-Dawley rats
weighing 150 grams assigned to one of three groups: 0.005 μΜ Epinephrine and 5 μΜ Amiodarone (ACLS);
ACLS + Ischemic Post Conditioning which received ACLS drugs as well as 4 cycles of 30 seconds pauses
during resuscitation; and SNPAL which received 10 μΜ sodium nitroprusside, 3.75 μΜ adenosine and 0.8 μΜ
levosimendan. Three samples were rejected due to time-to-cannulation exceeding 10 minutes or leakage in
or around the aorta leaving each group with an n=3. Rats were euthanized using isoflurane, hearts placed in
cold Krebs-Henseleit buffer then quickly cannulated and placed on a langendorff perfusion apparatus and
perfused with a modified, carbogenated K-H buffer for 15 minutes until they regained normal function. The
perfusion was then switched off for 15 minutes then switched back on and drugs administered. After 15
minutes of re-perfusion, buffer was switched to Ca++ free digestion buffer for 3 minutes and then 0.10% w/v
Type II Collagenase. Cells were isolated and stained with Annexin V, Propidium Iodide and for
phosphorylation of proteins. Results were collected by flow cytomet
er as % of total events above the first decade and analyzed by two-way ANOVA and chi-square statistics.
Results: Compared to the standard of care ACLS group, the SNPAL treatment group reduced necrosis by
29% ± 0.3% (p<.001) and apoptosis by 71% ± 0.1% (p<.001). Ischemic post conditioning reduced necrosis by
4% ± 0.2% (p=.05) and apoptosis by 3% ± 0.1% (p=.01) with ACLS drugs. SNPAL induced a five-fold
increase of Akt phosphorylation at T308 (p<.001) and a two-fold increase at S473 (p=.003), a two-fold
increase in phosphorylation of STAT3 (p=.05) and GSK-3β (p=.18), a 50% reduction in ERK1/2 (p<.001)
phosphorylation and a 25% reduction of P70S6k (p=.22) compared to ACLS. Ischemic preconditioning
induced a 13 fold increase in Akt phosphorylation at T308 (p<.001) and a 4.5 fold increase at S473 (p<.001).
IPC reduced P70S6k (p=.15) and ERK1/2 (p=.02) phosphorylation by 50%. Atrial heart-rates were similar in
all groups however both ACLS groups suffered from severe ventricular dysfunction upon resuscitation, the
group receiving SNPAL did not.
Conclusion: SNPAL significantly reduced necrosis as well as apoptosis compared to the standard of care.
Ischemic post-conditioning reduced necrosis and apoptosis but barely reached statistical significance. SNPAL
appears to trigger the same survival pathways as ischemic post-conditioning as well as additional survival
pathways. SNPAL not only appears to improve cell survival from cardiac arrest but mechanical dysfunction as
well. These results warrant further investigation into the cellular effects of ACLS drugs and suggest this drug
combination as a new treatment for cardiac arrest.
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August 5, 2014: 9:00 AM - 2:00 PM
Assessing the DNA damage response in p53 polymorphic variants present in different ethnic groups
Mario Capetillo, Andrés Santos, Robin Fuchs-Young
Department of Cellular and Molecular Medicine
Texas A & M University Health Science Center College of Medicine, College Station
Introduction: Breast cancer is a prevalent disease that affects many individuals throughout the world, but it is
predominantly present in females. The cause of this type of cancer can be attributed to things such as
environmental factors, genetic predisposition, or a combination of these. This study focuses on characterizing
the involvement of genetic factors, such as the hp53 gene, that contribute to the development of breast
cancer and seem to play an important role in pregnancy protection. This type of protection takes place only
after a woman carries a pregnancy to full-term early in her life. The full maturation of breast tissue due to
pregnancy has been proven to lower the occurrence of breast cancer in women of European decent.
However, African-American women experience a much lower level of pregnancy protection than Caucasians.
Hypothesis: We hypothesize that the molecular basis for this racial disparity is due to racially
disproportionate p53 variants with different apoptotic and DNA repair capabilities. In particular a nonsynonymous SNP that results in either a proline (P) or arginine (R) at position 72 of the p53 protein may be a
key player involved in orchestrating the molecular events involved in pregnancy protection.
Methods: The cells used in these experiments were MDA-MB-157. The cells were transfected with plasmids
either coding for the human p53 polymorph containing a Proline residue at position 72 (hp53 (72P)), or an
Arginine residue at position 72 (hp53 (72R)), or an empty vector as a negative control. 24 hours posttransfection the cells were treated with either a 250nM concentration of Doxorubicin or 6 Gy of ionizing
radiation every 3 hours for a total of 12 hours. After treatment the cell samples were collected and prepped
for a Western Blot. The antibodies used were Anti-p53 (p-S15), Anti-p53, and Anti-GAPDH.
Results: The various immunoblots indicated that the human p53 polymorphs (72R and 72P), have the similar
activation kinetics after a DNA damage response induced by Doxorubicin or ionizing radiation.
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August 5, 2014: 9:00 AM - 2:00 PM
Exploring the relationship between ambient temperature and aggression or impulsivity as measured
by criminal behavior
Christopher Chen, Pablo Ormachea, J.D., Gabe Haarsma, Ph.D., Sasha Davenport,
David M. Eagleman, Ph.D.
Department of Neuroscience
Baylor College of Medicine, Houston, TX
Introduction: The intuitive relationship between high ambient temperatures and increased criminal activity has been well supported
through a variety of methodologies (longitudinally, geographically, seasonally, daily, etc.). Yet, biological – neuroanatomical explanations
are particularly lacking. This study hopes to first re-affirm these previous assertions by using large, compiled data sets to statistically
show that a relationship between crimes of aggression or impulsivity and ambient temperature exists. Second, the specific types of crime
most affected by temperature will be determined. Finally, relationships uncovered by computational analysis will be explained from a
neuroanatomical stance by conducting a literature review.
Hypothesis: Crimes of aggression are hypothesized to vary with ambient temperature, while crimes of impulsivity are not expected to do
the same. Crimes of aggression should include assault (sexual and non-sexual), child sex crimes, murder, harassment/stalking, and
kidnapping. Non-sexual assault is predicted to be most affected by higher temperatures. Crimes of impulsivity should include theft, illicit
drug use (marijuana and non-marijuana), and alcohol-related violations. These crimes are not intuitively expected to be highly affected by
higher temperatures.
Methods: In order to establish the hypothesis, a literature review was performed using PubMed and Google Scholar to seek information
on potential mechanisms that could explain an association between heat and traits of aggression and impulsivity from a neuroanatomical
standpoint.
In order to test the hypothesis, weather and crime records from 1997-2010 were requested from the National Oceanic and Atmospheric
Administration’s National Climatic Data Center and the courts from Harris County, TX, Miami-Dade County, FL, and three boroughs of
New York City (New York, Queens, and Bronx Counties). A team of lawyers, programmers, and research assistants manually processed
the crime data into standardized, workable language fit for categorization. The team calculated medians for maximum (TMAX)
temperatures (C) per day across several weather stations per jurisdiction. This was converted into a monthly average and set against
calculated monthly crime rates per 100,000 for each jurisdiction in a linear regression model computed using R. Data was considered
significant at p<.001 with correction for multiple comparisons using Bonferroni correction. Average yearly rates for each selected crime
were also computed for comparison.
Results: Results of the computational analysis showed that the majority of selected crimes of aggression and impulsivity did not vary
according to ambient temperature. However, the data for non-sexual assault did support the hypothesis, as it was the most statistically
significant crime of either aggressive or impulsive types in both Harris County, TX and New York City, NY. Specifically, each increase in
1C was calculated to increase rates of non-sexual assault per 100,000 by 3.01 in Harris County, TX and 4.18 in New York City, NY.
Furthermore, nonsexual assault by far had the highest yearly rates across all selected crimes of aggression in all jurisdictions. Other
smaller but still significant crimes included alcohol-related violations (non-driving) in Harris County, TX and sexual assault crimes in New
York City, NY.
None of the selected crimes of aggression or impulsivity were statistically significant for Miami-Dade County, FL. A possible explanation
for this is Miami’s relatively constant annual temperatures due to its coastal location where neighboring bodies of water blunt temperature
fluctuations. Another possibility is that relative rather than absolute temperature changes may increase criminal activity, as the population
may be habituated to high but stable temperatures.
The literature review identified a constellation of various anatomical structures that studies linked to aggression and impulsivity. Findings
for impulsivity will not be discussed, as the review focused on crimes of aggression as represented by nonsexual assault, which was
found to be the only major crime to vary significantly with temperature. A general mechanism for aggression seems to involve structures
that either generate or inhibit aggressive impulses. The key structures reported to generate aggression (labeled “accelerators”) included
the amygdala, hypothalamus, hippocampus, and insular cortex. Key structures reported to inhibit behavioral output (labeled “brakers”)
included the anterior cingulate cortex and the prefrontal cortex. Studies on personality disorders and traumatic brain injury subjects
suggested that breakdowns in the balance between accelerators and brakers lead to aggressive behaviors like assault. Research
conducted into literature on the neuroanatomical mechanisms explaining heat affecting these brain regions yielded no studies. Instead,
several theories were found that would require further empirical confirmation. In general, results of the literature review found the need for
more research to be conducted to better explain aggressive behaviors and the mechanisms underlying them. This study hopes to
increase awareness for and encourage activity in research of the interplay of temperature and behavior.
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An Exploratory Study of Medications Prescribed to Individuals Diagnosed with Headache
Alvin Chi, Duren M. Ready
Headache Clinic, Scott and White, Temple, TX
Texas A & M University Health Science Center College of Medicine, Temple
Introduction: In the US, headache is a significant and increasing public health concern. The 2004 National
Headache Foundation Fact Sheet revealed that an estimated 45 million Americans suffer from chronic,
recurring headaches annually, making it more common than asthma, diabetes, and coronary heart disease
combined [1]. Although headache is common, headache remains undertreated and mistreated in the United
States [2]. As a result, headache’s direct cost to the healthcare system is substantial [3]. In light of migraine’s
significant public health burden, it is advantageous for Scott and White Health to assess its headache
prescribing habits. As an Accountable Care Organizations (ACO), Scott and White is strongly invested in
carefully managing chronic diseases such as migraine to reduce costs and increase effective care.
Objectives: The primary objectives of our study are to characterize the headache medication prescription
patterns in the Scott and White Health South and Central Divisions and to examine patient and provider
characteristics associated with headache prescription patterns.
Methods: A retrospective claims analysis was conducted using Baylor Scott and White (BSW) pharmacy and
medical claims data from the South and Central Divisions. Scott and White Health Plan claims were accessed
between June 2011 and June 2014 (the index period) through databases from both Scott and White Center
for Applied Health Research (CAHR) and Scott and White Health Plan. Patients included in the study had to
be at least 12 years of age or older on the index date and have 1 or more pharmacy or medical claims for
headache during the index period through Scott and White Health Plan. Patients seen at Scott and White
without Scott and White Health Plan insurance were excluded because pharmacy fill data was not available
for these patients. For each patient that met the above inclusion criteria, the following data were collected:
headache category at index date, prescription medications (if any) with a pharmacy claim for headache, the
number of prescriptions filled, comorbid conditions, the number of emergency department visits, and the
number and type of diagnostic procedures performed for headache (CT and MRI imaging, EEG, and lumbar
puncture). Multiple medications were recorded if patients were prescribed more than one medication.
Searched comorbid conditions included only those common to migraine. These included the following:
epilepsy, stroke, depression, bipolar disorder, anxiety disorders, asthma, narcolepsy, and sleep disorders
(restless leg, insomnia, sleep apnea, and chronic fatigue syndrome). Prescriber characteristics were also
collected and included the following: prescriber age, years in practice, specialty, and frequency of headache
patients seen. The number of symptom codes for headache (ICD-9-CM code 784.00) were also recorded, as
well as the number of diagnostic codes for headache filed. CAHR provided the average age and the
percentage of male and female patients for each medication class that was prescribed.
Results: Currently we are still working with both CAHR and Scott & White Data Management Group on the
best methods for obtaining this data through both medical claims and pharmacy claims data. No results have
been obtained at this time.
References
1.Devine, J.W., J.F. Farley, and R.S. Hadsall, Patterns and predictors of prescription medication use in the management of headache: findings from
the 2000 Medical Expenditure Panel Survey. Headache, 2005. 45(9): p. 1171-80.
2.Lipton, R.B., et al., Migraine prevalence, disease burden, and the need for preventive therapy. Neurology, 2007. 68(5): p. 343-9.
3.Stokes, M., et al., Cost of health care among patients with chronic and episodic migraine in Canada and the USA: results from the International
Burden of Migraine Study (IBMS). Headache, 2011. 51(7): p. 1058-77.
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August 5, 2014: 9:00 AM - 2:00 PM
The Role of Alpha Smooth Muscle Actin in Regulation Of Nuclear Translocation of Myocardin and
Serum Response Factor
David Cochran and Emily Wilson
Department of Medical Physiology Texas A & M University Health Science Center College of Medicine,
College Station
Introduction: Cardiovascular diseases such as atherosclerosis and hypertension are common conditions that can
predispose patients to more serious illnesses such as myocardial infarctions and cerebrovascular accidents. Some of the
pathology underlying these diseases can be attributed to a phenotypic switch in vascular smooth muscle from a
contractile state into a synthetic state mediated by serum response factor (SRF) and one of its co-activators, myocardin.
The purpose of this study was to investigate the role of the contractile status of the vascular smooth muscle on the
cellular localization of SRF and myocardin and any proliferation changes.
Hypothesis: We hypothesize that the contractile status of the smooth muscle cells may play an important role in
regulating the cytosolic vs. nuclear localization of SRF and myocardin and the cell proliferation rate. To test this
hypothesis we used aortic smooth muscle cells derived from ACTA-2 +/+ or Wild-type (WT) and ACTA-2 -/- or alpha
smooth muscle actin knockout (KO) mice to determine if this contractile protein plays a role in regulating localization of
these proteins and cellular proliferation.
Specific Aims: We determined if the translocation of SRF and myocardin to the nucleus was altered in cells in which
alpha smooth muscle actin was deficient compared to WT cells. We did this using immunofluorescence microscopy to
localize these proteins. We determined if there was altered proliferation of alpha smooth muscle actin KO cells compared
to WT cells. We used a MTT proliferation assay to determine the proliferation rates.
Methods: Immunofluorescence Microscopy: Cells were cultured in chamber coverslips in complete media. 24 hours
prior to initiation of the experiment, the growth medium was replaced with medium containing 1% FBS. The cells were
fixed and processed to determine the subcellular localization of SRF and myocardin. The dilutions for the mouse
monoclonal anti-myocardin antibody and the rabbit monoclonal anti-SRF antibody were 1:200 and 1:750 respectively.
An anti-mouse FITC conjugate was used as a secondary to the anti-myocardin antibody at a 1:200 dilution, and an antirabbit Alexa Fluor® 488 conjugate was used as a secondary to the anti-SRF antibody at a 1:200 dilution. The cells were
visualized by fluorescence microscopy. Cells treated with the secondary antibody only were used as the negative
control.
Proliferation Assay: Cell proliferation was quantified using the MTT Cell Proliferation Assay Protocol from ATCC®.
We compared the two different cell lines in 10% FBS and 1% FBS. The time points used were 0, 24, 48, and 72 hours.
Results: The localization of myocardin was not that much different; however, there was far more nuclear localization
of SRF in the alpha smooth muscle actin deficient cells. Further studies to understand the mechanism behind this would
be to determine the localization of myocardin related transcription factors (MRTF) A and B, which bind to the actin
cytoskeleton and localization is dependent on the actin polymerization state. In the proliferation assay, the WT cells
proliferated better than the KO cells, but near the end of the assay the KO cells seemed to show a faster rate of
proliferation compared to the WT cells. Further proliferation studies should be done after analysis of the differences in
integrins and other adhesion molecules at the cell surface on the alpha smooth muscle actin deficient cells because these
cells exhibited less adherence to the cell culture dishes. Proliferation studies on in vivo-like extracellular matrices could
prove useful in determining the proliferation differences in an in vivo environment.
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Impact of Shift Work Schedules and Circadian Clock Disruption on Estrus Cycles and
Neuroprotection in Response to Stroke in Adult Female Rats
Jason Coffman, Sam-Moon Kim, Nichole Neuendorff, Amutha Selvamani, Farida Sohrabji, and David Earnest
Department of NExT, Texas A&M University Health Science Center College of Medicine, Bryan Campus
Introduction: Increases in stroke risk or incidence are derived from a variety of environmental and
biological factors including poor dietary practices and related health problems (e.g., obesity, diabetes), age
and even gender. With regard to the compound impact of gender and aging, men are at greater risk for
stroke, but middle-aged and older women have a higher risk for ischemic stroke and poor prognosis for
recovery. The surge in the incidence of stroke among women occurs as early as mid-life near perimenopause
(age 45−54), suggesting that declining levels of ovarian steroid hormones (estrogens and progestins) may
trigger the pathological shift in neuroprotection, thus increasing the risk for neurovascular disease (Towfighi et
al., 2007). The impact of age-related loss of ovarian hormones on stroke risk, severity, and outcome is well
modeled by animal studies published by the Sohrabji lab (Selvamani and Sohrabji, 2010), where ischemic
injury caused by constriction of the middle cerebral artery (MCA) results in a much larger cortical and striatal
infarct volumes in middle-aged, reproductive senescent females (9-12 months), as compared to mature adult
females (5-7 months). Circadian clocks distributed in most tissues and individual cells throughout the body
may provide integral links between cycling levels of estrogen and neuroprotective responses. Recent studies
have identified circadian disturbances as contributing factors in the physiological processes coupling diabetes
and obesity with cardiovascular pathologies. One potential explanation for the decline in neuroprotection and
increased stroke risk in middle-aged, reproductive senescent females is that the decreases in ovarian
hormones and damped oscillations in their circulating levels negatively affects the coordination of peripheral
circadian clocks, leading to the altered regulation of growth factors, such as IGF-1, and decreased
neuroprotective responses to brain injury.
Hypothesis: The aim of this research was to determine whether circadian clock disruption in young
adult female rats exposed to rotating shift work schedules (i.e., 12hr advances in the light-dark cycle every 5
days) modulates the estrous cycle and induces corresponding increases stroke-induced brain injury and
functional impairment similar to that observed in middle-aged females.
Methods: Vaginal smears using a cotton swab were initially obtained from all animals for 15 days
during exposure to a standard 12 hour light:12 hour dark (LD 12:12) cycle. After this baseline determination of
estrous cyclicity, half of the animals were maintained on this standard LD 12:12 cycle whereas the other half
was exposed to a LD 12:12 cycle that was shifted by 12 hours every 5 days for 2 months (treatment groups:
fixed vs. shifted). Endothelin (ET)-1 was delivered to the MCA by stereotaxic placement of a Hamilton syringe.
Animals received a single dose of ET-1 (3µl; 0.5µg/µl solution, American Peptide Company INC), delivered at
the rate of 1.0µl/2min. All animals were terminated at 5 days post-MCAo.
Results: Circadian rhythms of wheel-running activity stably entrain to the fixed LD 12:12 cycle but
are severely disrupted during exposure to the shifted LD 12:12 cycle. During baseline determinations, regular
estrous cycles were observed in all animals, with an average cycle length of ~5 days. Following 2 months of
exposure to experimental lighting conditions, all rats maintained on the fixed LD 12:12 cycle exhibited estrous
cycles (length ~7 days) whereas cyclicity was abolished and smears were indicative of persistent estrous in
all animals exposed to the shifted LD 12:12 cycle. Exposure to the shifted LD 12:12 cycle had a significant
effect in increasing total infarct size (cortex and striatum) relative to that observed in rats maintained on the
fixed LD 12:12 cycle. Sensorimotor testing revealed a similar trend in which MCAo-induced functional deficits
were greater in rats exposed to the shifted LD 12:12 cycle than in those maintained on the fixed LD 12:12
cycle.
Conclusion: Similar to shift work schedules, shifted LD cycles disrupt circadian rhythmicity and
estrous cyclicity in mature adult female rats and produce a corresponding pathological shift in neuroprotection
(i.e., increased infarct size and sensorimotor deficits) similar to that observed in reproductive senescent
female rats.
These results suggest that the loss of ovarian hormones and damped oscillations in their circulating levels in
middle-aged females may precipitate circadian rhythm disturbances that link reproductive aging to
pathological changes in the regulation of growth factors and neuroprotection in response to injury in the aging
female brain.
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August 5, 2014: 9:00 AM - 2:00 PM
Bryan, Texas
A COMPARATIVE ANALYSIS OF HUMAN ORGANOPHOSPHATE POISONINGS USING
SOCIAL MEDIA
Ellen Colman and D. Samba Reddy*
Texas A&M University Health Science Center College of Medicine, Bryan TX 77807
Introduction: In the last year, two deadly chemical agent exposures have occurred in different countries. On
July 16, 2013, 23 children in Bihar, India died of food poisoning by monochrotopos pesticide. On August 31,
2013, approximately 1400 Syrian civilians died from exposure to the nerve gas sarin. Also, over a decade ago
on March 20, 1995 Tokyo civilians were exposed to sarin gas on the subway during their morning commute.
Sarin, soman and organophosphate (OP) chemical warfare agents act by inhibiting the enzyme
acetylcholinesterase, which is responsible for the breakdown of acetylcholine in synaptic sites. This causes
cholinergic crisis and hyperstimulation by ACh, eventually leading to death if left untreated. Symptoms of OP
poisoning include miosis, sustained muscle contraction, twitching, hypersalivation, excessive sweating,
fainting, vomiting, respiratory depression, and seizures. Current antidotes available are atropine, an antimuscarinic agent, and 2-PAM, which re-activates the acetylcholinesterase. Nerve agents cause seizures,
brain injury, and long-term neurological problems in survivors. Benzodiazepines such as diazepam or
midazolam are used to control such seizures or status epilepticus.
Objective: The aim of this population based, retrospective, epidemiological study is to use YouTube videos
and social media to assess the signs, symptoms, and treatments of each episode separately (Tokyo, Syria
and India), and to compare how they can be utilized to display the need for long-term follow up, access to
better antidotes, and education as a whole about the issue of chemical warfare worldwide.
Methods: YouTube videos were collected for each of the three exposure cases (Syria, Tokyo, and India). The
videos were downloaded, organized into groups based on symptoms of the victims. About 22 videos were
collected for Syria and 8 each for Tokyo and India, totaling 38 videos of various lengths, symptoms, and
numerous victims. Each symptom was tallied depending on how many times it occurred and in what incident.
A scale of severity was determined for the symptoms. Severe symptoms included: respiratory depression,
seizure activity, unconsciousness, neurological or psychological symptoms, convulsions, and
hypersecretions. Moderate symptoms included: fasciculations, miosis, vomiting, and any eye pain or blurred
vision. Mild symptoms included: weakness or limpness of the body, excessive sweating, coughing, headache,
and screaming, crying, or any visibly distressed state. For each episode, a pie graph was created to
determine the percentage of severe, moderate, and mild symptoms.
Results: From the analysis of the graphs and media data, Syria had the most percentage of severe
symptoms, which corresponds with the large number of deaths (1400). Syria was followed by Tokyo, with
46% severe symptoms, but still a notable percentage of mild symptoms as well (32%). India had over 50%
mild symptoms, but a still notable number of moderate and severe cases. Tokyo seemed to have the most
resources and variation of treatment, which may account for the little number of lives lost. Tokyo is also a
large metropolitan city, with easy and fast access to healthcare and medications, whereas Syria and India
were more rural areas which required travel to seek medical attention. Comparison of human and animal
symptoms shows very similar patterns of neurotoxicity in lab animals exposed to OPs such as GD or DFP.
Conclusions: Several key points have been noted on the effects of OPs on humans, including what has
been observed in humans poisoned with OP pesticides or nerve agents. It was noticed that seizures are more
likely to be observed in humans after exposure to nerve agents like sarin. The conclusions made from
observing the videos of the Syrian and Tokyo attack can be used as a model for the Indian episode or other
poisoning incidents. Because there are many survivors in India who are still young, follow up in this group is
possible, especially to assess chronic effect of OPs on the neurological health and cognitive function.
*Supported by NIH Grant U01-NS083460 (to Dr. Reddy)*
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August 5, 2014: 9:00 AM - 2:00 PM
Neuropsychological Evaluation in Spastic Paraparesis with SPG 11 mutations
Michlene Daniels, Joseph Masdeu, MD, PhD, Belen Pascual, PhD
Department of Neurology, Methodist, Houston, Tx
Texas A & M University Health Science Center College of Medicine, (Campus- Temple, College Station,
Houston)
Introduction: Mutations in spastic paraparesis gene (SPG11) are the most common cause of Autosomal
Recessive Hereditary Spastic Paraparesis with Thin Corpus Callosum (ARHSP-TCC). Patients commonly
report with cognitive decline and lower limb spasticity in infancy and up to early adulthood. Lesions that are
involved in this syndrome include the genu fibers of the forceps minor of the corpus callosum.
Methods: As an independent group, we performed several neuropsychological tests of four ARHSP female
patients with SPG11 mutations along with 10 controls. We tested for cognitive function, apraxia, and the
performance of tasks that involved callosal transfer. We evaluated simple response times, using eye tracking
software, of patients and controls that involved interhemispheric and intrahemispheric responses. Parcellation
and volumetric analysis of all regions of the corpus callosum were performed for all subjects. The corpus
callosum was segmented manually in the coronal plane along the entire rostrocaudal dimension. Volumes
were calculated for each region by multiplying the area measurement of the region on each slice by the slice
thickness and then summing all slices on which the region appeared.
Results: Testing of all four patients indicated significant cognitive decline, a greater degree of apraxia as
compared to controls, and a decrease in performance of tasks that involve interhemispheric transfer. For all
four patients, volumetric analysis revealed a significantly thinner corpus callosum as compared to controls in
all regions except for the isthmus. The most significant volume decrease was shown in the rostrum and genu
of the corpus callosum.
Conclusion: The impaired neuropsychological performances of ARHSP patients may represent the severe
corpus callosum volume loss, especially of the anterior portions including motor, premotor, and posterior
parietal areas. The results suggest that the speed of tactile, visual, and language information transfer is
significantly slower in ARHSP patients with SPG11 mutations as compared to the healthy population. The
disproportionally affected anterior region of the corpus callosum may explain conservation of certain
functions and deterioration of others such as motor findings in the extremities and delayed visual response
times. Marked volume loss in the anterior paracentral region would be expected in a disorder causing
prominent motor findings in the extremities. These differences suggest that spatacsin plays a larger role in
motor subsystems, understood in a wide sense and including premotor and prefrontal circuitry of the frontal
lobe.
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August 5, 2014: 9:00 AM - 2:00 PM
Aquaporin-4 and ICAM-1 Expression after Traumatic Brain Injury
1
1
2
1,2
Ashley Dawson , Sanjib Mukherjee MD, PhD , Sören Singel MD , Karen Newell-Rogers PhD
1
2
Texas A & M University Health Science Center College of Medicine, Temple, Baylor Scott and White
Hospital, Department of Neurosurgery
Introduction: Edema after brain injury is a serious and complicated problem that can cause uncontrollable
increases in intracranial pressure. Of the three main aquaporins in the brain, aquaporin-4 (AQP-4) has been
found to contribute most to pathology after brain injury and has been identified as a potential target for
treatment of edema.
Hypothesis: We believe that acute neuroinflammation caused by microglia activation and antigen
presentation after brain injury contributes to the up-regulation of aquaporin-4 around the injury site. As TPP is
a peptide that blocks the MHCII binding cleft, we hypothesize that pre-treatment with TPP will reduce
inflammation (marked by ICAM-1 expression) and AQP-4 expression after fluid percussion brain injury.
Methods: Five mice were sacrificed 1, 3 and 7 days after the injury. 3-day mice underwent either pretreatment with an injection of TPP or no pre-treatment. A saline-filled syringe was connected to a cannula
placed against the dura in an area correlating to 1.2 mm posterior and 1.2 mm lateral to the the bregma of the
skull. The syringe was hit once at a force of 1.7 atm in a 12 ms pulse. The sham mouse underwent the same
procedure without the actual FPI.
Brains were removed and sliced into 40 μm sections. Several slices from different areas in the brain (anterior,
posterior, peri-lesion area) were taken for each experiment. Slices were incubated in blocking buffer (10%
normal goat serum with 0.025% Tween-20) for 2 hours before transfer to the primary antibody solution (1:200
BioLegend hamster anti-mouse ICAM1-FITC and 1:200 Abcam rabbit anti-AQP4) for overnight incubation.
Slices were then washed 3 times for 5 minutes each and incubated in secondary solution (1:1000 AlexaFluor
647 goat anti-rabbit IgG) for 1 hour at room temperature. Slices were washed again as previously described
and placed on slides. Samples were cover-slipped with DAPI and visualized with confocal microscopy.
Results: These data indicate that aquaporin-4 and ICAM-1 expression is greatest three days after fluid
percussion injury (FPI). Confocal microscopy revealed the change in aquaporin-4 expression in astrocytes
after FPI. Prior to injury, AQP-4 is seen in concentrated areas that correlate with its expression in astrocyte
podocytes around the blood brain barrier. 3 days after FPI, AQP-4 is expressed in linear patterns that
suggest expression along the length of activated astrocytes. Finally, while it is seen that TPP acts to reduce
ICAM-1 levels, it does not definitively influence aquaporin-4 expression. Additional testing with a larger
sample of mice is needed to determine the effect of TPP on aquaporin-4 expression.
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August 5, 2014: 9:00 AM - 2:00 PM
Gulf War Illness Related Chemicals Adversely Affect the Proliferation of
Human Neural Stem Cells
Kurren Desai, Bharathi Hattiangady, Geetha Shetty, Ashok K. Shetty
Institute for Regenerative Medicine, Department of Molecular and Cellular Medicine, Texas A & M University
Health Science Center College of Medicine, Temple, TX
Introduction: Gulf war illness (GWI) affects nearly third of 700,000 military personnel who served in the
1991 Persian Gulf War. While the precise cause of this illness is unknown, epidemiological studies suggest
that exposure to several chemicals during the war underlie this multi-symptom illness. The chemicals include
acetylcholinesterase inhibitors such as pyridostigmine bromide (PB, used by service personnel as an antidote
against nerve gas attack) and pesticides such as permethrin (PM, used extensively during the war to combat
insects and rodents in the desert). Indeed, animal model studies have suggested that combined exposure to
these chemicals induces GWI-like central nervous system deficits such as cognitive and mood dysfunction
(Parihar et al., 2013, Hattiangady et al., 2014). Interestingly, these symptoms were allied with persistently
decreased proliferation of neural stem cells (NSCs) resulting in reduced neurogenesis (i.e. addition of new
neurons to the circuitry) in the hippocampus, a region of the brain vital for functions such as learning, memory
and mood. However, it is unknown whether NSCs in the brain of veterans afflicted with GWI have similar
dysfunction. Therefore, we examined the effects of low-doses of GWI-related chemicals (PB, PM) on the
proliferation and differentiation of human NSCs (hNSCs) derived from human induced pluripotent stem cells
(hiPSCs).
Hypothesis: We hypothesized that even low-doses of GWI-related chemicals such as PB and PM can
interfere with the proliferation and differentiation of hNSCs.
Methods: The hiPSCs used in this study were generated via reprogramming of fibroblasts obtained from
human skin biopsies. Reprogramming involved insertion of transcription factors such as Sox-2, Oct-4, Klf-4
and C-myc. This study utilized hiPSCs obtained from WiCell (Madison, WI) to generate NSCs. For
proliferation analyses, NSCs were plated onto geltrex-coated wells of a 24 well plate (10,000 cells/well) in a
neural expansion medium. A day after plating, the medium was replaced with a fresh medium containing
either GWI-related chemicals or vehicle. A total of 8 groups were prepared. Effects of two different
concentrations of PB (200µM, 300µM) and PM (20µM, 40 µM), alone or in combinations, were examined
along with control and vehicle-treated sister cultures. The culture medium was replaced on day 4, and all
cultures were terminated on day 5 with 2 % paraformaldehyde. Cultures were then processed with Ki-67
immunofluorescence (to detect proliferating cells) and treated with DAPI (to localize nuclei of all cells). Cells
were imaged and quantified using a Nikon confocal microscope and associated software. For characterization
of the effects of GWI-related chemicals on differentiation, NSCs were plated in a differentiation medium on
poly-ornithine/laminin coated wells. After two days, the medium was replaced with a fresh medium containing
chemicals. A total of 8 groups were prepared as described earlier. The culture medium containing respective
chemicals or vehicle was replaced every three days. All cultures were fixed in 2% paraformaldehyde on day
17, processed for TuJ-1 (a marker of neurons) immunofluorescence and DAPI treatment and examined and
quantified using a Nikon confocal microscope.
Results: Quantitative analyses demonstrated that both PB and PM have detrimental effects on the
proliferation of hNSCs even at low-doses. Proliferation of hNSCs was reduced in a dose-dependent manner.
Of the two GWI-chemicals examined, PM showed the greatest suppressive effect on the proliferative activity,
recovery and survival of NSCs. When PB and PM were combined, an additive adverse effect on hNSC
proliferation was clearly observed. A qualitative analysis of differentiation cultures also showed reduced
survival of NSC-derived TUJ1+ neurons and induced pathological changes such as beading of dendrites and
axons in surviving neurons in the drug treated groups. Additional quantification is in progress to ascertain the
definitive effects of these chemicals on the differentiation of NSCs.
Conclusions: Human NSCs are extremely vulnerable to GWI-related chemicals such as PB and PM, as
exposure to these chemicals even at low-doses can suppress their proliferation and survival.
21 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
EVALUATING EFFICACY OF NITRIC OXIDE RELEASING NANOPARTICLES
NAILA DHANANI, JOSEPH FERNANDEZ-MOURE, M.D., M.S.
Texas A & M University Health Science Center College of Medicine, Houston
Houston Methodist Research Institute, Departments of Nanomedicine and Surgery
Introduction: Nitric oxide (NO) is a fundamental endogenously derived cellular signaling molecule involved in
both physiological and pathological processes. In the human body, NO is synthesized when L-arginine is
converted to L-citrulline by nitric oxide synthase (NOS). NO can act as a vasodilator, antibacterial agent and
tumoricidal factor. As a vasodilator, NO inhibits endothelial apoptosis, platelet aggregation and adhesion,
vascular smooth muscle cell proliferation and migration, and leukocyte chemotaxis. NO stimulates
extracellular matrix changes, vascular smooth muscle cell apoptosis, and endothelial proliferation. NO also
plays a role in host defense as its signaling controls the differentiation, proliferation, and apoptosis of immune
cells. When secreted by activated immune cells, NO diffuses across cellular membranes and can exert
damage on invading pathogens via free radical production. Methicillin-resistant Staphylococcus aureus
(MRSA) is found to be susceptible to NO at levels non-toxic to fibroblasts.
Hypothesis: To integrate silica based nanoparticles with current surgical therapies for the development of
novel devices to overcome high infection rates in the current materials.
Methods: To synthesize nitric oxide releasing nanoparticles, tetraethoxysilane (TEOS) was cocondensed
with aminoalkoxysilane along with ethanol, water and ammonium hydroxide. The amine groups in the silica
particles were converted to N-diazeniumdiolate NO donors through the exposure to high pressures of NO.
1
The release of NO was quantified using Sievers 280 chemiluminescent NO analyzer then characterized
using SEM (scanning electron microscopy). Once release was established, the antibacterial properties of NO
were assessed. To determine in vitro efficacy, cultures of Staphylococcus aureus were inoculated with
varying concentrations of nitric oxide releasing particles (1x10^4, 1x10^6, 1x10^8) then plated overnight after
a two-hour interval. A computerized count of all colony-forming units (CFU) was performed in order to
2
visualize remaining bacteria. Following this, an in vivo model was established. In 36 rats, the linea alba was
incised and composite polyester mesh with NO releasing capabilities was fixed in underlay fashion. At the
time of closure, 1X10^4, 1x10^6, 1x10^8 MRSA USA300 strains were inoculated into the animals. 18 animals
served as controls and 18 were imparted with NO mesh. 6 rats in both the control and NO mesh group were
inoculated with MRSA 10^4, 10^6, 10^8 respectively. Tissues were harvested en bloc at 24 hours and
digested for CFU analysis.
Results: NO can be successfully released from a composite polyester mesh and can significantly reduce
bacteria growth in vitro. In an in vivo model for hernia repair in the infected field, NaNO mesh is capable of a
significant reduction in bacterial load.
Future Perspectives:
•Longer release profiles and integration with polymer matrix rather than coating
•Effects of mesh on fibroblasts, macrophages, and stem cells, surrounding tissue
•Larger animal model
1. Hong Jiang, Texas Therapeutics Institute at Brown Foundation Institute of Molecular Medicine for the
Prevention of Human Diseases.
2. Randall Olsen, M.D., Ph.D., Center for Molecular and Translational Human Infectious Diseases Research
Institute, Houston Methodist Research Institute.
22 | P a g e
Manganese Increases Proliferation within the Prepubertal Mammary Gland through Estrogen-Mediated
Pathways
Alina M. Hamilton1, Vinod Srivastava2, Jill Hiney2, Robert K. Dearth1, W. Les Dees2
1
Department of Biology, College of Science and Mathematics, University of Texas Pan American, Edinburg,
TX 78539
2
Department of Veterinary Integrative Biosciences, College of Veterinary Medicine, Texas A&M University,
College Station TX 77843
Texas A&M University Health Science Center College of Medicine, College Station, TX
Abstract
Manganese (Mn) is a trace element that is nutritionally required for normal physiological processes.
Prepubertal exposure to elevated levels of Mn can cause precocious pubertal development in female rats by
stimulating the release of puberty-related hormones LH, FSH and estradiol (E2). Recently we demonstrated
that elevated dietary Mn accelerates estrogen-regulated mammary gland (MG) development and ductal
differentiation. While we showed that the increase in MG ductal differentiation correlated with a significant
increase in proliferation, it is not known what specifically contributed to this phenotype. Thus, in the current
study we endeavored to elucidate key E2 -regulated markers that may be contributing to the increased
proliferation seen in Mn-treated females. Sprague Dawley female rats were exposed daily to 10mg/kg
manganese chloride (MnCl2) or saline (control) via gastric gavage from post-natal day (PND) 12 to PND 30, at
which point inguinal mammary gland #4 was harvested and processed. Western blot analysis revealed a
significant increase in protein expression of c-myc (p<0.05) and amphiregulin (p<0.05) in Mn-treated
females when compared to controls, but no change in cyclin D-1 protein expression. This precocious
increase is important because both c-myc and amphiregulin play fundamental estrogen-mediated roles in
mammary gland development. Thus, in the current study we identified two key estrogen-regulated proteins,
amphiregulin and c-myc, which could potentially, at least in part, drive the proliferative phenotype within
the mammary gland of Mn-treated female rats. Collectively, while our data is preliminary, it supports the
hypothesis that Mn centrally regulates advanced pubertal development through estrogen-mediated
pathways.
23 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Identification of the E2/E3 Ligase activity within the SpII spectrin protein
Carla Hayworth, Rachel Petrofes Chapa, Warren Zimmer Ph.D.
Department of Medical Physiology
Texas A & M University Health Science Center College of Medicine, (Campus- College Station)
Introduction and Background: Two isoforms of spectrin protein have been identified in human cells;
erythrocyte spectrin (αSpI/βSpI) and nonerythroid spectrin (αSpII/βSpII). Erythrocyte spectrin is a protein
essential in maintaining the shape, elasticity, and flexibility of erythrocytes by forming a scaffolding over the
cytoplasmic surface of the erythrocyte membrane (Hsu 2004). It is also now widely accepted that the
αSpI/βSpI serves an enzymatic role in the cell as well, and the amino acid sequence of the E2 site in αSpI
has been identified. To our knowledge, no studies have been published investigating the E2/E3 enzymatic
activity of αSpII. The αSpII/βSpII constitutes 2-3% of total brain protein and is found to be ubiquitinated, but
currently it is not known if a similar region surrounding Cys2071 of αSpII expresses E2/E3 ligase activity (Hsu
2004). A suspected E2 site has been compared to the αSpI E2 site and found that there is approximately 63%
identity between the sequences. If αSpI is found to have the properties of an ubiquitin enzyme, defects in its
functioning could have severe consequences for overall brain functioning considering it constitutes a
substantial proportion of total brain protein. Understanding the enzymatic activity of αSpII may be essential in
further discovery of mechanisms underlying several neurodegenerative diseases with ubiquitin
immunoreactivity in cystoplasmic inclusions including, but not limited, Alzheimer’s disease, Huntington’s
disease, Parkinson’s disease, various prion diseases, amytrophic lateral sclerosis, and motor neuron disease
(Reiderer 2011). In this study, our aim was to demonstrate expression of αSpII in BL21 bacterial cells and
subsequently evaluate the E2/E3 ligase activity of the αSpII c-terminal amino acid sequence that resembles
the αSpI E2 site.
Hypothesis: Based upon the analysis of the E2 site of αSpI, we hypothesize that the potential E2 site in αSpII
is capable of E2/E3 ligase activity.
Methods: Recombinant cDNA encoding αSpII fused to GST was purchased and transformed into bacterial
cells (BL21) that were engineered for expression of exogenous proteins. Expression of the full-length αSpII in
the bacteria was evaluated by lysing the cultures and performing a novel dot blot assay using αSpII antibody
and a secondary antibody developed using chemiluminescence. Attempts to purify the protein from the
bacterial lysate mixture were made using GS beads. These attempts were unsuccessful. Immunoprecipitation
using GST antibody also proved unsuccessful. Due to the failure of purification attempts, ubiquitin linkage
assays were performed on the whole bacterial lysates and evaluated by gel electrophoresis and western
blotting.
Results: The failed attempts to purify the protein made it difficult to draw conclusive results from the ubiquitin
likage assays. Further efforts in isolating the αSpII protein from the bacterial lysates need to be made.
Conclusion: Although results were inconclusive, the possibility that αSpII possesses E2/E3 ligase activity
was not ruled out. In the ubiquitin assays, there was ubiquitin present on some bands of the bacterial lysates
after western blotting. It could not be said for certain that αSpII is capable of self ubiquitination, it may
ubiquitinate other proteins or work in conjuction with αSpI.
24 | P a g e
Differential miRNA Expression in Highly Invasive Glioblastoma Cell Lines
1
1
2
Linda Herrera , Camille Duran , L. Gerard Toussaint , Raquel Sitcheran
1
1
2
Department of Molecular and Cellular Medicine, Department of Neuroscience and Experimental
Therapeutics Interdisciplinary Program in Neuroscience
Texas A & M University Health Science Center College of Medicine, (College Station)
Introduction: Glioblastoma multiforme (GBM) is the most common form of malignant brain cancer in adults
with only a 5-year survival rate in less than 3% of patients. GBM tumors are highly invasive, infiltrating
surrounding brain parenchyma. Invasive GBM cells are transcriptionally, and hence phenotypically, distinct
from the cells found at the tumor core. Subpopulations of glioblastoma cells with enhanced invasive
properties have been used as a model for examining genetic and epigenetic alterations necessary for
invasion. One potential mechanism of post-transcriptional regulation of gene expression contributing to the
invasive phenotype in glioblastoma is through microRNAs (miRNAs). In particular, miR-146a has been
reported to be involved in progression of several types of cancer by negatively regulating canonical NF-κB
pathway. On the other hand, miR-886-3p has been shown to regulate CXCL12 by directly binding to its
3’UTR. CXCL12 has been associated to the tumorigenesis process in breast, ovary, kidney and prostate
cancers. In this study, we isolated highly invasive cells in order to characterize pro-invasive and proproliferation signaling proteins in glioblastoma.
Hypothesis: We hypothesize that highly invasive cell lines will have differential miRNA expression when
compared to parental cells, uncovering potential mechanisms that drive cell invasion.
Methods: We isolated U87 glioblastoma invasive subpopulations by serially passing them through a matrigelcoated membrane. To analyze invasion, we performed an invasion assay in a three-dimensional collagen
matrix. We obtained the average number of invading cells and quantified the distance from the monolayer to
the deepest point of invasion. Moreover, we determined the expression of miR-146a and pre-miR-886 in
these cells and their possible targets using qRT-PCR and Western Blot analysis.
Results: We successfully selected highly invasive U87 glioblastoma cell subpopulations (IM2 and IM3). IM2
and IM3 cells displayed higher number of invasive cells, while also invading deeper into the collagen matrix
than the parental cell line. Moreover, IM2 and IM3 cells were transcriptionally different from the parental cell
line, shown by the differential expression of miR-146a and pre-miR-886. miR-146a downregulation appeared
to positively regulate NF-κB pathway activation. Pre-miR-886 was significantly upregulated but its potential
targets remain to be elucidated. Understanding the differential miRNA expression in highly invasive
subpopulations of glioblastoma cells provides an epigenetic mechanism to drive cell invasion and migration
and uncovers potential targets for therapeutic development.
25 | P a g e
Influence of ADRB2-79 and ADRB3 Metabolic Gene Single Nucleotide Polymorphisms on Body
Composition and Fitness
J Howell, B Sanchez, A Coletta, E Galvan, P Jung, R Dalton, K Levers, M Koozehchian, S Simbo, A
O’Connor, A Reyes, S Springer, C Goodenough, M Cho, C Rasmussen, RB Kreider.
Department of Health and Kinesiology, Exercise and Sports Nutrition Laboratory
Introduction: Numerous genetic single nucleotide polymorphisms (SNPs) have been suggested as
determinants of obesity, health, and biochemical modulation. This study focused on two specific metabolic
genes related to obesity (ADRB2-79 and ADRB3). The SNP allele variants of these genes were examined to
determine if overweight, sedentary women had inherent differences in body composition and/or markers of
health and fitness.
Hypothesis: Women with specific SNP allele variants are predisposed to possess healthier body
composition, and higher fitness markers than those with other allele variants, and also obtain more substantial
results from diet and exercise.
Methods: 118 sedentary women (38.9±12yr, 64.05±3.54 in,192.02±48.5 lbs, 42.2±6% body fat) were
measured for BMI, resting heart rate, blood pressure, resting energy expenditure, body composition, peak
oxygen uptake (VO2 max), and upper- and lower- body muscular strength and endurance prior to participating
in a 7 month weight loss program. Women who remained in the weight loss program returned each month for
st
the same testing until the end of the program. Buccal cheek swabs were obtained for genetic testing at the 1 ,
th
th
4 , and 7 testing sessions. Subjects were stratified by SNP genotype for the obesity related genes. Bivariate
correlation analysis and one-way ANOVA with LSD post-hoc analyses were performed. Data presented as
means ± standard deviation.
Results: For ADRB2-79 in comparison with VO2 max, First testing session data a trend favoring G allele
carriers with CG heterozygotes showing the greatest results (CG 2.03±0.36; CC 1.86±0.29; GG 1.92±0.43
L/min, p=0.03). CG heterozygotes also tested for greater upper body muscular strength during the bench
press exercise (CG 74.74±19.18; CC 64.37±15.65; GG 72.53±20.5 lb, p=0.01). The smaller sample size from
participants who have completed the weight loss program was insufficient to establish significance but there is
trend so far pointing to the probability that GG homozygotes have the highest percent increase in upper-body
strength (CC 69.5±19.07 (7.9%); CG 76.11±19.97 (1.8%); GG 79.0±27.7 (8.9%) lbs) and the greatest percent
decrease in fat percentage (CC 35.95±7.42 (12.0%); CG 37.89±5.82 (17.5%); GG 34.14±7.09 (20.3%) %).
Post hoc analysis was not able to be performed for ADRB3 because only one participant had the SNP type
CC. However, the gene still showed significant correlation with percent body fat (r= -0.232, p= 0.02), waist
circumference (r= -0.183, p=0.04), and relative VO2 max (r=0.19, p=0.04). One way ANOVA analysis showed
a trend that CT allele heterozygotes had a lower percent body fat (TT 42.63±5.9; CT 37.61±6.76 %, p=0.033).
Sample size was again too small to establish significance of T7 results but there was a noticeable trend that
showed TT homozygotes had a greater decrease in fat percentage (CT 34.97±9.39 (7.54%); TT 36.27±6.53
(17.55%) %) and a greater increase in VO2 max (CT 28.7±6.66 (12.28%); TT 27.8±5.78 (19.67%) mL/kg/min)
Conclusions: Analysis of data from the 1st testing session reveals a significant correlation between ADRB279 and ADRB3 and a number of health and fitness markers. The data from this study provides compelling
evidence that women may be genetically predisposed to have a certain body composition and athletic
capability. However, there is not enough data collected so far from women who have completed the weight
loss program to determine if they may also have a disposition which allows them to show better improvement
after diet and conditioning.
26 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Inhibition of histamine synthesis decreases cholangiocarcinoma progression, EMT and
metastatic potential
Victoria D. Huynh, Laura Hargrove, Sharon DeMorrow, Allyson Graf, Lindsey Kennedy, Quy Nguyen,
Brittany Ladd, Maryah Barker, Christopher Johnson and Heather Francis
Texas A & M University Health Science Center College of Medicine, (Temple), Central Texas Veteran’s Health
Care System and BaylorS&W Healthcare
Introduction: The tumor microenvironment of cholangiocarcinoma (CCA) is composed of numerous
cells including mast cells (MC). After activation, mast cells release numerous factors including
histamine, which increase tumor progression and angiogenesis. CCA tumors are also metastatic and
undergo epithelial to mesenchymal transition (EMT). We have shown that (i) CCA express histidine
decarboxylase (HDC) and secrete histamine; (ii) HDC inhibition reduces CCA growth and VEGF
expression via autocrine signaling and (iii) stimulation of the inhibitory H4 histamine receptor reduces
EMT and tumor growth.
Hypothesis: Recruitment of mast cells into the tumor microenvironment increases histamine release,
CCA growth and EMT/metastasis.
Methods: We evaluated the presence of mast cells by toluidine blue staining in human CCA tumors
along with the expression of mast cell markers: c-kit, chymase and tryptase by immunohistochemistry.
In tumors from nude mice treated with vehicle (NaCl), histamine or the HDC inhibitor, -methyl-dlhistidine (-methyl) we measured mast cell presence by toluidine blue staining and the expression of
PCNA, HDC and c-kit by qPCR. To evaluate EMT, we measured the expression of paxillin, vimentin
and MMP2/9 by qPCR in tumors from nude mice treated with vehicle (NaCl), histamine or a-methyl. In
vitro, human CCA cells were treated with MC supernatants (treated with the HDC inhibitor, -methyl)
and proliferation (qPCR for PCNA) and histamine secretion (by EIA) was measured. Vimentin, MMP2/9
and TIMP2 expression was measured by qPCR after stimulation with MC supernatants.
Results: Mast cells are present in human CCA tumors and the expression of c-kit, chymase and
tryptase are increased compared to non-malignant biopsies. In tumors from mice treated with
histamine there was increased PCNA, HDC and c-kit expression compared to vehicle-treatment. In
mice that were treated with the HDC inhibitor there was decreased PCNA, HDC and c-kit
expression compared to histamine or vehicle treatment. Further, the expression of paxillin,
vimentin, MMP2/9 was increased downregulated in tumors from animals treated with the HDC
inhibitor. In vitro, stimulation with MC supernatants treated with the HDC inhibitor decreased CCA
proliferation and CCA histamine release as well as vimentin and MMP2/9 expression, whereas
TIMP2 expression was increased.
CONCLUSION: MCs migrate into the tumor microenvironment of CCA and increase tumor
progression and EMT/metastasis. Inhibition of histamine synthesis decreases tumor growth and
EMT/metastasis and may be a potential therapy for patients suffering from CCA.
27 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Characterization of the molecular identity and functional roles of
store-operated calcium channels in human lymphatic endothelial
cells
Kate Kearney, Xu Peng, Shenyuan L. Zhang
Lymphatic endothelial cells (LECs) line lymphatic vessels in the human body and are
vital for the two main functions of the lymphatic system: collecting and transporting
interstitial fluid and supporting the immune system. Here, Store-Operated Calcium Entry
(SOCE) was studied in these cells by use of Western blotting and ratiometric calcium
imaging techniques. SOCE in human T cells is mediated by a Calcium Release-Activated
Calcium (CRAC) Channel, which is made up of two proteins, Orai1 and Stromal Interaction
Molecule 1 (STIM1). Orai1 is located in the plasma membrane (PM), and STIM1 is a
protein on the endoplasmic reticulum (ER) membrane. LECs have a presence of STIM1,
Orai1 and Orai2 (a homolog of Orai1) proteins. Thapsigargin (TG) is used to encourage
SOCE in LECs and this drug increases intracellular Ca2+ levels by depleting the
intracellular Ca2+ store, which should then open the CRAC channel. The use of CRAC
channel blockers, i.e. BTP2 and GSK5498A, inhibited TG-evoked SOCE in LECs,
indicating that this Ca2+ entry is conducted by CRAC channels. RNA interference (RNAi)
was then performed to determine the impact on SOCE by the loss of function in Orai1 and
Orai2. Interestingly, knocking down of Orai1, but not Orai2, shows decreased SOCE in
LECs. Histamine, which has been shown to stimulate SOCE in other types of cells by
releasing ER Ca2+ through IP3 receptors, also triggers Ca2+ entry in LECs in our study. In
contrast, we show that Estradiol (E2), a major component of Estrogen, did not significantly
influence intracellular Ca2+ homeostasis in LECs.
28 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Modeling TLR Agonist Protective Effects in Vitro
Mar’Kiffany Lane, Yifan Zhang, Danielle M. McGrath, Magnus Hook, and Dekai Zhang
Department of Inflammatory and Infectious Disease
Texas A & M University Health Science Center College of Medicine, (Houston)
Introduction:
Lower respiratory infections caused by influenza type A have continually caused many
fatalities worldwide. Recent epidemics have emphasized the importance of preventative
and containment strategies.
Hypothesis: We
hypothesize that the agonist TLR combination will inhibit viral replication and
by using this TLR combination we can produce a robust assay that can be probed in the
future.
Methods:
Here, we report the inhibition of viral replication following TLR agonist treatment in MLE-15
and A549 tissue cultures. Inhibition of viral replication was measured as previously
described. In brief: Human lung epithelial cells, A549 (ATCC) and MLE-15 cells were
seeded at 200,000 cells/well in a 24-well plate in 2ml of antibiotic containing normal growth
medium supplemented with 10% FBS. The cells were incubated at 37°C in a 5% CO2
incubator. The next day the media was removed and the TLR agonist combination (2.5 μM
ODN and 10 μM PAM-2) was added in serum free, RPMI 1640 medium. Four hours
following treatment cells were infected with Influenza A/HK/68 at an MOI of 1:1. 24 and 48
hours following infection cells and supernatant were collected, RNA was extracted as
described (OMEGA bio-tek, E.Z.N.ATM Total RNA Kit I) and reverse transcribed to convert
mRNA to cDNA. Lastly, qPCR was performed to determine the levels of viral mRNA were
normalized against b-actin.
Transfection efficiency was measured using 4 different transfection reagents using GFPplasmid methods previously described.
29 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Assessing the Flammability Characteristics of Alcohol Skin Prep Solution to Reduce Operating Room
Fire Risk
Sarah Luna, Aris J. Maguddayao, Ryan T. Wade, and William C. Culp Jr., M.D.
Depart of Anesthesiology
Scott and White Memorial Hospital, Temple, Texas
Texas A & M University Health Science Center College of Medicine, Temple, Texas
Background: Approximately 600 fires take place in the operating room each year, causing patient injury and/or
death. For a fire to occur, three sources must be present: an oxygen, fuel, and ignition source. There are many items
present in the OR that may act as the fuel source such as surgical drapes and alcohol prep solution. Alcohol prep
solutions serve as the fuel source in approximately 15% of all operating room fires, though this number may be
underestimated due to the lack of data published about operating room fires. Often, the alcohol prep solution will pool
underneath surgical drapes, trapping and keeping the flammable vapor close to the surgical site. The electrosurgical unit
(ESU), a high-voltage, high-temperature cutting device, is then activated and ignites the vapors, producing a dangerous,
colorless fire. Though there are recommendations to reduce the risk of fire on the packaging of most alcohol prep
solutions, fires still occur. Little to no data is available that clearly lists the flammability characteristics of 70% isopropyl
alcohol and the risk pooling under drapes presents. The purpose of this study is to thus evaluate the flammability hazards
of using an isopropyl alcohol prep solution in the operating room.
Methods: All testing was completed in an operating room. A petri dish was filled with 0.25g of alcohol by a
swabbing motion - the same technique nurses use to apply the prep solution in the OR. The petri dish was then placed on
a balance with a volatile organic compound (VOC) analyzer secured above, which measured the alcohol vapor
concentration above the solution. A surgical drape was then placed over the petri dish or left uncovered. The mass and
vapor concentration were then recorded every 10 seconds until the vapor concentration dropped below 0.01%. To
demonstrate flammability with the ESU, the device was activated in the presence of the alcohol prep solution. Different
drying times were tested in 30 second intervals until no fire was observed for 10 tests.
Results: Drying time and vapor concentration were measured and analyzed. The drying time increased and the
vapor concentration stayed above the flammability threshold for a longer period of time when a surgical drape was placed
over the solution. Ignition of alcohol with the ESU was possible until 150 seconds of drying time elapsed.
Discussion: Both the drying time and time for the vapor concentration to drop below the flammability threshold
increased with surgical drapes. Fire was not possible once the vapor concentration was below approximately 0.19%,
which took approximately 3 minutes. Placing surgical drapes over alcohol without allowing sufficient drying time to pass
increases the possibility of fire for a longer period of time. Of note, all testing was done in room air. A high oxygen
environment is often present in the OR, which increases flammability of most fuels present, including the prep solution, a
factor that should be further investigated. Though there are warnings on the label, OR fires still occur due to alcohol
vapors in the surgical site. Surgical staff should be aware of the dangers of using the alcohol prep solution with the ESU.
Care should be taken to not allow the prep solution to pool underneath the drapes.
30 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Prevention Operating Room Fires: Development of a Carbon Dioxide Suppression Device
Aris J. Maguddayao; William C. Culp Jr., MD; Bradly A. Kimbrough; Sarah Luna
Department of Anesthesiology
Scott & White Healthcare
Texas A & M University Health Science Center College of Medicine, Temple Campus
Introduction: Operating room (OR) fires present a real danger to surgical patients and is estimated to occur
over 600 times annually. There is intense attention from the American Society of Anesthesiologists, the
Anesthesia Patient Safety Foundation, and others in reducing the risk of OR fires. In order for fire to occur,
the three points of the fire triad must be present: an oxygen source, an ignition source, and a fuel source. The
electrosurgical unit (ESU) pencil serves as an ignition source and triggers the vast majority of OR fires.
Carbon dioxide (CO2) is a gas proven to prevent ignition and suppress fire by displacing oxygen.
Hypothesis: We hypothesize that a device can be created to produce a cone of CO 2 around an ESU pencil
tip, thus reducing this risk.
Methods: A fire prevention device (shroud device) was created by drawing 2D schematics, using computerassisted technology to create a 3D design, and working with a local manufacturing company to engineer a
prototype. CO2 was pumped through the shroud device and out of 12 angled shroud ports to create a fire
prevention cone surrounding the ESU tip. This device was then tested in a flammability testing chamber in
which the ESU pencil was activated for sustained current delivery to a copper test plate holding a laparotomy
sponge. The ESU was activated at 50W cut mode for 30 seconds or until the sponge ignited. The tests were
conducted in 21%, 50%, and 100% oxygen environments. Each test was performed five times with the device
turned “on” (CO2 flow at 10 L/min) and with the device turned off (control). Time to ignition was measured with
high speed videography. Additionally, mapping of CO 2 concentrations was performed with a micro-CO2
sampling device using a 6 x 6 x 3 cm matrix to depict CO2 concentrations in 3D space.
Results: The median ± SD [range] ignition time of the control group in 21% oxygen was 2.9s ± 0.44 [2.3 –
3.0], in 50% oxygen 0.58s ± 0.12 [0.47 – 0.73], and in 100% oxygen 0.48s ± 0.50 [0.03 – 1.27]. No fire was
observed when CO2 was applied in all concentrations of oxygen. The CO 2 concentration at the end of the
ESU tip was 83% while the average CO2 concentration one centimeter away from the end of the ESU tip on
the surface plane was 46%.
Discussion: The shroud device prevented fire during ESU use, even in super oxygenated environments as
seen in Figure 2. By comparing the time of ESU fulguration with and without the device, we are able to
understand the impact that oxygen displacement by a CO 2 flow of 10 L/min has on the ability of the
laparotomy sponge to ignite. Additionally, 4D contour mapping of CO 2 concentrations show that the shroud
device will effectively reduce fire risk in the active region around the ESU pencil tip as seen in Figure 3.
Limitations of the experiment include: the copper plate was not a good representation of human skin and the
experiment was performed in unrealistic oxygen environments. A fire prevention device was able to be
constructed using the shroud device with angled shroud ports to create a cone of CO 2 gas. Pumping CO2
through the shroud device effectively displaced enough oxygen around the ESU tip to prevent ignition from
occurring. Future testing should determine the optimum CO 2 flow rates and a more ideal device design. Use
of this device may substantially reduce the risk of patient injury caused by OR fires.
31 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Does Tannase from Intestinal Bacteria Modify the
Anticancer Effects of Tea Catechin in Colon Cancer Cells?
1,2
2
2
2
Tasneem Mahmood , Mohaiza Dashwood , Praveen Rajendran , and Roderick H. Dashwood
Center for Epigenetics & Disease Prevention (CEDP)
1,
2
Texas A&M University , College Station, Texas, Institute of Biosciences and Technology , Texas A&M Health
Science Center, Houston, TX
Introduction: According to the American Cancer Society, approximately 136,820 people will be diagnosed
th
with colorectal cancer (CRC) in 2014. It is the 4 most common cancer in both men and women and the
second most deadly cancer with 17% mortality in men and women. Green tea (Camellia sinensis, Theaceae)
and its major polyphenol constituent, epigallocatechin gallate (EGCG), have been reported to have many
health benefits including the prevention of CRC (Lambert et. al, 2007). Tannase, an enzyme known to be
produced in the colonic microflora, is known to catalyze the hydrolysis reaction of the ester bonds present in
the gallotannins, complex tannins, and gallic acid esters. Therefore, we sought to find out if Tannase (derived
from Streptococcus gallolyticus, Sg) treatment modifies the biological activity and anticancer effect of the
gallate containing tea polyphenol, EGCG.
Hypothesis: Since tannase can modify gallic acid esters, we hypothesized that the beneficial effects of green
tea gallate-containing compounds can be modified in the gut by tannase produced by the gut microflora.
EGCG metabolites generated by tannase were hypothesized to significantly alter anticancer activity and
certain biological effects (e.g. HDAC inhibition).
Methods: EGCG (1 mM in phosphate buffer pH 6.9) and Tannase (Sg-derived and dissolved in PBS) were
tested alone and in combination in a concentration range of 2.5, 5, 10, 20 and 40 µM. For cell viability assays,
HCT-116 colon cancer cells were cultured in McCoys growth medium containing 10% FBS/1% penicillinstreptomycin. CCK-8 cell viability and HDAC activity assays were performed as previously described
(Rajendran et al, 2013)
Results: EGCG and the combination (EGCG + Tannase) inhibited cell viability at 40 µM when incubated for
48 h with HCT116 colon cancer cells. We did not observe any additional effect of the combination treatment.
Notably, Tannase by itself slightly inhibited cell growth at all the concentrations tested. In the in vitro HDAC
activity assay, we found that neither EGCG nor the combination significantly inhibited HDAC activity.
32 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Quorum Sensing Molecule Identification in Mycobacterium Tuberculosis
Taylor G. Maloney, Ronak Tilvawala, Preeti Sule, Suat L. G. Cirillo, Jeffrey D. Cirillo
Department of Microbial Pathogenesis and Immunology
Texas A & M University Health Science Center College of Medicine, College Station, TX
Tuberculosis (TB) is the most common cause of mortality worldwide from a single bacterial infectious agent.
TB is due to airborne transmission of the acid fast, non-motile bacillus, Mycobacterium tuberculosis (MTB).
To compound this global pandemic, MTB has the ability to produce a surface that is rich in mycolic acids that
shields MTB from many of our first and/or second-line pharmaceutical treatments, creating drug-resistant
strains of MTB. Multiple drug-resistant TB (MDR-TB) is resistant to the two most useful drugs in our arsenal
against TB, rifampin and isoniazid. Improper treatment or management of patients with MDR-TB has led to
extensively drug-resistant TB (XDR-TB) which is resistant to additional TB treatments, including quinolones.
Furthermore, in India, Iran, and Italy, strains have arisen that are resistant to even more drugs than XDR-TB,
and have coined the term totally drug-resistant tuberculosis (TDR-TB). The current diagnostic tests for TB
are slow and costly, which convolutes our available treatment options. The need for more rapid diagnostics
and tests for TB drug resistance is critical in the prevention of future TB epidemics. The aim in this study is to
determine the structure of the autoinducer molecules produced by MTB that are involved in quorum sensing.
Quorum sensing in bacteria utilizes autoinducer molecules that regulate gene expression in response to
2
changes in bacterial population densities. Our project utilized an avirulent strain of MTB, mc 7000, to
produce two specific autoinducer molecules, Mycobacteria autoinducer 1 and 2 (MAI-1 and MAI-2,
2
respectively). MAI-1 and MAI-2 were collected from the mc 7000 strains, filtered, extracted with hexane,
dried with sodium chloride and sodium sulfate, and then isolated by high-performance liquid chromatography
(HPLC). Autoinducer molecules can sometimes signal across species, so in order to show that the isolated
peaks from HPLC contained autoinducers, we performed bioassays utilizing Streptomyces species. The
bioassay test showed that MAI-1 and MAI-2 could induce Streptomyces coelicolor to produce an antibiotic
2
that was easily identifiable by its bright red color. Thus far, we have extracted 690L of mc 7000 supernatant
and are working towards our goal of extracting 1,000L of supernatant. We are currently determining the
structure of MAI-1 and MAI-2 by NMR, IR, and mass spectrometry. It is our belief that obtaining this structure
will prove beneficial in the development of faster, more sensitive, and less expensive diagnostic tests as well
as advancing the field of pharmaceutical treatments in the fight against TB.
33 | P a g e
Smooth Muscle Gamma Actin Expression in Prostate Cancer Cells
Brandon McGhie, Georgina Kolcun, Warren Zimmer
Department of Physiology, College of Medicine
Texas A & M University Health Science Center College of Medicine, College Station
Prostate cancer is the most prevalent cancer and the second leading cause of cancer-related
death in men. The prostate, a male reproductive organ, functions mainly to secrete a slightly alkaline
fluid to aid the spermatozoa in the female vaginal tract. Some people with prostate cancer may be
asymptomatic, but those who are symptomatic may exhibit polyuria, hematuria, nocturia, and
dysuria. The most common screening methods are the digital rectal examination and prostatespecific antigen blood test. PSA is seen as controversial due to over diagnosing and possible
harmful consequences in some people.
Smooth muscle gamma actin (SMGA) is the major contributor in enteric smooth muscle cells and
composes microfilaments found in cytoskeleton and the contractile apparatus. Interestingly,
preliminary research done within the Zimmer lab has shown that smooth muscle gamma actin was
expressed in prostate epithelia. The previous results showed that the SMGA found in cancerous
prostate epithelia (PC-3) is different from the protein found in enteric smooth muscle cells. An
antibody that specifically binds to gamma actin at its amino terminus in smooth muscle cells was
used to probe cancerous prostate epithelial cells. No binding took place within the prostate, so an
antibody that can bind to an epitope of the 4 muscle actins was used and successfully bound to an
actin in the lysate. Now that it is known how to find muscle actin, we will now be able to (A)
localize smooth muscle gamma actin within the prostate cancer epithelial cells and (B) to sequence
the protein to find its amino terminal structure.
The results showed that the SMGA found in the prostate epithelia is localized in the cytoplasm,
not in cytoskeleton filaments, meaning that it most likely does not have any functions as support for
the cells. We will use Immunoprecipitation to further purify the actin contained within the lysates
and then use Mass Spectroscopy to find the amino terminus of the actin in the PC-3 cells to identify
which specific gamma actin is being expressed.
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August 5, 2014: 9:00 AM - 2:00 PM
Differential Vulnerability of Hippocampal GABA-ergic Interneuron Subpopulations in an Animal Model
of Gulf War Illness
T. Megahed, B. Hattiangady, B. Shuai, and A. K. Shetty
Institute of Regenerative Medicine and Department of Molecular and Cellular Medicine, TAMHSC
Texas A&M University Health Science Center College of Medicine, Temple Campus
Introduction: Approximately 250,000 out of the 697,000 of U.S. Gulf War Veterans suffer from Gulf War
Illness (GWI), a chronic illness affecting multiple symptoms including the central nervous system. Both animal
model and epidemiological investigations have indicated that these impairments in a majority of GW veterans
are linked to exposures to chemicals such as Pyridostigmine bromide (PB, an anti-nerve gas drug),
Permethrin (PM, an insecticide) and DEET (a mosquito repellant) encountered during the Persian Gulf War-1.
Our previous study in a rat model has shown that combined exposures to low doses of GWI- related (GWIR)
chemicals PB, PM, and DEET with or without 5-min of restraint stress (a mild stress paradigm) causes
hippocampus impairments in spatial and object recognition memory and increased depressive-like behavior
(Parihar et al.,2013; Hattiangady et al, 2014). Hippocampal interneurons synthesizing the inhibitory
neurotransmitter GABA play an important role in modulating hippocampal excitability, neurogenesis, and
cognitive and mood function. We quantified three subpopulations of interneurons (PV, NPY and SS) in the
hippocampus of animals exhibiting GWI-like symptoms.
Hypothesis: We hypothesized that cognitive impairments and declined neurogenesis seen in GWI is linked to
reductions in one or more of these subpopulations of GABA-ergic interneurons in the hippocampus.
Methods: Starting with two groups of rats, one group was exposed daily to a combination of chemicals DEET
(40 mg/kg/day), permethrin (0.13 mg/kg/day) and PB (1.3 mg/kg/day) & 5-minutes of restraint stress (i.e.
PCMS) for 28 days. The other group was not exposed to the above conditions with everything else remaining
constant. Three months after the above exposure regime, all rats underwent perfusion and GABAergic
interneurons were stained with Parvalbumin, Neuropeptide Y, and Somatostatin. Using light microscopy,
optical fractionator quantification (Stereo Investigator software: Microbrightfield) of PV, NPY and SS+
interneurons was used to quantify the GABAergic interneurons in two different regions of the hippocampus:
dentate gyrus and CA1+CA3. A student t-test was done for statistical analysis.
Results: Exposure to GWI-related chemicals (DEET, Permethrin & PB) and stress causes loss of
hippocampal parvalbumin (PV) and neuropeptide Y (NPY) positive GABA-ergic interneurons. Considering the
purported function of PV- and NPY- positive interneurons in the hippocampus, cognitive impairments and
declined hippocampal stem cell activity seen in GWI may be linked to reduced numbers of PV+ and NPY+
interneurons in the hippocampus.
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Depletion of CLIP Decreases Toll-Like Receptor-Induced Preeclampsia in Mice
Moheb Milad, Valorie Chiasson, Piyali Chatterjee, Olga Gasheva,
Kelsey Foster, Marcos Hernandez, Brett Mitchell
Texas A & M University Health Science Center College of Medicine, Temple TX
Introduction: Preeclampsia (PE) is a disease of pregnancy characterized by hypertension and proteinuria or
end organ damage in previously normotensive women. It affects 5-8% of all pregnancies and is responsible
for about 18% of maternal deaths. There is no treatment for PE except for the immediate delivery of the child,
often resulting in premature births and its associated complications.
We reported that activated Toll-like receptors (TLRs) are associated with PE in women. Activation of
TLR7 by ssRNA or TLR3 by dsRNA in pregnant mice leads to a pro-inflammatory immune response which
contributes to the elevation of blood pressure.
It has also been found that TLR activation leads to the expression of Class II-Associated Invariant
Peptide (CLIP) on γδ T cells, an important regulator of the immune system. The binding of CLIP to MHC
Class II receptors of these cells prevents their death and leads to persistent immune cell activation, which in
turn leads to the symptoms of PE.
We used a newly designed peptide (TPP) to eliminate CLIP binding. This is possible because TPP
has a higher affinity for the MHC Class II binding groove than CLIP. We wanted to determine whether TPP
can prevent and treat TLR-induced PE in mice. We also utilized CLIP KO mice to determine if they are
resistant to TLR-induced PE.
Hypothesis: It was our hypothesis that the presence of CLIP in the MHC class II binding groove of γδ T cells
is responsible for mediating TLR-induced inflammation and PE in mice.
Methods: To create our animal model, pregnant WT and CLIP KO mice were injected IP on gestational days
13, 15 and 17 with vehicle (Saline), a TLR3 agonist (Poly I:C), or a TLR7 agonist (R837) to induce PE. Some
WT mice were also treated with TPP on gestational days 13, 15 and 17 to see if this would prevent the
development of PE. Other WT mice were treated with TPP on gestational days 15-17 after PE was
established to see if this would treat PE. All mice were then euthanized on gestational day 18.
Blood pressure was measured on gestational days 13, 15 and 17 by tail-cuff plethysmography. Flow
cytometry was used to measure the splenic levels of CD3+/γδ+ T cells. Endothelial function was measured in
aortic rings and their responsiveness to acetylcholine or sodium nitroprusside. Western blots were run to
measure fibronectin and E-selectin, markers of endothelial injury and inflammation respectively. Aortic levels
of fibronectin and E-selectin were assessed using primary antibodies followed by secondary antibodies
conjugated to fluorescent dyes. A LiCor Odyssey scanner was used to image and quantitate the blots and
levels were normalized to actin.
Results: Pharmacologic and genetic depletion of CLIP normalized blood pressure, lowered levels of
CD3+/γδ+ T cells, prevented endothelial dysfunction, and decreased endothelial injury and inflammation.
Conclusions: The results of the experiment suggest that CLIP plays a significant role in the development of
TLR-induced PE. The results also show the potential that TPP has for the treatment of preeclampsia. We
found TPP to be effective as both a preventative measure and a treatment for TLR-induced PE in mice.
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August 5, 2014: 9:00 AM - 2:00 PM
Human Basophil Degranulation Mediated by the Anaphylatoxin C5a is IL3-dependent
M. Mohiuddin, D. Oweis, G. Tavana, P. Moore, DP Huston
Texas A & M University Health Science Center College of Medicine, Houston Campus
Introduction: Basophils make up 1-3% of peripheral leukocyte population and are important in the
pathogenesis of anaphylaxis and allergies. Upon activation, they can degranulate and release histamine,
leukotrienes, cytokines, and various proteolytic enzymes. Degranulation can occur upon allergen-induced
cross-linking of IgE bound to FcϵRI on basophils or upon stimulation by the anaphylatoxin, C5a, which is
generated during complement activation. Recent studies in this lab have demonstrated that FcϵRI-mediated
basophil degranulation is IL3 dependent.
Herein, we investigated the hypothesis that C5a-induced basophil degranulation is dependent on IL3.
Background: The C5a receptor is a seven transmembrane domain G-protein coupled receptor expressed on
the plasma membrane. Upon stimulation of the receptor, intracellular calcium is mobilized and PI3Kinase,
MAP Kinase, and ERK signaling paths are activated. The high affinity FcϵR expressed on basophils is a
tetrameric complex with one alpha, one beta, and two gamma chains, and when it is cross linked can cause
basophil degranulation with release of inflammatory mediators such as histamine. Lyn is activated upon FcϵRI
crosslinking and phosphorylates tyrosine resides on the FCϵRI gamma chains, subsequently recruiting syk.
Syk phosphorylation will activate other molecules The IL3 receptor is comprised of two peptide chains - an
alpha and common beta chain that it shares with IL5 and GM-CSF. With ligand binding, the two peptides form
a heterodimer. JAK activation causes phosphorylation of tyrosine residues on the common beta chain
creating further signal transduction of STATs and further intracellular proliferation and differentiation.
Methods: To test the hypothesis, basophils were isolated from human peripheral blood mononuclear cells
(PBMCs) by negative selection in a Robosep. Basophil purity was assessed through flow cytometry staining
for co-expression of CD123 (IL3Rα) and FcϵRI. Purified basophils were cultured for 24 hours in RPMI media
with or without recombinant human IL3 before stimulation the anaphylatoxin C5a, or the monoclonal antibody,
CRA-1, to crosslink FcϵRI. Basophil activation was determined by expression of the marker CD69 and
basophil degranulation by the CD63 marker.
Results: C5a alone did not activate or degranulate basophils, but caused robust degranulation of IL-3
activated basophils. Antibodies against the IL3 receptor or IL3 showed inhibition of basophil activation and IL3 plus C5a induced degranulation. Similar results were observed for FcϵRI crosslinking. These studies are the
first to demonstrate that addition of IL3 is necessary for activation and degranulation of basophils in response
to the anaphylatoxin C5a. IL3-activated basophils were a homogenous population of cells that degranulated in
the presence of C5a after IL-3 activation.
Discussion and Conclusions: CRA1 alone does not induce basophil-mediated activation. C5a alone does
not induce basophil-mediated activation. IL3 alone induces basophil-mediated activation. C5a does not
enhance IL-3 activation of basophils. Anti-IL3 and anti-IL3R mAb inhibit IL3-induced basophil activation.
Basophil activation is IL3-dependent. CRA1 alone does not induce basophil-mediated degranulation. C5a
alone does not induce basophil-mediated degranulation. IL3 alone minimally induces basophil degranulation.
IL3 and C5a induce basophil degranulation. IL-3 and CRA1 induce basophil degranulation. Anti-IL3 or antiIL3R mAb block IL3 plus either C5a or CRA1induced basophil degranulation. Basophil degranulation is IL3dependent.These studies demonstrate the potential use of IL3 inhibition for clinical therapy against C5amediated anaphylactoid reactions from complement activation, as well as anaphylactic responses to
allergens.
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August 5, 2014: 9:00 AM - 2:00 PM
3-Bromopyruvate Stresses Mouse GBM Cells to Express MHC Class-II in vitro
Yunib Munir; Sanjib Mukherjee; Sören Singel, M.D.; M. Karen Newell-Rogers, Ph.D.
Center for Cell Death and Differentiation, Department of Surgery
Texas A & M University Health Science Center College of Medicine, Temple Campus
Introduction: Glioblastoma Multiforme (GBM) is a highly malignant brain tumor of the astrocyte. With current
treatment the median survival of GBM patients is less than one year. Although the mechanism by which
these cancer cells evade detection by the immune system is unknown, this study evaluates how a metabolic
inhibitor, 3-Bromopyruvate (3-BP), can alter the expression of two important proteins involved in expressing
foreign antigens on the cell surface: Major Histocompatibility Complex Class II (MHC-II) and Class IIassociated Invariant Chain Peptide (CLIP). In this study, we show that treatment of mouse GBM cells (cell line
261) in vitro with 3-BP leads to an upregulation of MHC-II expression on the cell surface with no change in
CLIP expression. This ability to increase MHC-II expression while stressed may provide a treatment approach
for GBM by allowing tumor cells to express antigens and become detectable by the immune system.
Hypothesis: 3-Bromopyruvate will upregulate CLIP and MHC Class II expression in the mouse model of
GBM in vitro
Methods: Using cell line 261, a mouse model of GBM, a Proliferation Assay was performed to determine the
IC-50 of 3-BP on 261 cells. A 96-well flat-bottom plate was utilized; each well was filled with 2500 cells / 100
µL 10% RPMI and then treated with 50 µM, 25 µM, 12.5 µM, 6.25 µM, 3.13 µM, 1.56 µM, 0.78 µM, and 0 µM
concentrations of 3-BP for 72 hours. The cells were then treated with 125 µL of assay solution (5 mL 3X XTT
Reagent, 100 µL XTT Activator, 10 mL warm RPMI) and run through a plate reader to determine the optical
density for each treatment. Using that data, 150,000 cells / 2 mL 10% RPMI were plated into each well of a 6well plate and treated with 25 µM 3-BP for 24 hours and 48 hours. After treatment, we stained the cells using
anti-mouse antibodies for MHC Class-II and CLIP and ran them through the Flow Cytometer to determine the
level of staining, indicating the level of MHC Class-II and CLIP expression.
Results: From the Proliferation Assay we determined that the IC-50 of 3-Bromopyruvate on cell line 261 is
around 25 µM. Using this concentration of 3-BP to stress the mouse GBM cells, we found that cell line 261
upregulates MHC-II expression with no change in the expression of CLIP, relative to no treatment. This
increase in MHC-II expression suggests that stressed cells may be able to process and present foreign
antigen, thereby preventing those tumor cells from hiding from the immune system. This may further suggest
the importance of localized immunotherapy in treating GBM.
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August 5, 2014: 9:00 AM - 2:00 PM
Sensitivity and Specificity of a New Clinical Test for Anterior Cruciate Ligament Tears
1
2
Heather Naumann , Patrick McCulloch, MD and Patrick Massey, MD
1
2
Texas A&M College of Medicine , Houston Methodist Hospital Orthopedic Sports Medicine Surgery
2
Introduction: One of the most common surgeries performed by Sports Medicine Orthopedic Surgeons is
Anterior Cruciate Ligament (ACL) reconstruction. There are currently other tests used to diagnose ACL tears
in the clinic, such as Anterior Drawer, Lachman and Pivot Shift. The previously stated tests have different
specificities and sensitivities and we plan to perform the new test in the clinic, pre-operatively and postoperatively and calculate the sensitivity and specificity to compare to the tests already in use.
Hypothesis: The new Lelli test will have a higher sensitivity and specificity than the current tests making it a
better tool to diagnose ACL tears.
Methods: Patients will be evaluated in the clinic, pre-operatively and post-operatively. The inclusion criteria
for this study will be patients who report instability of their knee presenting after a twisting or contact injury to
their knee, unable to bear weight after the injury and have experienced significant swelling post-injury. The
exclusion criterion includes patients with multiple ligament tears, arthritis and any previous ACL surgery.
Patients will be examined with the standard ACL tests including: Lachman, Anterior Drawer, and Pivot
Shift. In addition, we will perform a new test, The Lelli Test, as follows: the patient will lay supine on an exam
table without shoes and a plastic board will be placed under the patient to provide a firm surface. While
testing the right knee, the examiner’s right hand in the form of a fist will be placed under the patient’s lower
right leg at the tibial tuberosity with the proximal side of the fist lined up with the most distal portion of the tibial
tuberosity. The examiner’s left hand will push from anterior to posterior on the femur cephalad to the
supracondylar region. If the patient’s heel passively rises off the board this is a negative test, while if the foot
does not rise passively this is a positive test indicating an ACL tear. The examiner will move to the patient’s
left side and evaluate the knee using the same 4 ACL tests. The results of all four ACL tests from both the
injured and uninjured knees will be recorded on the Lelli form. The new test will be recorded, as either
positive, if the foot does not come off the table with applied force, or negative, if the foot comes off the
table. In addition, the height of the foot displacement during the Lelli Test will be measured with a ruler and
recorded. A MRI will officially make a positive diagnosis of an ACL tear with subsequent arthroscopy, likewise
a negative diagnosis of an ACL tear will be made by MRI. Based on our findings we will calculate the
sensitivity and specificity of all 4 tests.
Results: The preliminary data from the first 8 patients is inconclusive. However, so far the data indicates
that the new test is of at least equal caliber to the other 3 standard tests in diagnosing an ACL tear. The gold
standard used in diagnosing the ACL tear is the MRI. The sensitivities and specificities of the 4 tests are
relatively the same at this point. As more patients are enrolled and tested, more discrepancies may be seen.
So far, the specificity of 0.75 and a sensitivity of 1.0 are seen with the Lelli, Lachman and Anterior Drawer
tests. The pivot Shift had a higher specificity of 1 and a sensitivity of 0.75.
Conclusion: This clinical study submitted for IRB approval is still currently in the process of enrolling
patients. The preliminary data from the first 8 patients shows that the 4 tests have similar findings, but with
the enrollment of more patients, hopefully 50 or more, more discrepancies in the statistics will be seen. As of
now, the Lelli test is not better or worse than the 3 standard tests currently in use.
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August 5, 2014: 9:00 AM - 2:00 PM
Evaluation of Two Neurosurgical Approaches to Cervical Spondylotic Myelopathy: Posterior and
Combined Posterior-Anterior
Quoc-Bao Nguyen, Jonathan A. Friedman, MD
The Texas Brain and Spine Institute, Bryan, Texas
Texas A & M University Health Science Center College of Medicine, Temple, Texas
Introduction: Cervical spondylotic myelopathy (CSM) is a progressive degenerative cervical spine disease
and a frequent cause of spinal dysfunction in patients older than 55 years of age. This disease can be
severely debilitating—potentially advancing to permanent paralysis, and even death. Treatments for CSM can
be divided into (1) conservative management, such as prolonged immobilization and anti-inflammatory
medications, and (2) surgical treatment for decompression. The current repertoire of surgical approaches for
CSM are: (1) anterior approaches, (2) posterior approaches, and (3) combined approach. However, the
optimal approach still remains controversial despite several factors that have been established to assist in
deciding on an approach.
Hypothesis: The combined anterior-posterior approach to cervical spondylotic myelopathy has better clinical
outcome than the posterior approach.
Methods: A systematic retrospective study of a population of 115 patients was performed for this study. In
order to compile all pertinent data, a patient database was compiled from St. Joseph’s Citrix server using
grouped terms for cervical myelopathy. The selected patients’ data was judiciously extracted based on
relevance to this study. After the proper data was gathered, we quantified patients’ cervical myelopathy
symptoms based on the McCormick Classification system. The clinical outcomes of the posterior approach
were then compared against those of the combined anterior-posterior approach using T-test and distribution
comparisons.
Results: The combined anterior-posterior approach demonstrates a statistically significant improvement in
cervical myelopathy recovery as compared to the posterior approach. In addition, combined anterior-posterior
approach shows a decrease in likelihood for additional myelopathy surgeries as compared to the posterior
approach. The results were unexpected because the selection bias in this study would suggest worse
outcomes for combined anterior-posterior patients. Patients undergoing the combined anterior-posterior
surgery generally have worse pre-operative clinical myelopathy and/or worse structural imaging findings. Our
study suggests that the combined anterior-posterior approach to cervical myelopathy has less complications
than the posterior approach. Utilizing the combined anterior-posterior approach could potentially improve the
patient’s surgical outcome and lower costs and time.
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August 5, 2014: 9:00 AM - 2:00 PM
Pupillary Reactivity to Cocaine Cues in Cocaine-Dependent Patients
Lillian Niakan, Ricky Savjani, David Eagleman, Ph.D.
Departments of Neuroscience and Psychiatry
Baylor College of Medicine, Houston, Texas
Texas A & M University Health Science Center College of Medicine, Houston, Texas
Introduction:
Processes other than the light reflex or a response to the sympathetic nervous system can
influence pupillary reactivity. Studies have shown that pupil dilation increases as cognitive load, arousal,
or attention allocated to a task increases. Our study investigates the reward system’s influence on
pupillary reactivity and its potential at accurately indexing cocaine craving in cocaine-dependent patients.
Hypothesis:
We hypothesize that participants dependent on cocaine will (1) yield a greater pupillary response
to cocaine images and that (2) the regions of the brain driving the pupillary response will be associated
with cocaine craving regions of the brain.
Methods:
Participants will perform a 3-stimuli, visual oddball task. Novel cocaine (target) images, natural
(non-target) images, and phase-scrambled (standard) images will be presented. There will be 4 runs of
the experiment. No images will be repeated between or within runs. Target stimuli detection is recorded
via left button presses while non-target stimuli detection is recorded via right button presses on a twobutton MRI-compatible button box. All stimuli will be presented for approximately 250 ms and have
been pseudorandomly dispersed throughout each run. The order in which each participant observes the
runs has also been randomized; however, each participant will have seen the same 4 runs by the end of
the oddball task experiment. Target stimuli will make up 25% of the total trials and non-target stimuli
will make up 25% of the total trials, as well. Luminance across all images has been equalized to control
for light reflex influence on the measured pupillary reactivity. The inter-stimulus interval has been varied
between 3 s and 6 s with a minimum inter-target interval of 6.5 s. The entire oddball task will take
approximately 30 minutes.
Prior to the start of the oddball task, participants will lie supine in the magnetic resonance
imaging (MRI) scanner as a high-resolution structural scan is acquired using a 32-channel head coil.
Functional MRI (fMRI) data will then be acquired throughout the oddball task for each participant.
Pupillary activity will be continuously recorded from the right eye using an MR safe camera –
LiveTrack AV for fMRI – with an infrared illumination source. The camera will sit directly behind the
Siemens 32 channel cold mirror that participants will look at in order to perform the visual oddball task
displayed on a CRS Boldscreen, inside the MRI scanner. Acquired data will be preprocessed to account
for gaps in data due to blinks or noisy and unreliable data. Linear interpolation calculations will be used
to correct for gaps in data due to eye blinks.
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August 5, 2014: 9:00 AM - 2:00 PM
H2O2 Induces HEK Cell Apoptosis By Disrupting the Interaction Between p38 and MKP-1.
Brennan Ochoa, Hao Feng, and David E. Dostal
Department of Molecular Cardiology
Introduction and Background: The stress-activated protein kinase (MAPK), p38 plays a major role in transmitting
extracellular signals from the cellular membrane to the nucleus, which leads to the phosphorylation of several transcription
factors and the regulation of genes involved in the control of many different cellular processes including survival,
proliferation, differentiation, apoptosis, and metabolism. MAPK phosphatases (MKPs) are responsible for deactivating
MAPKs through dephosphorylation of specific tyrosine and threonine residues. MKP-1 is the major member of MKPs that
inactivates MAPKs and has been implicated in the regulation of p38 and/or JNK signaling responses under a wide variety
.
of stress conditions Reactive oxygen species (ROS) are produced as a normal part of cellular metabolism and have the
potential to cause cellular damage. The ROS hydrogen peroxide (H 2O2) is a major contributor of oxidative damage within
the cell. It has previously been reported that H2O2 induces apoptosis through ASK1-MKK3/6-p38 signal cascade. In this
study we aimed to examine the effects of ROS on the up and downstream interactions of p38 leading to apoptosis.
Rationale and Hypotheses: Our lab has observed that p38 negatively regulates JNK phosphorylation through MKP-1
activation. While other phosphatases may be involved, only MPK-1 appears to be involved in p38 induced JNK dephosphorylation. This has led to the following hypotheses:
1.
2.
3.
4.
ROS induces apoptosis in HEK cells.
p38 and MKP-1 interact with each other.
180
182
The activation loop phosphorylation sites (Thr
and Tyr ) on p38 are necessary for p38 and MKP-1 to
interact.
The interaction between p38 and MKP-1 is related to the activity of p38.
Methods: To determine the temporal effects of ROS on apoptosis, HEK cells were treated with 100 or 500 µm H 2O2 for 15
or 60 minutes, followed by staining with annexin V and analysis with flow cytometry. To detect p38 activity induced by
H2O2, HEK cells were treated with 100 µm H2O2 for 15 or 60 minutes with or without NQDI (ASK-1 inhibitor). Protein
samples were then collected and Western blot analysis was used to quantify levels of P-p38, total-p38, P-MKK3, and
total-MKK3. HEK cell lines that overexpress p38 were generated by transfecting HEK cells with FLAG-tagged wild-type
p38 or FLAG- tagged dominant-negative p38 plasmid expression vectors. A cell line without any transfection was used as
a control. Co-immunoprecipitation (co-IP) studies were performed using WT p38 and DN p38 overexpressed cell lines
treated with 100 µm H2O2 for 15 minutes. After protein samples were collected, IP was performed using anti-FLAG
antibody conjugated to protein-A beads. Western blot analysis was performed to quantify expression levels of FLAG, p38,
and MKP-1, using respective antibodies.
Results:
1. H2O2 induces apoptosis in HEK cells that peaks at 15 minutes, with some recovery at 60 minutes.
2. H2O2 induces peak phosphorylation of p38 and MKK3 at 15 minutes.
3. Protein complexes occur between p38 and MKP-1.
180
182
4. Phosphorylation of p38 residues Thr and Tyr are not required for interaction with MKP-1.
5. H2O2 disrupts protein interactions between p38 and MKP-1.
Conclusion: In this study, ROS induced apoptosis in HEK cells was associated with increased p38 activation
(phosphorylation). ROS treatment was found to increase activation of MKK3, an upstream activator of p38, as well as
block the interaction of p38 with the inactivating phosphatase, MKP-1. Furthermore, interactions between p38 and MKP-1
180
182
did not require phosphorylation of p38 residues Thr
and Tyr , suggesting that MKP-1 forms a protein complex with
p38 before its activation. This study aimed to identify a potential clinically significant signaling cascade leading to
apoptosis in HEK cells that, when further confirmed in cardiac myocytes could open new avenues of treatment for
maladies related to cell death, like heart failure and ischemic injury.
42 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Regulation of the Renin-Angiotensin System in the Diabetic Heart
Authors: John R. Ogden, Qian Chen Yong, Kenneth Baker, Rajesh Kumar
Texas A & M University Health Science Center College of Medicine-Temple
Introduction: Diabetic cardiomyopathy is a major contributing factor to morbidity in diabetics. This disease
process occurs largely due to the dysregulation of the renin-angiotensin system (RAS). ACE inhibitors and
angiotensin type 1 receptor (AT1) blockers have proven effective in helping to regulate the systemic RAS.
However, recent studies show that there is a local, intracellular RAS (iRAS) which is not regulated by these
drugs. The cardiac iRAS in diabetic cardiomyopathy has been proven clinically to play a significant role in the
disease process. Several metabolic pathways have emerged as key regulators of signal transduction and
pathological mechanism of diabetic cardiomyopathy. High glucose (HG) conditions result in increased activity
of the hexosamine biosynthesis pathway (HBP). This increased activity results in excessive O-glycosylation,
and thus activation, of various transcription factors which may lead to increased iRAS activity. On the other
hand, hydrogen sulfide (H2S) was found to inhibit plasma renin activity, suggesting that cardiac H2S may
have a role in RAS activation in diabetic hearts.
Hypothesis: Hyperglycemia causes activation of HBP and reduction of H2S synthesis, resulting in the
intracellular RAS activation in the heart.
Methods: We treated neonatal rat ventricular myocytes (NRVMs) with normal glucose (NG), high glucose
(HG), with or without benzyl N-acetyl-alpha-D-galactosamine (BAG, an inhibitor of HBP), and PUGNAc (an
activator of HBP) for 24 hours. Protein and mRNA expressions of angiotensinogen (AGT), renin, and AT1
receptor in NRVMs were detected using western blot and real-time PCR, respectively. An H2S-specific probe,
SF-7-AM, will be used to measure H2S levels in NRVMs exposed to NG, HG, and DL-propargylglycine (PAG,
an inhibitor of H2S-synthesis enzyme), in which the protein and mRNA expression of the RAS-related protein
were also measured.
Results: PUGNAc increased AGT expression in NRVM, whereas HG treatment increased the expression of
AGT, renin and AT1 receptor, which were blocked by BAG. Hyperglycemic conditions inhibited H 2S
production in NRVMs; inhibition of cardiac H2S synthesis enzymes using PAG or siRNA increased AGT
expression in NVRMs cultured in normal glucose medium. Moreover, intracellular Ang II production generates
a positive feedback loop resulting in further production of AGT, renin and AT1. These data suggest that
activation of HBP and inhibition of H2S synthesis result in activation of the iRAS in NRVM; intracellular Ang II
further activates the iRAS, resulting in a sustained activation of the iRAS in the heart. Suppression of iRAS
activation using HBP inhibitors, H2S donors or direct renin inhibitors may provide potential treatment for
diabetic cardiomyopathy.
43 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Human basophil degranulation by bacterial-derived N-formylated peptides is IL-3
dependent
D. Oweis, M. Mohiuddin, G.Tavana, P.Moore, D.P.Huston
Texas A & M University Health Science Center College of Medicine, Houston
Introduction and Hypothesis
Basophils are the least abundant granulocytes in human peripheral blood ranging from 0.01% - 0.03%. Basophil cytoplasmic
granules contain heparin, histamine, platelet-activating factor (PAF) and other mediators of the immediate hypersensitivity response
causing allergies and anaphylaxis. These granules are released when the high affinity IgE Fc receptor (FcεRI) on the cell surface are
cross-linked. In addition to FcεRI crosslinking, basophils can degranulate in response to bacteria-derived N-formylated peptides
(formymethionine leucyl phenylalanine;fmet-leu-phe).
Recent studies from this lab have demonstrated the requirement for IL-3 activation of human basophils before they can
degranulate in response to FcεRI crosslinking. Herein, we investigated the hypothesis that N-formylated peptide-induced
basophil degranulation is dependent on IL-3 activation
Methods
Basophils were purified from peripheral blood by negative selection and cultured in media with or without IL-3 prior to
stimulation with fmet-leu-phe. Control cultures were stimulated with the FcεRI crosslinking mAb (CRA-1). Basophil purity was
confirmed by flow cytometry staining for co-expression of CD123 (IL-3Rα) and FcεRI. Basophil activation was demonstrated by
expression of CD69. Basophil degranulation was demonstrated by expression of CD63. IL-3 dependence of basophil activation and
degranulation was confirmed with anti-IL-3 and anti-IL3-R neutralizing antibodies.
Results and Conclusion
This study demonstrates that human basophils are activated by the cytokine IL-3. Furthermore, basophil degranulation in
response to either f-met-leu-phe or FcεRI crosslinking is dependent on prior IL-3 activation. Fmet-leu-phe alone could induce basophil
degranulation. In addition, there appears to be heterogeneity among IL-3 activated basophils for fmet-leu-phe responsiveness for
degranulation. However, as compared to the ability of IL-3 antagonism (with either anti-IL-3 or anti-IL-3R mAb) to inhibit IL-3dependent basophil degranulation in response to FcεRI crosslinking, IL-3-dependent degranulation induced by fmet-leu-phe is
refractory to antagonism of IL-3. These studies are the first to demonstrate that basophil degranulation in response to bacteria-derived
N-formylated peptide requires IL-3 activation. Furthermore, although all basophils could be activated by IL-3, only a subpopulation of
IL-3 activated basophils degranulated in response to fmet-leu-phe, suggesting heterogeneity among basophils for responsiveness to
fmet-leu-phe
In additional experiments (data not shown) ex-vivo production of IL-3 from T-lymphocytes stimulated with anti-CD3 and
anti-CD28 mAb conjugated beads were initiated. This will allow for future characterization of basophil responses with a model that
mimics in-vivo basophil-mediated inflammatory responses. Overall, this study demonstrates that IL-3 is a potential therapeutic target
to decrease human basophil-mediated allergic reactions to FcεRI crosslinking. In addition, fmet-leu-phe induced degranulation of
basophils is absolutely dependent on IL-3 activation, but only a subpopulation of basophils are susceptible to IL-3 plus fmet-leu-phe –
induced degranulation, and this susceptible subpopulation is refractory to IL-3 antagonism. Future studies need to be conducted in
order to characterize these basophil subpopulations.
Key Conclusions:
1) IL-3 is required for activation of human basophils before they can degranulate in response to FcεRI crosslinking or N-formylated
peptides.
2)Anti-IL-3 and anti-IL-3R neutralizing mAb inhibited IL-3 activation f basophils.
3)Only a subpopulation of IL-3 activated basophils degranulated in response to fmet-leu-phe.
4)Human basophils exhibit functional heterogeneity for responses to fmet-leu-phe
44 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
The Role of the Intracellular Renin Angiotensin System in Diabetic Cardiomyopathy
Deanne L. Phillips, Candice M. Thomas, Amanda L. Roth, Kenneth M. Baker, Rajesh Kumar
Division of Molecular Cardiology; Baylor Scott & White; Central Texas Veterans Health Care System
Texas A & M University Health Science Center College of Medicine, Temple, TX
Introduction: The renin angiotensin system (RAS) consists of systemic, local, and intracellular components
that are regulated independently. Studies from our laboratory have demonstrated that the intracellular RAS
(iRAS) is a major component of RAS activity in the diabetic heart. We reported that the over expression of
intracellular angiotensin II (Ang II) causes cell growth and cardiac hypertrophy. However, the specific role of
iRAS, independent of the extracellular RAS, in diabetic cardiomyopathy remains to be demonstrated. Since
angiotensin receptor blockers (ARBs) and angiotensin converting enzyme (ACE) inhibitors do not block iRAS,
a renin inhibitor, which blocks the iRAS, may provide a better therapeautic outcome in diabetes.
Hypothesis: iRAS contributes to the development of diabetic cardiomyopathy.
Methods: We produced diabetes by streptozotocin in AT 1a knockout (AT1aKO) mice, in which the
internalization and actions of Ang II from the extracellular space are prevented. Wildtype mice were also
studied for comparison. A sub-group of diabetic mice received a renin inhibitor aliskiren (20 mg.kg), AT 1
receptor antagonist valsartan (2mg/kg), or ACE inhibitor benazeprilat (10 mg/kg) by osmotic minipumps. The
doses were based on the effects of these drugs on cardiomyocyte Ang II levels in diabetic mice. Insulin and
vehicle-treated groups were included as controls. Blood glucose and cardiac function (by echocardiography)
were monitored in these animals starting from pre-diabetic stage and every 2 weeks thereafter up to 10
weeks of diabetes. The diastolic cardiac function was measured by mitral annulus flow (E/A ratio) while the
systolic cardiac function was measured by fractional shortening.
Results: Both diastolic and systolic functions were reduced in diabetic wildtype mice and AT 1aKO mice after
10 weeks of diabetes. All three RAS inhibitors equally prevented the development of diabetic cardiomyopathy
in wildtype mice and surprisingly, the AT1aKO mice.
This data demonstated that AT1aKO mice developed cardiac dysfunction to the same degree as wildtype
mice, proving that the iRAS activation is critical for the development of diabetic cardiomyopathy. It also
confirmed that ARBs and ACE inhibitors have off target effects. To be more conclusive about the role of iRAS,
our lab is currently generating heart specific AGT KO mice and AGT/AT 1a KO mice.
45 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Administration of CAP peptide ameliorates Theiler’s
virus-induced seizures
a
b
b
b
b
b
Lance C. Robinson , Megha Bijalwan , Andrew J. Steelman , Luke Bacon , Caleb Gottlich , Colin R. Young ,
d
d
b,c
Richard P. Tobin , M. Karen Newell-Rogers , C. Jane R. Welsh
a
Texas A&M University Health Science Center College of Medicine, College Station Campus
c
Department of Veterinary Integrative Biosciences, Department of Veterinary Pathobiology, College of Veterinary
Medicine, Texas A&M University
d
Center for Cell death and Cell differentiation, Scott and White Hospital, Temple, Texas
Introduction:
1
Epilepsy affects over two million people in the United States. Epilepsy is a neurological condition characterized
by the unpredictable occurrence of repeated seizures, which is the hallmark sign of this disorder and maybe caused by
genetic defects or acquired due to brain injury. Seizures can be classified as generalized, focal, or unclassified.
Generalized seizures can be further divided to absence seizures, tonic-clonic seizures, myoclonic seizures and atonic
seizures. Clinically each of these divisions presents with different symptoms. These symptoms vary due to the disruption
2
of electrical brain activity.
The pathogenesis of epilepsy is not completely understood. Theiler’s murine encephalomyelitis virus (TMEV)
3
infection in C57BL/6 mice serves as a mouse model to study the disease. Symptoms of Epilepsy develop during the first
3
week of infection and have been linked to damage of neurons in the temporal lobe .
The current research represents two experiments in order to explore the treatment and pathology of the disease.
First we explored the potential therapeutic actions of synthetic CAP peptide. CAP is computationally predicted to be a
competitive antagonist for CLIP peptide. In addition, we examined the changes in macrophage infiltration of the CNS in
TMEV-infected mice using flow cytometry.
Hypothesis:
The first hypothesis to be tested was that CAP peptides would be therapeutic in the TMEV model of virusinduced epilepsy and result in the decrease in the occurrence of epilepsy. Using flow cytometry we intended to identify the
major immune cells that participate in the disease process and test the hypothesis that CAP peptide therapy would reduce
CNS infiltration and activation status of inflammatory cells.
Methods:
CAP PEPTIDE ADMINISTRATION
C57BL/6 mice were assigned to one of four experimental groups with 5 mice per group. At 4 weeks of age, all
the mice were infected intracranially with BeAn stain of TMEV - day 0. Group 1 mice also received 1 x i.p. injection of
0.2ml of CAP peptide (day 0), Group 2 received 2 x peptide injections (day 0, day 1), and Group 3 received 3 x injections
(day 0, day 1, day 2). The control group received the i.c. TMEV injection and one i.p. injection of 0.2ml vehicle on day 0.
Mice were weighed and examined daily for signs of epilepsy and scored using the Racine scoring system.
FLOW CYTOMETRY
TMEV-infected and uninfected mice were terminated on day 3 p.i. Flow cytometry was used to examine the
effects of Theiler’s virus infection on cell populations in the CNS. The lymphocytes were isolated from the spleen and
4
5
extracted from the CNS tissues using a percoll gradient . 2x10 cells from each tissue were labeled with specific
4
antibodies to CD11b and CD45 -labeled with fluorescent dyes for 20mins at 4°C . Then cells were washed 2 times with
500μl and re-suspended in 250μl in PBS +2% FBS buffer and tested in a flow cytometer.
Results:
The administrations of varying doses of CAP peptide show a dose dependent response in the alleviation of
TMEV-induced epilepsy. Preliminary results indicate that activated macrophage populations in the CNS are increased by
3 days p.i with TMEV. Future studies include exploring the immune cell response and macrophage infiltration of the CNS
in mice that have been treated with CAP peptides.
Reference:
1. Hirtz D, Thurman DJ, Gwinn-Hardy K, Mohamed M, Chaudhuri AR, Zalutsky R. How common are the “common”
neurological disorders?. Neurology, 2007, 68:326-337
2. Epocrates
Premium.
“Overview
of
Seizure
Disorder”
Updated:
2013-05-23.
https://online.epocrates.com/noFrame/showPage.do?method=diseases&MonographId=112
3. Libbey JE, Kirkman NJ, Smith MCP, Tanaka T, Wilcox KS, White HS, Fujinami RS. Seizures following
picornavirus infection. Epilepsia. 2008, 49:1066-1074.
rd
4. Andrew J. Steelman, Dana D. Dean, Colin R. Young, Roger Smith 3 , Tomas W. Prentice, Mary W. Meagher, C.
Jane R. Welsh. Restraint stress modulates virus specific adaptive immunity during acute Theiler’s virus infection.
Brain, Behavior, and Immunity. 23. 2009. Pg. 830-843.
5. Murphy, K. (2012) Janeway’s Immunobiology. New York, NY: Garland Science
6. Cusick MF, Libbey JE, Patel DC, Doty DJ, Fujinami RS. Inflitrating macrophages are key to the development of
seizures following virus infection. Journal of Virology. 2013 Feb; 87(3): 1849-60.
b
46 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Specific Serotonin Reuptake Inhibitors Promote Cholangiocarcinoma Growth and Serotonin
Production
1
1
1
1
1,2,3
Hope Shin , Matthew McMillin , Cheryl Galindo , Gabriel Frampton , Sharon DeMorrow
1
2
Texas A&M University Health Science Center College of Medicine, Scott and White
3
Memorial Hospital, and Central Texas Veterans Health Care System (Temple)
Introduction: Cholangiocarcinomas, malignant tumors of the intrahepatic and extrahepatic biliary epithelium,
are aggressive cancers with devastating and often fatal consequence. The incidence and mortality of
cholangiocarcinoma have been increasing worldwide over the past two to three decades, and the prognosis
remains poor. CCA tumors have a neuroendrocrine phenotype and respond to neuropeptide Y, substance P,
dopamine, serotonin, and others. We have previously shown in human cell lines and in mice that the biogenic
amines serotonin and dopamine are not only elevated in CCA but also contribute to tumor growth and
progression. However, the expression and activity of serotonin transporters has not yet been explored in
CCA. These transporters are the target of many antidepressant medications, namely specific serotonin
reuptake inhibitors (SSRIs). The prevalence of depression in cancer patients is 3 to 5 times higher than the
general population, yet the effect of SSRIs on serotonin producing cancers such as CCA is unknown.
Hypothesis: We hypothesize that specific serotonin reuptake inhibitors (SSRIs), specifically fluoxetine and
sertraline, increase serotonin levels and have growth-promoting effects on cholangiocarcinoma.
Methods:
 In vivo model: Nude 8-week-old mice were injected with Mz-ChA-1 cholangiocarcinoma cells and
treated with 200 µl of fluoxetine, sertraline, or a saline solution. Tumor volumes were measured three
times a week for 25 days and subsequently excised. Tumors were either flash frozen or fixed in
formalin for molecular analyses, embedded in paraffin, and used for immunohistological staining.
 In vitro model: Six human cholangiocarcinoma cell lines (Mz-ChA-1, HuH-28, HuCC-T1, CCLP1,
SG231, TFK-1) and the human immortalized, nonmalignant cholangiocyte cell line H69 were obtained
from different origins. Serotnin EIA and real-time PCR were subsequently performed.
Results: Tumors injected into the mice that were subsequently treated with sertraline and fluoxetine showed
an increased growth curve over the 25 day time course compared to the control tumors. Moreover, after
excision, the fluoxetine and sertraline treated tumors also showed increased PCNA, a marker for proliferation,
through immunohistochemistry staining. After real-time PCR, the relative mRNA expression for VEGFC,
MMP2, and MMP9 were higher for fluoxetine and sertraline tumors. This demonstrates that SSRI treatment
leads to increased angiogenesis, remodeling and inflammation in the CCA tumors compared to the control.
Future studies will also assess the in vitro model and continue to examine the mechanism of SSRI treatment
on increased cholangiocarcinoma growth.
47 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Complete Inhibition of the SEC/SR axis Triggers Increased Biliary Damage
M. Sohner, A. Martínez, H. Francis, J. Venter, H. Standeford, M. Guerrier, J. Greene, G. Alpini, and S. Glaser
Digestive Disease Research Center, Veterans Administration, Texas A&M Health Science Center College of
Medicine (Temple Campus), Scott and White Healthcare
Introduction:
Secretin (SEC) is a peptide hormone that is secreted by S cells and cholangiocytes. SEC stimulates the
secretion of biliary bicarbonate (HCO3 ), forming a protective umbrella or barrier preventing biliary damage.
Lack of this protective umbrella is postulated to play a role in the pathogenesis of cholestatic livers diseases.
During bile duct ligation (BDL), large (but not small) cholangiocytes proliferate, which is characterized by
increased expression of the secretin receptor (SR). Proliferating cholangiocytes express and secrete VEGF-A
and NGF that stimulate proliferation via autocrine/paracrine mechanisms, which is regulated by Sec through
miR-125b and Let-7a, respectively. Lack of SEC or SR inhibits large cholangiocyte proliferation during
extrahepatic cholestasis induced by BDL. The individual knockdown of SEC or SR results in partial inhibition
of biliary proliferation and slight liver damage.
Hypothesis:
A lack of a the protective bicarbonate umbrella (provided by the SEC/SR axis) in bile ducts leads to increased
hepatobiliary damage
Methods:
In order to investigate the role of the SEC/SR axis in the prevention of biliary damage through the production
of the protective bicarbonate umbrella, a double knockout mouse model was created. SEC knockout (SEC
KO) mice were bred with SR knockout (SR KO); the resulting SEC/SR KO mice were genotyped by RT-PCR.
Fibrosis was evaluated by quantification of Picro Sirius Red staining of tissue sections from WT, SEC KO, SR
KO, and SEC/SR KO mice..
Immunohistochemistry (IHC) for CK-19 on liver sections was used to determine the Intrahepatic Bile Duct
Mass (IBDM). Cholangiocyte proliferation was measured by Western Blot analysis for PCNA and VEGF and
NGF gene expression was quantified by RT-PCR.
The expression of the apoptotic marker, Bax, was determined by RT-PCR and IHC. .
Secretory function was assessed by measuring the expression of CFTR and AE2 by RT-PCR and Western
Blot analysis and measuring bicarbonate levels in bile.
Results:
Biliary Mass, PCNA, VEGF-A, and NGF expression decreased in BDL SEC/SR KO mice compared to the
BDL WT control..
Picro Sirius Red staining increased in SEC/SR KO mice compared to either of the single KO mice (SEC KO
or SR KO).
Bax expression increased in SEC/SR KO mice compared to WT, SEC KO, and SR KO mice. CFTR and AE2
expression decreased in BDL SEC/SR KO mice compared to the BDL WT control. Bicarbonate levels in the
bile also decreased in the BDL SEC/SR KO mice compared to the BDL WT control.
Conclusion:
The complete inhibition of the SEC/SR axis triggers increased biliary damage due to the lack of a functional
SEC/SR/CFTR/Cl-HCO3/AE2 system, which causes a disruption of the protective bicarbonate umbrella
during extrahepatic cholestasis.
48 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
The role of the vagus nerve in the peripheral immune response following TBI
Ryan M. Stone, Sanjib Mukherjee, Richard Tobin, Susannah Rogers, Stephanie Henderson, Heather Motal,
Jessica Kain, Karen Newell-Rogers, Lee Shapiro
Department of Surgery and Neurosurgery
Introduction: Traumatic brain injury (TBI) damages the central nervous system (CNS) and causes acute and chronic
TBI-related syndromes, that severely affect quality of life. However, the systemic contributions to injury and to
consequent syndromes are relatively unexplored. The vagus nerve is thought to be involved in mediating inflammation,
including neuroinflammation, via the “cholinergic anti-inflammatory pathway.” Our preliminary data indicate that
following TBI, inflammation is observed in both the liver and the spleen. TBI also results in activation of the hepatic
acute phase response and bile acid system changes. Additionally, in the spleen, TBI causes expansion of CD3+, CD4+,
CD8+, activated CD4+, γδ and T regulatory cells after 24 hours. MHCII expression is decreased on γδ T cells 24 hours
after TBI. In addition, TBI causes expansion of B cells after 24 hours and a decrease in CLIP associated with MHCII on
B cells. Thus, there are clearly peripheral inflammatory consequences following a head injury. Previous studies have
indicated that acetylcholine (ACH), via the cholinergic vagus inputs, can mediate splenic lymphocytes and hepatic
inflammation (Rosas et al. 2011, Ji et al. 2013). However, the influence of the cholinergic vagus input into the spleen on
immune cell activation after TBI remains unknown. Understanding the role of parasympathetic cholinergic input to the
liver and spleen following TBI may enable new therapeutic targets to be identified.
Specific Aim: To determine if parasympathetic cholinergic input into the spleen mediates the effects on T cells and/or B
cells after traumatic brain injury (TBI).
Hypothesis: Vagotomized mice will have altered peripheral immune response as measured by number of immune cells
in the spleen.
Methods: Vagotomy – Vagotomy was performed on mice three days prior to FPI (n = 3). A small incision was made on
the ventral surface of the neck. The SCM was retracted, and the carotid sheath was penetrated. The vagus nerve was
isolated from the jugular vein and carotid artery, and cut. Fluid percussion injury (FPI): In order to induce a TBI, we
induced a FPI on mice, as previously described (Mukherjee et al. 2013). Tissue collection: Following FPI, spleens were
collected at 24 and 72 hours. Flow cytometry: Expansion of CD3+, CD19+, activated CD19+, γδ and T regulatory cells
was measured using flow cytometry.
Results: Our results indicate a statistically significant decrease in activated B cells and T regulatory cells in vagotomized
mice 24 hours post-FPI. Also, there is a trend for a decrease in peripheral inflammatory cells in vagotomized mice 24
hours post-FPI, and an increase in peripheral inflammatory cells in vagotomized mice 72 hours post-FPI. Analysis of the
liver and serum is ongoing.
49 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Analyzing a racially disparate polymorphism and its role in early onset breast cancer in AfricanAmerican women
Rose Thomas, Isabel Lambertz & Robin Fuchs-Young
Texas A & M University Health Science Center College of Medicine, (College Station)
Introduction: African Americans (AAs) women are significantly more likely than people of European
descent to develop early onset breast cancer (EOBC) that is often a triple negative (ER-, PR-, Her2-) breast
cancer with fewer treatment options and a worse prognosis. The mechanistic basis for this persistent health
disparity is largely unexplored, but it is highly likely that genetic susceptibilities contribute, both independently
and in collaboration with social, economic and environmental factors. EOBC is likely affected by etiologic
influences earlier in life, while late onset reflects accumulated lifetime exposures to environmental factors.
One possible contributor is the fact that the frequency of the P (proline) allele of codon 72 in the p53 gene of
AA women is increased, and this allele has been shown to be associated with increased incidence and early
onset of a variety of cancers, including those of the breast. We will determine the p53 genotype in AA breast
cancer patients and assess whether the P allele is disproportionately expressed in those with early onset
(premenopausal) disease. In addition, we will use an existing database and tumor bank to investigate the
interaction of p53 polymorphisms and other risk factors, including early parity and body weight, on the risk of
EOBC. In order to develop a genotyping protocol for these samples, we used samples from a mouse model
containing a “humanized” p53 gene, where the murine exon 4 was replaced with a human exon 4 encoding
either an R or a P at codon 72, resulting in the generation of a chimeric p53 mouse.
Hypothesis: A racially disparate SNP in position 72 of the p53 protein, encoding either a proline or
arginine, may be a possible contributor to increased susceptibility to EOBC in African-Americans.
Methods: Using the p53 chimeric mouse model, tissue samples from 28 mice were collected and
genomic DNA was extracted. Quantitative real-time PCR was then used to differentiate five different
genotypes in the chimeric mice: P/P, R/R, P/R, P/WT, R/WT, where WT represents the mouse wild type form
of p53. To successfully differentiate the genotype, custom TaqMan probes were created from exon 4,
containing the SNP, in human p53. The FAM reporter dye was used to represent the P genotype, the VIC
reporter dye was used to represent the R genotype. JOE/BHQ1 was used as a reporter dye on the
endogenous control probe on ApoB. The qPCR reactions were set up in a 96 well plate and loaded into an
ABI79000HT Real-Time PCR instrument. DNA quantification and genotype information was processed
through SDS 2.4 software. All DNA samples were tested in triplicate.
Results & Conclusion: DNA from 28 mice with humanized DNA and WT DNA were extracted and
®
successfully genotyped using a multiplex real-time QPCR assay with TaqMan probes. The protocol is ready
to be implemented in genotyping the human DNA samples, and the homozygote chimeric mouse samples
(P/P, R/R) will be used as controls in future qPCR reactions involving human genotyping. Genotype
differentiation of the P and R alleles in AA will hopefully provide insight into the increased susceptibility of AA
to EOBC.
50 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Bioinformatic Analysis of MSCRAMMs in S. Aureus
1,2
1
1
1
Dickson Tran , Ana Cohen , Vannakambadi K. Ganesh , Magnus Höök
1 Center for Infectious & Inflammatory Disease, Texas A & M Health Science Center
2 Texas A & M University Health Science Center College of Medicine, Houston, Texas
Introduction: Microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) are a group of surface
proteins implicated in the pathogenicity of Staphylococcus aureus by enabling Extracellular matrix (ECM) adhesion. A key
MSCRAMM, clumping factor A (clfA), is found to cause infection by binding to the host’s fibrinogen gamma chain at the Cterminal (last 17 amino acids). Two subdomains, N2 and N3, are the minimum required for ligand binding and crystal
structure data has revealed the hydrophobic trench formed between them is critical for fibrinogen interaction. Recent work
shows other sites may also contribute to clfA/fibrinogen interaction. Aurexis, a humanized monoclonal antibody that targets
clfA and has been tested in phase II trial for severe S. aureus infections, but failed to clear the infection better than the
placebo. Other MSCRAMMs such as clumping factor B (clfB), fibronectin-binding protein A (fnbpA) and B (fnbpB) have
also been found to interact with the fibrinogen. Here we analyze the variability of these MSCRAMMs amongst a collection of
S. aureus and speculate on the implications in clinical outcome.
Hypothesis: Polymorphisms in various MSCRAMMs and fibrinogen could contribute to altered ligand binding and
therefore explain clinical outcome.
Methods: We retrieved MSCRAMMs sequence information from the Patric database and fibrinogen alpha and gamma single
nucleotide polymorphism information from the SeattleSNPs database. We used MEGA to align the sequences and grouped
the variants to construct phylogenetic relationships. We screened the databases for polymorphisms at the
MSCRAMM/fibrinogen binding site, interdomain N2N3 trench of clfA, and the last 17 amino acids of fibrinogen gamma
chain (FGG), which are critical for ligand- protein interaction. Polymorphisms for clfA, clfB and fnbpA were plotted onto
their available crystal structures using PyMol.
Results: The loops in clfA and clfB had more polymorphisms than the sheets. This may explain the flexibility of the looped
regions to assume different conformations during ligand binding while the sheets are more conserved due to their essential
function of the protein. The humanized monoclonal antibody, Aurexis was developed based on the lab strain S. aureus
Newman. In examining the epitopes for Aurexis on over 350 strains of clfA, we find 7 out of the 18 contact residues are
variable. This information could explain why Aurexis phase II clinical trial failed and help design a better monoclonal to
target clfA. In addition, we found a common fibrinogen gamma-binding motif present in clfA, fnbpA and fnbpB amongst
our collection of isolates. These MSCRAMMs are known to bind to the same C-terminus site on the fibrinogen gamma
chain. This motif is also present in other well-studied fibrinogen binders MSCRAMMs. Future studies are needed to
determine the presence of this motif in other less studied MSCRAMMs.
51 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Estradiol suppresses cell growth of non-malignant colonocytes induced by
NF-κB signaling
Yuwei. V Tsai, Gyhye Yoo, and Clinton D. Allred
Department of Nutrition and Food Science, Texas A&M University, College Station Texas.
Texas A & M University Health Science Center College of Medicine, (Campus- Temple, College
Station, Houston)
Introduction:
A positive correlation between chronic inflammation and the incidence of colorectal cancer has been
shown in both epidemiological and clinical studies. Interleukin-6 (IL-6) is one of the
proinflammatory cytokines associated with inflammation, which leads to promoting NF-B (nuclear
factor kappa-light-chain-enhancer of activated B cells signaling) via IKK (IB kinase) expression.
The increased activity of NF-B pathway ultimately results in an increased of cell proliferation.
Recently, studies suggest that estrogen can provide a line of defense against colorectal cancer.
The goal of this study is to examine the ability of estradiol (E2) to reduce the activity of the NF-kB
signaling pathway activated by IKK in young adult mice colonocytes (YAMCs) based on utilizing a
cell growth assay as a physiological outcome.
Hypothesis and Aims:
We hypothesized that E2 would suppress cell proliferation of YAMCs triggered by NF-kB signaling
pathway via IKK. Our aims were to 1) to examine how E2 and IL-6 influence the cell growth, 2) to
examine the effect of IKK signaling on cell growth, and 3) to determine if E 2 decreases the cell
growth of cells transfected with IKK expression plasmid.
Methods:
Plasmid DNA (control vehicle and IKK expression vector) was transfected on chemically competent
E-Coli by heat shock. A single colony was isolated and inoculated in Luria Bertani broth. Plasmid
DNAs were purified and their concentration was examined using Nanodrop 2000
Spectrophotometer. YAMCs were raised on collagen-coated plates in RPMI 1640 medium at 33 ºC.
48 hours before cells were plated. Cells were stripped with charcoal dextrane treated FBS. For all
experiments, 4x104 cell/well were seeded on six-well plates. 1.5 mL/well of individual treatment was
applied after 24 hours of seeding. Treatments for cell growth assay were: 0.1% DMSO, 1nM E 2 and
15 ng/ul IL-6. For transfection assays, cells were transfected for 3 hours before treatment was
applied with 2 ug of plasmid DNA; vehicle as control and IKK expression plasmid. For all
experiments, cells were maintained under non-permissive conditions, 39 ºC and were trypsinized for
cell counting using hematocytometer under microscope after 72 hours of treatment.
Results/Conclusion:
E2 treatment suppressed the growth of YAMCs, whereas IL-6 treatment resulted in an increase of
cell growth. This similar effect of IL-6 was also evident in cells elevated with IKK expression
vector. But when E2 was applied to cells transformed with IKK expression plasmid, the cell growth
was reduced. Therefore, we concluded that estradiol suppresses cell growth of non-malignant
colonocytes induced by NF-kB signaling.
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UPREGULATION OF SUPEROXIDE DISMUTASE IN
OVARIAN CARCINOMA CELLS INCREASES THEIR
RESISTANCE TO DOXORUBICIN
Elizabeth Wiewiorowski, Seyed Hossein Mousavi-Fard, and Steve Maxwell
Department of Molecular & Cellular Medicine
Texas A & M University Health Science Center College of Medicine, (Campus- College
Station)
Introduction: Upregulation of cellular superoxide produces oxidative stress and damage that in
turn may lead the cell to undergo apoptosis. Superoxide dismutase is an antioxidant enzyme that
catalyzes the reaction of the harmful superoxide species into oxygen and hydrogen peroxide.
Chemoresistant or drug-refractory ovarian carcinomas typically express lower levels of superoxide
than those that are responsive to chemotherapies potentially because of an upregulation of the
SOD gene; just one of the many protective mechanisms that tumor cells have developed to fight
chemotherapies’ induction of oxidative stress. Therefore, chemotherapies that act to upregulate
cellular superoxides and counteract the increased levels of SOD may be able to cause drugresistant cancer cells to become re-sensitized to chemotherapeutics. For example, the anisomycin,
rifabutin, can upregulate mitochondrial superoxide and can also reverse drug-resistance.
Hypothesis: We hypothesized that the ability of chemosensitizing agents, such as rifabutin, to
sensitize drug-resistant cancer cells to chemotherapies is dependent on their capacity to increase
levels of apoptosis-inducing superoxides in the tumor cells. Therefore, an upregulation of cellular
SOD should result in increased resistance to doxorubicin and antagonism of rifabutin’s
chemosensitizing activity.
Specific Aim: The specific aim of this project was to establish a cell-based model to test our
hypothesis that superoxides mediate increased sensitivity of ovarian carcinoma cells to doxorubicin.
Moreover, cancer cells resistant to chemotherapies may have increased levels of SOD. Thus,
downregulation of SOD or upregulation of superoxide may lead to reversal of drug resistance in
ovarian carcinoma.
Methods: We transfected an SOD transgene into chemosensitive ovarian carcinoma cells, and
then measured the SOD activity in unlabeled and transfected ovarian carcinoma cells to confirm
upregulation of the enzyme. We then monitored the cells’ sensitivity to varying concentrations of
doxorubicin.
Results and Conclusion: As anticipated, the upregulation of SOD in previously sensitive cells
increased resistance to doxorubicin in a 48 hour growth curve. These SOD-transfected cells will
provide us with a cell-based model to further test our hypothesis that our novel rifabutin
chemosensitizer acts through oxidative stress to re-sensitize chemoresistant ovarian carcinoma
cells.
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August 5, 2014: 9:00 AM - 2:00 PM
Obesity Increases Cancer Risk by Dysregulation of CDK8-Cyclin C
Corinne Young, Jun-Yuan Ji
Texas A & M University Health Science Center College of Medicine, (Campus- College Station)
Introduction and Background: Dysregulated lipid metabolism contributes to human diseases and
pathological conditions such as obesity, diabetes, and cardiovascular complications, while mysteriously
present as a universal feature of human cancer cells. As such, epidemiological studies have established
obesity as an important risk factor for certain major cancers, such as breast, liver and endometrial cancers.
However, the molecular mechanisms that underlie this risk remain poorly understood. Cyclin-Dependent
Kinase 8 (CDK8) and its regulatory partner Cyclin C (CycC) are frequently amplified or mutated in many
human cancers, and dysregulation of CDK8-CycC is shown to promote tumorigenesis of melanoma and
colorectal cancers (Xu and Ji 2011). Studying the in vivo function and regulation of CDK8 and CycC in
Drosophila has led to the discovery of the unexpected role of CDK8-CycC in controlling lipid homeostasis:
CDK8 and CycC mutant animals accumulate more than 2-fold excess of fat than wild-type animals. We
recently reported that CDK8-CycC negatively regulates the expression of fatty acid biosynthetic genes by
directly inhibiting the transcriptional activity of SREBP in Drosophila and mammals. In addition, we found that
both CDK8 and CycC are destabilized by activation of the insulin-signaling pathway in cultured human liver
cancer cells (Zhao et al., 2012). This observation is important because the levels of insulin and insulin-like
growth factors (IGFs) are generally higher in obese and overweight population than those with normal bodymass index (Xu 2011). Remarkably, CDK8 is known to regulate the transcriptional activity of several targets
such as p53, -catenin, E2F1, Notch intracellular domain, and SMADs, all of which are commonly
dysregulated in a wide variety of human cancers. Furthermore, we have found that CDK8-CycC have tumor
suppressive roles in human endometrial cancer cells (Gu et al., 2013).
Hypothesis: We hypothesize that chronic hyperinsulemia and increased levels of IGFs in obese and
overweight people may increase the risks for endometrial cancer by down-regulating CDK8-CycC.
Methods: To determine the relationship between CDK8-CycC levels in endometrial cancers upon stimulation
by insulin and IGFs, we treated cultured the endometrial cancer cells with insulin and IGF-II for 24, 48, and 72
hours, and then monitored the levels of CDK8 and CycC by Western blot.
Results: The results demonstrated a reduction in CycC in the AN3 CA cell line with the addition of either
insulin or IGF-II at 48 and 72 hours. Interestingly, CycC was elevated in the HEC1A cell line. CDK8, on the
other hand, remained relatively constant in all experiments. Since CDK8 and CycC form the CDK8 submodule
with MED12 and MED13 in a 1:1:1:1 ratio, either reduction of CycC or an increase of CycC may result in the
formation of partial complexes, thereby having the same effects on the CDK8 submodule. If so, any change in
CycC may be just as detrimental to the transcriptional regulation and may pose the same risk as decreased
CycC or decreased CDK8.
Conclusion: We have obtained promising preliminary data that support the model that dysregulation of
CDK8-CycC by insulin or IGFs may contribute to increased risk of endometrial cancer. We will repeat the
experiment to examine the effect of insulin and IGF-II on CDK8 and CycC by collecting samples every 12
hours to determine the dynamic dysregulation of CycC that may be responsible for the differing results
between the AN3 CA and HEC1A cell lines. In addition, we would like to test the synergistic effects of
IGFs/insulin on both cell lines to simulate a more likely in vivo presence of other growth factors. Lastly, we will
determine whether insulin and IGF-II effect the expression of CycC by q-RT-PCR.
54 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Bryan, Texas
Quantification of Neuroinflammation by Area Fractionation Method in Epilepsy
Brain Injury Models
Iyan Younus, Jenessa Short, Victoria Dunlap, R. Kuruba, Xin Wu, and D. Samba Reddy*
Texas A&M University Health Science Center, College of Medicine, Bryan, TX 77807
Introduction: Stroke, seizures, trauma and other insults that injure the brain produce substantial
neuroinflammation characterized by elevated inflammatory biomarkers, cytokines and prostanoids.
Neuroinflammation plays a key role in epileptogenesis. Epilepsy and brain trauma are both associated with a
neuroinflammatory reaction that is thought to exacerbate the pathophysiology of the underlying condition.
Likewise inflammatory biomarkers are elevated following status epilepticus (SE) and neuroprotection
strategies are being targeted to non-neuronal components of inflammatory pathways in the brain. Notably,
neuroinflammation is known to cause proliferation and activation of astrocytes and microglia, the two key nonneuronal cells in the brain with a variety of homeostatic functions. SE is an emergency neurological condition
characterized by persistent seizures and high mortality due to massive neuronal cell death. Therefore, it is
thought that activation of the astrocytes and microglia by inflammatory stimuli induced by seizures or insults
causes long-lasting changes in the hippocampus and thereby enhances the risk of developing epilepsy.
Hypothesis & Objective: We hypothesize that neuroinflammation is a critical contributory event in
epileptogenesis and epilepsy. Astrogliosis and microgliosis have long been used as an index for inflammation
and neuronal damage. Astrogliosis is manifested as an increase in intermediate filaments and cellular
hypertrophy as well as an increase in the proliferation of astrocytes. The level of glial fibrillary acidic protein
(GFAP), an intermediate filament protein expressed by astrocytes in the brain, is utilized as a specific marker
for astrocyte activation and astrogliosis. Microglia, which acts as macrophage-like cells in the brain, responds
rapidly to inflammatory signals and this is manifested as microgliosis or the activation of microglia. Enhanced
expression of the calcium-binding protein IBA-1 (ionized calcium-binding adapter molecule-1), which is
specifically expressed in microglia, is utilized as an index of microglia activation or microgliosis. In the present
study we determined the extent of neuroinflammation following prolonged SE induced by OP compounds DFP
and GD in rats. We quantified the extent of astrogliosis and microgliosis by examining GFAP and IBA-1
expression, respectively, in various brain regions.
Methods: SE was induced by the OP AChE inhibitors DFP or GD in rats. Animals were perfused 72 hours
after SE induction for neuropathological studies. Brain slices were prepared for histology and immunostained
using specific antibodies GFAP and IBA-1. Optical images were acquired with an Olympus microscope using
20x settings. The extent of GFAP and IBA-1 immunostaining in the hippocampus subfields CA1, CA3 & DG
was measured by densitometry and quantified using the NIH ImageJ software. We utilized the area
fractionation method to calculate the percentage area of stained cells in relation to the total area for the entire
slice. The extent of neuroinflammation was analyzed in both SE models (DFP and GD).
Results: Astrocytes and microglia were identified with their characteristic morphology. In the hippocampus
subfields, greater number of hypertrophied GFAP+ve and IBA1+ve cells appeared at 72 h after SE.
Quantification by area fractionation method showed a significant (1.5 to 2-fold) increase in activation of
astrocytes and microglia in SE compared to control groups. Peak GFAP+ve expression was evident in the
amygdala followed by hippocampus dentate hilus in the GD model, while a greater IBA1+ve expression was
observed in the hippocampus CA1 and CA3 regions in both GD and DFP models.
Conclusions: These studies indicate that neuroinflammation is a devastating component of the neuronal
injury following exposure to OP chemical agents and seizures. The area fractionation approach appears to be
an optimum strategy for quantification of neuroinflammation using GFAP and IBA-1 as specific markers of
astrogliosis and microgliosis, reactively, in the brain.
**Supported by NIH Grant U01 NS083460 (to Dr. Reddy)**
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August 5, 2014: 9:00 AM - 2:00 PM
Bryan, Texas
THE TIME-COURSE OF ACETYLCHOLINESTERASE INHIBITION AND RECOVERY
FOLLOWING ORGANOPHOSPHATE INTOXICATION IN RATS
Marcus Zaayman, Ramkumar Kuruba, Ellen Colman and D. Samba Reddy*
Texas A&M University Health Science Center College of Medicine, Bryan, TX 77807
Introduction: Acetylcholinesterase (AChE) is an enzyme responsible for the normal hydrolysis of the
neurotransmitter acetylcholine (ACh). When this process is impeded by inhibition of AChE, a cholinergic crisis
results – producing a characteristic set of signs and symptoms. This is commonly seen with organophosphate
(OP) pesticides and nerve agents-- a set of substances that have long been used as chemical warfare
agents. It is well-established that seizure initiation is due to excessive cholinergic activity in the brain,
triggered by the OP intoxication-induced irreversible inhibition of AChE. The toxic effects of OPs and
subsequent rapid and sustained increase in the concentration of ACh at central and peripheral sites occurs
directly due to the inhibition of AChE. The aforementioned cholinergic crisis results as a consequence of ACh
accumulation in synapses and at post-synaptic sites in the brain, causing prolonged seizures, status
epilepticus (SE) and brain damage. AChE enzyme activity is a key biomarker for OP nerve agent exposures.
It can be detected within minutes to hours and up to several days after an attack.
Hypothesis & Objectives: We hypothesize that the time-course of OP exposure-induced AChE inhibition
correlates well with seizure activity in animals and humans. A rapid decrease will be followed by a period of
sustained depression for a prolonged period, before beginning to recover. This trend is expected due to the
irreversible inhibition of the enzyme, and as such, new molecules would first need to be generated before
activity can recover. Therefore, this study seeks to determine the time-course of brain AChE enzyme
inhibition and recovery following the OP intoxication induced by DFP in a rat model.
Methods: Experiments were conducted using adult rats. OP intoxication was induced by subcutaneous
injection of the diisopropylfluorophosphate (DFP). After DFP administration, peripheral and brain tissue
samples including the heart, diaphragm, frontal cortex, parietal cortex, amygdala, hippocampus and
cerebellum were harvested at several time points after exposure (0, 5, 10, 20, 40, and 60 minutes; 2, 4, 8, 24
and 72 hours; 1, 2 and 4 weeks after DFP). Samples were homogenized and a colorimetric assay based on
the Ellman protocol was conducted to measure the AChE activity (Ellman et al., 1961). Tissue supernatants
were added to either Tris buffer (no inhibitor control), eserine (an all-purpose cholinesterase inhibitor to
inhibits both butylcholinesterase (BChE) and AChE) or ethopropazine (a specific BChE inhibitor), all in the
presence of 1 mM acetylthiocholine (substrate) with 0.5 mM DTNB (coloring agent). Samples were read
kinetically in a plate reader every 60 seconds for 30 minutes. The triplicate absorbance values were averaged
and a curve for the kinetic measurements over the 30 minute time period was generated. The slope of this
curve represents the change in absorbance with time, and this was used to calculate the activity of AChE in
nmol/min/mg protein using Beer’s law. Protein levels were estimated using the Pierce BCA assay in
conjunction with a standard curve.
Results: The normal brain AChE activity was 0.2 to 0.5 nmol/min/mg protein. Our results show that DFP
caused an ultrafast irreversible inhibition of AChE within 5 to 10 minutes with maximal 95% inhibition within 40
minutes after exposure. AChE activity was lowest at 40 minutes and remained low up to 8 hours postexposure. AChE activity slowly begins to recover after 24 hours reaching near normal levels from 3 to 7 days.
This time-course was well correlated with the seizure activity scores in rats exposed to DFP.
Conclusions: These findings suggest the following: (1) DFP causes ultrafast inhibition of AChE that is well
correlated with the time-course of seizure activity, (2) Inhibition of AChE activity is spatially and temporally
distinct in various brain regions, and (3) The time-course of peripheral and brain AChE activity is very similar
following DFP exposure. Therefore, an AChE assay represents a valid biomarker for rapid diagnosis of OP
nerve agent exposures.
**Supported by NIH CounterACT Grant U01 NS083460 (to Dr. Reddy)**
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August 5, 2014: 9:00 AM - 2:00 PM
Developing a Model to Study MicroRNA-20a Function in Post-Stroke Astrocytes
L.H. Zingg, A. Selvamani, S. Bake, M.J. Park, and F. Sohrabji
Texas A & M University Health Science Center College of Medicine, College Station, Texas
th
Introduction: Stroke is the leading cause of disability and 4 leading cause of death in the U.S. Previous
studies from the lab have shown young females to consistently display smaller infarct volumes or affected
brain tissue when compared to males (Selvamani, et al., 2014). Previous work from the lab has also shown
aging astrocytes to directly contribute to infarct severity (Lewis, et al., 2012), and may therefore provide a
cellular target for improved stroke therapy among older females. In a recent profile study, it was found that
only one miRNA from 751 was significantly regulated by age and sex using FDR-corrected statistics.
Specifically, mir-20a-3p was significantly elevated in younger adult females when compared to middle aged
females, young adult males, and middle aged males. To assess the functional relevance of mir-20a-3p as a
target of MAP3K2 pathways in glial cells, we analyzed microRNA-predicted targets using bioinformatics
algorithms such as TargetScan and GeneCards. Additionally, in order to test these putative targets, we also
standardized a model of cultured astrocytes transfected with mir-20a mimetics and controls.
Hypothesis: The present study is designed to test the hypothesis of transfecting astrocyte cells in-vitro with
miR-20a-3p. We used real time PCR and Western blots to analyze miR-20a-3p in cell lysates and media of
astrocyte cultures.
Methods: Middle-aged female (10-12 months) Sprague Dawley rats were purchased from Harlan
Laboratories. Animals were anesthetized and subject to stereotaxic surgery to occlude the middle cerebral
artery (MCAo) using endothelin-1, as reported in Selvamani and Sohrabji (2014). 48 hours after stroke,
animals were anesthetized and sacrificed in order to collect astrocytes from the ischemic cortext of the brain.
o
Astrocyte cultures were incubated under 5% CO2 at 37 C. Cells were fed with Astrocyte Growth Media (AGM)
every 3 days. On day 10, cells were plated onto different dishes in prior to transfection. Cultures were
transfected when ~60-80% confluence was seen, using the Lipofectamine RNAiMAX Reagent with either the
microRNA mimetic or scrambled control. Three days later, cell lysates were collected for real time PCR and
Western blot analyses.
All animal procedures were in accordance with NIH guidelines for human care of laboratory animals and
approved by the Institutional Animal Care Committee.
Results:
 Astrocytes from middle aged female rats were successfully harvested from the stroke brain and
successfully transfected with the miR-20a-3p mimic.
 Post-stroke astrocyte QPCR analysis showed miR-20a-3p expression was significantly higher in adult
female astrocytes compared to middle-aged females. P<0.05.
 MiR-20a-3p expression was significantly higher in the media mimic group compared to the media
control group.
 MiR-20a-3p served as a viable model to target MAP3K2 pathways and foundation for future studies.
57 | P a g e
Texas A&M One Health Initiative: Investigating the Health Disparities of Humans and Animals on Isla
de Ometepe, Nicaragua
1
2
3
Christina A. Babu , Thomas A. Jeffreys , and Ashton J. Richardson
1
Texas A&M Health Science Center College of Medicine, Bryan, TX
2
Texas A&M Health Science Center School of Public Health, College Station, TX
3
Texas A&M College of Veterinary Medicine & Biomedical Sciences, College Station, TX
Overview: The purpose of this study was to investigate the human and animal health disparities of Isla de
Ometepe, Nicaragua, seeking to find a connection between the two in an environment where humans and
animals live in such close proximity. Texas A&M University and University of California at Davis partnered
together to develop a team of 6 students consisting of 2 veterinary medicine, 2 public health, 1 human
medicine, and 1 environmental science student. The team had 4 weeks of training at each university, followed
by 4 weeks in Nicaragua. The team worked closely with local physicians and veterinarians, planted
community gardens for a kid’s café program, and hosted 2 health fairs. The data obtained via human
diagnostics provided by local clinics, public health questionnaires, animal questionnaires/physical exams, and
observational data will be used to plan future health interventions on the island next year.
Human: Human diagnostic data was obtained from the clinics of a local physician, and a preliminary analysis
of results was conducted. Participants consisted of 11 males and 39 females. The average BMI of the island
2
was overweight (M 26.21, F 29.25 kg/m ). Males on the island were pre-hypertensive, and the systolic blood
pressures of females were pre-hypertensive as well. Fasting blood sugars were pre-diabetic, and non-fasting
were within normal range. The top 3 health disparities found in humans on the island were joint/muscle pain
(26%), hypertension (24%), and gastrointestinal disease (12%). Family histories showed a prevalence of
hypertension (36%), diabetes (28%), and cardiovascular disease (18%). Additionally, children of the
participants surveyed tended to suffer from gastrointestinal disease (14%).
A public health questionnaire was created, and 42 participants were surveyed. 93% of those
surveyed have lived on the island all their life, so future interventions could potentially have a significant effect
on the island. 54% reported losing a child with the most common cause of mortality being abortion (29%) and
diarrheal complications (14%). 95% of people obtained their water from a potable water source such as a
stream, with 90% performing no form of treatment (filter/boiling/chlorination) to the water. 60% stated that they
did not have good dental health with 90% stating money as the reason, and 63% believed that they did not
have a nutritious diet with 88% stating money as the reason. The primary pediatric concern was no
medication (91%), and this need for medication is similarly felt by many others (60%) in the community as a
whole. When not able to go to a doctor, 41% self-medicated and 23% used medicinal plants/natural remedies,
suggesting that planting medicinal gardens could potentially bring significant relief to a population with a lack
of medication and resources. Additionally, the discrepancies seen between parental illnesses and individual
health concerns suggest that the community does not realize the grave effects of hypertension on their health.
Animal: Data was collected on animal health and use via a questionnaire taken by 22 animal owners and
physical exams of 31 dogs, 5 horses, 1 cat, and 1 pig during a health fair in the community of Altagracia. The
questionnaire showed that 90% of respondents allowed their animals to roam freely, and 81% allowed their
animals to enter their homes, suggesting that these animals are acquiring as well as shedding pathogens into
the environment as well as the home. Upon examination, all horses were found to have ticks as well as signs
of internal parasites, and 20% were bitten by bats. The presence of ticks and fleas was found to be in a third
of the dog population as well. In regards to animal use, both livestock and small animals are, at minimum,
dual-purpose animals. Companionship, transportation, protection, pest control and traction were found to be
the top uses for animals. 90% of chickens are utilized as a food source, but inefficient collection techniques
result in only an average of 8 eggs being collected per day. The data collected from the questionnaires and
physical exams show that animals are an integral part of society and, therefore, carry an associated value in
terms of companionship and economics. Conversely, the data also highlights several management practices
that pose risks to animal health and, more importantly, the health of their owners. From a production
standpoint, the energy expenditure of livestock roaming makes meat production and traction use far less
efficient.
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August 5, 2014: 9:00 AM - 2:00 PM
The Effects of the Organochlorine Methoxychlor on the
Transcriptional Activation of Sox2
Rachel Bounds, Daria Muller, Michael Golding
Department of Veterinary Physiology & Pharmacology
Houston)
Introduction: Neuronal development in the fetus is a highly controlled process that can be negatively
influenced by prenatal exposure to environmental toxins. The organochlorine pesticide, methoxychlor, is an
endocrine disrupting chemical that has been shown to bio-accumulate and has been found at high
concentrations in women of reproductive age. One of the key regulators of mammalian neural development is
the transcription factor Sox2. Sox2 is a crucial regulator of neural patterning and stem cell lineage
commitment within the developing brain. Expression of this gene is regulated by thyroid hormone. Given the
essential role of the thyroid hormone in controlling Sox2 expression, we sought to examine whether
methoxychlor could influence the expression of Sox2.
Hypothesis: We hypothesized that methoxychlor would negatively influence normal Sox2 transcription levels
through activation or repression of the Sox2 regulatory enhancer region. Any variation in the normal
expression of Sox2 could affect the natural progression of neural stem cell differentiation.
Methods: Neural stem cells were transfected, via electroporation, with a Sox2 containing plasmid. Once the
transfected cells reached a confluent state, they were treated for 24 hours with 1nM, 10nM, 25nM, and 50nM
methoxychlor suspended in 0.1% DMSO. After treatment the cells were lysed and transcriptional activity
measured with a Luciferase Assay.
Results: We demonstrated a dose dependent increase in Sox2 transcriptional activity in response to
environmentally relevant concentrations of methoxychlor. This increase in transcriptional activity has the
potential to alter normal development and effect neuronal homeostasis in the fetus.
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August 5, 2014: 9:00 AM - 2:00 PM
Texas A&M University One Health Initiative: FDA Center for Veterinary Medicine Internship on
Antimicrobial Resistance
1
2
W. Jacob Cobb and Sarah Genzer
1
2
Texas A&M University College of Veterinary Medicine & Biomedical Sciences, College Station, TX
Antimicrobial resistance continues to be an ongoing issue of both human and veterinary
health concern. Many different pathways contribute to antimicrobial resistance on both the human
and animal side of this imperative topic, however the Food and Drug Administration (FDA) believe
that human exposure to resistant microbes through the food chain is the most significant contributor
to antimicrobial-resistant infections in humans. Much scrutiny and debate has been focused on the
use of antimicrobial compounds in agricultural applications, but there is a paucity of data showing
actual causation of resistance development due to these practices. Over the past 11 years, the Food
and Drug Administration’s Center for Veterinary Medicine has put forth three guidelines for
industry that are aimed at curbing the development of resistance to currently used and newly
developed antimicrobial compounds used in veterinary practice. Currently the Food and Drug
Administration is partnering with Cornell University to develop a method that would facilitate
tracking of antimicrobial resistance trends across the foodborne disease pathway. By combining
current antimicrobial resistance monitoring systems with other sources of data such as agricultural
censuses and antimicrobial usage data from farms, this new methodology being developed will allow
for population-level analyses of the effects of policy changes on antimicrobial usage with the
development of antimicrobial resistance across the foodborne disease pathway. Herein are described
the efforts two Texas A&M University students made to assist and facilitate this project so that it
may reach its eventual goal.
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August 5, 2014: 9:00 AM - 2:00 PM
Effect of Pediatric Vaccines on the Hippocampus: Relevance to Autism
1
1
1
1
1
J.B. HAMMOND , B. GADAD , W. LI , U. YAZDANI , T. JOHNSON ,
1,2
1
L. HEWITSON , D.C. GERMAN
1
2
Psychiatry, UT Southwestern Med. Ctr., Dallas, TX; Johnson Ctr. for Child Hlth. & Develop., Austin, TX
Introduction:
Autism Spectrum Disorders (ASDs) are characterized by impairments in social and
cognitive abilities, speech communication dysfunction, symbolic play, and repetitive
behavior. Some reports suggest that exposure to ethyl mercury (EtHg), in the form of
thimerosal, in childhood vaccines may have played a causative role in ASDs (Barile et al,
2011). Post-mortem brains from people with ASDs can exhibit significant reductions in in
the size of CA1 hippocampus cells (Kemper & Bauman, 1993).
Hypothesis:
The 1990s vaccine schedule for human infants, which contains the mercury-preservative
thimerosal, plays a role in the development of autism.
Methods:
Rhesus Macaque monkeys were treated with the 1990s schedule of vaccines (n=12) and
compared with untreated control monkeys (n=16). The area of the CA1 neurons was
measured by two investigators who were blind to the experimental condition of the animals.
Results:
There is no significant reduction in CA1 soma area for subjects exposed to the 1990s
vaccine schedule. These data do not support the hypothesis that thimerosal-containing
vaccines play a role in autism.
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Effect of zinc exposure before and during methylmercury toxicity in zebrafish embryos
Jeannie M. Nguyen, Angela M. Harrington, Louise C. Abbott
Texas A&M Health Science Center, College of Medicine
Department of Veterinary Integrative Biosciences, College of Veterinary Medicine and Biomedical
Sciences, Texas A&M University, College Station, TX 77843-4458, USA
Introduction: Mercury, particularly the organic form of methylmercury (MeHg), is a known
developmental teratogen that readily crosses the blood brain barrier and blood placental barrier.
Smaller eyes, enlarged pericardial sac, delayed development and adverse neurologic effects such as
cerebral palsy have been associated with this heavy metal toxicant. MeHg levels in the environment can
be found at concentrations of 10 ppb or higher, and due to its lipid soluble properties, MeHg
accumulates in the food chain, particularly in fish. Despite the environmental impact and threat this
toxic heavy metal imposes on human health, little is known about treatment of the adverse effects of
methylmercury toxicity.
Aim: Studies show that zinc pretreatment may decrease toxic effects of heavy metals by inducing
protective enzymes in antioxidant pathways. We tested the hypothesis that zinc supplementation
before and during methylmercury exposure decreases adverse effects of methylmercury toxicity on
zebrafish embryo development.
Methods: Adult AB strain zebrafish bred and embryos collected. Two or three embryos were placed
per well in 24 well culture plates. Control embryos included untreated embryos in embryo medium and
MeHg exposed embryos at a concentration of 50 ppb in embryo medium. Four experimental groups
examining effects of zinc on MeHg toxicity included: embryo medium + 50 ppb MeHg + 0.25 µM or 4.0
µM zinc for 24 hours or for 72 hours of exposure. Four groups assessing effects of zinc alone included:
embryo medium + 0.25 µM zinc or 4.0 µM for 24 or for 72 hours. Prior to MeHg exposure, all
experimental groups were pretreated with embryo medium + 0.25 µM or 4.0 µM zinc, and after
incubating for 4 to 5 hours all groups were changed into their respective experimental group solutions.
All groups were incubated for 72 hours with 24 hour exposure groups changed to fresh embryo
medium for the last 48 hours. Embryos were observed every 24 hours after exposure to MeHg and dead
embryos were removed. Starting at 48 hours after exposure to MeHg, embryos were observed for
hatching from the chorion. After 72 hours, hatched embryos were tested for elicited movement. At 72
hours all embryos were anesthetized with MS-222, and then euthanized on ice followed by fixation
with 10% neutral buffered formalin. A blinded study examined embryo morphology including body
length, yolk sac size and eye size.
Results: The data suggest zinc exposure significantly reduced the effects of MeHg toxicity on length,
yolk sac area, and eye area for certain experimental groups. Hatch rate did not improve with zinc
exposure. Conversely, experimental groups exposed to MeHg + 4.0 µM zinc showed a statically
significant decrease in hatch rate from both control groups, untreated and MeHg only embryos. While
not significant, zinc exposure showed a trend to improve adverse effects on elicited movement.
Conclusion: Our study showed that prophylactic zinc exposure and zinc exposure during MeHg
exposure are beneficial for zebrafish development with respect to embryonic length, yolk sac area, and
eye area. However, the effects are not equally distributed. The beneficial effects of zinc are
concentration and time dependent. There were no beneficial effects of zinc exposure on death rate,
hatch rate, and elicited movement.
62 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Neurosteroid-Independent Synaptic Antiseizure Activity of Midazolam
Sandesh Reddy, Iyan Younus, and D. Samba Reddy*
Introduction & Background: Midazolam is a benzodiazepine anticonvulsant with rapid onset and
intermediate duration of action. The antiseizure activity of midazolam is a result of the drug increasing the
frequency of synaptic GABAA receptor chloride-channel opening in the brain. Midazolam is the drug of choice
for persistent acute seizures and status epilepticus, especially those caused by nerve agents and
organophosphate pesticides. There are indications that midazolam elevates endogenous neurosteroid levels
via TSPO, a cholesterol transporter. Therefore, the primary goal of this study was to confirm or disprove the
theory that midazolam’s anticonvulsant activity is mediated by endogenous neurosteroids such as the GABA A
receptor-modulating allopregnanolone (AP).
Hypothesis & Objective: We hypothesized that midazolam’s anticonvulsant property does not result from
neurosteroid interaction. To test this hypothesis, we used both the hippocampus kindling and 6-Hz models of
epilepsy in mice and determined midazolam mechanisms using three specific blockers: PK11195, a TSPO
blocker; finasteride, a 5-reductase neurosteroid inhibitor; and flumazenil, a benzodiazepine antagonist. We
compared the seizure protection of midazolam alone with the seizure protection of midazolam pretreated with
flumazenil, PK11195, or finasteride.
Materials and Methods: In the 6-Hz model, midazolam was injected in increasing doses (0.05-3 mg/kg, i.p.)
in different groups of mice, and seizure duration and symptoms were recorded during corneal stimulation 15
min after drug administration. These parameters were compared with the resulting parameters when 15
mg/kg PK11195 was administered 30 min before midazolam, 50 mg/kg finasteride 1 hour before, and 2 mg/kg
flumazenil 30 min before. Inhibitors were also administered without midazolam. ED 50 and ED99 values were
calculated for all dose combinations. The time-course consisted of stimulations 15 min, 1 h, 4 h, and 24 h
after midazolam. Dose-response and time-course studies were conducted in the same manner in the kindling
model, except seizure duration, seizure stage, and afterdischarge duration were recorded following
hippocampus stimulation. Plasma and brain AP levels in mice treated with midazolam were measured by LCMS assay.
Results: Midazolam produced a dose-dependent protection against seizures in both models. In the 6-Hz
model, pretreatment with PK11195 or finasteride (ED50 of 0.36 mg/kg each) did not elicit any significant
change in seizure protection compared to midazolam alone (ED 50 of 0.4 mg/kg). Animals pretreated with
flumazenil, however, displayed a dose-dependent blockade of midazolam’s anticonvulsant effects, leading to
drastically worsened seizure protection (ED50 >3 mg/kg). When given alone, the inhibitors did not affect
seizures. The data from the kindling model displayed a similar trend, showing negligible difference in the
antiseizure responses between the control midazolam group and groups pretreated with PK11195 or
finasteride. However, flumazenil pretreatment significantly inhibited the antiseizure effect of midazolam.
Inhibitors alone also yielded no protection in the kindling model. Additionally, plasma and brain levels of AP
were similar between vehicle and midazolam groups.
Conclusions: The results indicate that the antiseizure effects of midazolam are not mediated by endogenous
neurosteroids such as AP. Instead, they are directly mediated by synaptic GABA A receptors through specific
benzodiazepine binding sites. Midazolam is a powerful antiseizure agent for controlling acute seizures
including those induced by nerve agents and organophosphate pesticides.
*Supported by NIH grant U01-NS083460 (to Dr. Reddy)
63 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Bryan, Texas
AN OPTIMIZED BRAIN STEREOLOGY PROTOCOL FOR NEURON COUNTING
THE REDDY LAB GROUP
Jenessa Short, Jonathan Brewer, Victoria Dunlap, Megan Swonke, Iyan Younus, Divya Raju, Ramkumar
Kuruba, Xin Wu and D. Samba Reddy*
Introduction & Background: Neuronal injury and neurodegeneration is a hallmark pathology observed in a
variety of acute and chronic neurological conditions such as epilepsy, stroke, TBI, Parkinson’s disease and
Alzheimer’s disease. Quantification of absolute neuron numbers and interneuron counts in various brain
regions is essential to understand the impact of neurological insults or disease progression on neuronal
survival and neurodegeneration. However, conventional qualitative scoring-based protocols are superficial
and less sensitive for use in studies of neuroprotection treatment evaluations. Here we describe optimization
of a sensitive, medium-throughput protocol of stereological quantification of neurons and interneurons in the
brain.
Materials and Methods: Rats were perfused with 4% paraformaldehyde and brains were cryoprotected with
0.1 M phosphate buffer (PB, pH 7.4) containing 30% sucrose for 72 hours and rapidly frozen with tissue tek
on dry ice. Serial sections (30 μm) were cut coronally through the cerebrum containing the amygdala and the
hippocampus. Every 20th section in each series of 20 sections (interval: 600 μm) were processed for NeuN
(principal neurons) and parvalbumin (interneurons)-immunoreactivity. The newCAST stereology system
consisted of a color digital camera interfaced with an Olympus BX51 microscope and motorized stage driven
by the Visiopharm stereology software. The absolute number of principal cells and interneurons were
separately quantified in each hippocampus subfield. The program was set for calculation of the cell density,
using the 60X oil immersion lens, and cell volume, using the 10X lens in a protocol considered as 3D
counting.
Results: Our extended protocol allows an unbiased quantification of total neurons in the hippocampus
subfields in less than 3 hours, with the possibility of the completion of up to 4 animals in a single day. The
protocol was utilized for quantification of NeuN-positive principal neurons and parvalbumin-positive
interneurons in the hippocampus subfields CA1, CA3, dentate gyrus, and dentate hilus. In the control group,
our counting yielded a total of 1.4 million principal neurons and 60,000 interneurons in the hippocampus.
These numbers represents absolute total cells and are consistent with the literature reports of total neurons in
the hippocampus in age-matched rats.
Conclusions: The optimized stereology protocol allows sensitive throughput counting of total neurons in any
brain region. Important quantitative descriptions of the geometry of 3D structures can be completed from
measurements made on 2D images. This is accomplished by using formulas that take into account pertinent
structural frameworks that are lost during the formation of brain sections. Thus providing an eloquent
quantitative tool for studies of neuroprotection or neurodegeneration in a variety of neurological and brain
injury models.
*Supported by NIH Grant U01-NS083460 (to Dr. Reddy)*
64 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
The Incidence and Outcome of DVT after Endovenous Laser Ablation
William P Shutze, Sr., Katherine Kane, Taylor Hicks, John Kedora, Steve Hohmann, Toby Dunn,
Brad Grimsley, Dennis Gable, Greg Pearl, Bertram Smith, Richard Lueking, Elizabeth Nguyen,
Grace Lassiter
Cardiovascular Surgery
Baylor University Medical Center, Dallas, Texas
Introduction: Endovenous ablation of the saphenous vein (EVLT) has become the preferred treatment for
treating saphenous reflux, which results in symptomatic lower extremity venous insufficiency or varicose veins.
This office based ambulatory procedure was noted during initial FDA trials to have a low incidence of
postoperative deep vein thrombosis (DVT). Later clinical experience suggested that the actual incidence of DVT
may be much higher. The study also looked at the difference in rate of DVT between an 810 and 1470 wavelength
laser. The 810 targets hemoglobin to cause thrombosis of the vessel and the 1470 targets water molecules in the
vessel wall.
Hypothesis: The incidence of DVT after EVLT with the 1470 laser is lower than the incidence of DVT with the 810
laser.
Methods: We reviewed the office records and the pre and post treatment ultrasounds of patients undergoing
EVLT in our office from 2005 -2014 to determine the frequency of postoperative DVT in patients we had treated
and then graded them according to a previously published classification.
Results: There were 1470 veins treated in 445 males and 1025 females. There were 1165 patients on the 810
laser and 305 on the 1470 laser. The CEAP class for these patients was 1(0), 2(629), 3(529), 4(150), 5(51) and
6(101). The greater saphenous vein was treated in 1346, the lesser saphenous vein (LSV) in 71 and both were
treated in 23. There were also 18 cases where branches or accessory veins were treated. DVT occurred in 78 of
legs treated for an incidence of 5.3% overall. The 810 laser had an incidence rate of 6.0% and the 1470 had a rate
of 2.6%. The DVTs in the femoral vein were of level 2(21), 3(26), 4 (11), 5(10) and 6 (8). Six patients developed
DVT in the popliteal vein after EVLT of the LSV. Treatment for the postop DVT consisted of observation (44),
anticoagulation (24), antiplatelet therapy (8), and nonsteroidal anti-inflammatory agents (1). Duration of therapy
was usually one week but several patients were treated for periods ranging from 1 to 7 weeks. No pulmonary
emboli occurred in any of these patients. The DVTs resolved completely in all patients. The incidence of DVT after
EVLT using the 810 laser is higher than previously reported but is not associated with PE and mainly consists of
low risk level 3, 4 and 5 DVT. The DVT typically resolves after one week and can be treated with a short course of
antiplatelet or anticoagulation therapy although observation appears to be sufficient as well. The data also shows
that the 1470 laser has a lower incidence of DVT, due to targeting water molecules in the vessel walls in lieu of
hemoglobin targeting by the 810 laser.
65 | P a g e
August 5, 2014: 9:00 AM - 2:00 PM
Intraoperative Ultrasound and Isolated Lumbar Schwannomas
1
1
2
Clarence Thomas Jadye Kee, M.S. , Quoc-Bao D. Nguyen , Jason M. Hoover, M.D. , Daniel J. Lamont,
2
PA-C
1
Texas A & M University Health Science Center College of Medicine, Bryan, Texas
2
The Texas Brain and Spine Institute, Texas A&M Health Science Center, Bryan, Texas
Houston)
Introduction
Intraoperative Ultrasound (IOUS) is an underutilized asset in neurosurgery. It provides real-time
information that is easily repeatable with no irradiation of the patient. We present a unique case of an
isolated intradural and extradural schwannoma at the level of the lumbar spine. The intradural mass
was obscured on the MRI but was notably identified by IOUS. The case presents the value of IOUS
during spinal tumor surgery and another possible subset of Schwannomas not currently described in
the literature.
Presentation
A 67-year-old, left-handed male with a history of intermittent low back pain had an acute onset of
low mid-back pain radiating into his legs for 3-4 weeks. A thorough neurological examination was
negative except for obvious weakness in his right ankle with dorsiflexion. Magnetic Resonance
Imaging (MRI) without contrast of the lumbar spine demonstrated a large spinal mass (2.5 x 1.9 x
1.4 cubic centimeters) in the central spinal canal between L4 and L5. Review of his cervical spine
computerized tomography (CT) imaging of his chest, abdomen, and pelvis provided no evidence for
a metastatic tumor to the spine.
Operative Course & Pathology
The patient underwent L4-L5 laminectomies and a fleshy, fibrous mass was excised from the central
and right lateral recess region. Subsequently, intraoperative ultrasound detected a nodular area on
one of the nerve roots in the intradural space. Using a microscope, we incised the dura midline and
retracted it. Histopathological studies of the two extracted masses (extradural and intradural) were
both identified as benign schwannoma. These specimens revealed spindle-shaped cells with areas of
compact elongated cells and occasional nuclear palisading (Antoni A pattern) as well as hypocellular
areas of loosely textured cells (Antoni B pattern). Further immunostaing patterns supported the
diagnosis of schwannoma and ruled out meningioma.
Conclusion
The information gathered from imaging is powerful within the operating room. Intraoperative
Ultrasound provides an effective means to delineate masses within the spinal canal that is currently
underused. In our unique case, the use of IOUS allowed us to identify an obscured intradural mass.
Thus, improving the patient’s outcome by eliminating the risks of further monitoring and
reoperation.
66 | P a g e
Effect of Secretin in Cardiac Contractile Regulation
Kindra N. Walker, Syeda H. Afroze, Damir Nizamutdinov, Micheleine Guerrier, Allyson Martinez, David E.
Dostal, Shannon Glaser
Scott and White Healthcare Digestive Disease Research Center
Central Texas Veterans Health Care System
Background: Secretin is a neuroendocrine hormone produced by S cells in the duodenum as well as
cholangiocytes in the liver. Secretin’s main function is to regulate gastrointestinal pH through bicarbonate
production and gastric acid inhibition. It has been reported that the mRNA of secretin and its membranebound receptor are expressed in the heart, lung, kidney, testes, brain, and duodenum. Secretin has a role in
water homeostasis of the body through its actions on the kidneys. It has also been shown to increase cardiac
output and heart rate. Previous studies have shown that secretin works through a G protein coupled receptor
activating cAMP synthesis. It is believed that increased cAMP will lead to an increase in the amount of
calcium available for contraction of the heart leading to a positive inotropic effect.
Goals: The primary goal of this study is to evaluate secretin’s effects on neonatal rat cardiac myocytes,
cardiac fibroblasts, and isolated right ventricle papillary muscle performance.
Methods: The cardiac myocytes, cardiac fibroblast, and papillary muscles were isolated from 2 day-old
neonatal rats. Isolated cells were plated and after 48 hours of incubation were starved overnight with serum
free media. Prior to experimentation, cells were pretreated with 2μM Fura 2-AM in SFM at 37°C for 15
minutes. Cells were washed once and temporal effects of 100nM secretin were recorded at various time
points (0-30 minutes). After that calcium ion influx was measured, using ion optics, at various time points.
Intracellular Calcium measurements were recorded using ion optics Wizard 6.2 software. Similarly, the
isolated papillary muscle was treated with 100nM secretin for different time points and the contractility was
measured by ion optics.
Results: Secretin treatment of cardiac fibroblasts increases systolic and diastolic intracellular calcium levels
over 30 minutes of incubation. However, secretin caused little to no effect on cardiac myocytes calcium
exchange. The effect of secretin on contractile performance of isolated papillary muscles showed stable
increase in force generation within 40 minutes of recordings with approximately 22% increase in twitch
amplitude.
Conclusion: This study demonstrates the effect of secretin on function and regulation of contractile
performance in the heart. Results suggest that secretin increases cardiac contractility through a calcium
independent regulation pathway and thus elevated secretin might have a potential role in cardiomyopathy.
67 | P a g e
College of Medicine
2014
Summer Research Program Participants
Bryan/College Station
Medical Students
Babalola, Ebunoluwa
Herrera, Linda
Maloney, Taylor
Nguyen, Quoc-Bao
Thomas, Rose
Zaayman, Marcus
Undergraduates
Capetillo, Mario
Colman, Ellen
Hamilton, Alina
Howell, John
McGhie, Brandon
Tsai, Yuwei
Younus, Iyan
Zingg, Leonardo
Volunteers
Abbas, Darren
Bosques, Kimberly
Cochran, David
Coffman, Jason
Hayworth, Carla
Robinson, Lance
Young, Corinne
Houston
Medical
Daniels, Michlene
Mohiuiddin, Maha
Oweis, Divina
Undergraduates
Lane, Mar'Kiffany
Mahmood, Tasneem
Volunteers
Chen, Chris
Dhanani, Naila
Naumann, Heather
Niakan, Lillian
Tran, Dickson
Wiewiorowski, Elizabeth
Mentors
Dr. Robin Fuchs-Young
Dr. L. Gerard Toussaint
Dr. Jeffrey D. Cirillo
Dr. Jonathan A. Friedman
Dr. D. Samba Reddy
Dr. D. Samba Reddy
Dr. W. Less Dees
Dr. RB Kreider
Dr. Warren Zimmer
Dr. Clinton D. Allred
Dr. D. Samba Reddy
Dr. Farida Soharbji
Dr. D. Samba Reddy
Dr. Marcella D. Cervantes
Dr. Emily Wilson
Dr. David Earnest
Dr. Warren Zimmer
Dr. Jane R. Welsh
Dr. Jun-Yuan Ji
RISE
Prairie View
Medical
High School
Medical
Medical
Medical
Medical
Medical
Mentors
Dr. Belen Pascual
Dr. David Huston
Dr. David Huston
Dr. Dekai Zhang
Dr. Roderick H. Dashwood
Dr. David M. Eagleman
Dr. Joseph Fernandez-Moure
Dr. Patrick McCulloch
Dr. David Eagleman
Dr. Magnus Hook
Dr. Steve Maxwell
Medical
Medical
Medical
Medical
Medical
Medical
68 | P a g e
Temple
Medical
Chi, Alvin
Dawson, Ashley
Desai, Kurren
Huynh, Victoria
Megahed, Tarick
Milad, Moheb
Ochoa, Brennan
Ogden, John
Shin, Hope
Sohner, Mark
Stone, Ryan
Undergraduates
Kearney, Kate
Luna, Sarah
Maguddayao, Aris
Phillips, Deanne
Westmorland, Jessica
Volunteers
Bowman, Jason
Lowe, Jordan
Munir, Yunib
Weber, David
Dr. Duren M. Ready
Dr. Soren Signel
Dr. Ashok Shetty
Dr. Heather Francis
Dr. Ashok Shetty
Dr. Brett Mitchell
Dr. David E. Dostal
Dr. Kenneth Baker
Dr. Sharon DeMorrow
Dr. S. Glaser
Dr. Karen Newell-Rogers
Dr. Shenyuan L. Zhang
Dr. William C. Culp, Jr
Dr. William C. Culp, Jr
Dr. Kenneth Baker
Dr. Vincent VanBuren
THBD
THBD
THBD
THBD
Prairie View
Dr. David E. Dostal
Dr. Soren Signel
Dr. Soren Signel
Dr. Carl Tong
Medical
Medical
Medical
UnderGrad
69 | P a g e
2014 Summer Research Program Series
Bryan: MREB 1151
Temple: MEC 214
Houston: ALKB 1114F
Date
Time
Topic
Speaker
* Change Locations
Record Keeping
Dr. Van Wilson
Houston : ALKB 1105
CST*R Grand Rounds
Dr. Rod Dashwood
Houston : ALKB 412
Commercialization in Research
Dr. Joe Jilka
Houston : ALKB 1105
Temple: MEC 214
Scientific Method
Dr. David McMurray
9:00 AM
Scientific Misconduct
Dr. Vernon Tesh
Tuesday, June 17
12:00 PM
Exercise and Medicine
Dr. Richard Kreider
*Friday, June 20
8:30 AM
MD/PhD
Dr. Julian Leibowitz
Tuesday, June 24
12:00 PM
TBA
Dr. Paul Hicks
Friday, May 30
9:00 AM
Tuesday, June 3
12:00 PM
Friday, June 6
Tuesday, June 10
Friday, June 13
9:00 AM
12:00 PM
Friday, June 27
9:00 AM
Using Animal Models
Dr. James Elliott
Tuesday, July 1
12:00 PM
CST*R Grand Rounds
Dr. L. Garry Adams
Friday, July 4
12:00 PM
Friday, July 11
9:00 AM
Biotechnology/Ethics
Dr. James Samuel
12:00 PM
Translational Science
Dr. David Huston
Friday, July 18
Tuesday, July 22
Friday, July 25
Tuesday, July 29
Temple: MEC LH (210)
Houston : Classroom
HOLIDAY
Tuesday, July 8
Tuesday, July 15
Temple: MEC 213
Houston : Classroom
9:00 AM
12:00 PM
9:00 AM
12:00 PM
Center for Applied Health Research Dr. Laurel Copeland
Science Microbe
Houston : Classroom
Dr. Samuel Shelburne III, MD PhD
TBA
Dr. Zimmer
Research
Dr. Jane Welsh
Undergraduate Students Presentations: Houston/Temple
Friday, August 1
Undergraduate Student Presentations Bryan
Friday, August 5
Poster Presentations
70 | P a g e
Dr. Warren E. Zimmer
Program Director
Scott Exter Professor
Texas A&M Health Science Center
College of Medicine
Rm. 310B Reynolds Medical Building
College Station, TX, 77843-1114
Email: [email protected]
Phone: 979-845-2896
71 | P a g e

tamhsc summer research program

Transcription

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