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Supplement 1/2010
Supplement 1/2010 Editor in Chief Maria Puiu Department of Medical Genetics University of Medicine and Pharmacy „Victor Babes” Timisoara, Romania Tel/fax: +40 256 220479 E-mail: [email protected] Assistant Editors Cristina Rusu Cristina Skrypnyk Vlad Gorduza Secretary Vlad L. David Editorial Board Dorica Dan Adela Chirita-Emandi Amelia Dobrescu Mihai Gafencu Simona Farcas Diter Atasie Radu Popp Cristina Popa Corina Duncescu Honorary Members Ségolène Aymé, France Gheorghe Benga, România Romanian Journal of Rare Diseases ISSN 2068 – 5882 [email protected] www.rjrd.ro Publisher: „Victor Babes” Printing House Address: P-ta Eftimie Murgu 2, 300041 Timisoara, Romania Tel./fax: 0256 / 495 210 [email protected] www.evb.umft.ro Scientific board Adrian C. Nicolescu, Kingston, Canada, Adrian Lupescu, Tuebingen, Germany Alex Almasan, Cleveland, Ohio, USA Alexander Rodewald, Germany Alice C. Ceacareanu, New York, USA Aurelia Szekely, Zalau, Romania Bogdan Iorga, Köln, Germany Calin Popoiu, Timisoara, Romania Cassian Sitaru, Freiburg, Germany Claudia Cosmineanu-Halaby, New York, USA Cristina Rusu, Iasi, Romania Cristina Skrypnyk, Oradea, Romania Curocichin Ghenadie, Chisinau, Moldavia Danalache Bogdan Alexandru, Montreal, Canada Dorin Bogdan Borza, Nashville, USA Dumitru Andrei Iacobas, New York, USA Dumitru Moldovan, Tg. Mures, Romania Emilia Severin, Bucuresti, Romania Eugen Boia, Timisoara, Romania Gabriela Anton, Bucuresti, Romania Gabriela Doros, Timisoara, Romania George-Lucian Moldovan, Boston, USA Gertrude-Emilia Costin, Maryland, USA Igor Cemortan, Chisinau, Moldavia Ioan Simedrea, Timisoara, Romania Ionel Sandovici, Cambridge, UK Katalin Csep, Tg. Mures, Romania Leonard Girnita, Stockholm, Suedia Marcel Ionita, Birmingham, USA Margit Serban, Timisoara, Romania Maria Puiu, Timisoara, Romania Marius Bembea, Oradea, Romania Mihai D. Niculescu, Chapel Hill, USA Minodora Dobreanu, Tg. Mures, Romania Mircea Covic, Iasi, Romania Mircea Ivan, Bloomington, USA Natalia Cucu, Bucuresti, Romania Peter Manu, New York, USA Sanda Marchian, Sibiu, Romania Sergiu P. Pasca California, USA Sorin Ursoniu, Timisoara, Romania Tamás Jánossy, Szeged, Hungary Tudor Oprea, New Mexico, USA Valerica Belengeanu, Timisoara, Romania Victor I. Pop, Cluj-Napoca, Romania Vlad Gorduza, Iasi, Romania Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 ABOUT The Romanian Journal of Rare Diseases© is an international journal addressing rare diseases and orphan drugs from the perspectives of basic and clinical genetics, molecular genetics, cytogenetics, epigenetics, population genetics, biotechnology, neurogenetics, cardiogenetics, oncogenetics, pharmacogenetics and related fields, by publishing original works, review articles, clinical reports and other contributions from all areas covered by The Romanian Journal of Rare Diseases©. This publication is the international official journal of the National Committee for Rare Diseases, founded and started as part of the project of the Romanian Prader Willi Association, „The Norwegian-Romanian Partnership (NoRo) for progress in Rare Diseases” funded by the Norwegian Government granted by the Norwegian Cooperation Program for growth and sustainable development in Romania. The Romanian Prader Willi Association and the „Victor Babes” Publishing House, E. Murgu Square 2, 300041, Timisoara, tel./fax.0256220479, publish the journal quarterly PURPOSE & AREA OF INTEREST For the journal are of interest articles from basic and clinical research. The journal publishes original articles, short reports, special communications, provided that they are based on adequate experimental evidence, clinical studies, case reports, images in rare diseases, letters to the editor, book reviews, reports of congresses and other articles that will be brought to Editorial Board’s attention based on the public’s for the journal. Editorials are published by invitation but we look forward to be offered such material from researchers with experience and results in the study of rare diseases. Requirements for publication in the Romanian Journal of Rare Diseases are in accordance with the requirements of „Uniform Requirements for Manuscripts Submitted to Biomedical Journals” 5th Edition. JAMA 1997, 277:927-934. LEGAL DISCLAIMER The entire contents of the Romanian Journal of Rare Diseases© are protected under international copyrights©. The authors bare the entire responsibilities for the content of the article. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Table of contents REPORTS (FOLLOWING CONGRES PROGRAM) R1. THE ORIGIN OF ROMANIANS FROM MITH TO REALITY: GENETIC EVIDENCE FROM OWN STUDIES ON OLD AND MODERN HUMAN POPULATIONS FROM ROMANIA ................ 14 Alexander Rodewald, Calin Tesio, Georgeta Cardos, Dorina Banica R2. Y HAPLOTYPES – IDENTIFICATION, DIVERSITY, MEANING ................................... 15 Marius Bembea, Attila Patocs, Kinga Kozma, Claudia Jurca, Cristina Skrypnyk R3. PRENATAL CYTOGENETIC TESTING IN A PROSPECTIVE STUDY ON 1,000 PREGNANT WOMEN .............................................................................. 15 V. Pop, M. Militaru, A. Popp, F. Stamatian R4. THE SHORT STATURE – HOW IMPORTANT IS THE GENETIC ETIOLOGY ..................... 16 Eusebiu Vlad Gorduza, Lacramioara Butnariu R5. BOALA GAUCHER TIP 1 ÎN ROMÂNIA: EPIDEMIOLOGIE, GENOTIP, FENOTIP ŞI EVOLUŢIA ................................................................................... 17 Paula Grigorescu–Sido R6. THE CLINICAL CHARACTERISTICS AND OUTCOME IN PATIENTS DIAGNOSED WITH MUCOPOLYSACCHARIDOSES IN ROMANIA .................................... 17 Camelia Al-Khzouz R7. BAC-FISH- A TARGETED TOOL FOR ACCURATE CYTOGENETIC ANALYSIS-FROM BASICS TO DIAGNOSIS .............................................................. 18 Eli Ormerod R8. FIRST PATIENT WITH LATE ONSET TYPE II GLYCOGENOSIS IN ROMANIA: DIAGNOSIS, TREATMENT, EVOLUTION ............................................................... 18 Ioana Nascu, Camelia Al-Khzouz, Simona Bucerzan, Paula Grigorescu-Sido R9. MOLECULAR DIAGNOSIS OF PROGRESSIVE MUSCULAR DYSTROPHIES ..................... 19 France Leturcq R10. SOMETHING BORROWED AND SOMETHING NEW: IGF-1R AND UBIQUITIN ON THE LIST OF TARGETED THERAPY IN CANCER ................................................. 19 Prof. Dr. Doc. Leonard Girnita, MD, PhD R11. TRANSLATIONAL TOOLS FOR RARE DISEASES: THE SOLUTIONS DEVELOPED BY ORPHANET ............................................................................. 20 Ségolène Aymé R12. HAEMOPHILIA A CHALLENGE FOR HEALTH CARE IN ROMANIA ............................ 21 M. Serban, M.Puiu, H. Ionita R13. SYNDROMIC MENTAL RETARDATION – DYSMORPHIC FEATURES SUGGESTIVE FOR THE DIAGNOSIS ..................................................................................... 21 Cristina Rusu, Mihail Volosciuc, Elena Braha, Lacramioara Butnariu, Monica Panzariu, Mircea Covic R14. THE ROLE OF ENVIRONMENTALLY-INDUCED EPIGENETIC CHANGES IN THE ETIOLOGY OF TYPE 2 DIABETES ...................................................................... 22 Ionel Sandovici R15. ESTABLISHING DIAGNOSTIC TESTING SCHEMES FOR PRADER WILLI SYNDROME IN ROMANIAN POPULATION. FOCUS ON THE EPIGENETIC MECHANISMS UNDERLYING THE IMPRINTING DEFECTS CAUSING THE PWS ..................................... 22 Natalia Cucu, Gabriela Anton, Cosmin Arsene, Anca Botezatu, Maria Puiu, Corin Badiu, Vasilica Plaiasu, Danae Stambouli (Cytogenomic, Bucuresti) R16. EPIGENETIC CHANGES AS MARKERS OF EARLY TUMOR DEVELOPMENT ................. 23 Dumitrescu RG, Liu Z, Marian C, Bebu I, Yang Y, Spear SL, Kallakury BVS, Seillier-Moiseiwitsch F, Freudenheim J, Mason J, Shields PG Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 R17. SINGLE NUCLEOTIDE POLYMORPHISMS ASSOCIATED WITH C-REACTIVE PROTEIN LEVELS IN A COHORT OF YOUNG FILIPINO ADULTS .................................... 24 Ghenadie Curocichin,Thomas W. McDade, Christopher Kuzawa, Judith Borja, Li Qin, Damien C. Croteau-Chonka, Amanda F. Marvelle, Ethan M. Lange, Linda S. Adair, Karen L. Mohlke, Leslie A. Lange R18. APPROACH TO RARE DISEASES AT THE NATIONAL AND EUROPEAN LEVEL .............. 25 Ségolène Aymé R19. PROJECT “PARTNERSHIP NORVEGIAN- ROMANIAN (NORO) FOR PROGRESS IN RARE DISEASES” FUNDED THROUGH A GRANT RECEIVED FROM THE NORWEGIAN COOPERATION PROGRAMMES ......................................................... 28 Dan Dorica, Maria Puiu R20. TELEMEDICINE AND E-LEARNING – MODERN APPROACH IN RARE DISEASES TOPIC .... 30 M. Gafencu, D. Dan, M. Puiu ORAL PRESENTATIONS (FOLLOWING CONGRES PROGRAM) C1. PRENATAL DIAGNOSIS – CYTOGENETIC FINDINGS IN 750 AMNIOCENTESIS ................ 31 Cristina Gug, G. Budau, N. Hrubaru, B. Mureşan, T. Cioată, Dana Chiriac, C. Olaru, D. Grigoraş, G. Furău, Liana Antal C2. STRUCTURAL ABERRATIONS OF CHROMOSOME 16 PRESENTED IN 12 CASES ............ 32 Cristina Ionescu, Luminita Roibu, Lorand Savu C3. GENETIC HEARING LOSS – PRENATAL SCREENING ............................................. 32 Cristina Dragomir, Adriana Stan, Madalina Badila, D Stefanescu, Emilia Severin, L Savu C4. DIAGNOSTIC PROBLEMS IN CASE OF PRIMARY AMENORRHEA ASSOCIATED WITH DWARFISM DISARMONIC ................................................................................ 33 Otilia Marginean, Ioan Simedrea, Cristina Gug, Belei Oana, Craciun Adrian, Tamara Marcovici, Ioana Maris, Camelia Daescu, Laura Olariu, Daniela Cioboata C5. THERAPEUTIC STRATEGY FOR AMELIORATION METABOLIC DISEASES IN INBORN ERRORS OF METABOLISM ............................................................................... 34 Jurca C, Bembea M, Kozma K, Skrypnyk C, Iuhas O, Harbuz R, Jurca A C6. WILLIAMS SYNDROME - CLINICAL FEATURES AND CARDIAC INVOLVEMENT IN CHILDREN .............................................................................................. 34 G. Doros, A. Popoiu, I. Anca, M. Gafencu, B. Zoica, C. Laudacescu, M. Puiu C7. MULTIPLE MALFORMATIONS IN INFANT - CASE REPORT ................................... 35 T.Marcovici, I. Simedrea,, L. Tunea, A. Militaru,, O.Belei,, D.Chiru,, G. Brad, M.Puiu, C8. HEREDITARY ONYCHO-OSTEODYSPLASIA (HOOD; NAIL-PATELLA SYNDROME) – A CASE REPORT .......................................................................................... 35 Vasilica Plaiasu, Diana Ochiana, Cristina Skrypnyk, Dorica Dan C9. GENETIC EXPLORATION – DECISIVE FACTOR IN THE DIAGNOSIS AND MANAGEMENT OF THE PRIMARY IMMUNODEFICIENCIES .......................................... 36 M. Bataneant, M.Serban, M.Cucuruz C10. ANGIOGENIC GENE THERAPY – FIRST CLINICAL TRIAL IN ROMANIA ...................... 36 Andrei Anghel C11. GENERATION OF NOVEL RECOMBINANT FACTOR IX VARIANTS WITH ENHANCED CLOTTING ACTIVITY ...................................................................... 37 Laura Marusciac, Carmen Bunu C12. CHARCOT-MARIE-TOOTH DISEASE TYPE 2C: PART OF THE TRPV4 RELATED CHANNELOPATHY SPECTRUM .............................................................. 38 Horia C. Stanescu, Guida Landoure, Anselm A. Zdebik, Barrington G Burnett, Tara L Martinez, Hitoshi Inada, Yijun Shi, Addis A Taye, Lingling Kong, Clare H Munns, Shelly S Choo, Christopher B Phelps, Reema Paudel, Henry Houlden, Christy L Ludlow, Michael J Caterina, Rachelle Gaudet, Charlotte J Sumner, Kenneth H Fischbeck, Robert F. Kleta C13. RAPID PRENATAL DIAGNOSIS BY QF-PCR IN 2010 .......................................... 38 Adriana Stan, Daniela Tudor, Cristina Dragomir, Emilia Severin, Lorand Savu Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 C14. MOLECULAR DIAGNOSTICS IN HEMOCHROMATOSIS TYPE 1 AND WILSON’S DISEASE ... 39 E. Seclaman, L. Tamas, A. Anghel, Valerica Belengeanu C15. FAST AND SENSITIVE MUTATION SCREENING USING HIGH RESOLUTION MELTING ANALYSIS ...................................................................................... 40 CB Iancu, D Iancu, C Constantinescu, E. Neagu, A Constantinescu, G Girbea, L Barbarii C16. DYSTROPHIN GENE MUTATION ANALYSIS IN PATIENTS WITH DUCHENNE MUSCULAR DYSTROPHY AND COGNITIVE IMPAIRMENT ........................................... 40 D. Iancu, E. Neagu, N. Butoianu, C.B. Iancu, C. Constantinescu, C. Burloiu, A. Constantinescu, G. Girbea, C. Iliescu, D. Craiu, L. Barbarii C17. DETECTION OF K-RAS POINT MUTATIONS IN CODONS 12 AND 13 IN COLON TUMORS BY THE PCR-RFLP METHOD ................................................................ 41 Georgeta Cardos, Florina Cionca, Georgeta Butur, Dana Terzea, Mihaela Mihai, Florin Andrei, Simona Enache, Florina Vasilescu, Cristina Iosif and Carmen Ardeleanu, C18. MOLECULAR DIAGNOSIS IN THE MANAGEMENT OF DUCHENNE DISEASE – THE EXPERIENCE OF UMF TIMISOARA ............................................................... 41 Liviu Tamas, Maria Puiu, Andrei Anghel, Edward Seclaman, Oana Mitrasca, Mirela Mititelu C19. MEDICAL GENETIC SERVICES, BETWEEN REAL AND IDEAL ................................. 42 Marius Bembea C20. SUPERNUMERARY CHROMOSOME IN MOSAIC IN A DYSMORPHIC CHILD WITH DEVELOPMENTAL DELAY ........................................................................ 43 Valerica Belengeanu, Simona Farcas, Cristina Popa, Monica Stoian, Alina Belengeanu, Nicoleta Andreescu C21. THE FOLLOW-UP OF A DE NOVO CASE WITH “CRI DU CHAT” SYNDROME .............. 43 Valerica Belengeanu, Simona Farcas, Cristina Popa, Monica Stoian, Dragos Belengeanu, Nicoleta Andreescu, Carmen Radulescu C22. NATIONAL REGISTRY OF DUCHENNE AND BECKER MUSCULAR DYSTROPHIES – IMPROVEMENT IN PATIENT’S STANDARD CARE ..................................................... 44 E. Neagu, D. Iancu, CB Iancu, A. Constantinescu, C. Constantinescu, G. Girbea RA Nicolae, N. Butoianu, D. Craiu, L. Barbarii C23. PALLIATIVE MEDICINE IN PEDIATRIC ONCOLOGY - A CHALLENGE IN MODERN MEDICINE. III RD PEDIATRIC CLINIC TIMISOARA EXPERIENCE .................................... 44 E. Boeriu, M. Cucuruz, S. Arghirescu, R. Costa, G. Brad, I. Ursache, M. Serban C24. CARDIAC DISTURBANCES IN GENETIC DISEASES ............................................. 45 Anca Popoiu, Gabriela Doros, Maria Puiu C25. A TOXICOLOGICAL SCREENING ON IN VITRO AND IN VIVO EXPERIMENTAL MELANOMA AND SKIN CARCINOMA MODELS ........................................................ 45 Cristina A. Dehelean C26. THE INTERRELATION BETWEEN MAXILA-PALLATA CLEFT AND CONGENITAL CORD MALFORMATION ................................................................................. 46 Balan Cristina Daniela, Mihai Voloşciuc, Braha Elena, Monica Pânzaru, Vlad Gorduza C27. „SAVE THE CHILDREN” WITH RARE DISEASES ............................................... 47 Iulia-Simina Jurca, Florin Jurca, Simina, Ionela Moaca, Pop Norbert, Stefan Berci, Iulia Popa, Cristina Irimia, Oana Rosca, Graziella Ecob, Adrian Juverdeanu, Carmen Dumitranoiu, Narcis Dobre, Mihai Gafencu, Maria Puiu POSTERS P1. Identification of microdeletions in the AZF regions of Y chromosome in infertile patients in west Romania ......................................................... 48 Anda Alexa, Andrei Anghel, Edward Seclaman, Liviu Tamas, Mirela Mititelu, Oana Mitrasca Monica Stoian, Valerica Belenganu P2. MOLECULAR CYTOGENETIC FISH TECHNIQUE VERSUS CONVENTIONAL CYTOGENETIC ANALYSIS FOR PRENATAL CYTOGENETIC DIAGNOSIS ........................... 49 Nicoleta Andreescu, Valerica Belengeanu, Monica Stoian, Simona Farcas, Cristina Popa Miruna Muntean Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P3. ROBINOW SYNDROME, DOMINANT AUTOSOMAL FORM - CASE REPORT .................. 49 Nicoleta Andreescu, Valerica Belengeanu, Marioara Boia, Monica Stoian, Simona Farcas, Cristina Popa P4. CYTOGENETIC STUDY IN TURNER SYNDROME-THE EXPERIENCE OF MEDICAL GENETICS CENTER IN IASI DURING 2005-2009 ........................................ 50 G.Andron, R.Popescu, C. Rusu, M. Volosciuc, L. Butnariu, E. Braha, M Panzaru, L Caba, M. Macovei, R. Cojocariu, M. Gramescu, C.Bujoran, I. C. Ivanov, E. V. Gorduza P5. TRISOMY 8 IN PHILADELPHIA-NEGATIVE CELLS OF CHRONIC MYELOID LEUKEMIA PATIENTS RECEIVING TYROSINE KINASE INHIBITORS: REPORT OF TWO CASES ............. 50 Aurora Arghir, Sorina Mihaela Chirieac, Andreea Tutulan-Cunita, Oana Ciocan, Carmen Saguna, Marioara Cristea, Agripina Lungeanu P6. INFERTILITY DUE TO GENETIC ANOMALIES IN BOTH PARTNERS OF THE COUPLE. CASE REPORT ................................................................................ 51 Atasie D, Chicea R, Ispasoiu F, Corina Ispasoiu P7. PREGNANCY WITH EDWARDS SYNDROME IN A COUPLE WITH ADVANCED REPRODUCTIVE AGE. CASE REPORT ................................................... 51 Atasie D, Chicea R, Ispasoiu F, Corina Ispasoiu P8. BIOMOLECULAR SUBSTRATE OF PAEDIATRIC ACUTE LEUKEMIAS AND ITS IMPACT ON OUTCOME ....................................................................... 52 L. Balint-Gib, A. Oprisoni, O. Ciocârlie, S. Arghirescu, M. Puiu, A. Isac, V. Ordodi, L. Ritli, M. Serban P9. THE FIRST CASE WITH HYPERIGM SYNDROME IN ROMANIA ................................ 52 M.Bataneant, B.Toth, L.Marodi, M.Baica, R.Mihart, M.Radulescu, M.Serban P10. CLINICAL, MORPHOLOGICAL AND IMMUNE-GENETICAL CORRELATIONS IN CELIAC DISEASE CHILDREN ............................................................................ 53 Belei Oana, Simedrea Ioan, Tamas Liviu, Marginean Otilia, Daescu Camelia Marcovici Tamara, Brad Georgiana, Puiu Maria P11. CORNELIA DE LANGE SYNDROME – CLINICAL AND EVOLUTIVE ASPECTS ............... 54 Marioara Boia, Maria Puiu, Anca Popoiu, Aniko Manea, Mirabela Dima P12. CLINICAL AND EVOLUTIVE ASPECTS IN TAR SYNDROME-CASE REPORT ................. 54 Marioara Boia, Valeria Belengeanu, Anca Popoiu, Dana Iacob, Aniko Manea, Lucica Stoica P13. MICROSATELLITES STUDY IN TURNER SYNDROME – PARENTAL ORIGIN OF X CHROMOSOME ........................................................................... 55 Bolca D, Carel JC, Grigorescu-Sido P, Pop IV P14. MOSAICISM IDENTIFICATION BY MOLECULAR CYTOGENETICS IN PATIENTS WITH TURNER SYNDROME ............................................................................. 55 Bolca D, Carel JC, Grigorescu-Sido P, Pop IV P15. HIPERMETHYLATION OF MIR-124A GENE PROMOTER IN CERVICAL ONCOGENESIS ... 56 Anca Botezatu, Demetra Socolov, Cristina D.Goia, Iulia V. Iancu, Irina Huica Adriana Plesa, Gabriela Anton P16. FACIAL MALFORMATIONS IN AMNIOTIC BAND SYNDROME ................................. 56 Elena Braha, M. Voloşciuc, Lacramioara Butnariu, Monica Panzaru, Roxana Popescu, Cristina Rusu P17. CYTOGENETIC RESPONSE OF BONE MARROW TO FIRST LINE THERAPY IN CHRONIC MYELOGENOUS LEUKEMIA ......................................... 57 Cornel Bujoran, Mircea Covic, Iuliu Ivanov, Gabriela Dorohoi, C.Catalin, Ecaterina Hanganu–Turtureanu, Cristina Burcoveanu, Angela Dascalescu Eusebiu Vlad Gorduza P18. CLINICO-GENETICAL CORRELATIONS IN DARIER AND HAILEY-HAILEY DISEASE ....... 57 Elena Buteica, Florin Burada, Cristina Angelescu, Alice Buteica, Anca Riza, Irina Stoicescu P19. ROBERTSONIAN TRANSLOCATIONS – IMPORTANT CHROMOSOMAL ANOMALIES ....... 58 Caba L, Covic M, Rusu C, Volosciuc M, Butnariu L, Braha E, Bujoran C, Gramescu M, Ivanov I, Panzaru M, Popescu R, Sireteanu A, Gorduza EV P20. IDENTIFICATION BY DNA MICROARRAY TECHNOLOGY OF CANDIDATE GENES INVOLVED IN MAINTAINING INTERSTITIAL CAJAL CELLS (ICC) AND INTERSTITIAL CAJAL-LIKE CELLS (ICLC) PHENOTYPES IN C-KIT MUTANT MICE ............................... 59 Georgeta Cardos, Carmen Ardeleanu,, Georgeta Butur and Mihail Eugen Hinescu Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P21. PRELIMINARY RESULTS: ALPHA 1 ANTITRYPSIN DEFICIENCY AND LUNG CANCER ..... 59 Catana Andreea, Prof.Dr. Pop Ioan Victor, Dr. Popp Radu, Prof Dr. Pop Monica, Petrisor Felicia P. 22. DISCUSIONS ABOUT A NEW CASE OF WILLIAMS SYNDROME -EVALUATION AND MANAGEMENT .................................................................... 60 A. Chelmus, R.Popescu, C. Rusu, M. Volosciuc, C.Bujoran, I. C. Ivanov, V. E. Gorduza P23. CYTOGENETIC AND CLINICAL FEATURES IN CRI DU CHAT SYNDROME – REPORT OF TWO CASES ............................................................................... 60 Sorina Mihaela Chirieac, Magdalena Budisteanu, Aurora Arghir, Andreea Tutulan-Cunita, Ioana Borcan & Agripina Lungeanu P24. PRADER WILLI SYNDROME IN INFANCY ....................................................... 61 Adela Chirita, Ramona Giurescu, Maria Puiu, Corina Duncescu, Gabriela Doros, Boia Mariana ,Valeria Belengeanu, Ioana Micle P25. TESTICULAR ADRENAL REST TUMORS IN AN ADOLESCENT BOY WITH 2-HYDROXYLASE DEFICENCY ........................................................................ 61 Adela Chirita, Monica Marazan, Ramona Cojocaru, Duncescu Corina, Ioana Micle, P26. TESTING OF ASSOCIATION BETWEEN INSULINA-IGF2 REGION AND DIABETES MELLITUS, OBESITY AND BREAST CANCER ......................................................... 62 Danut Cimponeriu, Pompilia Apostol, Mihai Toma, Stavarachi Monica, Irina Radu, Anne Marie Craciun, Cristian Serafinceanu, Rusu Lavinia, Traean Burcos, Popa Emil, Popa Ileana, Stanilescu Sorin P27. K-RAS GENE MUTATIONAL STATUS AND ITS CORRELATIONS WITH THE HISTOPATHOLOGICAL FINDINGS IN THE COLORECTAL CARCINOMAS .......................... 62 Florina Lucia Cionca, Georgeta Cardos, Mihaela Mihai, Alina Georgescu, Mihai Stoicea, Simona Enache, Valentin Enache, Georgeta Butur, Carmen Ardeleanu P28. RISK FACTORS OF CYSTIC FIBROSIS LIVER DISEASE ........................................ 63 Ioana M. Ciuca, L. Pop, I. Popa, Z. Popa, L. Tamas, P29. OPTIMISATION OF THE NEUROFIBROMATOSIS TYPE MANAGEMENT 63 R.Cojocariu, M.Macovei, E.Braha, M.Panzaru, L.Butnariu, R.Popescu, L.Caba, M.Volosciuc P30. MTHFR ENZYME POLYMORPHISMS (C677T AND A1298C) AND THE RISK FOR DOWN SYNDROM ............................................................... 64 Ruxandra Cretu, Daniela Neagos, L.C. Bohiltea P31. EPIGENETIC MARKERS IN DISEASE: DIAGNOSIS, PREVENTION AND NEW THERAPIES ........................................................................................ 65 Natalia Cucu, Anton Gabriela, Arsene Cosmin, Daniela Nedelcu, Maria Puiu, Pavel Chirila Radu Stefanescu P32. CHROMOSOME INSTABILITY AND PRIMARY IMMUNODEFICIENCY SYNDROME .......... 66 M. Cucuruz, E. Boeriu, M. Puiu, I. Ursache P33. BTK – GENE IN AGAMMAGLOBULINEMIA ...................................................... 66 M. Cucuruz, E. Boeriu, Z. Ellenes, M. Bataneant, E. Ursu, G. Brad P34. VESICO-URETERAL REFLUX IN PLURIMALFORMATIVE SYNDROMES UNFAVORABLE PROGNOSTIC FACTORS ............................................................. 67 Camelia Daescu, Ioana Maris, Ioan Sabau, Ioan Simedrea, Oana Belei,Tamara Marcovici, Adela Emandi-Chirita, Maria Puiu, P35. IS PECTUS EXCAVATUM A GENETIC DISEASE? ............................................... 67 David VL, Popoiu MC, Anca Popoiu, Puiu M, Boia ES P36. GENETIC COUNSELING IN DUCHENNE MUSCULAR DYSTROPHY, A MULTILEVEL APPROACH ............................................................................. 68 Amelia Dobrescu, Maria Puiu, Lavinia Sîrbu, Elena Buteica, Daniela Tache, Dorina Iovanescu P37. CONSIDERATIONS OF REPRODUCTIVE COUNSELLING IN A MOSAIC 45, X/46, X, I(XQ) TURNER SYNDROME ............................................................. 69 Amelia Dobrescu, Maria Puiu, Popa Cristina, D. Iliescu P38. NON-COMPACTION LEFT VENTRICLE, DIAGNOSTIC FEATURE FOR 1P36 MONOSOMY SYNDROME ............................................................................... 69 G. Doros, V. Stan, A. Popoiu, M. Gafencu, J. Puiu, G. Miclaus, B. Zoica Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P39. EVALUATION OF TROMBOPHILIC FACTORS AND MTHFR GENE POLYMORPHIMS IN RECURRENT PREGNANCY LOSS ............................................... 70 P40. ASPECTS OF MOLECULAR GENETICS IN PROSTATE CANCER .............................. 71 Dumache Raluca, Miclea Florin, David Dana, Kaycsa Adriana, Negru Serban, Ionescu Daniela, Puiu Maria P41. PROBLEMS IN ETIOLOGY DIAGNOSIS AND INTERVENTION IN COGNITIVE REGRESSION IN CHILDREN ............................................................................. 71 Dumitriu Simona, Ageu Luminita P42. A RARE CASE OF CONGENITAL ARTHROGRYPOSIS: THE PENA-SHOKEIR I PHENOTYPE ................................................................... 72 Corina Duncescu, Elena Pop, Ramona Giurescu, Adela Chirită, Monica Mărăzan, Ioana Micle P43. PERSISTENT SEVERE HYPOGLYCEMIA SINCE INFANCY: CASE REPORT ................... 72 Corina Duncescu, Monica Mărăzan, Adela Chirită, Elena Pop, Ramona Giurescu, Ioana Micle P44. GENETIC EVALUATION OF SEX DEVELOPMENT DISORDERS – CASES REPORT .......... 73 Simona Farcas, Valerica Belengeanu, Nicoleta Andreescu, Monica Stoian, Dorina Stoicanescu Anca Muresan, Dana Amzar, Marius Craina P45. A CASE OF SEPTO-OPTIC DYSPLASIA ......................................................... 74 Simona Farcas, Cristina Popa, Nicoleta Andreescu, Monica Stoian, Alina Belengeanu, Marioara Boia, Elena Bernad P46. TREACHER COLLINS SYNDROME- CASE REPORT ............................................ 74 Valeria Filip, Cristina Skrypnyk, Elena Popescu, Radu Galis. P47. OCULO-AURICULO-VERTEBRAL SPECTRUM – UN ENTITY EASY TO DIAGNOSE ......... 75 Cerasela Munteanu, Marius Galiţă, Laura Ionescu, Mihail Voloşciuc P48. CLEIDO – CRANIAL DYSPLASIA – CLINICAL STUDY ........................................ 75 Madalina Grigoras, Alexandru Vlad, Cristina Rusu P49. PRENATAL DIAGNOSIS AFTER ESTABLISHED CARRIER STATUS OF BALANCED STRUCTURAL CHROMOSOME ABNORMALITY ....................................................... 76 Cristina Gug, T. Cioată, D. Grigoraş, D. Chiriac, G. Budău, N. Hrubaru, C. Mureşan, A. Creţu, V. Karadja, V. Gorduza, G. Furău P50. PRENATAL DIAGNOSIS OF IPEX SYNDROME AND IDENTIFICATION OF 2 NEW FOXP3 MUTATIONS ............................................................................. 77 Radu Harbuz, James Lespinasse, Stéphanie Boulet, Christine Francanet, Isabelle Crevaux, Pierre-Simon Jouk, Joël Lunardi, Marius Bembea, Pierre F Ray P51. PSYCHOLOGICAL TREATMENT IN GENETIC DISEASES ...................................... 77 Hogea Lavinia P52. PARACETAMOL USAGE DURING EARLY PREGNANCY CAN CAUSE CONGENITAL ABNORMALITIES? ...................................................................... 78 Daniela Iacob, RE Iacob, C Ilie, Marioara Boia, Aniko Manea, Maria Julieta Puiu P53. PREMATURE NEW BORN WITH HEART CONGENITAL MALFORMATIONS ................. 78 Daniela Iacob, RE Iacob, C Ilie, Marioara Boia, Aniko Manea, Mirabela Dima P54. CHROMOSOMAL ABERRATIONS DETECTED BY PRENATAL CYTOGENETIC ANALYSIS OF 3000 CASES ............................................................................. 79 Cristina Ionescu, Luminita Roibu, Lorand Savu P55. A PRELIMINARY EVALUATION OF MANGANESE SUPEROXIDE DISMUTASE (MNSOD) GENETIC POLYMORPHISMS, DIETARY ANTIOXIDANTS, AND RISK OF BREAST CANCER ...... 79 Daniela Ionescu, Raluca Dumache, Maria Puiu, Cristina Dehelean P56. IMPACT OF ACQUIRED POSTPARTUM HEMOPHILIA ON THE NEWBORNS ................ 80 A. Isac, L.Pop, M.Puiu, H. Ionita, D. Savescu, A. Balan P57. CONGENITAL MANDIBULAR ANOMALIES – CLINICAL CONSIDERATIONS .................. 80 Oana Iuhas, Claudia Jurca, Radu Harbuz, Kinga Kozma, Marius Bembea P58. MULTIPLEX RT-PCR AS A USEFUL AND COST EFFICIENT METHOD FOR FUSION GENE TRANSCRIPTS DETECTION IN ACUTE LEUKEMIA ................................. 81 D. Jardan, C. Jardan, R. Talmaci, D. Coriu P59. INTERPHASIC FISH ON PERIPHERAL BLOOD IS A SENSIBLE METHOD FOR EVALUATION OF MINIMAL RESIDUAL DISEASE IN CML ............................................. 81 C. Jardan, D. Jardan, R. Talmaci, D. Coriu 10 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P60. CHIMERISM ANALYSIS – ELECTIVE METHOD FOR THE OUTCOME PREDICTION OF ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION ............................. 82 C. Jinca, S. Arghirescu, M. Puiu, A. Oprisoni, L. Balint-Gib, V. Ordodi, M. Serban P61. CHEDIAK-HIGASHI SYNDROME-CLINICAL AND EVOLUTION ASPECTS ..................... 82 E. Boeriu, M. Cucuruz, M. Lelik, G. Brad, I. Ursache, A. Botiz, S. Tamas, M. Serban P62. COMBINED EFFECT OF DIET AND THE RS1799883 POLYMORHISM OF THE INTESTINAL FATTY ACID BINDING PROTEIN 2 GENE ON THE RISK OF DEVELOPING THE METABOLIC SYNDROME .......................................................... 83 Katalin Csep, Gyongyi Dudutz, Claudia Banescu, Anamaria Butila Todoran, Laszlo Koranyi P63. RARE FAMILIAL CASE OF LISSENCEPHALY ................................................... 84 Kinga Kozma, Marius Bembea, Claudia Jurca, Radu Harbuz, Cristina Skrypnyk, Oana Iuhas, Marius Ivascu P64. DIAGNOSIS AND MANAGEMENT OF OSTEOGENESIS IMPERFECTA ......................... 84 M. Macovei, R. Cojocariu, M. Panzaru, R.Popescu, L.Caba, L.Butnariu, E. Braha M.Volosciuc, C. Rusu P65. DOWN SYNDROME-CLINICAL AND IMAGISTIC CORRELATION .............................. 85 A.Manea, C. Ilie, M. Boia, J. Puiu, D. Iacob, M. Dima P66. DOWN SYNDROME, DIAGNOSIS AND PREVENTION .......................................... 86 Marchian Sanda P67. DISCUSSION OF A CASE STICKLER SYNDROME ............................................... 86 Otilia Marginean, Ioan Simedrea, Maria Puiu, Belei Oana, Bochean Camelia, Tamara Marcovici, Camelia Daescu, Daniela Chiru P68. EARLY TREATMENT MANAGEMENT IN HYPODONTIA ........................................ 87 Popa Malina, Dinu Stefania, Luca Magda, Lazar Cristina, Bratu Cristina, Szuhanek Camelia, Balan Raluca, Ogodescu Emilia, Ogodescu Alexandru P69. HLA ROLE IN CHRONIC AUTOIMMUNE THYROIDITIS PREDISPOZITION ................... 87 Alina Martinescu, Irina Durbala, Eduard Circo P70. 13P CHROMOSOME POLYMORPHISM IN A COUPLE WITH RECURRENT MISCARRIAGE – CASE REPORT ......................................................................... 88 Anca Mitroi, Iuliana Dimofte, Mariana Aschie P71. MOLECULAR PROFILING OF ADAM12 IN BREASTCANCERS ................................ 88 Diana Narita, Andrei Anghel, Edward Seclaman, Razvan Ilina, Natalia Cireap P72. ADAM17: SIGNALING SCISSORS IN THE TUMOR MICROENVIROMENT ..................... 89 Diana Narita, Andrei Anghel, Edward Seclaman, Razvan Ilina, Natalia Cireap P73. CLINICAL STUDY REGARDING THE LINK BETWEEN MTHFD POLYMORPHISM AND TRISOMY 21 ........................................................................................ 89 Daniela Neagoş, Ruxandra Creţu, L.Camil Bohiltea P74. REPORT OF CYTOGENETIC ANALYSIS FOR 68 CASES WITH ANOMALIES OF SEXUAL DEVELOPMENT ............................................................................ 90 Diana Ochiana, Vasilica Plaiasu, Gabriela Motei, Amalia Costin P75. CONGENITAL HEART DEFECTS IN CHAR SYNDROME ........................................ 91 Monica Panzaru, Mihail Volosciuc, Cristina Rusu „Elena Braha” Lavinia Caba, Lacramioara Butnariu, Mihai Macovei, Roxana Cojocariu, Cristina Rusu P76. MILD HYPERHOMOCYSTEINEMIA SECONDARY TO HOMOZYGOUS C677→T MUTATION IN THE METHYLENETETRAHYDROFOLATE REDUCTASE GENE, ASSOCIATED WITH PORTAL-VEIN THROMBOSIS AND PORTAL CAVERNOMA .................................... 92 Petrescu Carmen, Grigorescu-Sido Paula, Bârsan Mircea, Zoica Bogdana, Scridon Traian P77. RARE CHROMOSOMAL DELETIONS WITH BREAKPOINTS OVERLAPPING THOSE OF THE WILLIAMS-BEUREN SYNDROME ................................................................. 92 V. Plaiasu, D.Ochiana, G.Motei, A.Costin, T.Ciomartan, I.Anca P78. CHROMOSOMAL CHANGES OF THE ACROCENTRICS IN ACUTE LEUKEMIA ............. 93 Popa Cristina, Ionita Hortensia, Surducan Dan, Nicoleta Andreescu, Alina Belengeanu, Maria Puiu, Margit Serban Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 11 P79. Cytogenetic issues in „CRI DU CHAT” syndrome - The experience of Iasi medical genetics center ................................................................ 93 R. Popescu, C. Rusu, M. Volosciuc, L. Butnariu, E. Braha, M Panzaru, L Caba, M. Macovei, R. Cojocariu, M. Gramescu, C.Bujoran, I. C. Ivanov, E. V. Gorduza P80. ETHICAL CONSIDERATIONS REGARDING GENETIC MODIFIED ORGANISMS .............. 94 Pusta Dana, Mariela Militaru, I. Paşca, Rodica Sobolu P81. ULTRASOUND SOFT MARKERS IN SECOND TRIMESTER AND FETAL ANEUPLOIDY ..... 95 Viorica Radoi, Dana Mierla, Luminita Neagu, Andreea Mustata, Silviu Predoi, L.C.Bohiltea P82. CORRELATIVE STUDY OF HLA ANTIGENS (A, B AND C LOCI) IN SOLID BRAIN TUMORS ............................................................................... 96 A.Rodewald, A.Koenig, Angelika Kroll, Georgeta Cardos P83. EHLERS – DANLOS SYNDROME – DISCUSSIONS CONCERNING DIAGNOSIS AND MANAGEMENT ..................................................................................... 96 Alexandra Rusu, Mihail Volosciuc, Lacramioara Butnariu, Cristina Rusu P84. CLINICAL VARIABILITY IN VELO - CARDIO- FACIAL SYNDROME .......................... 97 Cristina Rusu, Mihail Volosciuc, Monica Panzaru, Constantin Iordache, Florentina Cucer, Vlad Gorduza P85. KABUKI SYNDROME – IASI MEDICAL GENETICS CENTER’S EXPERIENCE ................ 98 Cristina Rusu, Mihail Volosciuc, Adriana Sireteanu, Elena Braha, Lacramioara Butnariu Constantin Iordache, Florentina Cucer P86. OCCURENCE OF HIGH TITER INHIBITORS IN A PATIENT WITH MILD HEMOPHILIA A – ANALYSIS OF RISK FACTORS .......................................................................... 98 M. Serban, L. Pop, D. Mihailov, E. Ursu, M. Puiu, D. Savescu P87. PARTIAL MONOSOMY 9P22-PTER - CLINICAL AND CYTOGENETIC STUDY ............... 99 A. Sireteanu, M. Voloşciuc, E. Braha, V. Gorduza, C. Rusu P88. FETAL GENDER DETERMINATION BY ANALYZING THE FREE FETAL DNA CIRCULATING IN MATERNAL BLOOD - POSTER ..................................................... 100 Adriana Stan, Cristina Dragomir, Emilia Severin, Lorand Savu P89. DISMORFISM PHENOTYPE WITH MINOR AND PARTIAL AGENESIS OF BODY CALOS .... 100 Dana Metea Stefanescu, Liviu Pop, Cristina Dragomir, Simona Farcas P90. DELINEATION OF CHROMOSOMAL ANOMALIES IN METAPHASE AND INTERPHASE CELLS BY FISH AND ARRAY CGH TECHNIQUES ..................................................... 101 Monica Stoian, Valerica Belengeanu, Eli Ormerod, Nicoleta Andreescu, Simona Farcas, Sorin Iurian P91. PHENOTYPE AND CYTOGENETIC CHANGES IN A TRISOMY 8 IN MOSAICISM CASE - FROM NEWBORN TO INFANCY ................................................. 102 Monica Stoian, Valerica Belengeanu, Nicoleta Andreescu, Simona Farcas, Cristina Popa, Marioara Boia P92. MEDICAL REHABILITATION FOR LIFE QUALITY IMPROVEMENT IN A CASE WITH ARTHROGRYPOSIS ........................................................................ 102 Dorina Stoicanescu, Mariana Cevei, Valerica Belengeanu, Lucia Stoican, Monica Stoian Nicoleta Andreescu P93. IMPROVED FUNCTIONAL PROGNOSIS AFTER MEDICAL REHABILITATION IN A CASE WITH CRI DU CHAT SYNDROME .......................................................... 103 Dorina Stoicanescu, Mariana Cevei, Valerica Belengeanu, Lucia Stoican, Simona Farcas, Cristina Popa P94. RARE VARIANT E19A2 BCR-ABL FUSION TRANSCRIPT IN TYPICAL CHRONIC MYELOID LEUKEMIA: CASE REPORT ..................................................... 103 Andreea Tutulan-Cunita, Sorina Mihaela Chirieac, Gabriela Mocanu, Catalina Luca, Marieta Costache, Agripina Lungeanu, Aurora Arghir P95. SEVERE MICROGNATHIA, QUESTION MARK EARS – A RELATIVELY RECENT SYNDROME, AURICULO-CONDYLAR SYNDROME ................................................................... 104 M. Volosciuc, Elena Braha, Aurica Rugină P96. THE KLIPPEL FEIL SYNDROME: DIFFICULTIES OF GENETIC COUNSELING .............. 104 Andra But, Corina Duncescu, Adela Emandi-Chirita, Loredana Covle, Maria Puiu, Narcis Dobre P97. HUNTER SYNDROME: THE HISTORY OF A DIAGNOSIS ...................................... 105 Maria Puiu, Cristian Jinca, Laura Pop, Margit Serban 12 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P98. LOBSTEIN’S DISEASE – CASE REPORT ........................................................ 105 Boia ES, Marioara Boia, Popoiu MC, David VL P99. MUSCULOSKELETAL DISORDERS IN PATIENTS WITH PRADER WILLI SYNDROME ....... 106 Popoiu MC, David VL, Boia ES, Maria Puiu P100. PRADER WILLI SYNDROME – FROM RESEARCH PROJECT TO MULTICENTRIC APROACH ............................................................................. 106 M. Puiu, D. Dan, M. Serban, M. Gafencu, G. Anton, N. Cucu P101. SOLITARY MEDIAN MAXILLARY CENTRAL INCISOR SYNDROME – CASE REPORT ...... 107 Dinu Stefania, Popa Malina, Bratu Cristina, Jianu Rodica, Szuhanek Camelia, Lazar Cristina, Ogodescu Emilia P102. TREATMENT MANAGEMENT OF OLIGODONTIA ............................................ 107 Dinu Stefania, Bratu Elisabeta, Glavan Florica, Schiller Eleonora, Popa Malina, Ogodescu Alexandru, Dinu Cristian Dorin, Luca Magda, Balan Raluca P103. GENETIC DETERMINISM OF CHRONOLOGIC AGING SKIN ................................. 108 Anca Dragomirescu P104. KLINEFELTER SYNDROM – CASE REPORT ................................................... 108 Rodica Urtila, Valeria Belengeanu, Eulalia Boceanu, Sonia Tanasescu, Patricia Urtila P105. SURGICAL APPROACH OF A CHILD WITH CONGENITAL ANOMALIES OF THE KIDNEY AND URINARY TRACT ........................................................................ 109 Vasilie D., Puiu M., Poclid I., Cristina, Mejdi R. P106. SPECIAL NEEDS CHILDREN’S DAY ............................................................ 109 Ionela Alina Moaca, Iulia Jurca-Simina, Florin Jurca-Simina Iosif Reascu, Norbert Pop, Stefan Berci, Iulia Popa, Irimia Cristina, Oana Rosca, Graziella Ecob, Adrian Juverdeanu, Carmen Dumitranoiu, Mihai Gafencu, Maria Puiu, Narcis Dobre P107. THE IMPORTANCE OF EARLY DIAGNOSIS IN MARFAN SYNDROME ...................... 110 Anca Popoiu, Gabriela Doros, Bogdana Zoica, Popoiu MC, David VL, Boia ES, M Puiu P108. NEED SURGERY ON GENETIC DISEASES ..................................................... 111 Jurca Simina Florin Ioan, Jurca Simina Iulia Elena, David Vlad, Puiu Maria P109. LIMITS OF STANDARD KITS FOR CYSTIC FIBROSIS GENETIC TESTING IN ROMANIA ...... 111 L.Pop, I.M. Ciuca, L.Tamas, Z.Popa, Gh. Budau, I.Popa P110. THE DIAGNOSTIC VALUE OF SERUM CARBOHYDRATE ANTIGEN LEVELS 19-9 IN DIABETIC PATIENTS .......................................................................... 112 Claudia Borza, Rodica Mateescu, Germaine Savoiu, Corina Serban, Lelia Susan P111. SERUM PROSTATE-SPECIFIC ANTIGEN (PSA) LEVELS IN MEN WITH TYPE 2 DIABETES ....................................................................................... 112 Claudia Borza, Rodica Mateescu, Germaine Savoiu, Corina Serban, Lelia Susan P112. Rare Diseases Week in Timisoara ........................................................ 113 Oana Rosca, Ionela Alina Moaca, Iulia Jurca-Simina, Florin Jurca-Simina, Iosif Reascu Norbert Pop, Stefan Berci, Iulia Popa, Irimia Cristina, Graziella Ecob, Adrian Juverdeanu, Carmen Dumitranoiu, Narcis Dobre, Mihai Gafencu, Maria Puiu Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 13 Reports (Following Congress Program) R1. THE ORIGIN OF ROMANIANS FROM MITH TO REALITY: GENETIC EVIDENCE FROM OWN STUDIES ON OLD AND MODERN HUMAN POPULATIONS FROM ROMANIA Alexander Rodewald1, Calin Tesio2, Georgeta Cardos3, Dorina Banica4 1 Institute of Human Biology-University of Hamburg, Germany 2 Faculty of Biology, University of Bucharest, Romania 3 „Victor Babes” National Institute of Pathology, Bucharest, Romania 4 „Marius Nasta” National Institute of Pneumology, Bucharest, Romania The origins of Romanians are stil under debate, there are more theories excluding each other, interpreting different archeological and historical sources, no one being able to exhaustively explain toponimical, linguistic and etnographic data. The aims of our populational and molecular paleogenetic study were: • To analyze the genetic structure of old human populations from the Bronze and Iron Ages from Romania, their genetic variation and the degree of their genetic kinship with today’s Romanian population and other modern European populations, • To elucidate the genetic structure of modern Romanian population and their genetic relationship with other European populations. The biological material was represented by: • Ancient (aDNA) extracted from skeletal remains (bones and teeth) of 50 old individuals from the Bronze and Iron Age excavated from different archeological sites from Romania, • Modern DNA isolated from 300 blood samples collected from modern Romanian people, after their inform consent. Mitochondrial (Hypervariable Regions – HVR-I and II) and nuclear (the VWA31A microsatellite and the Amelogenin Gene) DNA markers were analyzed from aDNA. From modern DNA were also analyzed mitochondrial DNA markers, more nuclear microsatellite DNA markers (D1S80, D3S1358, TH01, FGA, D5S818, D7S870, D13S317, VWA31A, CSF1PO, TPOX) and the Amelogenin Gene. Biostatistical analysis and phylogenetic networks of mtDNA types were constructed by means of the Median-Joining Network Method. Genetic distances were counted by both Cavalli-Sfortza and Edwards Method and Principal Component Analysis. The phylogenetic trees were constructed based on Fst distances counted by Kimura 2-parameter and pairwise differences formulas. Our results revealed as follows: • Old human populations from the Bronze and Iron Ages from Romania have shown closer genetic relationship to Turks of indo-European origin, but also with modern Romanian, Greek and Italian populations, • Modern Romanians were genetically closer to modern Bulgarians and Greeks. Based on our results, we may conclude that old human populations from the Bronze and Iron Ages from Romania have might contributed to the foundation of modern Romanian genetic pool. 14 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 R2. Y HAPLOTYPES – IDENTIFICATION, DIVERSITY, MEANING Marius Bembea1, Attila Patocs2, Kinga Kozma 1, Claudia Jurca1, Cristina Skrypnyk1 1 University of Oradea, Faculty of Medicine and Pharmacy; Municipal Clinic Hospital; Genetics Department, C. Coposu str. 12 , 410469 Oradea, Romania; e-mail: [email protected] 2 Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, 46 Szentkiralyi str, H-1088 Budapest, Hungary; e-mail: [email protected] The Y chromosome is transmitted only by paternal lineage and it passes thrue from father to son as one piece, most of all unchanged. Small changes may occur, which are called polymorphism. Different combinations of polymorphism realize haplotypes. The DNA copy from one generation to another, although rare, mistakes can occur. As Y chromosome does not change the acid nucleotides during crossing-over (except the pseudo-autosomal region), these errors are the only ones passing to the next generation. There are 4 types of changes that can occur from a generation to another: indel, SNP (single nucleotide polymorphisms), microsatellites (Y-STR) and minisatellites. STR test determine the haplotypes while haplogroups are predicted based on haplotypes. Identification of Y haplotypes in different populations of the world has gained importance in recent years and its expansion is growing. This led to the formation of an International Reference Database (Y-chromosome Haplotype Reference Database - YHRD) which is gathering the frequency of haplotypes Y dissemination mode in populations worldwide. The aim of this reference database is to offer information for analysis in forensic science, in research and anyone who is interested of family or population genetics history. The authors present the results of a research about Y haplotype diversity in 4 distinct populations of Bihor County: a control group from the general population of Bihor County (n=73 male individuals) and three isolated ethnic population: Slovak (n=35), German (n=36) and Roma (n=36). Haplotype data has been deposited in the Y-chromosome haplotype reference database (YHRD-Accession Numbers: YA3602, YA3603, YA3604, YA3605, release 33). Key words: Y chromosome , haplotype, haplogroup, Y-STR, YHRD. R3. PRENATAL CYTOGENETIC TESTING IN A PROSPECTIVE STUDY ON 1,000 PREGNANT WOMEN V. Pop, M. Militaru, A. Popp, F. Stamatian Department of Medical Genetics, U.M.F. Cluj-Napoca Introduction. Chromosomal syndromes and congenital malformations are a public health problem because they have a high global incidence and because they are serious, chronic-disabling health problems that can be prevented, which is why prenatal cytogenetic testing is required. Objectives. This study aimed at different aspects of pregnancy associated chromosomal abnormalities, such as prenatal screening, frequency assessment, prevention, cytogenetic analysis issues, correlations between indications and results. Materials and methods. The study was performed on 932 cases, from 04/01/2002 to 12/31/2008. Inclusion criteria of pregnant women in the study group were well defined and standardized. Genetic consult and counseling were performed in cases with genetic risk for chromosomal abnormalities. Fetal cells obtained through amniocentesis were cultured for 10 days in 5% CO2 atmosphere on nutrient medium (HAM F10 and amniochrome) at 37°C and pH 7.3. Visualization of chromosomes was performed by Giemsa staining and metaphase G-banding; cytogenetic analysis was performed on camera-captured images, on 8 different mitoses using a 100x-immersion objective. Results and discussions. Distribution of chromosomal abnormalities reveals that the autosomal aneuploidies are the most common, representing three fifths of total (59%), while other types Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 15 represent only two fifths of total (41%), of which heterosomal abnormalities are 17%, genome abnormalities are 5%, and structural anomalies are 19%. In this group, the autosomal chromosomal abnormalities had the following distribution: trisomy 21 - 56.82%, trisomy 18 - 25%, trisomy 13 - 4.54% rare autosomal aneuploidies - 13.64%. Gonosomal aneuploidies were present in 17.33% of cases with the following distribution: monosomy X - 6 cases (46.15%), heterosomal trisomy XXY - 4 cases (30.77%), heterosomal trisomy XXX - 3 cases (23.08%). Genome abnormalities have been detected in four cases, of which 3 had homogeneous triploidy and one had mosaic tetraploidy, with an XX euploid cell line and an XX/XY tetraploid cell line. Structural chromosomal abnormalities were very diverse, the following types of chromosomal rearrangements being identified in this group: marker chromosomes in 5 cases (35.71%), chromosomal deletions in 4 cases (28.57%), chromosomal translocations in 3 cases (21.43%), and rare structural abnormalities in 2 cases (14.29%). Male to female ratio was 1.27 (42/33 or 56%/44%, 1/0.79 respectively). Homogeneous aneuploidychromosomal mosaicism distribution was 58/17 (77%/23%, 1/0.23 respectively). Conclusions. Statistical analysis of the global share of the inclusion criteria or their combinations, showed the following: the individual share of each criterion decreases in the direction OSH> GA> SEA >AP+ > IF+; the combined criteria have a 40% share; in positive cases, the combination of 2 or 3 criteria accounts for 57.89% and 42.11%, respectively; in positive cases, the combinations of 2 criteria are: the advanced gestational age-maternal serum screening (54.6%), advanced gestational age- ultrasound markers (18.2%), ultrasound markers-maternal serum screening (18.2%), maternal serum screening-positive parental history (9.0%). Keywords: genetic risk pregnancy, prenatal screening, numerical chromosomal abnormalities, structural chromosomal abnormalities, genome abnormalities, homogeneous aneuploidy, chromosomal mosaicism, autosomal trisomies, gonosomal trisomies, recurrent miscarriage, cytogenetic test, chromosomal analysis, genetic counseling, informed consent, genetic consult, genetic advice. R4. THE SHORT STATURE – HOW IMPORTANT IS THE GENETIC ETIOLOGY Eusebiu Vlad Gorduza, Lacramioara Butnariu Medical University „Gr. T. Popa” Iasi The short stature is define by a retardation of stature growth over than -2SD in rapport with age’s medium height. A pathologic short stature is identified in 80% of cases with height deficit >-3DS. The prevalence of short stature is 2,5% of population, the main causes being: chronic infections, the endocrine diseases, malnutrition and genetic factors. The pathologic short stature can be dividing in disproportionate and proportionate (with prenatal or postnatal onset). The disproportionate short stature is the consequence of an osteocondrodysplasia. It is about 200 osteocondrodysplasias, the majority are rare and are produced by anomalies of growth and remodeling of bone (cartilage). The most well-known osteocondrodysplasias are: osteogenesis imperfecta (I-IV) the diseases produced by mutations of fibroblastic growth factors receptors (achondroplasia, hypocondroplasia etc.) the syndrome Ellis canCreveld, the syndrome McCune Albright. The proportionate short stature represents a cardinal sign in many genetic diseases. In general, a syndrome with short stature presents also the anomalies in other systems, usually being identified the changes at level of members, heart or skin. The growth retardation represents a characteristic of many unbalanced chromosomal anomalies. From these diseases can be listed the syndromes: Turner, Down, Edwards, Patau, Wolf Hirschhorn, cri du chat, Prader-Willi, Williams. The proportionate short stature can be detected also in some monogenic diseases, like the syndromes: Cornelia de Lange, Noonan, Rubinstein-Taiby, Russell-Silver or Smith-Lemli-Opitz. In conclusion, in any case of pathologic short stature, after the exclusion of another etiology, it is important to study the genetic component, by genetic consultation and analysis. 16 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 R5. BOALA GAUCHER TIP 1 ÎN ROMÂNIA: EPIDEMIOLOGIE, GENOTIP, FENOTIP ŞI EVOLUŢIA Paula Grigorescu–Sido Spitalul Clinic de Urgenţă pentru Copii – Centrul de Patologie Genetica Cluj Obiectiv: Prezentarea epidemiologiei, a caracteristicilor clinice şi moleculare ale pacienţilor cu boală Gaucher tip I din România ca şi a evoluţiei acestora cu şi fară terapie de substituţie enzimatică. Pacienţi şi metode: 50 pacienţi (F/M=1,63/1) cu boala Gaucher tip 1 (diagnosticaţi enzimatic şi molecular) au fost evaluaţi clinic şi prin determinarea iniţial şi la interval de 6 luni a: hemoglobinei; numărului de trombocite; volumului splenic şi hepatic; chitotriozidazei; densităţii minerale osoase (BMD – scor Z) la nivelul coloanei lombare şi a scorului de severitate. 39 de pacienţi (78%) au fost trataţi cu imiglucerase (44,4 ± 13,6 U/kg i.v., la interval de 2 săptămâni) pe o perioadă de 3,1±1,4 ani. Rezultate: Lotul reprezintă 22,7% din numărul posibil de pacienţi cu boala Gaucher din România. Vârsta medie a acestora a fost de 15,48 ani la debutul clinic; şi 28,89 ani la confirmarea specifică a diagnosticului. Există o frecvenţă importantă (35,9%) a genotipului N370S/L444P, predictiv pentru un fenotip sever. 30% dintre pacienţi au fost splenectomizaţi, înainte de ERT. Anemia, trombocitopenia, splenomegalia şi boala osoasă au fost prezente la 38%; 70%; 100% şi respectiv 84% dintre pacienţi. Corelaţia genotip – fenotipa fost prezentă, dar imperfectă. ERT a dus la normalizarea hemoglobinei, a numărului de trombocite, a volumului hepatic şi a chitotriozidazei după 0,5; 1,5; 2,5 şi respectiv 3 ani de tratament. În patru ani, splenomegalia s-a redus de la 14,4 x N, la 3,06 x N. Evoluţia clinică a bolii osoase a fost favorabilă, dar BMD s-a redus la pacienţii trataţi cu 30U/kg la 2 săptămâni. Pacienţii netrataţi s-au agravat progresiv iar doi au decedat. Concluzii: Boala Gaucher tip 1 este nediagnosticată în ţara noastră, mai ales la bărbaţi. Există o frecvenţă mare a genotipului N370S/L444P şi un fenotip sever la majoritatea pacienţilor. ERT a fost eficientă asupra manifestărilor hematologice şi a visceromegaliei şi mai puţin asupra densităţii minerale osoase. Pentru obţinerea unor rezultate terapeutice optime, este esenţială iniţierea terapiei cât mai aproape de debutul clinic al bolii cu o posologie adaptată gradului de severitate a acesteia. R6. THE CLINICAL CHARACTERISTICS AND OUTCOME IN PATIENTS DIAGNOSED WITH MUCOPOLYSACCHARIDOSES IN ROMANIA Camelia Al-Khzouz Pediatric Departament of University of Medicine and Pharmacy. „Iuliu Haţieganu” Cluj Emergency Childrens Hospital Cluj Napoca – The Genetics Pathology Center Introduction. Mucopolysaccharidoses (MPS) are lysosomal diseases caused by intralysosomal storage of acid mucopolysaccharides, characterized by dysmorphic cranio-facial features, somatic retardation, organomegaly, progressive mental retardation up to demence, bone dysplasia, osteoarthropathy with contractures in flexion and corneal opacities. Taking into account the perspectives of specific treatment, the identification of the deficient lysosomal enzyme is essential. Patients and methods. The study group included 21 patients (4 girls and 17 boys), aged between 1 – 14 years, with suggestive clinical MPS findings. The study methods consisted in: nonspecific assessment; clinical examination, anthropometrical measurements, imagistic procedures, biochemical assays, respiratory function tests, ophtalmological, ENT, neurological and psychological assessment; specific examinations: measurement of lysosomal enzymes involved in glicosaminoglican (GAG) metabolization: a) α-L-iduronidase, α-glucosaminidase, β-galactosidase, B arilsulphatase, β-glicuronidase (Biochimical Department of Univ. of Medicine and Pharmacy Cluj) b) iduronateRomanian Journal of Rare Diseases 2010 | Supplement 1/2010 17 sulphate-sulphatase and c) assessment of urine elimination of GAG (Univ.Sahlgrenska Suedia; Univ. Mainz). Results. The complete diagnostic assessment revealed: a) dysmorphic cranio-facial osteoarthrophaty, umbilical hernia, important hepatomegaly and mild splenomegaly (12 patients): severe moderate and mild hypostature in 11,6 and 4 patients respectively: b) multiple dysostosis 17 cases; c) degenerative keratopathy - 5 cases; d) severe, medium severity and mild mental retardation or normal intellect - 6, 6, 2 and 7 cases, respectively. The specific diagnosis was established in 17 patients: Type I MPS, type II B MPS, type III B MPS, type IV B MPS and type VII MPS - 3, 6, 6, 1 and 1 patient, respectively. Conclusions. Measurement of lysosomal enzymes allows establishment of specific diagnosis, a diagnosis of certainty, compulsory for starting enzyme replacement therapy. Two of type I MPS patients are on Aldurazyme treatment, while another patient with type II B MPS is on iduronatesulphatase ( Elaprase) therapy as a part of an international trial. R7. BAC-FISH- A TARGETED TOOL FOR ACCURATE CYTOGENETIC ANALYSISFROM BASICS TO DIAGNOSIS Eli Ormerod Department of Medical Genetics, Oslo University Hospital, Oslo, Norway BAC-FISH is now widely used as a targeted tool for confirmation of array-CGH findings in patients with suspected chromosomal imbalances. The technique is also used for parental follow up studies and for analysis of de novo inversions, „balanced” translocations or cytogenetically visible chromosome aberrations with normal aCGH findings. BAC-FISH is of significant importance for an accurate clinical interpretation. In our laboratory we have performed 73 BAC-FISH analyses on samples from 40 patients selected on aCGH (Agilent Technologies, Santa Clara, CA, 44k to 244k) or cytogenetic findings from conventional metaphase spreads. In cases where aCGH analysis revealed telomere involvement, we have used commercially available subtelomere probes. FISH analysis was performed with BAC clones selected to cover most of the human genome and derived from RPCI-11 and 13 libraries (30,388 clones), (Osoegawa et al., 2001) and from „CalTech” Human BAC CIT-D library (2062 clones). The selected BAC clones were grown in LB medium overnight and isolation was performed with BACMAX DNA Purification kit from EPICENTRE. Labelling and nick translation of the DNA was performed with a FISH-Tag DNA Kit (Invitrogen). Of the 40 patients BAC FISH analysis or subtelomere FISH 31 were informative. In particular, the microduplications can be difficult to validate by FISH unless the duplication is a result of a translocation to another chromosome or to another position on the same chromosome. R8. FIRST PATIENT WITH LATE ONSET TYPE II GLYCOGENOSIS IN ROMANIA: DIAGNOSIS, TREATMENT, EVOLUTION Ioana Nascu, Camelia Al-Khzouz, Simona Bucerzan, Paula Grigorescu-Sido Pediatric Departament of University of Medicine and Pharmacy „Iuliu Haţieganu” Cluj Emergency Childrens Hospital Cluj Napoca – The Genetics Pathology Center Introduction: Type II glycogenesis (Pompe disease) is a metabolic storage disease, autosomal recessive inhereted, determined by acid glucosidase deficiency which determines glycogen storage in the myocitic lysosoms. The disease is severe and prograssive towards the chronic respiratory failure and wheelchair restraint. Material and methods. We present a 10 years 11 months male, diagnosed with late onset type 18 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 II glycogenosis, treated with Myozime (20mg/kg/dose i.v. perfusion each 2 weeks) since december 2007, the first patient from Romania treated. Methods included: a) for diagnosis: clinical examination, biochemical tests: transaminases, muscular enzymes, electromyography, muscular biopsy and specific tests: acide glucosidase, DNA analysis (Bologna Neurophisiopathology Department and „Erasmus” University Rotterdam); b) evolution monitoring each 6 months (clinical examination, muscular enzymes). Results: the patients presented walking fatigue, muscular hypotonia, progressive motor deficiency. Other tests showed: hepatocytolisis, increased muscular enzymes, myogenic electromyography, muscular biopsy (vacuolary myocites). Specific tests confirmed the diagnosis and DNA analysis revealed 2 mutations on GAA gene IVS1 (-13T→G) (2104 C → T p.R702C). Evolution during the treatment was favourable. Conclusions: Enzyme replacing therapy has completely changed the prognosis of this disease. R9. MOLECULAR DIAGNOSIS OF PROGRESSIVE MUSCULAR DYSTROPHIES France Leturcq Institut Cochi , Université Paris Descartes; Laboratoire de Biochimie Génétique et Moléculaire, Hôpital Cochin, PARIS, France Muscular dystrophies (MD) are a heterogenous group of monogenic inherited diseases that primarily affect skeletal muscle and are characterized by progressive muscle wasting and weakness. Since the historical discovery of the defective gene involved in Duchenne and Becker Muscular dystrophy (DMD gene, 1986), 24 distinct genes have been identified as possible cause of MD when mutated. To enable adequate genetic counselling (carriers and prenatal diagnosis) it is mandatory to characterize the gene involved and its mutation(s) in every patient with an MD phenotype. This is obtained thru molecular explorations of the gene and its products. The choice of the gene to investigate is oriented by clues derived from clinical features, mode of inheritance and above all by muscle biopsy analysis. We will describe the molecular diagnostic strategies in use in our laboratory for some important MD such as : DMD/BMD (dystrophin), Emery-Dreifuss (emerin and lamin A/C), limb-girdle muscular dystrophies (calpain-3, dysferlin, sarcoglycans, glycosylation defects). Finally we will briefly depict the present progress of massive multigene mutation analysis by CGH-array, and the new avenues opened by allele-specific therapeutic strategies. Mots clés : muscular dystrophy, gene, DMD, diagnosis. R10. SOMETHING BORROWED AND SOMETHING NEW: IGF-1R AND UBIQUITIN ON THE LIST OF TARGETED THERAPY IN CANCER Prof. Dr. Doc. Leonard Girnita, MD, PhD Associate Professor of Pathology Karolinska Institutet Department of Oncology-Pathology, Cellular and Molecular Tumor Pathology Cancer Center Karolinska, CCK R8:04 Karolinska Hospital, S-17176, Stockholm, Sweden The result of receptor mediated signal transduction is the propagation of a cascade of protein conformational changes in response to agonist stimulation. These signals are, in part, generated by the growth factor receptors. Among them, IGF-1R is one of the most important players in cancer development. Phosphorylation is known as being the central process governing IGF-1R signaling. However, during the last years we described the involvement of ubiquitination on IGF-1R function. We could prove that Mdm2 and -arrestin acts as a crucial component in the ubiquitination and consequently, down-regulation and signaling of the IGF-1R receptor. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 19 Recently, we found that the cyclolignan picropodophyllin (PPP) inhibits phosphorylation of IGF1R and activation of downstream signaling without interfering with the highly homologous insulin receptor (IR). Furthermore, PPP treatment caused strong regression of tumor grafts and prolonged survival of animals with systemic tumor disease. Furthermore we demonstrated that PPP also down regulates the IGF-1R, without affecting other receptors. PPP-induced IGF-1R downregulation required expression of the MDM2 E3 ligase, which recently was found to ubiquitinate and cause degradation of the IGF-1R. In addition knockdown of -arrestin1, the adaptor molecule known to bridges MDM2 and IGF-1R, prevented downregulation of the receptor and significantly decreased PPP-induced cell death. In conclusion the present study demonstrates that an IGF-1R inhibitor causes receptor downregulation, probably through Mdm2/-arrestin -induced ubiquitination of IGF-1R. Taken together, we proposed a model for clinical development of the receptor tyrosine kinases targeted therapy in cancer, in which the central objective is to achieve RTK downregulation through increased ubiquitination rather than barely inhibition of the receptor activity. R11. TRANSLATIONAL TOOLS FOR RARE DISEASES: THE SOLUTIONS DEVELOPED BY ORPHANET Ségolène Aymé INSERM, SC11, Orphanet – Plateforme Maladies rares – 102 rue Didot – 75014 Paris – France [email protected] To resolve the issue of information dispersion, Orphanet gathers an inventory of rare diseases. Each disease has a unique identifier and is placed in a poly-hierarchy classification system. All the classifications of diseases can easily be displayed on the website. Orphanet has also developed an encyclopaedia published in an electronic, open-access journal, the Orphanet Journal of Rare Diseases. To help physicians diagnose rare diseases, Orphanet provides a query system of signs and symptoms. The possible diagnoses are listed in order of probability. To support appropriate referrals, Orphanet has developed a continuously updated directory of expert clinical centres and expert clinical laboratories in 38 countries. To promote quality services, data on quality management of clinical laboratories are available on the website. Distinct logos indicate which laboratories are certified, accredited and/or participate in external quality assessment. This information is gathered and validated in partnership with EuroGentest. To facilitate collaboration between researchers and between researchers and Industry, Orphanet lists all ongoing national and European-level funded research projects by type of research and by disease. The licensing opportunities are displayed, as well as the patient registries, biobanks and highly specialised platforms and know-how, which may be of interest in R&D. To help patients establish contact with other patients, Orphanet furnishes information on existing patient organisations. To build up the rare disease community, Orphanet publishes a free-access electronic bimonthly newsletter which has over 12,000 subscribers. To support policy-makers, Orphanet regularly publishes reports in a collection entitled „Orphanet Report Series”. This set of services has been instrumental in contributing to the establishment of research networks and has a measurable effect on the referrals of patients to expert clinics and of samples to expert laboratories. 20 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 R12. HAEMOPHILIA A CHALLENGE FOR HEALTH CARE IN ROMANIA M. Serban, M.Puiu, H. Ionita University of Medicine and Pharmacy „Victor Babes” Timisoara Haemophilia, together with at least 8,000 other illnesses, is part of rare diseases, defined by their frequency as below 1 in 2000 persons, corresponding to a prevalence of less 500/million inhabitants. In light of these figures, the incidence of haemophilia accumulated approximately 100/million inhabitants, may be rated as „very rare disease”. Fortunately, it is placed, among the approximately 40 diseases in this group, which benefits of specific therapies. Unfortunately, like other rare diseased, the therapy is extremely expensive, requiring a continuous use throughout life, resulting in major difficulties for countries with limited resources. They are expressed in limited life expectancy (35-40. vs. 65-75 years) and mobility impairments in up to 75% of patients with the debilitating impact (severe forms of disease) reflected limited social and professional inclusion. Quality of care of haemophilia can be defined by several distinguishing features: • Existence of a network of haemophilia treatment centers (HTC), with 24 hours a day, 7 days a week availability in hospital or ambulatory conditions; Infrastructure competency for proper diagnosis of all forms of hemophilia and inhibitors of anti-F VIII or IX • Treatment: native blood products (fresh plasma or cryoprecipitate) are out of usage. Exclusiveuse of industrial preparations, safe and effective concentrated factors (CF) allowing to solve the bleeding (on demand). • Emergency assistance operators, life-saving interventions and orthopedic invasive surgery enable mobility recovery and allow initiation of home therapy. Optimum quality represents prophylactic substitution with CF at least until the age of 18 years, solving the appropriate therapeutic inhibitors of anti-F VIII or IX and ensuring normal life style and socio-professional integration. Comparing the numbers of reference which quantify these features in different European countries, we illustrate the reality for Romania. Unfortunately we have a bad situation even in comparison to other former communist countries of Europe. We are still dominated by excessive centralization of resources, limiting on one hand, the quality of diagnosis and on the other, the therapeutic availability. R13. SYNDROMIC MENTAL RETARDATION – DYSMORPHIC FEATURES SUGGESTIVE FOR THE DIAGNOSIS Cristina Rusu, Mihail Volosciuc, Elena Braha, Lacramioara Butnariu, Monica Panzariu, Mircea Covic Medical University „Gr. T. Popa” Iasi Mental retardation is defined as an IQ below 70, with dysfunction in more than two developmental areas (communication, self care, home living, social skills etc) and onset during childhood. It is classified as mild, moderate and severe according to the IQ level and specific and nonspecific according to the presence or absence of birth defects associated to mental retardation. The paper aims to present cranio-facial defects that may be associated to developmental delay and could be suggestive for a certain specific diagnosis. Evaluation, normal and abnormal variants are discussed for every criteria. Abnormal skull contour, general facial appearance, ocular, nasal, oral and auricular defects are discussed successively. The presentation will be illustrated with many cases from Iasi Medical Genetics Center’s experience. Finally, the evaluation protocol for a child with syndromic mental retardation will be presented – family and personal history, anthropometric measurements, physical examination and photographs, genetic tests. In conclusion, we present some of the most suggestive dysmorphic elements associated to learning disability to facilitate recognition in practice of specific mental retardation cases. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 21 R14. THE ROLE OF ENVIRONMENTALLY-INDUCED EPIGENETIC CHANGES IN THE ETIOLOGY OF TYPE 2 DIABETES Ionel Sandovici Metabolic Research Laboratories, Department of Obstetrics and Gynaecology University of Cambridge, Cambridge CB2 2SW, United Kingdom Type 2 diabetes (T2D) is a disease of increasing prevalence resulting from an interaction between genetic and environmental factors throughout life. The elucidation of several genetic etiologies of both monogenic and polygenic T2D has revealed key regulators of glucose homeostasis and insulin secretion in humans. Genome-wide association studies (GWAS) have been instrumental in most of these recent discoveries. The T2D susceptibility genes identified so far are mainly involved in pancreatic beta-cell maturation or function. However, common DNA variants in those genes only explain approximately 10% of T2D heritability. Emerging evidence suggests that epigenetic mechanisms such as DNA methylation and histone modification changes may also play important roles in the etiology of T2D. Importantly, these epigenetic tags can be modulated by the environment, resulting in stable changes that can be passed through many cell divisions during the lifetime of an individual and even transgenerationally. In this presentation I will review the existing evidences that support the involvement of epigenetic changes in the etiology of T2D. Then, I will present our recent results that led to the discovery of abnormal epigenetic regulation at the T2D susceptibility gene Hnf4a in pancreatic islets of aged rats and rats exposed to an altered diet during early development. Finally, I will also present preliminary data from our recent genome-wide transcriptional and DNA methylation profiling of aged versus young rat islets. These results show that aging associates changes in DNA methylation that impact transcription of a large number of genes, some of which have been previously linked with an increased risk for T2D development. R15. ESTABLISHING DIAGNOSTIC TESTING SCHEMES FOR PRADER WILLI SYNDROME IN ROMANIAN POPULATION. FOCUS ON THE EPIGENETIC MECHANISMS UNDERLYING THE IMPRINTING DEFECTS CAUSING THE PWS Natalia Cucu1, Gabriela Anton1, Cosmin Arsene1, Anca Botezatu1, Maria Puiu2, Corin Badiu1, Vasilica Plaiasu1, Narcis Dobre3, Danae Stambouli4 1 University, Bucuresti, National Institut of Virusology, Bucuresti, 2 Medical University, Timisoara 3 VitroBioChem 4 Cytogenomic, Bucuresti. Prader Willi syndrome (PWS) is a complex disorder whose diagnosis may be difficult to establish on clinical grounds and whose genetic basis is heterogenous. Clinical diagnosis of the Prader-Willi and Angelman syndromes remain difficult in many instances because of the individual variations in the phenotype and because the phenotype develops only with age. Knowledge of the basic genetic defect and its possible inheritance, but also the recently discovered epigenetic defect and its inheritance is also necessary for genetic counseling. About 70% of cases are due to a 15q11-q13 deletion in the paternally contributed chromosome. These deletions are optimally detected by FISH method . The other aproximately 30% of cases of PWS are due to maternal uniparental disomy (UPD), that can best be documented using microsatelllite pobes. Less then 2% of cases have an imprinting defect, which causes nonexpression of paternal genes in PWS critical region. This latter group is detectable through identification of parent-oforigin differences by using methylation sensitive SNRPN, a process called “methylation analyses”. Chromosome analysis by classical karyotyping method is usually a routine part of the evaluation of these patients, in order to rule out other abnormalities, and will also detect rare instances of translocations or other chromosome rearrangements. From the point of view of genetic counceling, 22 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 none of the recurrence of PWS and its sister syndrome, AS, have involved the typical deletion or UPD, but rather have involved translocations and imprinting defects. Also, the methylation analysis, which confirm about 99% of cases without indicating the exact molecular mechanism, may be used in prenatal diagnosis, providing the knowledge of the correct developmental stage when the imptin is established in the germ line. Imprinting and epigenetic reprogramming in mammalian germ cells, the zygote and early embryos play crucial roles in regulating genome functions at critical stages of development . Aberrant chromatin states leading to aberrant gene expression patterns are determined by specific DNA methylation and histone modification processes (epigenetic modifications). They can occur secondary to a DNA sequence modification or mutation in a cis- or trans-acting factor or as true or primary epimutation in the absence of any DNA sequence change. Primary epimutations often occur after fertilization and lead to somatic phenotype mosaicism. It has been estimated that the rate of primary epimutaions isgreater than DNA mutations and actually may be underestimated. Knowledge of theso called “open windows” of vulnerability of the genome during the crucial stages of development and their interaction with the environment would be beneficial for the activities of establishment of optimal diagnosis and therapeutic or preventive schemes. R16. EPIGENETIC CHANGES AS MARKERS OF EARLY TUMOR DEVELOPMENT Dumitrescu R.G.1,2 , Liu Z. 3, Marian C.2 , Bebu I. 2, Yang Y. 2, Spear S.L. 2, Kallakury B.V.S. 2, Seillier-Moiseiwitsch F. 2, Freudenheim J. 4, Mason J. 3, Shields P.G 2 1. Saba University Medical School, Saba Island, Netherlands Antilles 2. Georgetown University Medical Center, Lombardi Cancer Center, Washington DC 3. Tufts University, School of Nutrition Science and Policy, Boston, Massachusetts 4. State University of New York, School of Public Health, Buffalo, New York Introduction: There is increasing evidence that changes in DNA methylation are important in breast carcinogenesis. The purpose of this study is to determine the hypermethylation status of the promoter regions of several tumor suppressor genes in normal breast tissues and identify the determinants of these epigenetic changes. Changes in one-carbon metabolism where folate is a critical substrate might affect gene expression through methylation or DNA synthesis. Methods: One hundred forty one healthy women with no history of cancer who were undergoing reduction mammoplasty were recruited to our study. All participants completed a structured interview and donated breast tissue. Methylation for p16INK4, BRCA1, ERα and RAR-β promoter regions from normal breast tissues were determined by methylation specific PCR. Subsequently, p16INK4 hypermethylation was analyzed using pyrosequencing. Genetic variants of MTHFR and MTR genes were determined by TaqMan assays. We analyzed folate concentrations in lean breast tissues, using pretreatment with folate conjugase followed by the standard microbiological assay. Spearman correlation coefficients, t-tests and logistic regression models assessed the associations between methylation phenotypes, genetic variants and folate levels in relationship with breast cancer risk factors. Results: p16INK4, BRCA1, ERα and RAR-β hypermethylation were identified in 31%, 17% 9% and 0% of the women, respectively. Women with BRCA1 hypermethylation had an eight-fold increase in the risk of ERα hypermethylation (p=0.007). p16INK4 hypermethylation was present in 28% of African-Americans, but 65% in European-Americans (p=0.02). DNA methylation phenotypes were significantly associated with race and family history of any cancer or breast cancer in particular. p16INK4 methylation phenotype were associated with the MTR AG/GG genotypes. p16INK4 promoter hypermethylation was also associated with breast folate levels (p-value= 0.02). Furthermore, the relation between p16INK4 promoter hypermethylation and breast folate was modulated by race (p-value= 0.03), family history of breast cancer (p-value= 0.04), and current smoking (p-value= 0.01). Conclusions: Gene promoter hypermethylation is commonly found in healthy breast tissues from women without cancer, indicating that these events are frequent and early lesions. Race and family Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 23 history of cancer increase the likelihood of these early events, DNA methylation phenotypes in normal breast tissue are associated with tissue folate levels and may be influenced by a genetic variant of the MTR gene. These associations appear to be modulated by risk factors such as race, smoking and history of breast cancer in the family. These findings may help elucidate gene-nutrient interactions that may play a role in the determination of breast cancer susceptibility. R17. SINGLE NUCLEOTIDE POLYMORPHISMS ASSOCIATED WITH C-REACTIVE PROTEIN LEVELS IN A COHORT OF YOUNG FILIPINO ADULTS Ghenadie Curocichin1,2 ,Thomas W. McDade3, Christopher Kuzawa3, Judith Borja4, Li Qin1, Damien C. Croteau-Chonka1, Amanda F. Marvelle1, Ethan M. Lange1,5, Linda S. Adair6, Karen L. Mohlke1, Leslie A. Lange1 1 Department of Genetics, University of North Carolina, Chapel Hill, NC 2 Department of Family Medicine, Moldova State Medical and Pharmaceutical University, Chisinau, Moldova 3 Department of Anthropology, Northwestern University, Evanston, IL, 4 Office of Population Studies, University of San Carlos, Cebu City, Philippines 5 Department of Biostatistics, University of North Carolina, Chapel Hill, NC, 6 Department of Nutrition, University of North Carolina, Chapel Hill, NC Introduction. C-reactive protein (CRP) is a reliable hallmark of chronic inflammation that may play an important role in atherosclerosis. CRP levels are associated with risk of type 2 diabetes, obesity and cardiovascular events [1-5]. Genome-wide association studies (GWAS) in middle-age and older populations predominantly of European descent have demonstrated associations of SNPs at several loci with serum CRP concentrations [6-8]. Substantial differences in CRP concentrations exist across populations [6]. Goal: Determine whether GWAS-identified variants are associated with CRP in a Filipino young adult sample. Materials and Methods. Population: Young Filipino adults (21-23 years old) , 931 males and 828 females, from the Cebu Longitudinal Health and Nutrition Survey (CLHNS) [9], examined in 2005. SNP selection. Based on available published data regarding association with CRP concentrations Exclusion of SNPs in high linkage disequilibrium (r2> 0.8) Potential functional significance based on SNP annotation. Genotyping. Genotyping was performed using TaqMan assays (Applied Biosystems). 11 SNPs in 7 loci: rs1205, rs3091244, rs1892534, rs2228145, rs1260326, rs10778213, rs1169288, rs7310409, rs769449, rs429358, rs7412. Triallelic SNP rs3091244 was genotyped using two separate assays (C/T and C/A); final genotype was called based on combining the two assays [10]. SNPs rs429358 and rs7412 were selected to define APOE haplotypes ε2, ε3, and ε4. Statistical analyses. 32.7% of samples had CRP concentration below the detectable level 0.1 mg/L Distributions of CRP and log-CRP concentrations were non-normal. For this reason, linear regression on log(CRP+0.1) and tobit regression on log-CRP (left-censored) were applied. Consistency between the results obtained using both models was treated as proof of validity. Outcome: Log-CRP levels adjusted for a measure of pathogen exposure and gender. Samples with CRP concentration >10 mg/L were excluded from analysis. Mean concentrations of CRP were calculated for quintiles of genotype score, where genotype score was calculated using the number of copies of each risk allele, weighted by the fitted linear regression model coefficients. The regression model included 6 loci that were significant when all SNPs were included in the model. 24 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Results. SNPs at five loci show significant association with CRP levels SNP Locus Annotation Risk allele Linear regression model Tobit model Estimate (SE) P-value Estimate (SE) P-value 0.0015 0.0046 1.8x10-9 4.1x10-8 0.050 0.335 (0.057) 0.433 (0.078) 0.296 (0.113) 0.169 (0.079) rs1205 CRP 3’UTR C rs3091244 CRP 5’ near gene A/T* rs1892534 LEPR - C 0.286 (0.043) 0.340 (0.060) 0.309 (0.086) 0.142 (0.061) rs2228145 IL6R Asp358Ala A 0.180 (0.050) 0.0013 0.273 (0.066) 0.0003 rs7310409 HNF1A Intronic G 0.169 (0.047) 0.0003 0.282 (0.061) 2.6x10-6 rs1169288 HNF1A Ile27Leu A 0.152 (0.046) 0.0010 0.249 (0.060) 3.5x10-5 rs1260326 GCKR Pro446Leu T 0.036 (0.045) 0.43 0.079 (0.058) 0.18 Rs10778213 12q23.2 Intergenic T 0.030 (0.054) 0.59 0.030 (0.071) 0.67 Rs769449 APOE Intronic G 0.206 (0.076) 0.0068 0.337 (0.10) 0.0007 Rs429358 APOE Cys130Arg T 0.213 (0.075) 0.0078 0.346 (0.099) 0.0008 rs7412 APOE Arg176Cys N/A 0.041 (0.070) 0.56 0.049 (0.091) 0.59 0.045 *2 variables for number of copies of A and T allele, respectively (2-degree of freedom test) Risk allele: the allele that is associated with elevated CRP levels in previously reported GWAS No SNPs showed statistically significant interaction with pathogen exposure score Conclusions. 1. For five out of seven CRP loci identified in European-based studies significant associations were demonstrated in a young adult Filipino population 2. Six SNPs explain 5.5% of variance of CRP concentrations 3. These results strongly support the feasibility of genetic analysis of inflammatory biomarkers associated with cardiovascular risk in younger populations R18. APPROACH TO RARE DISEASES AT THE NATIONAL AND EUROPEAN LEVEL Ségolène Aymé INSERM, SC11, Orphanet – 96 rue Didot – 75014 Paris – France Tel/+33 1 56 53 81 37 Fax:+33 1 56 53 81 38 [email protected] Member States’ (MS) policies and actions in the field of rare disease are rapidly evolving. Up till now, several countries have taken action to adapt their health care system to meet the needs of the RD patient community, or have planned to do so. With regard to centres of expertise, there are three categories of countries in Europe: those which have a policy regarding RD and have established centres of expertise within this framework (Denmark, France, Italy, Norway and Sweden); those which have established centres of expertise, though not specifically for RD (Belgium, Croatia, Czech Republic, Finland, Greece, Ireland, Portugal and the UK) and those which have no centres with this denomination, although they have centres Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 25 with all the characteristics of a centre of expertise (this is the case for almost all of the other countries). In addition, several research and/or public health networks have de facto identified centres of expertise in their own field. The Orphanet database lists 378 centres of expertise, mainly concentrated in Belgium, France, Germany, Italy, Spain and the UK. They cover very different realities as the qualifying criteria to define a centre as an expert centre differ from one country to another, both in terms of the mission and in terms of resources. Recently, some principles have been agreed on by the High Level Group of Health Services and Medical Care but various professional groups are not yet sufficiently aware of these principles, which where certainly not considered when designating the current expert centres. As a consequence, it is still difficult for health care users to understand what services they can obtain from these centres. In addition some experimental European reference networks of centres of expertise have been established and funded for a three year period only, a time period too short to allow any assessment of the added-value of these networks. With regard to genetic tests, they are now offered internationally, through both public and private sector genetic testing services. Currently, 956 laboratories offering tests for 1,559 genes are registered with Orphanet in Europe at large. The test offer differs greatly from one large country to another: Germany (1,141 genes), France (874 genes), Italy (625 genes), Spain (582 genes), the UK (414 genes). The test offer in medium and small-sized countries ranges from 1 to 233 genes. According to available data, only testing for Cystic fibrosis is provided by every country and 121 diseases are testable in only one country in Europe. This situation explains the large cross-border flow of specimens, highlighting the need to provide access to services in other countries when necessary, especially for very rare diseases. Legal and financial issues concerning cross-border testing are not yet fully addressed. With regard to the provision of information to patients and professionals, several MS have established web-based information services (Sweden and France) and telephone help lines (Bulgaria, Denmark, France, Italy, Netherlands, Norway, Spain, Sweden and the UK) although their resources vary considerably. Orphanet has expended its data collection on expert resources to almost all European countries. The difficulty is to maintain updated information about several thousands of diseases and to provide this information in languages understandable by the end users. So far information is only available in English, French, German, Italian and Spanish. Translation into Portuguese is on the way, and is planned for Flemmish/Dutch, Polish and Romanian. With regard to funding for research on rare diseases, the multinational common calls for proposals E-Rare now covers Austria, France, Germany, Greece, Israel, Italy, Spain, Turkey, The Netherlands and Portugal. Patient registries and databases constitute key instruments to develop clinical research in the field of rare diseases, to improve patient care and healthcare planning. They are the only way to pool data in order to achieve a sufficient sample size for epidemiological and/or clinical research. They are vital to assess the feasibility of clinical trials, to facilitate the planning of appropriate clinical trials and to support the enrolment of patients. Currently, 402 patient registries in the field of rare diseases are listed by Orphanet, of which 47 are regional, 295 national, 34 European and 26 global. The main problem faced by these registries is their sustainability as they are long term projects and most funding sources only support short term projects. The way forward is to establish a public/private partnership in this area with the support of regulatory agencies. Among MS, major disparities in access to treatment are observed. Although all Orphan Medicinal Products (OMP) receive market authorisation at the EU level, their accessibility at MS level depends both on marketing decisions by the company and on the willingness of health authorities in each MS to quickly establish OMP prices and reimbursement rules. Monitoring this situation is necessary to be able to assess the impact of measures taken at MS and EU level. An agreement on indicators is now necessary. When a disease is rare, the expertise for the disorder is also rare and existing information and resources tend to be scattered, making it difficult for those willing to develop research as well as for the health care professionals in charge of caring for patients. It is why European countries have joined forces to develop Orphanet, the European portal for rare diseases. To resolve the issue of information dispersion, Orphanet gathers an inventory of over 6,000 rare diseases. Each disease is described by its name and synonyms, prevalence, age of onset, mode of in- 26 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 heritance, list of signs and symptoms, ICD10 code, MIM and MeSH terms and linked causative genes with direct access to SwissProt and GenAtlas. Each disease has a unique identifier and is placed in a poly-hierarchy classification system. All the classifications of diseases can easily be displayed on the website. Orphanet has developed an encyclopaedia in English which is published in an electronic, open-access journal, the Orphanet Journal of Rare Diseases. The abstracts are also available in French, German, Italian and Spanish. The encyclopaedia is searchable by either disease name and/or category of diseases. Orphanet gathers clinical practice guidelines produced by reputable institutions or organisations, in any language. Orphanet produces emergency guidelines in collaboration with learned societies in the field and translated in five languages. To support physicians trying to diagnose a rare disease, Orphanet provides a query system of signs and symptoms. Five key-words can be combined. The possible diagnoses are listed in order of probability. To combat unnecessary delays to obtaining a diagnosis or appropriate care and disease management, Orphanet has developed a continuously updated directory of expert clinical centres and expert clinical laboratories. These can be searched by disease name, disease category, or gene(s) implicated, as well as by region or country. To promote quality services, data on quality management of clinical laboratories are available on the website. Distinct logos indicate which laboratories are certified, accredited and/or participate in external quality assessment. This information is gathered and validated in partnership with EuroGentest (www.eurogentest.org). To facilitate collaboration between researchers and between researchers and Industry, Orphanet lists all ongoing national and European-level funded research projects by type of research and by disease. These projects are also searchable by institution and by funding agency. The licensing opportunities are displayed, as well as the patient registries, biobanks and highly specialised platforms and know-how, which may be of interest in R&D. To help patients establish contact with other patients, Orphanet furnishes information on existing patient organisations in Europe. For disorders that do not have a patient organisation to date, Orphanet has developed a service through which patients can register to be put in contact with others with the same disease. To build up the rare disease community, Orphanet publishes a free-access electronic bimonthly newsletter. This newsletter communicates political news as well as scientific developments in the field of rare diseases and orphan drugs to over 12,000 subscribers. To support policy-makers, Orphanet regularly publishes reports in a collection entitled „Orphanet Report Series”. These publications, which include the themes of „rare disease prevalence”, „orphan drugs”, „patient registries”, „Research in Europe”, „Genetic testing offer in Europe” are regularly updated. All these services are freely at the disposal of the 20,000 daily users of the website. Two-third are health professionals and one-third are patients and their relatives. This set of services has been instrumental in contributing to the establishment of research networks and has a measurable effect on the referrals of patients to expert clinics and of samples to expert laboratories. Each review articles is accessed over 500 times per month and the encyclopaedia articles for lay public are downloaded 40,000 times per month. The next service to be developed is an encyclopaedia for paediatric anaesthesia which could save lives as the emergency guidelines have already done so. Bibliography: Orphanet website: www.orpha.net Website of the Rare Disease Task Force: www.eucerd.eu Financial support: Orphanet is supported by grants from the European Commission (RDPortal-2066119 and RDPlatform-Health-F2-2008-201230), by grants from the French Ministry of Health and from INSERM. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 27 R19. PROJECT „PARTNERSHIP NORVEGIAN- ROMANIAN (NORO) FOR PROGRESS IN RARE DISEASES” FUNDED THROUGH A GRANT RECEIVED FROM THE NORWEGIAN COOPERATION PROGRAMMES Dan Dorica, Maria Puiu RoNard, Romania Title: PROJECT „PARTNERSHIP NORVEGIAN- ROMANIAN (NORO) FOR PROGRESS IN RARE DISEASES” funded through a grant received from the Norwegian Cooperation Programmes Author: Dorica Dan, president of Romanian Prader Willi Association, Romanian National Alliance for Rare Diseases, BoD member of IPWSO and EURORDIS, EU CERD member; Partners: Romanian Prader Willi Association, Ministry of Health Romania, The Norwegian Prader Willi Association, Frambu – Center for Rare Diseases Norway, „St. Family” Church Zalau, „Acasa” Foundation, City Hall Zalau (DASC), County Council Salaj (DGASPC, ISJ Salaj), Romanian Medical Genetics Society, University of Medicine and Pharmacy „Victor Babes” Timisoara and Romanian National Alliance for Rare Diseases, established a partnership for long-term international co-operation. Project Aim: To contribute to the improvement of the quality of life for people affected by rare diseases in Romania by providing equal access to early diagnosis, quality treatment and rehabilitation services through a comprehensive and accessible network of facilities and resources as set forth by the National Plan for Rare Diseases. Objectives: 1. To define a team of professionals and patients’ representatives (NATIONAL RARE DISEASES TASK FORCE) to design, implement, monitor and evaluate the National Plan for Rare Diseases in Romania. 2. To contribute to the development of new high quality services for rare diseases in Romania on national level through the creation of a pilot reference center for personalized intervention for those affected by rare diseases.) 3. To enhance the training capacity within the country for the prevention, diagnostic, treatment and rehabilitation of rare diseases A. To design and implement a training network for specialists and staff B. To create accredited online training courses (euniversity) about rare diseases for professionals: social workers, psychologists, nurses, teachers, doctors, etc. 3. Permanent development and maintaining of a shared best practice network on RD (noro) Activities: Ob.1 • Trimestrial meetings with the members: to asses and evaluate the needs and establish the priorities; • Creating curricula for the different training courses; • Monitoring the implementing of the National Plan for Rare Diseases in Romania, assessing the needs and establishing the priorities; Ob.2. A. Developing and implementing a training network for specialists and staff involved in the diagnosis, treatment and rehabilitation of people with rare diseases. • Organizing workshops, seminars, and training on specific areas of RD; • Selection of a training team of trainers; • Creating curricula on specific jobs involved in RD, ex: genetic councilor; • Performing trainings; 28 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 B. The creation of accredited online training courses (euniversity) about rare diseases for professionals: social workers, psychologists, nurses, teachers, doctors, etc. • Creating curricula for specific training themes; • Purchasing the necessary soft to perform this trainings; • Authorization/ accreditation the trainings; • Organizing the first training courses; • Organizing video conferences on specific topics in RD; C. Pilot Centre for Rare Diseases • A residential part: • To provide help for families after experience of diagnosing a RD; • A place to learn how to face the new situation and to dial with the disease in their every day life; • A place to organize meetings and trainings for patients; • Support groups and counseling activities for patients and families; • Hospitalization of the RD patients in ACASA hospital for rehabilitation (in the same town); (ACASA is a foundation, founder member together RPWA in establishing the National Alliance for Rare Diseases in Romania) • Establishing a data base of patients that needs rehabilitation program in ACASA; • Specializing services of the center for RD patients: physical rehabilitation, weight management, individualized therapeutically approach; • Day Care component: • Individualized therapeutically approach of the diseases; • Behavior intervention therapy; • Occupational Therapy and Educational activities; • Recreational activities; • Weight management • Support groups and counseling activities; Ob.3 • Organizing a workshop: organizational and management aspects in a rare diseases centre – frambu model; best practice and shared values; involving norwegian and romanian partners; • Organizing presentation of the project’s results in the pwee conference (april 2009 romania), eurordis membership meeting greece 2009, ipwso conference taiwan 2010; • Permanent updating of the project results on the websites, ensuring visibility and transparency of the project; • Writing articles in the newsletters and rd journals to promote the project’s results; • Organizing work visits for partners in both countries; - one at the beginning of the project - one during the implementation of the project; - one in the last months of the project; Conclusion: The initiative to form this partnership was taken based on the background of the organisations, a common mission and goals in the field of rare diseases and their commitment to contribute to the definition of the Romanian National Care Plan for Rare Diseases. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 29 R20. TELEMEDICINE AND E-LEARNING – MODERN APPROACH IN RARE DISEASES TOPIC M Gafencu1, D Dan2 M Puiu1 1 Medical University, Timisoara 2 RoNARD Background: The need to obtain the diagnosis of genetic diseases as soon as possible incited teams of experts to search for solutions. One of these solution is offered by the new technologies. Thus the image in electronic form can be transmitted in real time, allowing a rapid clarification of the diagnosis. Information about diseases can be disseminated through web solutions. AIM: Illustration of footsteps taken by medical teams from ANBRARO demonstrating that telemedicine is a viable solution for informing and training of various medical specialists. Method: Using the Internet as a source of information and collecting information from centers carrying out research in this field, our team has managed to form dedicated free source website on information about rare genetic diseases. Because of the interest shown and the evident need for early diagnosis for this kind of pathology, we continued with publication of reviews an translation of medical documents. Results: The number of people visiting and downloading medical information on our webpage is continuously growing. Also, people contacted the specialists which wrote the documents. The number of patients and medical organizations that are addressing us is increasing. Conclusion: Modern times ask for modern methods. Diagnosis from distance and dissemination of information to large number of medical personnel is not possible without internet. Capacity offered by mobile telephony and high quality incorporated cameras may offer rapid diagnosis. The processing of data in real time will help the patient reach a degree of independence to bring the life quality at an acceptable level. 30 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Oral Presentations (Following Congress Program) C1. PRENATAL DIAGNOSIS – CYTOGENETIC FINDINGS IN 750 AMNIOCENTESIS Cristina Gug1,2, G. Budau1, N. Hrubaru1, B. Mureşan3, T. Cioată1, Dana Chiriac1, C. Olaru4, D. Grigoraş1, G. Furău5, Liana Antal6 1 University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania, 2Genetics Medical Center „Dr. Cristina Gug”, Timisoara, Romania, 3 Obstetrics-Gynecology „Bega” Hospital, Timisoara, Romania, 4 Genesium CO SRL, Timişoara, România, 5 Department of Obstetrics-Gynecology „Vasile Goldis” West University, Arad, Romania, 6 Department of Obstetrics-Gynecology, Faculty of Medicine, University of Oradea, Romania, The aim of the study was to evaluate the frequency and structure of prenatal detected chromosome abnormality. In this paper we report results of 750 fetal karyotypes performed by cultured amniocytes. First and second trimester ultrasonographic findings of aneuploidies include structural abnormalities and fetal sonographic markers. The cytogenetic analysis with GTG banding of amniotic fluid cells revealed 18 aneuploidies (2,4%) and 25 structural abnormalities (3,3%). Each case has received genetic counseling. Maternal age over 35 years was found in 8/18 cases of aneuploidy. We detected 3/18 aneuploiodies (poliploidy in 2 cases and trisomy 13 in one case) in fetuses with structural abnormalities and 8/18 aneuploidies in fetuses with sonographic markers. Biochimical screening in cases with aneuploidies showed high risk established according to double testing (6/18 cases), triple testing (6/18 cases), quadruple testing (1/18 cases) and low AFP (2/18 cases). We found trisomy 21 (13 cases) that includes: omogen trisomy 21 (9 cases), mosaic trisomy 21 (3 cases) one of which with 2 celular clones 47,XX+21/46,XX, another with 3 celular clones 47,XY+21/ 48,XYY+21/46,XY and the other of chimera 47,XX+21/46,XY and trisomy 21 in de novo Robertsonian translocation trob(21;21) (1 case). Other autosomal trisomies that were found are the following: omogen trisomy 18 (1 case), mosaic trisomy 18 (1 case), omogen trisomy 13 (1 case) and mosaic trisomy 22 (1 case). Poliploidia was found in the form of the triploidy 69,XXX (2 cases). We found 1 balanced Robertsonian translocation: maternal trob(13;22). We also found balanced translocations: maternal t(7;10)(p22;p12.1) 2 cases in the same family, maternal t(17;20)(q12;q11) 1 case and paternal t(15p;19p) 1 case. Pericentric inversions were showed in 12 cases: maternal inv(10) 1 case, maternal inv(9) 4 cases, paternal inv(9) 6 cases and de novo inv(9) 1 case. Other structural abnormalities include: duplication dup(10)(q22.2-ter) due to maternal balanced translocations (4;10)(q22.2;q22.2-ter) 1 case, ring chromosome r(6) 1 case, marker chromosome 2 cases. We also found 4 cases of deletions: del(7)(q21-ter) 1 case, del(1)(p31.2-ter) 1 case, del(1)(q32.1-ter) 1 cases, del(X)(q26-ter) 1 case. The karyotype using standard banding is an informative method able to detect fetal chromosomal abnormalities. Key words: fetal karyotype, prenatal diagnosis, amniocentesis. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 31 C2. STRUCTURAL ABERRATIONS OF CHROMOSOME 16 PRESENTED IN 12 CASES Cristina Ionescu, Luminita Roibu, Lorand Savu Genetic Lab, Bucuresti, Romania Trisomy 16 is estimated to occur in more than 1% of pregnancies, making it the most common trisomy in humans, and also the most common chromosomal cause of miscarriages, as the condition is not compatible with life. Unbalanced chromosome 16 structural abnormalities are rarely observed in live born or older children, but few patients survive to adulthood. The frequency of chromosome 16 deletion/duplication disorders is not known, and estimates of their occurrence vary widely. Duplication of a segment of chromosome 16 may result in similar symptoms as the deletion of the same region, in some individuals. The duplication appears to have a milder effect than the deletion, with a higher proportion of individuals with this chromosomal change showing no apparent disability. These structural abnormalities may cause delayed development, anomalies of the face, head, and internal organs, hypotonia, intellectual disability, and autism spectrum disorders. Duplication of the heterochromatic region 16q11-q12 is probably without clinical significance, but duplication of large regions of the long arm of chromosome 16 (from 16q13 to qter) results in severe malformations and early lethality. Partial trisomy 16q21-qter may be associated with a nonlethal phenotype in some cases. Although some people have these chromosome 16 structural aberrations, without serious consequences, they can still pass it to their children, who may be more severely affected. Over the past 10 years, in over 6000 karyotypes, we found 12 cases with chromosome 16 structural aberrations, 2 of which in prenatal diagnosis, one in a male newborn and 10 in patients with the age between 28 and 42 years. We identified 5 cases of 16q duplication, 4 cases of 16q deletion, two 16p deletions and one 16p duplication. In none of these cases we could obtain information about the patients, neither if the two pregnancies were delivered to term nor if the babies were affected. Accurate phenotype prediction can only be achieved after precise description of the cytogenetic defect in the affected patients (using very high-resolution microarrays) and its correlation with existing genetic and physical maps of chromosome 16. Accordingly, patients with relatively smaller defects involving chromosome 16 aberrations are useful in defining chromosomal regions associated with particular phenotypes. C3. GENETIC HEARING LOSS – PRENATAL SCREENING Cristina Dragomir1, Adriana Stan1, Madalina Badila1, D Stefanescu1, Emilia Severin2, L Savu1 1 Genetic Lab, Bucharest, Romania 2 „Carol Davila” Univ Med Pharm, Bucharest, Romania New born hearing screening has been instituted in many countries since the identification of congenital hearing loss in the neonate period, followed by intervention within the first few months, is critical to overcome the following difficulties: limited communication, low school achievements and life long dependency. Approximately 90% of children with deafness are born to hearing parents. Most cases with non-syndromic deafness inherited the disorder in an autosomal recessive manner. The GJB2 and GJB6 genes mutations - DFNB1 locus on chromosome 13 - account for approximately 85% autosomal recessive nonsyndromic hearing loss. 35delG GJB2 gene mutation has been reported to be the most common mutations among Caucasians. Two deletions in the DFNB1 locus outside the GJB2 gene, but truncating the neighboring GJB6 gene are associated with non-syndromic hearing loss. These mutations, named del(GJB6-D13S1830) and respectively del(GJB6-D13S1854) are found in both monogenic and digenic GJB2/del(GJB6-D13S1830) or GJB2/del(GJB6-D13S1854) patterns of inheritance. 32 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Considering all these aspects and the fact that currently in Romania there are no reports of GJB2 and GJB6 gene mutations we engaged in a prenatal screening investigation. The aim of prenatal screening was to estimate the prevalence of the three mutations namely 35delG, del(GJB6D13S1830) and del(GJB6-D13S1854) and also to evaluate the screening feasibility and acceptability in pregnant woman who were undergoing chorionic villi sampling or amniocentesis for chromosomal abnormality investigation. We analysed fetal samples from pregnant women who approached our laboratory for prenatal diagnosis testing - classic cytogenetic techniques, FISH or QF-PCR - after amniocentesis or chorionic villi sampling. Participation of patients in the study was done based on informed consent which assured the identity and confidentiality of the obtained results. All DNA fetal samples were investigated for 35delG GJB2, del(GJB6-D13S1830) and del(GJB6-D13S1854) mutations using ARMS PCR and PCR multiplex. The screening revealed a rather large frequency of the healthy carriers with the 35delG GJB2 mutation of 3,14% in Romania. This is consistent with the frequency of healthy carriers of 35delG mutation in the general Caucasian population (2-4%). None of the fetuses were found to be homozygous for 35delG GJB2 mutation. All subjects were found to be negative for the GJB6D13S1830 and GJB6-D13S1854 deletions. Also, our study was well received by pregnant women and appears to be feasible and highly acceptable. The genetic test for hearing loss is highly accurate and is available as prenatal and postnatal diagnosis method in our laboratory since last year. C4. DIAGNOSTIC PROBLEMS IN CASE OF PRIMARY AMENORRHEA ASSOCIATED WITH DWARFISM DISARMONIC Otilia Marginean 1, Ioan Simedrea1, Cristina Gug2, Belei Oana1, Craciun Adrian1, Tamara Marcovici1, Ioana Maris1, Camelia Daescu1, Laura Olariu1, Daniela Cioboata1 1 st I Pediatric Clinic of Clinical Children ‘s Emergency Hospital ”Louis Turcanu” Timisoara, Romania 2. Genetic department of University of Medicine and Pharamacy „Victor Babes ” Timisoara Romania Introduction: Turner Syndrome (TS) is a complex issue involving dismorfic phenotype and gonadal disgenezie due to qualitative and / or quantitative X chromosome changes. Aim: Presentation of diagnostic problems raised by the case of ST to a child who only accessed the health system in order to elucidate the etiology of primary amenorrhea. case Case presentation. The authors present patients Florentina S., 15 years, from rural areas, admitted in our service in November 2009 for primary amenorrhea. Clinical examination has revealed the Real height = 134cm, age height = 158.4 cm, short stature - 15.41% greater than - 3 SD, striking discrepancy between the upper and lower rail, mild ptosis, outlines pterigium coli, dorsocervical low implantation of hair, primary amenorrhea biological investigations have shown the value of the age parameters. Hormonal investigations hypogonadism hipergonadotrop : Estradiol = 22pg/ml, 12.3mIU/mL FSH = LH = 17mIU/mL. Gynecological Exam: vulva look infantile, infantile uterus. GenitalUltrashall: small right ovary, left ovary not see, uterus infantil.Xray bone age: chronological age appropriate 15 years, toe IV (right hand) soon. Xray of leg: patelară bilateral lateral dislocation, Genoa varum by tibiale.Xray medial proximal epiphyseal inclination. Spine: upper thoracic scoliosis dextroconvexa, lower lumbar sinistroconvexa. Cytogenetic examination: karyotype is 46, Xi (xq). Conclusions: 1. Turner syndrome is subdiagnosticat. 2.Toate cases of primary amenorrhoea associated with short stature should be carefully investigated and prenatal diagnosis is a necessity carotipate.3. Implementation of prenatal diagnosis is a necessity. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 33 C5. THERAPEUTIC STRATEGY FOR AMELIORATION METABOLIC DISEASES IN INBORN ERRORS OF METABOLISM Jurca C1, Bembea M1, Kozma K1, Skrypnyk C1, Iuhas O1, Harbuz R, Jurca A1 Faculty of Medicine and Pharmacy Oradea E-mail: [email protected] In the coming decades, molecular biology, protein engineering, and the Human Genome Project will have an enormous impact on the treatment of genetic and other disease. The optimal treatment of single gene defects requires an unusual degree of diagnostic precision, often at the level of the affected molecule. For proper management, it is often critical not simply to treat a biochemical abnormality but to identify precisely the basic biochemical defect. The therapeutic tools and approaches which are presently available fall into a number of general diseases, including replacement of needed metabolites, removal of toxic metabolites or interference with their accumulation, replacement of damaged organs, and restoration of the normal form of mutant gene products. Each of these approaches has been applied effectively to a number of genetic diseases, and it is the purpose of this report. C6. WILLIAMS SYNDROME - CLINICAL FEATURES AND CARDIAC INVOLVEMENT IN CHILDREN G. Doros1, A. Popoiu1, I. Anca2, M. Gafencu1, B. Zoica1, C. Laudacescu1, M. Puiu1 1 University of Medicine and Pharmacy „Victor Babes” Timisoara, IIIrd Pediatric Clinic 2 University of Medicine „Carol Davilla” Bucharest Aim: To present four patients, with clinical features for Williams syndrome, admitted in the Cardiology Department of the III rd Pediatric Clinic, between 2007-2010, to be evaluated for cardiac murmur. Williams syndrome is caused by the deletion of genetic material from the region q11.2 of chromosome 7, including more than 20 genes, several of these contributing to the characteristic features of this disorder. Material and method: The patients were young children, two girls and two boys, with age between 2-7 yo., extremely friendly, with moderate mental retardation, short stature, characteristic elfin face, blue eyes, stellar iris, joint laxity, systolic murmur and attraction to music. They performed clinical examination, ECG, echocardiography, cardiopulmonary X ray and angio CT, and after that they were referred to the genetic, ENT, ophthalmology, neurology and surgery department for evaluation. Results: Supra-valvular aortic stenosis was confirmed in all cases, associated with hypoplasia of aortic arch and large aortic coarctation in one girl and severe coarctation, hypertension, and carotid artery stenosis associated with pulmonary artery stenosis in one boy. The girl mentioned had from birth irreducible inguinal hernia, operated in newborn period, followed by cardiopulmonary arrest, resuscitated. The inghino-labial hernia reoccur and was operated this year. She also still have feeding problems for semisolid food. Barium examination reveal large esofageal stenosis in the 1/3 inferior part. All the patients have feeding problems. Characteristic for all were proeminent lips with an open mouth, defective tooth enamel and spaced teeth. Two of the patients associated hypercalcemia. The boy mentioned have nephro-calcinosis. Conclusions: After clinical examinations, all patients are able to be considered Williams syndrome. The FISH test confirmed the diagnose. They are in follow up programe and became members of the Williams Syndrome Children Association. 34 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 C7. MULTIPLE MALFORMATIONS IN INFANT - CASE REPORT T.Marcovici1,2, I. Simedrea1,2, L. Tunea1,2 A. Militaru1,2, O.Belei1,2, D.Chiru1,2, G. Brad2, M.Puiu1,2 1 „Victor Babes” University of Medicine and Pharmacy, Timisoara, Romania 2 „Louis Turcanu” Children’s Emergency Hospital Timisoara, Romania Background: Posterior urethral valves (PUV) are the most common cause of urethral obstruction in males. They are produced by mucosal folds in the posterior urethra. The pathological consequences of PUV are determined by the location and severity of obstruction. Increased resistance to urine outflow causes bladder muscle hypertrophy. Clinical presentation can be very diversified: from mild signs to end stage renal failure. Nearly 50% of the cases are diagnosed before the second month of life. The treatment is surgical. Methods: We report a 3 months old male infant admitted in our clinic for fever and rhinopharyngitis. He is the second child of a young, healthy and unrelated couple. No malformations are noticed in the family. Patient presented abdominal mass and difficulty voiding. Complete evaluation (anamnesis, clinical examination, imagistic and laboratory studies) was made. Results: The patient manifested straining during micturition and dribbling of urine, mild cardiac murmur and club feet. Imagistic exams (ultrasonography, intravenous urography, voiding cystourethrography) demonstrated crossed non-fused renal ectopia, both kidneys situated on the right side of the body; bilateral hydrouretheronephrosis; enlarged bladder, with irregular, grossly trabeculated walls and dilation of the posterior urethra. No vesico-ureteral reflux was found. Normal renal function and sterile urine were present. Cardiac ultrasonography revealed small atrial septal defect. Surgical ablation of the valves was performed in the pediatric urology department with good outcome. Conclusions: Posterior urethral valves are a component of a complex malformative syndrome in this patient. The ablation of the valves may preserve kidney function. Long term monitoring is required. Key words: posterior urethral valves, renal ectopia, multiple malformations, infant C8. HEREDITARY ONYCHO-OSTEODYSPLASIA (HOOD; NAIL-PATELLA SYNDROME) – A CASE REPORT Vasilica Plaiasu1, Diana Ochiana1, Cristina Skrypnyk2, Dorica Dan3 1 Genetics Department, Institute for Mother and Child Care ”Prof.dr.Alfred Rusescu”, Bucharest, Romania 2 Municipal Clinical Hospital „Dr. Gavril Curteanu”, Oradea, Romania 3 Prader-Willi Association, Zalau, Romania Introduction: Nail-patella syndrome (NPS), also referred to as Hereditary Onycho-Osteodysplasia (HOOD), is a rare autosomal dominant disorder characterized by a classic clinical tetrad of changes in the nails, knees, and elbows and the presence of iliac horns. Renal involvement is inconstant. The incidence has been estimated at 22 per million inhabitants’ live births with a variable expression within and among families. This condition is caused by mutations in the LMX1B gene on chromosome 9q34.1. Case report: We present a female patient, 25 years old, who visited our clinic for the evaluation of deformity in one knee, difficulty in standing and walking, with frequent subluxations of the left knee joint, spondylolisthesis, abnormality of pectus, hypotrophy of arms and legs, reduced temperature in the hands, numerous lipothymic episodes. Her sister and her mother were also known to have had similar nails and skeletal symptoms, but milder. For our proband pelvic radiography confirmed iliac horns with accompanying skeletal features. Conclusion: The family history and clinical signs confirmed the clinical diagnosis of HOOD. Molecular genetic testing was not yet performed. Early diagnosis of NPS is important to prevent early Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 35 secondary arthrosis and severe renal damage. Considering the different aspects of this syndrome, it is necessary to have a multidisciplinary approach for diagnostic and treatment of all the symptoms. Genetic counseling is also recommended. C9. GENETIC EXPLORATION – DECISIVE FACTOR IN THE DIAGNOSIS AND MANAGEMENT OF THE PRIMARY IMMUNODEFICIENCIES M. Bataneant, M.Serban, M.Cucuruz University of Medicine and Pharmacy „Victor Babes” Timisoara, Romania Primary immunodeficiencies (PID) are a heterogeneous group of hereditary disorders characterized by one or more intrinsic defects of the immune system. Although the first description was performed more than 50 years ago, today there are known approximately 200 of primary immunodeficiencies, in more than 150 of them there were identified the genetic substrate. The prevalence of PID is not well established worldwide. They are rare genetic disorders (Eurordis, Nord) but more frequent than cystic fibrosis or hemophilia. It is consider that they have a prevalence of 1/10.000 persons, with variation from a country to other. But recent data show a higher frequence, even of 1 / 250 persons. Earlier diagnosis in PID is very important because it reduce the hospitalizations and aggressive antiinfectious treatments. Today a lot of PID have specific treatment and if it start earlier it can prevent severe complications ( chronic suppurative, autoimmune, malignant, allergic). But the diagnosis of PID needs sofisticate and expansive explorations which limit the access to an earlier and correct diagnosis in countries with a low income as Romania is. Due to missing of access to the genetic explorations, the most cases of PID in Romania have only a diagnosis of probability in accordance with international criteria doing imposible a correct management and genetic council but also the prophilaxy of the disease through prenatal diagnosis. C10. ANGIOGENIC GENE THERAPY – FIRST CLINICAL TRIAL IN ROMANIA Andrei Anghel Biochemistry Department, UMF „Victor Babes” Timisoara Critical limb ischemia (CLI) is an ideal research target, in that its treatment outcome is clearcut, ending in either limb amputation or functional limb salvage. Surgical and endovascular revascularization is the treatment of first choice for CLI. Unfortunately, with current therapy, it can be expected that >25% of CLI patients will require major limb amputation within 12 months. Therapeutic angiogenesis is a new strategy that attempts to improve the perfusion of ischemic vascular beds by promoting the formation of new blood vessels using vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), angiopoietins, etc. A double-blind, placebo-controlled, VEGF and HGF trial was conducted in patients with claudication demonstrating ankle/brachial index <0.5. Patients were randomly assigned to placebo and gene therapy. Study drug (VEGF and HGF pBLAST plasmids) was injected directly into the muscles of the ischemic limb. The clinical evolution has been monitored by ankle-brachial index, walking distance and the relief of rest pain. Intramuscular administration was safe and well tolerated. Six months after therapy 70% of gene therapy patients show partial or complete relief of rest pain, improvement of ischemic ulcer lesions and increased walking distance on the rolling carpet. The improved perfusion to the distal ischemic limb was demonstrated by a mean increase of 0.1 in the ankle-brachial index. Therapeutic angiogenesis with VEGF and HGF gene transfer is a safe and non-inflammatory procedure with a good effectiveness in the re-vascularisation of the ischemic limb. 36 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 C11. GENERATION OF NOVEL RECOMBINANT FACTOR IX VARIANTS WITH ENHANCED CLOTTING ACTIVITY Laura Marusciac1, Carmen Bunu1 1 Department of Physiology-Immunology, „Victor Babes” University of Medicine and Pharmacy Timisoara Background: Factor IX is a plasma glycoprotein belonging to the serine protease family, having a role in the intrinsic coagulation pathway, and being involved in normal hemostasis. A deficiency of factor IX results in the clinical syndrome hemophilia B, which is an X-linked inherited bleeding disorder. Factor IX-deficient patients are a heterogeneous group, as defined by their plasma protein levels (normal, reduced, undetectable), or by their gene mutations. Our objective was to generate recombinant factor IX variants with enhanced clotting activity, for use as replacement therapy, in order to improve the quality of life for hemophilia B patients. Material and methods: For the generation of the variants we used as template a factor IX variant which already presented 3 mutations: G4Y/V86A/R338A. Nucleotide substitutions were introduced with the QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA), followed by transformation of XL1-Blue Supercompetent Cells and culture overnight in LB medium. We then extracted the Plasmid DNA was then exracted from the overnight cultures using the peqGOLD Plasmid Miniprep Kit I (PEQLAB, Erlangen, Germany) and we performed restriction analysis and sequencing for plasmid verification. Overnight E.coli cultures were then prepared in LB medium in order to obtain DNA for cell transfection, and the plasmid DNA was extracted with JETstar The Novel Plasmid Purification System (Genomed, Loehne, Germany). The plasmid consisted of a citomegalovirus vector, with an adeno-associated virus promotor. HEK293T cells were then transfected, using the calcium-phosphate method. In order to assess the activity of the factor IX variants, aPTT tests were performed using the secretion medium from the transfected cells, and the factor IX concentration was determined using ELISA. Results: We obtained the following recombinant factor IX variants: G4Y/V86A/R338L/S377W, G4Y/V86A/R338L/S377A, G4Y/V86A/R338L/S377G, G4Y/V86A/R338L/S377K, and G4Y/V86A/ R338L/S377Y. The transformation of supercompetent cells was successful in 60% of the cases, and we obtained quantities ranging from 340 to 900 µg of DNA from 200 ml E.coli overnight cultures. APTT showed consistent reductions in clotting time for our factor IX variants, and the ELISA assays showed a lower antigen expression for our variants, when compared with both the wild-type DNA and the template DNA (pAAV-CMV-hFIX-G4Y/V86A/R338A). By correlating the results of the aPTT with the ones from the ELISA assays, we found that our most efficient variant, the pAAV-CMV-hFIXG4Y/V86A/R338L/S377W, shows a 10-fold increase in clotting activity, when compared with the wild-type, and a 2-fold increase when compared with our template DNA. Conclusions: Our results indicate that our recombinant pAAV-CMV-hFIX-G4Y/V86A/R338L/S377W variant exhibits significantly enhanced clotting activity compared with both wild-type factor IX, and the template DNA, as demonstrated in vitro. Therefore, this recombinant factor IX variant is a good candidate for further evaluation in protein replacement therapy for hemophilia B, as well as gene-based therapeutic strategies. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 37 C12. CHARCOT-MARIE-TOOTH DISEASE TYPE 2C: PART OF THE TRPV4 RELATED CHANNELOPATHY SPECTRUM Horia C. Stanescu1,2, Guida Landoure1,2, Anselm A. Zdebik 1,Barrington G Burnett3, Tara L Martinez4,5, Hitoshi Inada6, Yijun Shi3, Addis A Taye3, Lingling Kong4, Clare H Munns7,8, Shelly S Choo6, Christopher B Phelps6, Reema Paudel9, Henry Houlden9, Christy L Ludlow10, Michael J Caterina7,8, Rachelle Gaudet6, Charlotte J Sumner2, Kenneth H Fischbeck2, Robert F. Kleta1,2 Institute of Child Health / University College London / Great Ormond Street Hospital, London, UK National Institutes of Health, Bethesda, MD, USA 3 Neurogenetics Branch, National Institute of Neurological Disorders and Stroke (NINDS), National Institutes of Health (NIH), Bethesda, Maryland, USA 4 Department of Neurology, Johns Hopkins University, Baltimore, Maryland, USA. 5 W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA. 6 Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA 7 Department of Biological Chemistry and 8 Department of Neuroscience, Center of Sensory Biology, Johns Hopkins University, Baltimore, Maryland, USA. 9 University College London, Institute of Neurology, London, UK 10 Laboratory of Neural Bases of Communication and Swallowing, James Madison University, Harrisonburg, Virginia, USA 1 2 Background. Charcot Marie Tooth disease is one of the most polymorphic and extensively studied neural disorders. Type 2C of this disorder (characterized by vocal fold paresis and an autosomal dominant transmission pattern with incomplete penetrance) was first described in 1994. We have studied two unrelated families afflicted with CMC2C. The affected subjects show evidence of a motor neuropaty causing progressive weakness of distal limb, dyaphragm and laryngeal muscles. Methods. Finemapping via linkage analysis using polymorphic markers and haplotype reconstruction; mutation detection by direct sequencing of all the genes in the region of interest; expression studies at mRNA level; functional studies of the identified mutations (toxicity, subcellular localization, electrophysiology) in experimental overexpression studies (SDR neurons, HEK cells, Xenopus oocytes). Results. We have been able to refine the region on chromosome 12q (first published in 2003) through linkage analysis (combined LOD score for the two families = 10); two heterozygous missense mutations of TRPV4 (C805T and G806A, leading to amino acid substitutions R269C respectively R269H) were identified; TRPV4 mRNA expression predominantly in cartilage but also at the level of peripheral nervous tissue was confirmed; the nature (gain of channel function) and the toxic effect of the identified mutations were assessed in different expression systems. Conclusion. CMT2C is shown to be caused by mutations affecting one ankyrin repeat of TRPV4. It is a genetically heterogeneous disorder with incomplete penetrance, highly variable in terms of its clinical manifestations, being part of an unusual spectrum of TRPV4 related channelopathies (which include skeletal dysplasias and degenerative neuropathies). Therefore, this study underscores the potential importance of TRP channels in neurodegeneration. C13. RAPID PRENATAL DIAGNOSIS BY QF-PCR IN 2010 Adriana Stan1, Daniela Tudor1, Cristina Dragomir1, Emilia Severin2, Lorand Savu1 1 Genetic Lab, Bucuresti, Romania 2 Facultatea de medicina si farmacie „Carol Davila”, Bucuresti, Romania The rapid prenatal diagnosis of trisomy 13, 18, 21 and sex chromosome aneuploidies by QF-PCR is currently requested by women with high risk pregnancies after the screening tests - double test and triple test - thanks to the short period of time till the results is obtained (24 hours). 38 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 For the last 3 years the rapid prenatal diagnosis techniques performed in our laboratory included 17 STR markers - 4 STR markers for each chromosome 13, 18 and 21 and 5 markers for sex chromosomes. From January 2010, Genetic Lab’s analysis included 22 STR markers - 7 markers for the sex chromosomes, 6 for chromosome 21, 5 for chromosome 18 and 4 for chromosome 13. Concurrently we elaborate a new method based on international published data (K. Mann, E. Petek, B. Pertl) obtained after almost 15 years of research; this method included 2 multiplexPCR reactions - A modulus: 5 STR markers for each chromosome 13 and 21, 6 for chromosome 18 and B modulus: 10 markers for sex chromosomes. This new method proved to be better then the other two thanks to the highest STR number which raised the probability of having at least two informative markers - heterozygous markers - for each chromosome, needed for test validation; the advantage is the short period of time for reporting the result (almost 24 hous) by eliminating the back-up reactions in case of obtaining a single informative marker for a specific chromosome, the rest being uninformative (homozygous). At the moment our team is working to extend the QF-PCR based analysis because the standard test is useful just for the triploid samples or for complex mosaics cases that includes chromosomes 13, 18, 21, X and Y; however analyzing only these 5 chromnosomes, the actual cause of miscarriage remains mostly unknown. The cytogenetic analysis also meets serious impediments in obtaining a result when the product of conception is received in the laboratory after several days since the fetus evolution has stopped. The first extra chromosomes included were 15, 16 and 22 - because these chromosomes are frequently involved in the first trimester miscarriage; trisomy 16 have the highest frequency, around 15% of all cases of miscarriages, followed by trisomy 22. Conclusion: We are confident that the QF-PCR approach still remains the alternative to conventional cytogenetic testing, considering the possibility to include more STR markers for extra chromosomes. C14. MOLECULAR DIAGNOSTICS IN HEMOCHROMATOSIS TYPE 1 AND WILSON’S DISEASE E. Seclaman, L. Tamas, A. Anghel, Valerica Belengeanu Biochemistry Department, UMF „Victor Babes” Timisoara Hemochromatosis type 1 is a hereditary disease characterized by excessive absorption of dietary iron resulting in a pathological increase in total body iron stores. One of the better characterized genes responsible for hereditary hemochromatosis is human hemochromatosis protein (HFE) gene. HFE gene has three common alleles,. Mutations of the HFE gene account for 90% of the cases of non-transfusional iron overload, three of them (C282Y, H63D and S65C) are more comon. Wilson’s disease is an autosomal recessive genetic disorder in which copper accumulates in tissues; this manifests as neurological or psychiatric symptoms and liver disease. Although 300 mutations of Wilson disease protein gene (ATP7B) have been described, in most populations the cases of Wilson’s disease are due to a small number of mutations specific for that population. In Western populations the H1069Q mutation is present in 37-63% of cases. Molecular diagnostic of both diseases is based on hybridization probes (melting curve analysis) for known mutations and HFE and ATP7B genes sequencing and. Several types of mutations were detected in UMF Timisoara, Biochemistry Laboratory; the H1069Q mutation was found in one family (two homozygous and one heterozygous). Heterozygous H63D genotype (wildtype C282) was identified in one patient. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 39 C15. FAST AND SENSITIVE MUTATION SCREENING USING HIGH RESOLUTION MELTING ANALYSIS CB Iancu, D Iancu, C Constantinescu, E. Neagu, A Constantinescu, G Girbea, L Barbarii Institutul National de Medicina Legala „Mina Minovici”, Bucuresti Introduction Mutation scanning requires the use of techniques able to detect sequence variants with high specificity and sensitivity, in a cost-effective manner. High resolution melting analysis (HRMA) is a PCR-based method which detects DNA sequence variation by measuring fluorescence intensity differences during the melting transition of a DNA duplex. Method HRMA was performed on two sets of DNA samples obtained from 25 patients with Duchenne muscular dystrophy (DMD) and 15 patients with Crohn disease, all with known genotype. The samples were amplified in triplicate with specific primers using MeltDoctor™ HRM Master Mix and a 7900HT Fast Real-time PCR System(Applied Biosystems). Melting curve analysis was performed on the same equipment, using the SDS v2.4 and HRM v2.0 software (Applied Biosystems). Variants were identified relative to wild type control samples, based on differences in melting curve shape and melting temperature shifting. Results All mutations existing in the DMD sample set were sensitively identified (misense mutation in exon 45 and splicing mutation in exon 56). The NOD2/CARD15 gene polymorphisms associated with increased risk of Crohn disease necessitated optimization in order to detect the C>G transversion in exon 8. A second primer pair designed to transform a two-melting domain amplicon into a single melting domain one allowed us to increase sensitivity and specificity in this case. Conclusions HRMA is a sensitive method with good specificity for mutation scanning. HRMA is a closed tube system, which both reduces the contamination risk and increases sample throughput. With good optimization, HRMA offers significant savings in cost and turnaround time when compared with other scanning methods. C16. DYSTROPHIN GENE MUTATION ANALYSIS IN PATIENTS WITH DUCHENNE MUSCULAR DYSTROPHY AND COGNITIVE IMPAIRMENT D. Iancu1, E. Neagu1, N. Butoianu2, C.B. Iancu1, C. Constantinescu1, C. Burloiu2, A. Constantinescu1, G. Girbea1, C. Iliescu2, D. Craiu2, L. Barbarii1 1 National Institute of Legal Medicine „Mina Minovici”, Bucharest 2” Alexandru Obregia” Clinical Hospital, Bucharest Introduction. Variable alteration of cognitive functions is found in many patients with Duchenne muscular dystrophy (DMD). Therefore a large number of researches have been done in order to identify the genetic basis of this clinical feature. It was found that mutations affecting the exons in the second half of the DMD gene, particularly those of the Dp140 and Dp71 isoforms, represent a significant risk factor for the intellectual disability of the DMD patients. Materials and Method. 12 patients with DMD and variable levels of cognitive disability were selected from the DMD National Registry. Each patient was clinically and psychologically evaluated and then a blood sample was obtained for DNA extraction. Mutation detection was initially performed by Multiplex Ligation-Dependent Probe Amplification (MLPA - MRC Holland) using SALSA P034 and P035 kits. The cases without large mutations detected on MLPA - were subjected to Sanger sequencing of the whole coding region of the dystrophin gene. Rezults. Large deletions involving exons 46 – 50 were detected in 5 cases with mild to moderate mental retardation (2 related and 3 isolated cases). One case of DMD and autism presented a frameshift mutation consisting of two bases insertion in exon 37. Three samples are currently under evaluation and for the last three cases no mutation was detected. Conclusions Detected deletions affect the Dp140 isoform of the dystrophin gene, concordant with previously found association between these mutations and cognitive impairment. The two related cases with the same mutation presented with different degrees of mental retardation, suggesting the action of other modulating factors. 40 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 C17. DETECTION OF K-RAS POINT MUTATIONS IN CODONS 12 AND 13 IN COLON TUMORS BY THE PCR-RFLP METHOD Georgeta Cardos1, Florina Cionca1, Georgeta Butur1, Dana Terzea1, Mihaela Mihai1, Florin Andrei1, Simona Enache1, Florina Vasilescu1, Cristina Iosif1 and Carmen Ardeleanu1,2 1 „Victor Babes” National Institute of Pathology, Bucharest, Romania 2. „Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania The novel anti-Epidermal Growth Factor Receptor (EGFR) therapy has improved the prognosis for patients with metastatic colorectal carcinomas (CRC). Gain-of-function K-RAS point mutations, present in 25–45% CRC, maintain the active form of the K-RAS protein and lead to EGFR-independent activation of intracellular Ras/Raf/mitogen-activated protein kinase (MAPK) signaling pathways, making the anti-EGFR tumor therapy ineffective. Therefore, KRAS mutation status is a crucial parameter for selecting patients for anti-EGFR therapy; only patients without mutant tumors will benefit from this new therapy. In this context, the aim of our study was to detect the K-RAS mutational status in codons 12 and 13 (about 99 % of all mutations in the K-RAS gene) in CRC patients in order to contribute to the selection of patients for the new therapy with Avastin (an anti-EGFR agent) in our country. DNA was isolated from fixed and paraffin embedded primary and/or metastatic tumors from 117 CRC patients and K-RAS mutational status analyzed by the PCR-Restriction Fragment Length Polymorphism (RFLP) method implemented in our laboratory, with Mva I (for codon 12) and Hae III (for codon 13) restriction enzymes. Preliminary results include 35 patients (29,9%) with gain-of-function mutation in codon 12 out of 117 analyzed at molecular level, and 2 patients with mutation in codon 13 out of 10 analyzed. We have validated our PCR-RFLP method by successfully participating in the „Ring trial Molecular-Pathological KRAS Mutationtest of Colorectal Carcinoma”, organized by the German Society of Pathology ([email protected]) and the German Federal Association of Pathologists ([email protected]). This international validation and a high level of addressability offer the opportunity to assess the K-RAS mutation status in our laboratory at a lower cost (in contrast with more expensive diagnosis abroad) and a European analysis quality level. C18. MOLECULAR DIAGNOSIS IN THE MANAGEMENT OF DUCHENNE DISEASE –THE EXPERIENCE OF UMF TIMISOARA Liviu Tamas1, Maria Puiu2, Andrei Anghel1, Edward Seclaman1, Oana Mitrasca1, Mirela Mititelu1 1 Biochemistry Department, UMF „Victor Babes” Timisoara 2 Medical Genetics Department, UMF „Victor Babes” Timisoara Dystrophinopathies are a group of muscle diseases which have as common cause mutations in the DMD gene, which encodes the protein dystrophin. Dystrophinopathies diseases are fatal, with X-linked inheritance, most patients are men and women are carriers. Frequency of the diseases is high - 1:3500 newborns (male). In Duchenne muscular dystrophy (DMD), DMD gene mutations cause a complete absence of dystrophin synthesis. Early childhood onset is before age 5 and at age 12 - 13 years the patient needs a wheelchair. The dilated cardiomyopathy (CMD) appears after age 18 and together with respiratory problems will be the cause of death of patients before the age of 30 years. Method. Genomic DNA was extracted from 24 blood samples using QIAamp DNA Blood Mini Kit kit (Qiagen). Samples were collected from DMD patients with clinical diagnosis of DMD, from their relatives and in one case amniotic fluid was collected for prenatal molecular diagnosis. Isolated DNA was quantified and analyzed by MLPA technique (Multiplex Ligation-dependent Probe Amplification), which can identify extensive mutations (deletions or duplications of gene fragments). The results which were obtained by capillary electrophoresis were quantified and interpreted using Coffalyser program. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 41 Results. Extensive deletions were identified in 12 patients (66.6%) and duplications only in two patients (11.1%). 6 female patients with heterozygous genotypes confirmed the previously identified mutations in male patients. In one case prenatal diagnosis was performed on a patient having a child with DMD, confirmed by molecular diagnosis. The MLPA analysis identified an extensive deletion (deletion of exons 34, 35, 36, 37, 38, 39, 40, 41, 42, 43), mutation found in the mother. After genetic counseling, the mother decided to terminate the pregnancy. In four patients (22.2%) no deletions or duplications were identified. Conclusions. The results confirm literature data that indicate a high frequency of large deletions and a lower frequency of duplications, the two types together representing over 77% of mutations identified in patients with clinical diagnosis of DMD. MLPA technique is particularly effective if Duchenne disease, allowing the detection of mutations in most cases and both prenatal diagnosis and diagnosis in children with characteristic symptoms of disease. The only limitation is represented by the fact that you can not identify point mutations, and complementary techniques (gene sequencing) are required, which are however more expensive and laborious. C19. MEDICAL GENETIC SERVICES, BETWEEN REAL AND IDEAL Marius Bembea Faculty of Medicine and Pharmacy Oradea, Municipal Hospital Dr. Gabriel Curteanu „Oradea Pediatric Clinic - Department of Genetics Genetic diseases are a heavy burden in both health and economic terms. This burden affects not only the family, but also the society. As exogenously determined diseases become better controlled, the endogenous (including genetic) become increasingly important. Although individually, each is a rare genetic disease, genetic diseases taken as a whole are important firstly because they are numerous (several thousand!) and secondly because most of them are severe and incurable. Some have an effective treatment but most of them do not. That’s why prevention is extremely important, either by preventing the emergence of new cases in the family, either by preventing complications in those affected. In most European countries there are genetic services within public health structures (Ministry of Health, National Institute of Public Health, National Public Health Service, etc.). Genetic services normally operate incorporated in university clinics and regional centers, being in close liaison with academic departments of human genetics (institutes, research centers, etc.) and with genetics offices in departmental hospitals. In our country, without a strategy for genetic diseases, structures of genetics services are missing (obviously, without equipment) which places us in an unwanted backward place in Europe. Experience of the Genetics Department in Oradea presented a retrospective of 25 years and nearly 4,000 patients observed in this period, on the one hand emphasizes the usefulness of these services and on the other hand on perfecting them. 42 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 C20. SUPERNUMERARY CHROMOSOME IN MOSAIC IN A DYSMORPHIC CHILD WITH DEVELOPMENTAL DELAY Valerica Belengeanu1, Simona Farcas1, Cristina Popa1, Monica Stoian1, Alina Belengeanu2, Nicoleta Andreescu1 1 Department of Medical Genetics, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania 2 Department of Cellular and Molecular Biology, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania Chromosomal anomalies can also be present in Mosaicism, defined as the presence of more than one genetically distinct cell in a single organism. From a human genetics perspective mosaicism may not be readily apparent in individuals with various clinical presentations such as mental retardation or multiple congenital anomalies because phenotypic variations may be subtle and can often be overlooked during examination or interpreted as normal variation in the population. On the diagnostic side, low-level mosaicism for clinically significant chromosome abnormalities can be missed if masked by a high percentage of normal cells or dismissed as an artifact of the culturing process that is required for conventional cytogenetic testing. The mosaicism may also evade the detection of conventional chromosome analysis based on the culture of blood lymphocytes because the culturing process may introduce a selection bias that distorts the percentage of abnormal cells and that a different percentage of the abnormal chromosome complement may exist in distinct cell lineages. We report a 24-month-old female with developmental delay, dysmorphic features, hypertelorism, broad nose, wide palpebral fissures and deep set eyes, dysplastic and posteriorly rotated ears, vultuous cheeks, large mouth, short and wide neck, eczema, bilateral simian crease. Cytogenetic analysis was performed and the analysis of 50 mitoses revealed 3 clones with trisomy. For elucidation we performed FISH analysis and we detected mosaicism, trisomy was observed in 25% of the cells evaluated. When the diagnosis of a chromosomal mosaicism is suspected additional analysis to culture of lymphocytes from peripheral blood should be done for studying other tissues using as well the classic chromosomal studies or completed by the new molecular techniques FISH, array-CGH to attest mosaic trisomy. C21. THE FOLLOW-UP OF A DE NOVO CASE WITH „CRI DU CHAT” SYNDROME Valerica Belengeanu1, Simona Farcas1, Cristina Popa1, Monica Stoian1, Dragos Belengeanu2, Nicoleta Andreescu1, Carmen Radulescu3 1 Department of Medical Genetics, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania 2 Department of Oral Rehabilitation, Dental Techniques College, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania 3 Pediatric Clinic, Pediatric Clinic Hospital Sibiu, Romania The majority of deletions of the short arm of chromosome 5 are associated with Cri du chat syndrome and patients show phenotypic and cytogenetic variability. The syndrome has an estimated incidence of between 1:15 000 and 1:50 000 live births. The existence of a critical region responsible for the phenotype indicates that a region in 5p15.2-15.3 must be deleted for the typical phenotype of the syndrome to be present. In about 90% of patients the deletion of the short arm of chromosome 5 (5p-) occurs de novo, while in 10% of cases 5p deletion results from parental balanced translocations. The proband was investigated at birth for cat-like cry, including the other syndromic features such as microcephaly, round face, micrognathia, flattening of occiput, high-arched palate, hyperRomanian Journal of Rare Diseases 2010 | Supplement 1/2010 43 telorism, low set ears, transverse palmar crease, short-webbed neck and widely spaced nipples. No other associated malformations were noted. The cytogenetic analysis was performed for the proband and her parents, on GTG (G-bands by trypsin using Giemsa) banded chromosomes. The cytogenetic analysis of the proband’s parents showed normal karyotypes. The karyotype of the proband was 46,XX,del(5)(p14→pter), confirmed by FISH analysis using Vysis Cri-du-Chat Region Probe. Reevaluated at age of 2 years, the patient had severe delayed developmental milestones. Larger deletion is reported to be associated with a severe phenotype and cognitive impairment. Our patient will be followed-up and efforts are concentrated on a social insertion. C22. NATIONAL REGISTRY OF DUCHENNE AND BECKER MUSCULAR DYSTROPHIES – IMPROVEMENT IN PATIENT’S STANDARD CARE E. Neagu1, D. Iancu1, CB Iancu1, A. Constantinescu1, C. Constantinescu1, G. Girbea1, RA Nicolae1, N. Butoianu2, D. Craiu2, L. Barbarii1 1 National Institute of Legal Medicine – Genetics Laboratory 2 „Prof. Dr. Al. Obregia” Neuropediatric Hospital National Registry of Duchenne and Becker muscular dystrophies, part of the fourth National Program for rare diseases founded by the Ministery of Health, allows, besides forming a patients database, also the improvement of care and diagnostic standards. Registration in the database is made after a previous genetic investigation, which involvs also an obtained informed consent and genetic counseling session(s), as well as patient participation in an investigation algorithm permitting access to therapies that improve quality of life and delay complication.The registry is the result of sustained efforts of a large number of specialists, pediatric neurologists, pediatricians, cardiologists, geneticists, etc.The National Registry is part of the global registry of Duchenne muscular dystrophy and Becker database, with the main goal of speeding up finding an effective etiological therapy. C23. PALLIATIVE MEDICINE IN PEDIATRIC ONCOLOGY - A CHALLENGE IN MODERN MEDICINE. III RD PEDIATRIC CLINIC TIMISOARA EXPERIENCE E. Boeriu 1,2, M. Cucuruz 1,2, S. Arghirescu 1,2, R. Costa 1, G. Brad 1, I. Ursache 1, M. Serban 1 Children’s Emergency Hospital „Louis Turcanu”, Timisoara 2 rd 3 Pediatric Clinic, Hemato-Oncology Department University of Medicine and Pharmacy „V. Babes „Timisoara 1,2 Introduction: Despite the progress in the Pediatric Oncology in the recent decades, it remains a field with diseases that have unfavorable and progressive evolution, fact that justify the implementation of the palliative medicine principle. Objectives: We aimed to analysis the cases with unfavorable evolution from the last two years (2008-2010) from the Pediatric Oncology Department of Children Emergency Hospital „Louis Turcanu”, Timisoara. A local record was done and the role of palliative care measures in the Pediatric Oncology was underlined. Materials and method: The study group consisted in 31 patients aged 1 month-31 years old (51.6% girls and 48.4% boys) from Timis and surrounding villages. We analyzed the study lot clinical and paraclinical, the cause of death, the evolution of symptoms and their control. Statistical analysis of data was performed. Results: Acute lymphoblastic leukemia was present in 28.5% of cases, followed by sarcoma (25.8%) and brain tumors (19.35%). The time interval from the onset of malignity to death was between 1-3 years in 51.6% of cases and over 3 years in 16.2% of patients and less than 3 months of onset was observed in 16.2% of patients. Curative therapy was followed by the palliative one (74.19%) and in 6.45% of the cases only palliative therapy was done. 44 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Conclusions: Palliative care address to all patients with low life expectances in order to improve their life quality until death. The period between the diagnosis and death justifies the application of palliative care even from the beginning of Oncology diseases. The pain was the most frequent symptom, which was well controlled, with the exception of neuropathic pain. In Oncology, it is important to have symptoms control for increasing life quality of these patients. C24. CARDIAC DISTURBANCES IN GENETIC DISEASES Anca Popoiu, Gabriela Doros, Maria Puiu University of Medicine and Pharmacy „Victor Babes” Timisoara Cardiac malformations present at birth are an important component of pediatric cardiovascular disease and constitute a major percentage of clinically significant birth defects, with an estimated prevalence of 4 to 50 per 1000 live births. For example, it is estimated that 4 to 10 liveborn infants per 1000 have a cardiac malformation, 40% of which are diagnosed in the first year of life. The true prevalence, however, may be much higher. The approach to the newly diagnosed patient with CHD should include routine examination of all relatives for a potential genetic contribution. Identification of some genetic causes of CHD has highlighted the importance of obtaining an accurate medical history of other family members and documenting an extended pedigree. In some forms of cardiovascular disease, for example, hypertrophic cardiomyopathy and Marfan syndrome, the familial nature (autosomal dominant inheritance) is well recognized; however, for other problems, for example, bicuspid aortic valve, family clustering has not been widely appreciated in the past. Recent studies have shown that a familial bicuspid aortic valve is likely to be inherited as an autosomal dominant condition with reduced penetrance. There is a 24% prevalence of bicuspid aortic valve in first-degree relatives of patients with left ventricular outflow tract obstruction.Increasingly, medical practice is evolving toward a recommendation that other family members undergo clinical evaluation, which may include an electrocardiogram and echocardiogram. Patients with CHD require multidisciplinary care. Their families deserve up-todate genetic information as it relates to their child’s prognosis and to the kindred’s risk for future inheritance of genetic abnormalities associated with cardiac defects. Families of these children will need information about recurrence risk. Pediatric cardiologists and pediatric cardiac surgeons are currently well equipped to care for patients with CHD, but they need to constantly update their understanding of the contribution of genetic abnormalities to these birth defects. As children grow into adulthood, internists, obstetricians, cardiologists, and thoracic surgeons will step in to care for CHD as it is superimposed on adult medical issues. C25. A TOXICOLOGICAL SCREENING ON IN VITRO AND IN VIVO EXPERIMENTAL MELANOMA AND SKIN CARCINOMA MODELS Cristina A. Dehelean University of Medicine and Pharmacy „Victor Babes” Timisoara, Faculty of Pharmacy, Toxicology Department, 300041-Timisoara, Eftimie Murgu Square no.2, Romania Corresponding author: email: [email protected] Melanoma and skin carcinoma are pathologies with an increased incidence in the last decade and their deep evaluation including genetic profiles are considered important scientific aims. All the aspects regarding description of pathology in experimental models are important for early prevention and treatment surveillance. New trends in pharmaco-toxicological evaluations consist in application of the modern techniques such as vibrational spectroscopy (FT-Raman, SERS etc) and other types of measurements including melanin, erytema, transepidermal waterless measurement using a Mexameter, TEWA-meter for early pathologic observations. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 45 The devices and techniques applied are indicated to be related with in vitro and in vivo studies such as MTT assay angiogenesis and specific experimental in vivo evaluation and if it is possible with gene detection. Our work in last period includes reproducible experimental models for melanoma and skin carcinoma. Some important strains in evaluation of skin pathology are mice: CD1 Nu/Nu, C57BL/6J, B6D2F1 that can easy develop skin pathology such as carcinoma and melanoma using a chemical or biological model. The first one model use chemicals as tumor initiators and promoters: 7,12-dimethylbenzanthracene and phorbol esters. The most acceptable and fast model in this case is a double application of 7,12-dimethylbenzanthracene and the application of phorbolesters between these and/or after these applications. The second biological model consists in inoculation of B16 cells (mouse melanoma cells) in tail vein, intrasplenic or subcutaneous in 0.5x105 dosages. An imunohistochemical evaluation can complete these data (melan A and S100). All these data are correlated with in vitro tests on specific cells including B16 (mouse melanoma cells). Future work will complete all these with a gene profile evaluation. The main conclusion is that in vitro and in vivo experimental evaluations can offer important knowledge about pathology evolution and all the particularities and possible therapy surveillance and also some important toxicological data. Applications of modern and non invasive methods for early diagnosis are new trends in cancer exploration. Key words: carcinoma, melanoma, diagnosis, treatment, toxicological, in vitro, in vivo. Acknowledgements: This work was supported by CNCSIS-UEFISCSU, project number PNII-IDEI code 1257/2007. C26. THE INTERRELATION BETWEEN MAXILA-PALLATA CLEFT AND CONGENITAL CORD MALFORMATION Balan Cristina Daniela, Mihai Voloşciuc, Braha Elena, Monica Pânzaru , Vlad Gorduza, Department of Human Genetics, U.M.F. „Gr.T.Popa”, Iasi, Romania Labio-maxillo-palatal cleft represents a congenital malformation present in certain syndromes, which are signs of other abnormal form or structure. Cleft are quite common: 1 / 1000 in Europe. It can be determined by genetics and environment and is difficult to predict, during pregnancy, whether the child will have cleft. If there are risks, they are shown by karyotype analysis and a fetoscopy, which can sometimes be difficult to bear by the mother. Baby heartbeat irregularities and anomalies can be determined fairly quickly by echocardiography, but there is little chance to be sure before birth. The aim was to assess the possibility of finding an interrelation between these two anomalies in syndromes or isolated. Previous studies of the literature showed a frequency rate of the two anomalies between 4.6 and 63.5%. We made a detailed analysis regarding the cases which have both cleft and congenital heart malformations. In some specific syndromes we found a clear interrelationship. We studied in detail special cases and we did various statistics using the diagnostic program Possum and the site www. orphanet.net (a laborious basis for rare diseases), reaching certain results in terms of both organ damages during embryogenesis. As conclusion we can say that in the study group, there was a clear increase of both these malformations in trisomy 13 and 18, Kabuki and Aarskog syndromes, presenting cases which reveal the seriousness of the association of the two malformations in a single patient. 46 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 C27. „SAVE THE CHILDREN” WITH RARE DISEASES Iulia-Simina Jurca, Florin Jurca, Simina, Ionela Moaca, Pop Norbert, Stefan Berci, Iulia Popa, Cristina Irimia, Oana Rosca, Graziella Ecob, Adrian Juverdeanu, Carmen Dumitranoiu, Narcis Dobre, Mihai Gafencu, Maria Puiu “Save the Children” organization, Timis office Introduction: Rare diseases are a major public health problem and in recent years they have become a global priority in health strategies. Their rarity makes them little known by the medical staff, which disrupts the proper diagnosis and healthcare. In our projects we intend to contribute, both by involving as many volunteers as possible and by providing information, coming directly into contact with patients and their families, and change their perception and even civil society perception towards the rare diseases’ issue. Objectives: Improving children’s quality of life whom disabilities caused by rare genetic diseases and information quality in identification, prevention and counseling. Methods: Activities based on specialized voluntary -patient-family direct relationship making use of playing games, art and group therapy. Results: Our project has started three years ago by forming and training a team of volunteers consisting of medical students and young professionals eager to engage both in actions to support children with rare diseases and their families as well as community awareness and information activities regarding this topic. We have made our activities known by applying, winning and carrying out projects collaborating with the City of Timisoara, Timis County Council, “For Community” foundation and MOL gas stations, projects developed through partnerships between Save the Children, Timis Branch, University of Medicine and Pharmacy „Victor Babes“ Timisoara, Children’s Emergency Hospital „Louis Turcanu“ – Timisoara, Clinical Evaluation and Rehabilitation for Children and Adolescents „Cristian Serban“ Medical Center - Buzias, Prader-Willi Association of Romania- Zalau, National Alliance of Rare Diseases in Romania – Zalau, Faculty of Arts- Timisoara, School of Music „Ion Vidu“ , General Direction of Social Assistance and Child Protection Timis, “Beyond words „Association etc. We used different games and puzzles to interact with children, materials purchased to overcome feelings as embarrassment and disbelief, and which also promoted interaction of children in order to create friendship and support, communication and experience exchange between them. Also, we held drawing, modeling clay, music contests, during which children were stimulated to be competitive but also to help each other. With the psychologist volunteers from Timis branch of Save the Children organization , we held meetings with families of children with rare diseases (support groups), thereby sharing their experiences in providing answers not put to questions till then, solutions for various situations and most importantly, to accept the child as a different disease entity. One of our most important activity consists of the organization of Rare Diseases Week in Timisoara, a very dear event to us, held in late February since 2008. Rare Diseases Day is a public information campaign made in most European countries regarding the existence of rare diseases. In Timisoara, we share information leaflets and we are available to all those who want information on rare diseases, for a week, in Liberty Square. Conclusions: Sustained actions have proven to be effective both for children with rare diseases and their families, and volunteer group participants. We have the joy that our efforts be appreciated by direct beneficiaries and general public positive feedback, the media and authorities which support us. Our satisfaction as volunteers is maximum when we receive as a reward a child sincere smile, and even if most are students in the final year at the Faculty of Medicine, we hope the time will allow as that „voluntary smiles” in the future, too. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 47 Posters P1. Identification of microdeletions in the AZF regions of Y chromosome in infertile patients in west Romania Anda Alexa, Andrei Anghel, Edward Seclaman, Liviu Tamas, Mirela Mititelu, Oana Mitrasca, Monica Stoian, Valerica Belengeanu Department of Biochemistry, UMF „Victor Babes” Timisoara Genetic factors cause about 10% of male infertility. Some of the genetic causes are chromosomal abnormalities or gene mutations. Recent extensive molecular studies have revealed that deletions in the azoospermia factor region of the long arm of the Y chromosome are associated with severe spermatogenic impairment. Our study has evaluated the frequency of Y chromosome microdeletions in infertile patients from west Romania. The frequency of Y chromosome microdeletions has been evaluated in 59 infertile men, using the Y Chromosome Deletion Detection System kit, Version 2.0 (PROMEGA), which amplifies 20 specific sequences of the Y Chromosome. Genomic DNA was isolated from blood cells, using a standard extraction procedure. The multiplex polymerase chain reaction (PCR) was performed starting from 50 ng of genomic DNA, in a final volume of 25 μL. PCR products were separated by agarose gel electrophoresis, visualized with an UV transilluminator and photographed. (Figure 1) Of the total 59 infertile men analyzed, two individuals (3,4 %) ) showed Y chromosome microdeletions, of which deletion in AZFc region was the most common (83.34%) followed by AZFd (16.66%). (Figure 2). No microdeletions were observed in AZFa and AZFb regions. At both patients, the complete deletion of AZFc region was detected (including DAZ gene). This is the most frequent cause of male infertility. 48 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P2. MOLECULAR CYTOGENETIC FISH TECHNIQUE VERSUS CONVENTIONAL CYTOGENETIC ANALYSIS FOR PRENATAL CYTOGENETIC DIAGNOSIS Nicoleta Andreescu1, Valerica Belengeanu1, Monica Stoian1, Simona Farcas1, Cristina Popa1, Miruna Muntean² 1 Department of Medical Genetics, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania ²Department of Obstetrics and Gynecology, „Dr. Popescu” Clinic Hospital of Obstetrics and Gynecology, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania Rapid and safe prenatal diagnosis has become a standard of care in high-risk pregnancy. Rigorous analysis of the results of prenatal diagnosis and the possibility of interruption of anomalous pregnancies at low gestational age is the key to facilitate a decision during the pregnancy. Impact of prenatal diagnosis on the incidence of severe birth defects is important because a significant number of anomalies can be diagnosed since the late first trimester. In the Laboratory of Medical Genetics, at the University of Medicine and Pharmacy Timisoara, during November 2006-March 2010, 227 pregnant women were evaluated. The methodology included molecular FISH technique for rapid screening of common aneuploidies performed in parallel with conventional cytogenetic analysis. For the study group, conventional cytogenetic analysis and interphase FISH allowed the identification of 28 chromosomal abnormalities, representing 12.3% of the total number of pregnancies investigated. The results support the implementation of molecular analysis of interphase FISH for rapid screening of common aneuploidies and usefulness of this technique to detect chromosome abnormalities in cases in which ultrasound abnormalities were found specific for a chromosomal syndrome. In prenatal cytogenetic diagnosis, FISH analysis should be considered an effective alternative to full karyotype. P3. ROBINOW SYNDROME, DOMINANT AUTOSOMAL FORM - CASE REPORT Nicoleta Andreescu¹, Valerica Belengeanu¹, Marioara Boia², Monica Stoian¹, Simona Farcas¹, Cristina Popa¹ ¹Department of Medical Genetics, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania ²Department of Neonatology, „Louis Turcanu” Children Hospital, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania Robinow syndrome is characterized by mild to moderate short stature due to postnatal growth retardation, craniofacial abnormalities, skeletal malformations and genital abnormalities. In literature are reported cases with autosomal dominant and autosomal recessive transmission suggesting the genetic heterogeneity of this syndrome. We report a case of a boy, 10 months old, born of non-consanguineous couple. Head circumference at age of 6 months was 46 cm, anterior fontanel 9 cm/5 cm. Examination revealed dysmorphic facial features including macrocephaly, frontal bossing, hypertelorism, epicanthal folds, prominent eyes, long eyelashes, depressed nasal bridge small nose, anteverted nares, posterior rotated ears, prominent philtrum, thin upper lip, triangular mouth, down turned corners of the mouth, highly arched palate, micrognathia. The clinical examination also found: brachydactyly, clinodactyly of the fifth finger, large first toe, dysplasia of toenails, cryptorchidism, micropenis. Radiographic examination showed slightly shortening of forearms and calf bones, cervico-thoracic spina bifida. Head radiography revealed macrocephaly and thin skull bones. The patient presented has the cardinal sign for Robinow Syndrome: characteristic facies, brachymelia, short stature, and genital hypoplasia. These are symptoms found in both forms of Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 49 Robinow Syndrome. The Robinow Syndrome autosomal recessive form may appear more sever abnormalities of the bones and spinal column than in the dominant form. In this case there the shortening of forearms and calf bones was slightly and the height is not under the second standard deviation. The phenotypical appearance of the patient presented in this paper fits within the clinical criteria for Robinow Syndrome, autosomal dominant form. P4. CYTOGENETIC STUDY IN TURNER SYNDROME-THE EXPERIENCE OF MEDICAL GENETICS CENTER IN IASI DURING 2005-2009 G.Andron*, R.Popescu1, C. Rusu1, M. Volosciuc2, L. Butnariu1, E. Braha1, M Panzaru1, L Caba1, M. Macovei1, R. Cojocariu1, M. Gramescu1, C.Bujoran2, I. C. Ivanov3, E. V. Gorduza1 1 „Gr. T. Popa” University of Medicine and Pharmacy, Iasi- Department of Medical Genetics 2” St. Mary” Children Hospital, Iasi- Medical Genetics Center; 3„ St. Spiridon” Hospital ,Iasi- Immunology and Genetics Laboratory „Gr. T. Popa” University of Medicine and Pharmacy, Iasi- student Turner syndrome (determined by total or partial monosomy of X chromosome) is one of the most encountered chromosomal anomalies in female patients. Most often, phenotypic aspects include limphoedema during the neonatal period, short stature, abnormal development of female secundar sexual features, and primary amenorrhea at puberty. This study proposes the cytogenetic evaluation of Turner syndrome cases diagnosed during the period of 2005-2009 in the Medical Genetics Center, Iassy. The diagnosis was based on chromosomal analysis - lymphocyte culture from peripheral blood and G-banding, and FISH (fluorescent in situ hybridization) - in some cases. During this time, 2105 karyotypes were made, 30 of them being make on patients with Turner syndrome.16 cases were identified with homogenous chromosomal anomalies (53,33%): 13 cases with X monosomy (45,X), 1 case - 46,X i(Xq), 1 case - 46,X del(X) and one with a complex chromosomal anomaly - 44,X t.rob(13,14)(q10,q10). The rest of 14 cases (46,66%) presented as mosaics: X monosomy with X trisomy – 45,X/47,XXX (3 cases), X monosomy with normal line – 45,X/46,XX (2 cases), X monosomy with ring chromosome X – 45,X/46,X,r(X) (2 cases), X monosomy with X isochromosome for the long arm - 45,X/46,X,i(Xq) (4 cases). Two of the mosaics are complex presenting 3 cellular lines: 46,XX/46,X,i(Xq)/45,X and 46, XX/47,XX+m/45,X. The last mosaic case 46,XX/46,X,i(Xq) – have not a full X monosomy, but the Turner syndrome phenotype being explained by the presence of X isochromosome for the long arm. The presented study proves the cytogenetic complexity of Turner syndrome and shows similar aspects with specialty literature that states X homogenous monosomy as encountered in 55% of the cases, the rest of them being represented by chromosomal mosaics or partial X monosomies. P5. TRISOMY 8 IN PHILADELPHIA-NEGATIVE CELLS OF CHRONIC MYELOID LEUKEMIA PATIENTS RECEIVING TYROSINE KINASE INHIBITORS: REPORT OF TWO CASES Aurora Arghir1, Sorina Mihaela Chirieac1, Andreea Tutulan-Cunita1, Oana Ciocan2, Carmen Saguna2, Marioara Cristea1, Agripina Lungeanu1 1 „Victor Babes” National Institute of Pathology, Bucharest, Romania 2 „Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania The introduction of tyrosine kinase inhibitors (TKI) dramatically changed the therapy options in chronic myeloid leukemia (CML). The high rates of cytogenetic and molecular responses observed in TKI treated patients were unprecedented. However, clonal cytogenetic anomalies are observed in the Philadelphia-negative cell population in 2-17% of patients receiving TKI, and some of these patients bear a high risk for myelodysplastic syndrome or acute myeloid leukemia. 50 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 While most patients received imatinib mesylate, recent reports revealed Philadelphia-negative cytogenetic abnormalities acquired during dasatinib therapy. We report on two CML patients which developed trisomy 8 in Philadelphia-negative cells during TKI treatment. At the time trisomy 8 was detected, the first patient received imatinib for 24 months and had a major cytogenetic response, while the second patient had a complete cytogenetic response obtained after 18 months of dasatinib. Detailed clinical characterization and systematic follow-up of these patients are necessary in order to define the biological impact of cytogenetically abnormal Philadelphia-negative clones. Genetic studies of a growing cohort of patients will allow a better understanding of the pathogenetic mechanisms underlying the emergence of clonal chromosomal anomalies in Philadelphia-negative cells. P6. INFERTILITY DUE TO GENETIC ANOMALIES IN BOTH PARTNERS OF THE COUPLE. CASE REPORT Atasie D1, Chicea R1, Ispasoiu F1, Corina Ispasoiu2 1 Polisano IVF Center Sibiu 2 Obstetrics&Gynecology Clinic Cluj-Napoca Introduction: Genetic abnormalities are one of the causes of infertility but they are found rare in both partners. Case report: This article presents an interesting case of an infertile couple with repeated IVF failure. The genetic testing revealed abnormalities in both partners. The female cariotype was a mozaicism 46XX/47XXX(83%/17%). The male cariotype showed a balanced translocation between chromosomes 9 and 15 and an inversion of the chromosome 9: 46,XY,t(9;15)(q21.2:p11.2),inv(9). The couple was counseled regarding the risks of these genetic abnormalities and decided to appeal to a sperm donor. Conclusions: although genetic abnormalities are implicated in infertility, the presence in both of the partners of the couple is a rare situation. Key words: infertility, IVF procedures, balanced translocation, mozaicism P7. PREGNANCY WITH EDWARDS SYNDROME IN A COUPLE WITH ADVANCED REPRODUCTIVE AGE. CASE REPORT Atasie D1, Chicea R1, Ispasoiu F1, Corina Ispasoiu2 1 Polisano IVF Center Sibiu 2 Obstetrics&Gynecology Clinic Cluj-Napoca Introduction: Among autosomal trisomies, the Edwards syndrome is the 2nd as frequency after the Langdon –Down syndrome, having an incidence of 1/3000-1/7000 of births, affecting especially the elder couples. Being a severe disorder, the syndrome associates more than 130 different malformations and a bad prognostic, less than 10% of the newly-born survive at the end of their first year of life. Case-report: This article takes into discussion the case of an 40-year-old primiparae during her 23 week of gestation, to whom several malformations were detected during the morphological fetal ultrasound: fetal hypotrophy, bilateral choroid plexus cysts, diaphragmatic hernia, bilateral clubfoot, unique umbilical artery. The amniocentesis revealed the Edwards syndrome (47XX+18). Taking into account the vital risks for the fetus, a therapeutically abortion was suggested to the patient. The anatomopathological exam showed fetal hypotrophy, bilateral clubfoot, low-set years, hydrocephaly, diaphragmatic hernia, left pulmonary hypoplasia, preductal coarctation of the aorta. The cariotypes of both parents were normal. Conclusions: This article presents a case of Edwards syndrome in an elder couple. The incidence of this syndrome is higher in elder parents so the follow-up of the pregnancy by non-invasive means such as fetal ultrasound, double, triple tests, is mandatory. Any suspicion of genetic abnormality must be followed by an amniocentesis and cariotype. Key-words: Edwards syndrome, amniocentesis, cariotype Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 51 P8. BIOMOLECULAR SUBSTRATE OF PAEDIATRIC ACUTE LEUKEMIAS AND ITS IMPACT ON OUTCOME L. Balint-Gib1, A. Oprisoni2, O. Ciocârlie2, S. Arghirescu2, M. Puiu2, A. Isac2, V. Ordodi2, L. Ritli2, M. Serban2 1 Children’s Hospital, Oradea 2 Medical University, Timisoara Introduction. Risk adapted therapy based on prognostic factors raised the survival rates of children with acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). Biomolecular anomalies are important predictive factors included in all risk classifications. With this study we aimed to analyze the biomolecular substrates of acute leukemia and their impact on the outcome. Material and method. This retrospective (2000-2008) observational study was conducted on 104 children with ALL and 23 children with AML. In ALL we analyzed the presence of BCR-ABL1, MLL-AF4, TEL-AML1, E2A-PBX1, SIL-TAL1 rearrangements and in AML the presence of AML1-ETO, CBFβ-MYH11, PML-RARα rearrangements. Results: Out of 104 ALL cases 36(34.61%) presented rearrangements: 5(4.8%) were BCR-ABL1 positive, 4(3.84%) MLL-AF4 positive, 5(4.8%) E2A-PBX1 positive, 18(17,3%) were TEL-AML1 positive and 4(3.84% out of ALL cases, and 44.44% out of T-ALL cases) were SIL-TAL1 positive. BCR-ABL1 and MLL-AF4 rearrangements were associated with a very bad prognosis (5-years pEFS=20% and pOS=40% for BCR-ABL1 positive ALL; 1-year pEFS and pOS was 0% for MLL-AF4 positive ALL). TELAML1 positive ALL was associated with a good evolution (5-years pEFS=83.3% and pOS=89.9%) The E2A-PBX1 rearrangement conferred an intermediate prognostic (5-years pEFS and pOS of 40%). For SIL-TAL1 positive T-ALL, 5-years pEFS and pOS were 25%. As far as AML, 5(21.73%) patients presented rearrangements: AML1-ETO was detected in 3 cases (13.04%) and PML-RARα in 2(8.69%) cases of acute promielocytic leukemia (APL). 2/3 AML1-ETO positive AML patients relapsed and died due to resistant disease; both PML-RARα positive APL patients died early due to hemorrhagic complications. Conclusions: Despite modern therapy protocols, in developing countries, BCR-ABL1 and MLLAF4 rearrangement represents unfavorable prognostic factors. Intensive therapy protocols that can control aggressive disease are frequently associated with lethal complications, leading to decreased survival rates. Biomolecular analysis in childhood acute leukemia remains an important diagnostic tool for optimal disease management. P9. THE FIRST CASE WITH HYPERIGM SYNDROME IN ROMANIA M.Bataneant1, , B.Toth2, L.Marodi2 ,M.Baica1, R.Mihart1, M.Radulescu1, M.Serban1 1 University of Medicine and Pharmacy „Victor Babes” Timisoara, Romania 2 Department of Infections and Pediatric Immunology, Medical and Health Science Center, University of Debrecen, Hungary Introduction. Hyper IgM syndrome is a rare primary immunodeficiency with X-linked inheritance, characterized by hypogammagloblinemia with normal or increased IgM and neutropenia in over 60% of patients. Definitive diagnosis is exclusive confirmed by the molecular biology exploration. Case presentation. A 10 months old male was admitted in IIIrd Pediatric Clinic Timisoara for susspicion of Bruton disease. There were 4 infants deaths on maternal line. At 6 months of age he had Pneumocystosis and needed mechanical ventilation for 33 days. Explorations showed L= 12.160/mmc, Ne=48,8%, Eo=0,5%, Ly=40%, Mo=11%, IgG=1,97g/l, IgA=0,1g/l, IgM=0,72g/l. Number of B lymphocytes was normal and T cell subpopulations were increased. At that moment despite we thought at the hyperIgM syndrome we couldn’t perform CD40 ligand expression and genetic test and we considered it a case of transitory hypogammaglobulinemia of infancy. 52 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 There were excluded cystic fibrosis, HIV infection, tuberculosis. At 2 years and 10 months he was admitted for oral and perianal ulcers, bloody diarheea and pneumonia,. Hemoleucogram revealed severe neutropenia - 0,2 % and immunogram IgG = 0,03 g/l, IgA= < 0,57g/l, IgM= 1,24 g/l. Molecular analysis showed a c.614 T>C missense mutation in exon 5 of the CD40LG gene and mother is carrier. Conclusion. In all cases with hypogammaglobulinemia with normal IgM, pneumocystosis and neutropenia we must exclude hyper IgM syndrome, the decisive role in diagnosis it has the genetic exploration which permits not only a correct diagnosis and management of therapy but also a genetic council and profilaxy of the disease through prenatal genetic testing. P10. CLINICAL, MORPHOLOGICAL AND IMMUNE-GENETICAL CORRELATIONS IN CELIAC DISEASE CHILDREN Belei Oana 1, Simedrea Ioan 1, Tamas Liviu 2, Marginean Otilia 1, Daescu Camelia 1, Marcovici Tamara 1, Brad Georgiana 1, Puiu Maria 1 1 First Pediatric Clinic, University of Medicine and Pharmacy Victor Babes Timisoara 2 Biochemistry Department, University of Medicine and Pharmacy Victor Babes Timisoara Introduction: Celiac disease (CD) is an immune –mediated entheropaty, determined by gluten ingestion in genetical predisposed subjects (haplotype DQ2 or DQ8). Aim: To perform a screening study on a pediatric group that associates certain risk factors for CD and to assess the correlation between different forms of celiac disease, immunological, morphological parameters and haplotypes. Methods:The study developed between January 2008 - November 2009. The serological tests used for screening included IgA anti-tissue transglutaminase (IgA tTG) and anti-endomysial antibodies (EMA). In case of positive result for at least one test, we performed intestinal biopsy. Interpretation was done according to Marsh criteria. The study included 2 groups. The first lot was made of 30 celiac patients diagnosed with CD during screening.The second lot was composed by 30 control subjects matched for age and sex. We assessed the HLA DQ2 and DQ8 using PCR technique to all celiac patients and in control lot. Statistical testing was done by using SPSS 16 software. Multivariate conditional logistic analyze was performed. Results: From 30 celiac patients, 18 patients presented atypical forms of disease, 4 patients presented silent forms of disease, and only 8 associated the classical forms of disease. Alleles distribution in group of celiac patients was: 28 were positive for HLA DQ2 and from them 7 patients associated haplotype HLA DQ2 homozygous and 21 associated haplotype HLA DQ2 heterozygous. 2 cases presented HLA DQ8. In the control lot, 2 subjects from 30 associated haplotype HLA DQ2 heterozygous.The rest were negative for HLA DQ2 or DQ8. We tried to correlate the clinical forms of disease with IgA tTG antibodies serum level, severity of villous injury and haplotype. We obtain a significant correlation between IgA tTG serum level and severity of villous injury (r = 0,621092). We also established a positive correlation only between subgroup Marsh IIIc and the severe classic form of celiac disease. The forms of disease and the haplotypes did not correlate. Conclusions: The polymorphism of CD presenting forms as well as the lack of concordance between clinical symptoms and the type of intestinal injury, make the intestinal biopsy the gold standard for diagnosis when the clinical suspicion of gluten intolerance exists.The HLA polymorphism seems to have no impact on clinical forms of disease.The presence of molecules DQ2 or DQ8 is mandatory, but not sufficient for development of CD. Due to its high negative predictive value, the assessement of haplotype must be used in clinical practice only at uncertain cases. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 53 P11. CORNELIA DE LANGE SYNDROME – CLINICAL AND EVOLUTIVE ASPECTS Marioara Boia, Maria Puiu, Anca Popoiu, Aniko Manea, Mirabela Dima University of Medicine and Pharmacy Timisoara, Romania Introduction: Cornelia De Lange syndrome is a rare disease in medical practice characterized by facial dimorphism associated with multiple congenital malfunctions with lead to failure in growth. Method: This study took place at the Department of Neonatology in the University of Medicine Timisoara and contains an analysis of the clinical and biological symptoms of three patients. Facial dimorphism associated with multiple congenital malfunctions and pre and postnatal growth deficiency are the criteria for including them in the study. Results: All three patients have shown normal gestational age with intrauterine growth deficiency that have persisted after birth. The postnatal clinical symptoms that have given us the correct diagnosis are: microcephaly(60%), hypertonia(100%), respiratory and feeding difficulties in the newborn period and infancy(100%), psychomotor retard, hirsutism(100%), micromielia(100%). Evolution: one patient died at the age of 2 months with cardio-respiratory insufficiency. The other 2 patients have major growth retard. Conclusions: The clinical aspect has been sufficient to establish the correct diagnosis. The most important clinical signs were: facial dimorphism, hirsutism, microcephalia, cardiac and bone malformations. Key words: malformations, syndrome. P12. CLINICAL AND EVOLUTIVE ASPECTS IN TAR SYNDROME -CASE REPORT Marioara Boia, Valeria Belengeanu, Anca Popoiu, Dana Iacob, Aniko Manea, Lucica Stoica University of Medicine and Pharmacy Timisoara, Romania Introduction: TAR syndrome is an rare autosomal recessive disorder characterized by severe thrombocytopenia and bilateral absent radii. TAR syndrome represents an association of malformations that includes many tissues and organs. Material and Method: A case of a newborn was presented with a complex malformed syndrome: skull and facial dysmorphism, bilateral radial aplasia, intense systolic murmur and severe thrombocytopenia. Results: The historical record of the newborn was not containing any special/unusual elements: newborn female with the gestational age of 38 weeks, birth weight of 2750 g, APGAR score was 8/9. There are not other malformations in her family. The physical examination showed generalised purple. Dismorfic facies, highly arched palate, low-implanted ears, micrognathia, short neck. Hypoplasia of the shoulder girth, short forearms, bilateral simian line Conclusions. The clinical aspects were suggestive for the orientation of the diagnosis. The association between severe and persistent thrombocytopenia and the skeletal modifications being certain elements for positive diagnosis. The caryotype modifications were not present and also no other cases with TAR syndrome in this family. 54 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P13. MICROSATELLITES STUDY IN TURNER SYNDROME – PARENTAL ORIGIN OF X CHROMOSOME Bolca D1, Carel JC2, Grigorescu-Sido P3, Pop IV1 1 Chair of Medical Genetics, UMF „Iuliu Hatieganu” Cluj 2 Robert-Debre Hospital, Research Laboratory INSERM CIC-EC5, Paris 7 University 3 Chair of First Pediatric Clinic, UMF „Iuliu Hatieganu” Cluj Turner syndrome is due to complete or partial absence of the second sex chromosome. Clinical picture is characterized by a large heterogeneity, being sometimes correlated with a mosaic of cell populations or with the presence of structural abnormalities of X chromosome, but more often being unexplained by these karyotype differences. The data obtained by recent studies, sometime discordant, suggest that the imprinting of the genes on X might explain this phenotypic variability, particularly for: cognitive function, height growth and lipid metabolism. Aim: evaluation of the parental origin of X chromosome in a lot of patients with Turner syndrome, in aim to identify his role in Turner syndrome phenotype. Material and method: In study it was included 208 couples mother-daughter with 45,X karyotype, without mosaicism detectable by Flourescence in Situ Hybridisation (FISH), from France Hypophyse database, organized from1973 until 1997, in aim to selection and oversee the patients treated with growth hormone in France. Parental origin of X chromosome was determined after the study of 9 microsatellites localized on X chromosome, by comparing the allele length of the couple mother-daughter. The techniques of molecular biology were: ADN extraction (from the peripheral blood lymphocytes or from the oral mucosa cells), ADN pre-amplification, amplification by PCR of the selected microsatellites, control electrophoresis on agarose gel, passage on ABI PRISM Linkage Mapping Sets v2.5 and the analysis of the results by Gene Mapper soft. Paternal origin was established if at least 2 markers presented alleles of different length, and maternal origin if all the alleles were similar between mother and daughter, being confirmed thereafter by the calculus of the discriminant power (0,001). Results: For 208 couples who were analyzed, it was established the parental origin of X chromosome in 186 couples, thus: 136 (73%) patients present a maternal origin of the X chromosome and 50 (27%) had a paternal origin of X chromosome. Beginning with this finding we will analyze further the correlation with the phenotype for these patients. Conclusions: This multicentric study is the first which analyzes the phenotypic variability in Turner syndrome on such a great sample, allowing thus a more correct evaluation of the physiopathologic role of the parental origin of X chromosome in patients with 45, X homogeneous karyotype. P14. MOSAICISM IDENTIFICATION BY MOLECULAR CYTOGENETICS IN PATIENTS WITH TURNER SYNDROME Bolca D1, Carel JC2, Grigorescu-Sido P3, Pop IV1 1 Chair of Medical Genetics, UMF „Iuliu Hatieganu” Cluj 2 Robert-Debre Hospital, Research Laboratory INSERM CIC-EC5, Paris 7 University 3 Chair of First Pediatric Clinic, UMF „Iuliu Hatieganu” Cluj Turner syndrome is frequently associated with chromosomal mosaicism, some studies showing the presence of sex chromosomes mosaicism at 40-45% of Turner syndrome patients. The cell populations in mosaic may explain in some situations the very heterogeneous clinical picture in Turner syndrome. It was observed at 5% of patients some mosaicisms with Y chromosome, his identification being important due of the high risk of gonadoblastoma in these patients. Aim: Mosaicism identification by Fluorescence in Situ Hybridization (FISH) in patients with Turner syndrome who had a 45, X homogeneous karyotype in conventional cytogenetics. Material and method: In study it was included 231 patients diagnosed with 45,X homogeneous karyotype, from the France Hypophyse database organized from 1973 to1997, in aim to selection Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 55 and oversee the patients treated with growth hormone in France. It was done the FISH analysis of peripheral blood lymphocytes, using centromeric probes for X and Y chromosomes. Results: It was identified 23 patients (9,9%) with sex chromosomes mosaicisms, the second cell line being observed at 19 patients (8,2%): 17(7, 35%)-45,X/46,XX; 2(0,85%)-45,X/47,XXX and the third cellular line at 4 (1,7%) – 45,X/46,XX/47,XXX. The level of mosaicism was between 2 and 32%. None of the patients studied by FISH have the Y chromosome. Conclusions: FISH analysis of sex chromosome may be an useful instrument, associated to the techniques of conventional cytogenetics, the identification of a mosaicism of X and Y chromosome being important for the clinical management. P15. HIPERMETHYLATION OF MIR-124A GENE PROMOTER IN CERVICAL ONCOGENESIS Anca Botezatu1, Demetra Socolov2, Cristina D.Goia1, Iulia V. Iancu1, Irina Huica1, Adriana Plesa1, Gabriela Anton1 1 Institute of Virology „Stefan S Nicolau”, Bucharest, Romania 2 University of Medicine and Pharmacy„Gr.T.Popa”, Iasi MicroRNAs (miRNAs) are small, noncoding RNAs that can contribute to cancer development and progression by acting as oncogenes or tumor suppressor genes. Identification of genes that undergo cancer-specific CpG island hypermethylation and correlation of these data with pre-neoplastic lesions, tumor stage, progression, and long-term prognosis are becoming increasingly common. Subsequent analysis proved that miR-124a is also frequently methylated in colon, breast and lung carcinoma cell lines, as well as in leukemias and lymphomas. The hypermethylation of miR-124a promoter in cancer cells results in an increased expression of its target CDK6. This study was conducted to investigate the promoter methylation status of the miR-124a gene promoter in pre-neoplastic cervical lesions and cervical cancer and his possible implication in regulation of p21 mRNA levels, and the CDK6 partner-cyclinD1. Promoter methylation was evaluated using a methylation-specific polymerase chain reaction for bisulphite treated DNA (EpiTect Bisulfite Kit – Qiagen) samples. The DNA samples were isolated from 50 cervical swabs (High Pure PCR Template – Roche). We found significantly higher methylation frequencies for the miR-124a gene promoter in preneoplastic lesions and in cervical cancer lesions. CylinD1 gene expression level is increased in cervical squamous carcinomas and adenocarcinomas, p21 gene is dowregulated in squamous carcinomas in adenocarcinomas, therefore we can conclude that in gene expression regulation are involved other factors. The miRNAs promoter gene hypermethylation is a frequent event in cervix carcinogenesis and holds a great prognostic value P16. FACIAL MALFORMATIONS IN AMNIOTIC BAND SYNDROME Elena Braha, M. Voloşciuc, Lacramioara Butnariu, Monica Panzaru, Roxana Popescu, Cristina Rusu Amniotic band syndrome (ABS) is a set of congenital anomalies in which the major feature is the congenital ring constriction affecting all parts of the embryo (mainly the limbs, craniofacial area). The facial anomalies are polymorphous and asymmetric, the type and the severity depend on band location and time of onset. The disease is rare, the prevalence is between 1/1200 – 1/15000 newborns and 1/3 could have facial malformations. We report 10 cases with clinical features strongly suggestive for ABS. The cases were evaluated in the last 3 years and diagnosed relatively precocious (age between 1 day and 2 years 7 months). The facial malformations are polymorphous from simple asymmetric face, broad nasal root to severe anomalies (lip or/and palate cleft, oblique facial clefts, ossification defects of the calvaria). The limb anomalies (ring constriction, digital amputations), are constantly present. 56 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 The clinical features of the disease are easy to recognize. This lead, usually, to a precocious diagnosis. The disease assessment has to be multidisciplinary. The patients’ prognosis depends on the location and severity of the anomalies, one of these could be disabling and recuired psychological support even after surgery. We present a brief review of literature in pathogenic concepts (embryologic, mechanic, vascular and genetic hypothesis), the clinical features and medical assessment. The prenatal diagnosis is difficult and could be made by fetal echography. P17. CYTOGENETIC RESPONSE OF BONE MARROW TO FIRST LINE THERAPY IN CHRONIC MYELOGENOUS LEUKEMIA Cornel Bujoran1, Mircea Covic2, Iuliu Ivanov3, Gabriela Dorohoi4, C.Catalin4, Ecaterina Hanganu– Turtureanu4, Cristina Burcoveanu4, Angela Dascalescu4 Eusebiu Vlad Gorduza2 1 Spitalul clinic de copii „Sf.Maria” Iasi, Laboratorul de Citogenetica, 2 Universitatea de Medicina si Farmacie „Gr. T. Popa” Iasi, Disciplina de Genetica, 3 Spitalul clinic „Sf.Spiridon” Iasi, Laboratorul de Imunologie si Genetica, 4Clinica de oncohematologie al Spitalului clinic „Sf. Spiridon”Iasi The aim of our retrospective study (2002-2009) is to identify the cytogenetic response of bone marrow to first line therapy, with imatinib, in chronic myelogenous leukemia (CML). The chromosomic analysis of bone marrow cells was obtain after 2 days cell culture. Between 2002 and 2009 we analyzed 336 new cases and the cell culture succeeded at 287 patients (85,4%). The clinical reasons for analyses were: chronic myelogenous leukemia (CML) (152 cases) and other blood diseases (135 cases). In cases with CML the chromosomal analysis indicated: a normal karyotype (KN) – 31 cases (20,4%); the presence of a Philadelphia chromosome in all cells (PhO) - 102 cases (67,1%); the presence of a chromosomal mosaics with a cell line with Philadelphia chromosome - 12 cases (7,9%) and the presence of a minor clones (dup crs Ph, 8 or 19 chromosome trisomy) - 7 cases (4,6%). At patients with chromosomal changing was applied a therapy for control of hematological features and to stop the progression of disease to blastic phase. Until 2003, in CML we used an unspecified therapy (hydroxiurea, busulfan or interferon) and from the end of 2003 year we introduced a specific therapy with imatinib (Glivec) that inhibit the tirosinkinase. At 102 patients with homogenous CML Ph(+), we applied a treatment with imatinib 400mg/zi, and the control of therapy implied the repeat of cytogenetic analysis of bone marrow. At periodic controls we identified two types of responses: 1 – the decrease of Ph chr, without cytogenetic remission (CI-37;CV-17) and 2 – complete cytogenetic remission at all periodic controls (CI-12; CV-50). Our study indicates the utility of cytogenetic analysis of bone marrow in diagnosis (the evidence of Ph chromosome) and the monitoring of cytogenetic response at therapy with imatinib in CML. P18. CLINICO-GENETICAL CORRELATIONS IN DARIER AND HAILEY-HAILEY DISEASE Elena Buteica, Florin Burada, Cristina Angelescu, Alice Buteica, Anca Riza, Irina Stoicescu University of Medicine and Pharmacy from Craiova Introduction: Darrier disease (DD), also known as Keratosis Follicularis, and Hailey-Hailey disease (HHD) or Familial Benign Pemphigus, are rare genodermatosis, with autosomal dominant inheritance pattern, being caused by mutations in ATP2A2 gene, encoding for a Ca pump, sarco/endoplasmatic protein SERCA in DD, and ATP2C1, a Ca/Mn pump, hSPCA1 protein, in HHD. Material and methods: Clinical investigation and laboratory analysis was conducted on two patients: a man, 33 years-old, with DD, and a woman, 43 years-old, with HHD. Results: The patient with DD presents mucosal lesions and dental modifications associated with mild mental retardation. In the HHD case skin lesions are associated with neuropsychiatric and endocrinologic disorders. In both cases, the mutation is inherited from parents, as family background Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 57 shows. Histologically, both HHD and DD have acantholytic suprabasal keratinocytes; however, DD also has dyskeratotic keratinocytes, and in HHD the acantholysis is incomplete . Conclusion: In DD and HHD, the genetic mutations alter calcium and manganese homeostasis, leading to abnormal desmosomal adhesion between keratinocytes. Even if these diseases have similar features, clinically, genetically and histopathologically they are distinct entities, requiring a multidisciplinary approach in diagnosis. P19. ROBERTSONIAN TRANSLOCATIONS – IMPORTANT CHROMOSOMAL ANOMALIES Caba L1, Covic M1, Rusu C1, Volosciuc M2, Butnariu L1, Braha E1, Bujoran C2, Gramescu M3, Ivanov I3, Panzaru M1, Popescu R1, Sireteanu A1, Gorduza EV1 1 University of Medicine and Pharmacy „Gr.T.Popa” Iasi, Medical Genetics Department 2 Pediatrics Hospital „Sfanta Maria” Iasi 3 „Sfantul Spiridon” Hospital Iasi Introduction: Robertsonian translocations are frequent chromosomal rearrangements (incidence 1:1000) involved two acrocentric chromosomes, homologous or nonhomologous, that fusion near the centromer with lack of the short arms. Presence of robertsonian translocation in balanced or unbalanced form required karyotype of the first degrees because of the risk of transmission to descendant and prenatal diagnosis for the balanced robertsonian translocation. Material and method: Our lot is formed by 35 patients with robertsonian translocations diagnosed in Medical Genetics Center Iasi between January 2001 – June 2010 from a total number of 2133 karyotypes. Chromosomal analysis was done using culture of lymphocytes and GTG bands. Clinical reasons for karyotype were : first degrees for an unbalanced robertsonian translocations (trisomy 21 trisomy 13) or balanced one, patient with suggestive phenotype for autosomal trisomy of an acrocentric chromosome (Down syndrome in 20 cases, Patau syndrome in 1 case), couples with reproductive problems and in a single case mieloblastic acute leukaemia. Results: There were 20 robertsonian translocations in unbalanced form and 15 in balanced form. Robertsonian translocations identified are from the three groups: D;D (13/14), D;G (13/21, 13/22, 14/21, 15/21) and G;G (21/21, 21/22). The highest frequency is for 14/21 robertsonian translocation (14 cases) followed by 13/14 and 13/22 (5 cases each), 13/21 and 21/21 (4 cases each). Less frequent were 21/22 robertsonian translocation (2 familial cases) and 15/21 (one case). The karyotype was made to the parents for 10 patients. 3 from 10 were de novo anomalies (14/21-one case, 21/21two cases). In 7 cases the anomaly was found at one of the parents (predominant in mother – 6 cases)- robertsonian translocation 13/14, 13/21, 13/22, 14/21, 21/22- and in 3 of these situations, the anomaly was transmitted in balanced form. Just in three cases we found a complex anomaly : 44,X,der(13;14),(q10;q10) (the translocation is from the father) ,XY,rob(13;21)(q10;q10),+21/ 47,XXY,rob(13;21)(q10;q10),+21 (the translocation is transmitted from the mother), 46,XX,+21,ro b(13,22)(q10;q10). Conclusions: Identification of robertsonian translocation type is important for calculating the risk of recurrence and providing genetic counseling, given that these anomalies are on a different acrocentric chromosome, and, on the other hand, these translocations may be transmitted from a parent with balanced anomaly to a child with unbalanced one. 58 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P20. IDENTIFICATION BY DNA MICROARRAY TECHNOLOGY OF CANDIDATE GENES INVOLVED IN MAINTAINING INTERSTITIAL CAJAL CELLS (ICC) AND INTERSTITIAL CAJAL-LIKE CELLS (ICLC) PHENOTYPES IN C-KIT MUTANT MICE Georgeta Cardos1, Carmen Ardeleanu1,2, Georgeta Butur1 and Mihail Eugen Hinescu1,2 1 „Victor Babes” National Institute of Pathology, Bucharest, Romania 2 „Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania DNA Microarray technology is a new and valuable tool used in high-throughput expression gene studies, allowing complete genome monitoring. We used this technology for a comparative investigation of gene expression in normal and c-kit mutant mice, in order to study the ICC and ICLC phenotypes with physiologic implications. ICCs were discovered by Ramon y Cajal, at the end of the 19th century, as networks in the gastrointestinal tract walls, being involved in digestive motility and neurotransmission, but also in GIST (gastro-intestinal stromal tumors) pathogenesis. Other extra-digestive organs (such as: gall bladder, pancreas, heart and some blood vessels, uterus, fallopian tubes) have been shown to contain ICLCs, whose functions are yet unknown. Both ICCs and ICLCs express C-Kit protein, a tyrosine-kinase receptor with essential role in differentiation and maintenance of these cellular types. To conduct our comparative gene expression study, different digestive and extra-digestive organs from control and mutant mice (WBB6F1/J-KitW/KitW-v/J strain) were sampled. Total RNA was extracted using the AllPrep DNA/RNA Mini Kit (Qiagen) and analyzed by Bioanalyzer and the RNA 6000 Nano assay Kit (Agilent Technologies). DNA microarray chips from the Whole Mouse Genome Microarray Kit (Agilent Technologies) were hybridized and scanned by the Agilent DNA Microarray Scanner. The DNA microarray data analysis was performed with the Feature Extraction 5.1.1 and the GeneSpring GX 10.Expression Analysis Software (Agilent Technologies). More than 2000 genes demonstrated differential expression by >2 fold in mutant versus control mice. Certain genes were up-regulated, such as: Dmbx1, Eif4a1, Zfp593 and Zfp69 (involved in transcription regulation), Abi2, CaCng8, Hbb-b1 (cellular transport and cell junctions) and Calcr, Cyp3a44, Mcpt1 (signal transduction and metabolic process regulation). Other genes were downregulated, such as Wnk1, Sema5a, Nf1, Tnfrsf21 (involved in apoptosis, intra-cellular transport and inter-cellular communication). Some of these genes, validated by RT-Quantitative PCR, may become candidate biomarkers for studying ICCs and associated pathology in humans. Financial support: PN 06.26 – 02.18 and PN 09.33.02.09 Projects. P21. PRELIMINARY RESULTS: ALPHA 1 ANTITRYPSIN DEFICIENCY AND LUNG CANCER Catana Andreea1, Prof.Dr. Pop Ioan Victor1, Dr. Popp Radu1, Prof Dr. Pop Monica2, Petrisor Felicia1 1 Medical Genetics Department, University of Medicine and Pharmacy Cluj, Romania 2 Leon Daniello Pneumology Hospital Cluj, Romania Background. Alpha 1 antitrypsin S and Z deficiency alleles of Serpina 1 gene, were reported to potentially increase the risk of lung cancer development amog heterozygous carriers. Objectives. This is a cross-sectional, randomized, case control study, for the evaluation of the frequency of MS and MZ alleles among patients with lung cancer. Subjects The study included 30 cases of lung cancer diagnosed patients (histopathological examination), recruted from the Pneumology Hospital Leon Daniello Cluj and 30 healthy unrelated controls, selected among patients observed in the Internal Medicine department. Methods. PCR amplification of relevant gene segment was followed by restriction enzyme digestion Taq1. Detection of A1AT gene S and Z alleles was determined through analysis of resulting restriction fragment length polymorphism (RFLP). Rezults. The molecular analysis identified the genotype MS in 4 of the patients with lung cancer and 3 of the controls. The heterozigous MZ state was detected neither among cases nor in controls. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 59 Conclusions. The preliminary results of the study did not reach statistical significance, so we consider presently insufficient information regarding the distribution of A1AT mutant alleles among individuals with lung cancer. Future studies, with larger number of case-controls are required to establish the appropiate connection betweenn MS and MZ carrier status and risk of lung cancer development. P. 22. DISCUSIONS ABOUT A NEW CASE OF WILLIAMS SYNDROME EVALUATION AND MANAGEMENT A. Chelmus*, R.Popescu1, C. Rusu1, M. Volosciuc2, C.Bujoran2, I. C. Ivanov3, V. E. Gorduza1 1 „Gr.T.Popa” University of Medicine and Farmacy, Iasi - Department of Medical Genetics, 2 „Sfanta Maria” Hospital for children, Iasi – Centre of Medical Genetics; 3 „Sfantul Spiridon” Hospital, Iasi - Imunology and Genetics Laboratory * „Gr.T.Popa”University of Medicine and Farmacy, Iasi - student Williams syndrome is a rare genetic disorder, caused by a microdeletion on the shoart arm of chromosome 7, region q11.23. The phenotype is characterized by a distinctive ,,elfin face’’ and includes: delay neurodevelopment, congenital heart defects, neonatal hypercalcemia, ophtalmo logical anomalies, hypothyroidism, delay in growth, hyperlaxity in joints. Also the neuropsychological profile includes visuo-spatial deficits and hypersociability. We will present a new case of William syndrome diagnosed in the Centre of Medical Genetics Iasi, in order to illustrate a rare disease, to discuss about the genetic counselling and the management of the patient and her family. The female patient (1 year old) is the third child of an young, unrelated and apparently healthy couple. The birth was natural, at term; the child presented a slightly delay in the intrauterine development (weight - 2800 g; hight - 49 cm).The clinical evaluation revealed : normal morphometry, coarse appearance, distinctive ,,elfin face’’, stellate iris pattern, slightly protruding eyes, small nose with flat nasal bridge and upturned nostrils, ears with low insertion, square hands with slightly divergent fingers, tendency for valgus and bilateral flat foot, mild hipotonia, heart murmur, a mild delay in psychomotor development. The echocardiography – supravalvular aortic stenosis, pulmonary branch stenosis, malformed aortic valve. Because the chromosomal analys was normal, we performed FISH analys, which confirmed the diagnosis.Parental karyotypes – in work. The patient deceased during an anesthesia for an ophtalmologic intervention. In conclusion we present a new case of William syndrome to discuss the evaluation and management within this syndrome. P23. CYTOGENETIC AND CLINICAL FEATURES IN CRI DU CHAT SYNDROME – REPORT OF TWO CASES Sorina Mihaela Chirieac1, Magdalena Budisteanu2, Aurora Arghir1, Andreea Tutulan-Cunita1, Ioana Borcan1 & Agripina Lungeanu1 1 „Victor Babes” National Institute of Pathology, Bucharest, Romania 2 Clinical Hospital of Psychiatry „Prof. Dr. Alex. Obregia”, Bucharest, Romania Cri du Chat syndrome is a rare genetic condition resulting from a deletion in the short arm of chromosome 5. The significant clinical features are the characteristic high-pitched „cat-like” cry, distinct facial dysmorphism, abnormal dermatoglyphics, and severe psychomotor and mental retardation. Classical and molecular cytogenetic studies showed 5p deletions of variable size in association with a wide clinical spectrum. In this paper, we present two cases with large deletions, comprising the entire short arm of chromosome 5, and characteristic, though distinct clinical features. There is no specific therapy for Cri du Chat syndrome; however early diagnosis and estimation of the deletion size improves prognostic and patient management. 60 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P24. PRADER WILLI SYNDROME IN INFANCY Adela Chirita1, Ramona Giurescu1, 2, Maria Puiu 2, Corina Duncescu 1, Gabriela Doros1, 2, Boia Mariana3 ,Valeria Belengeanu2, Ioana Micle1,2 1 „Louis Turcanu” Clinical Emergency Hospital for Children Timisoara. 2 „Victor Babes” University of Medicine and Pharmacy Timisoara 3 Premature infants - „Louis Turcanu” Clinical Emergency Hospital for Children Timisoara Background: Prader Willi Syndrome (PWS) is a genetic disorder characterized, depending on age, by two distinct phenotypes. In the first year of life infants associate hypotonia, feeding difficulties and failure to thrive. Mental retardation, hyperphagia and short stature are the main features after one year. Usually the diagnosis is established when obesity occurs. PWS is a complex disease with metabolic and neurological implications; it needs a complex approach with a multidisciplinary team, begun as early as possible. Early diagnosis may help prevent obesity and short stature (growth hormone therapy). Aim: To present a six month male infant, who was admitted in the emergency care unit with severe respiratory syndrome, somnolence, feeding difficulties and a particular phenotype: severe hypotonia, small hands and feet, hypogonadism, undescended tests, characteristic facial features (narrow bifrontal diameter, almond-shaped palpebral fissures and down-turned mouth) and skin folds with obese appearance. On basis of clinical examination PWS was suspected. Methods: After clinical and biological investigations, he was directed to interdisciplinary consults. The patient performed complex cardiological examination: clinical, ECG, Echocardiography, cardiopulmonary X ray and biological investigations, followed by genetic exploration. Results: The following diagnoses were established: primary pulmonary hypertension with severe cardio-respiratory failure, patent foramen ovale with reversed shunt, bacterial pneumonia. The clinical suspicion of PWS was confirmed by genetic testing, detection of microdeletions by FISH method. He was treated with iv antibiotics, Sildenafil, diuretics and Digoxin. Because of the associated pathology, the evolution was unfavorable, death occurring at 8 months of age. For this case the early diagnosis of PWS led to genetic counseling in terms of future pregnancies. Conclusion: A high degree of susceptibility is needed to establish early in infancy the diagnosis of PWS, based on individual phenotype, the dominant trait being hypotonia, somnolence and feeding disorders. The particularity of the case was the obese appearance and the severe associated pathology. Keywords: Prader Willi syndrome, infancy, hypotonia, pulmonary hypertension P25. TESTICULAR ADRENAL REST TUMORS IN AN ADOLESCENT BOY WITH 21 -HYDROXYLASE DEFICENCY Adela Chirita1, Monica Marazan1,2, Ramona Cojocaru1, Duncescu Corina1, Ioana Micle1,2 Spitalul de Urgenta pentru Copii”Louis Turcanu” Timisoara Universitatea de Medicina si Farmacie „Victor Babes”Timisoara Congenital adrenal hyperplasia (CAH) is an inherited disorder resulting in impaired production of cortisol and aldosterone. Testicular adrenal rest tumors (TART) are known complications in male patients with CAH. AIM: We discuss the embryological, clinical features and treatment options of TART in a patient with CAH. PATIENT AND METHODS: We present a case of TART in a 16 years old boy with salt-wasting type of CAH (21-hydroxylase deficiency). The patient had periodic clinical evaluations, complete metabolic and endocrinological profile which guided the treatment. RESULTS: He was diagnosed at 6 weeks, when he begun substitution treatment with hydrocortisone and fludrocortisone. He was a compliant and well controlled patient until age 15, when he went away to college. At 16 years scrotal ultrasound sowed bilateral non-homogeneous, extensive masses. Now, the hormone profile revealed increased ACTH and 17-OHprogesterone levels. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 61 Treatment was made by changing the night dose of hydrocortisone into dexamethasone to prolong ACTH suppression. CONCLUSIONS: Puberty has a negative impact on treatment compliance, so increasing the risk for TART. They are not malignant but can result in gonadal dysfunction and infertility. It is important to screen for these tumors in CAH patients, to inform, discuss and offer cryopreservation of semen as soon as possible. Key words: Congenital adrenal hyperplasia, testicular adrenal rest tumors, adolescent P26. TESTING OF ASSOCIATION BETWEEN INSULINA-IGF2 REGION AND DIABETES MELLITUS, OBESITY AND BREAST CANCER Danut Cimponeriu1, Pompilia Apostol1, Mihai Toma1, Stavarachi Monica, Irina Radu1, Anne Marie Craciun2, Cristian Serafinceanu2, Rusu Lavinia3, Traean Burcos4, Popa Emil4, Popa Ileana4, Stanilescu Sorin4 1 University of Bucharest 2 IDNMD „N Paulescu” Bucharest 3 Institute of Food Bioresources, Bucharest 4 Coltea Clinic Hospital, Bucharest Genes from Insulina-IGF2 region (11p15.5) are candidates for some complex diseases, like type 1 diabetes mellitus (T1DM), type 2 diabetes mellitus (T2DM), obesity and breast cancer. In this study we have tested the potential association between three markers from Insulina-IGF2 region and diseases mentioned above. Clinical data and biological samples have been colected from unrelated patients with T1DM (n=204), T2DM (n=215), obesity (n=200) and breast cancer (n=100). For the control lot we selected 850 clinically healthy subjects, which have no relatives with diabetes, obesity and cancer. All subjects selected for this study are Caucasians and live in the South part of Romania. Insulin -23Hph, Insulin +1127Pst1 and IGF2 Apa polymorphisms have been genotyped in all blood samples by PCR RFLP method. Results obtained have shown that all three polymorphisms are distributed in acorance with Hardy-Weinberg equilibrium in all lots and that -23Hph and +1127Pst polymorphisms are in strong linkage. The -23Hph AA and +1127 Pst -/- genotyes increase the risk for T1DM (OR: 3,22, 95%CI: 2,09-4,98, p<0,0001), especially in patients without macroalbuminuria (OR: 4,32, 95%CI: 2,54-7,45, p < 0,0001). No other associations between genotypes or alles of investgated markers and T2DM, obesity and breast cancer have been identified. In conclusion -23Hph AA si +1127 Pst -/- genotypes increase the risk for T1DM onset. Acknowledgments. This study was supported by Romanian Ministry of Education and Research (Research Project PNII Partnerships 42-161). P27. K-RAS GENE MUTATIONAL STATUS AND ITS CORRELATIONS WITH THE HISTOPATHOLOGICAL FINDINGS IN THE COLORECTAL CARCINOMAS Florina Lucia Cionca, Georgeta Cardos, Mihaela Mihai, Alina Georgescu, Mihai Stoicea, Simona Enache, Valentin Enache, Georgeta Butur, Carmen Ardeleanu „Victor Babes” National Institute of Pathology, Bucharest, Romania Background: Gain-of-function K-ras point mutations, present in 20–40% of the colorectal carcinomas, maintain the active form of the ras p21 protein and lead to epidermal growth factor receptor (EGFR) independent activation of intracellular signaling pathways, making the anti-EGFR tumor therapy ineffective. The aim of the present study is to identify possible correlations between the mutational status of the K-ras gene and the histopathological findings in patients with colorectal adenocarcinoma. 62 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Methods: We studied 34 patients with colorectal adenocarcinoma (31) or liver metastases of colorectal adenocarcinoma (4). (male/ female:19/15, age range: 34-87, average age 61). The formalin-fixed paraffin-embedded tissue samples were analyzed using an indirect bistadial immunohistochemical (IHC) technique, performed with a Dako EnVision+ Dual Link System-HRP, with antibodies for the EGFR. The mutations in exon 2, codon 12 of the K-ras gene were detected by PCR-Restriction Fragment Length Polymorphism method, with MvaI restriction enzyme. Results: A K-ras mutation in exon 2, codon 12 was present in 23, 52 % of the cases, 7 adenocarcinomas and 1 liver metastasis. EGFR was positive in 10 cases in tumor cells and in 7 cases in the vessels. The relationship between the presence of a K-ras mutation in exon 2, codon 12 and the positive immunohistochemical reaction for EGFR did not reach statistical significance. Conclusions: There were no significant correlations between the mutational status of the K-ras gene and the IHC reaction for EGFR, proving once more the major role of the molecular analyses in colorectal carcinoma anti-EGFR therapy. P28. RISK FACTORS OF CYSTIC FIBROSIS LIVER DISEASE Ioana M. Ciuca1, L. Pop1, I. Popa1, Z. Popa2, L. Tamas3, 1 Pediatric II Department, UMF „Victor Babes” Timisoara, Romania 2 National Cystic Fibrosis Centre, Timisoara, Romania 3 Biochemistry Department, University of Medicine and Pharmacy, Timisoara, Romania Introduction: Cystic fibrosis(CF) associated liver disease is the second cause of death in CF and may be the first disease expression in CF. It seems that many recognized risk factors like severe mutations, history of meconium ileus and male gender could suggest the occurrence of the liver disease. Aims & Methods: Aim study was to asses liver disease`s frequency and its correlation with recognized risk factors. Study was prospective for a five years period; 158 patients were followed up by clinical assessment, liver function tests (LFTs), abdominal ultrasound examinations (US) and CFTR tests. In some cases liver biopsy, MRI and elastogramme was performed. Results: Cystic fibrosis associated liver disease (CFLD) was diagnosed in 51 patients (32.27%), slightly predominance of boys. The disease occurred more frequently in adult patients and among children age 7-14 years, most of cases being diagnosed after 10 years. Class I and II mutation were present in 56.87% CF CFLD patients. Meconium ileus was a risk factor (OR=1.12) for developing CFLD, being present in 21% from CFLD patients. Pancreatic insufficiency was strongly associated with LD, certified to be risk factor (OR=1.25). Conclusion: The frequency of CF associated liver disease is rising. CF children older then 10 year, with severe mutation, history of meconium ileus, pancreatic insufficiency and are more likely predisposed to develop liver disease. In witch way the disease evolution or the risk factors control can be influence remain to be determine. P29. OPTIMISATION OF THE NEUROFIBROMATOSIS TYPE 1 MANAGEMENT R.Cojocariu, M.Macovei, E.Braha, M.Panzaru, L.Butnariu, R.Popescu, L.Caba, M.Volosciuc Neurofibromatosis type 1 (NF) is a monogenic, multisystemic disease, with an autosomal dominant inheritance, caused by mutations in the NF1 gene, that encodes a protein with tumor suppression activity called neurofibromin. Clinically, the disease is manifested by café au lait spots and an increased predisposition to develop multiple benign or malignant tumors originating in cells of the nervous system. NF is characterized by a marked variable expression noticed even in the patients with the same mutation, in the same family. Therefore, it is difficult to anticipate the complications and prognosis and, therefore, the monitoring of the evolution of the disease, depending on patient age and the severity of the clinical manifestations, is required. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 63 NF patient management requires: • Monitoring of tumor progression and changes in neurocutaneous lesions, considering the increased risk of malignancy; • Assessment of the neuropsychological development in the context of frequent association with a moderate mental retardation, learning and speech difficulties; • Neurological assessment: subtle abnormalities of coordination, gait and balance, cortex hiperexcitability and seizures; • Regular checking of blood pressure considering the risk for renal artery or aortic stenosis and pheochromocytoma; • Assessing the existence of bone abnormalities such as scoliosis and pseudoarthrosis; • Periodic ophthalmologic evaluation because of the possibility of an optical glioma and the risk of blindness. We will present the case of the patient SSA (13 years), with short stature and low weight, café au lait spots, axillary freckles, psychomotor retardation. The family history is positive: the mother, SE (49 years), diagnosed with the same occasion, shows multiple cafe au lait spots, neurofibromas and vision abnormalities; a brother (18 years) diagnosed with brachial plexus palsy, another brother who died at 20 years of brain tumor (cases with possible diagnosis of NF-1). The clinical manifestations are very different for the members of the same family and justify the need for an individualized plan of management / hospitalization, according to the age and type of abnormalities presented, and a careful clinical and biological evaluation of all family cases. P30. MTHFR ENZYME POLYMORPHISMS (C677T AND A1298C) AND THE RISK FOR DOWN SYNDROM Ruxandra Cretu, Daniela Neagos, L.C. Bohiltea Genetic Department, UMF Carol Davila, Bucharest Introduction: Folic acid belongs to B vitamins group and is used for prevention of severe congenital malformations and Down Syndrome (DS). Numerous studies suggested that folate metabolism alteration is the consequence of specific polymorphisms that occur in the genes involved in the regulation of metabolism. These alterations can increase the risk of women having babies with DS. Methylenetetrahydrofolate Reductase (MTHFR) is involved in folate metabolism and catalyzes the conversion of 5,10-methylentetrahydrofolate to 5-methylTHF, the methyl donor for the remethylation of homocysteine to methyonine. MTHFR gene was mapped on humane chromosome 1p36.3 and it contains 11 exons. Multiple polymorphisms have been reported in the general population, some of them with altered function in the homozygous individuals. Two polymorfisms with an increased frequency in the population were described: C677T and A1298C; they are thermolabile variants and determine the accumulation of homocysteine in circulation and the decrease of folic acid concentration. Materials and methods: The study was performed on a total number of 72 women split in two groups: one group of 26 women (ages 22-40) who gave birth to at least 1 DS baby and the other group comprises 46 women (ages 30-50) who gave birth only to healthy children (the control group). The whole peripheral blood samples were collected and the DNA was extracted from the blood leucocytes using perGold blood DNA mini kit. The polymorphysm of the MTHFR gene was studied with the PCR-RFLP technique. Results: The allele frequencies of MTHFR 677C>T and 1298A>C in DS mothers (case) and control mothers were analyzed. Thus, in case mothers, the frequency of 677T allele was 26.9% (14/52 allele) (λ2: 1.83), while the frequency of 1298C allele was 44.2% (23/52 allele). The variant allele frequencies among control mothers were 48,04 % (35/92 allele) for the MTHFR 677T allele and 31,52% (29/92 allele) for the MTHFR 1298C allele (λ2: 2,32, ), respectively. Conclusions: The frequency of 1298C allele and the CC homozygous genotype was increased for the group of the women that gave birth to DS babies comparative with the control women with 64 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 normal children, suggesting that this MTHPR polymorphysm plays a role in the SD pathogenesis. Preconceptional screening can identify the women with increased risk of having SD babies. Women of childbearing age under 30 can reduce the incidence of DS in their babies if they intake folate before pregnancy. P31. EPIGENETIC MARKERS IN DISEASE: DIAGNOSIS, PREVENTION AND NEW THERAPIES Natalia Cucu, Anton Gabriela, Arsene Cosmin, Daniela Nedelcu, Maria Puiu, Pavel Chirila, Narcis Dobre, Radu Stefanescu, University Bucharest National Institute of Virusology, Bucharest Naturalia, Bucharest Genexplore, Bucharest VitroBioChem There is a growing awareness in the medical field that having the correct epigenotype is essential for health cells and organs. If epigenetic signature or patterns are not properly established and maintained, disorders as diverse as mental retardation, immune deficiencies, metabolic dysfunctions and sporadic or inherited cancer may follow through aberrant or inappropriate silencing of growth regulating genes and simultaneous abnormal oncogenes activation. DNA methylation and histone modifications are crucial chromatin remodelling processes that together with associated proteinprotein and protein-DNA interactions and the so called “ interference ARN” molecules represents actually the epigenetic domain targets for the establishment of new biomarkers in diagnosis, prevention and new therapies. As fundamental principle, destabilizing chromosomal structures methylation defects help create an instable genomic state from which cancer cells evolve. Such methylation defects are present in cells before the onset of obvious malignancy and therefore cannot be explained simply as a consequence of a deregulated cancer cell. It is a tremendous need for sensible detection of methylation phenotypes in cells during months or years before the time when cancer may be clinically detectable by specific, mutation based screening approaches and by using modern sensible, even highthroughput techniques. Suchaberrant methylation of certain genes has been linked with the tumour response to chemotherapies and patient survival and presently constitutes a cancer progression or remission indicator. As for the plasticity of the epigenome, it has been well documented and proved that the chromatin state defining a specific transcriptional activity may be modified by environment and moreover is heritable. Also, a genetic approach has to be implied in the entire epigenetic research picture, which includes genes controlling methyl donor metabolism, familial cancer risk genes, genes controlling different metabolic functions (trigligerides metabolism, hormone metabolism etc) and genes controlling nuclear receptor functions. The so called “methylome” combined with genotyping approach permits the integration of an ever growing repertoire of candidate genes and methylation defects with the genetic alterations (mutations) catalogued during the past two decades in medical molecular genetics. With the advent of modern equipment and recently established specialized facilities for highthroughput approaches, an “-omic” approach can be envisaged that would complete the entire picture of environmentgenome interaction for understanding the real, subtle harmful changes in our genome stability. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 65 P32. CHROMOSOME INSTABILITY AND PRIMARY IMMUNODEFICIENCY SYNDROME M Cucuruz, E. Boeriu, M. Puiu, I. Ursache IIIrd Clinic of Pediatrics, Emergency Children Hospital „Louis Turcanu” Timisoara Aim: The study is proposing as goal the cytogenetic and immunological evaluation of four children, diagnosed with Ataxia telangiectasia (AT) in our service. Material and method: The diagnosis of AT was achieved on the basis of clinical data: appearance of the ataxic syndrome, of the conjunctival telangiectasia and of the predominantly humoral immunodeficiency syndrome. Immunologic data specify severe hypogammaglobulinemia in three cases, significant decrease of the lymphocyte population, especially lymphocytes B-CD19+; NK-CD56+ and NK-CD16+ cells were highly increased. Results: Cytogenetic data performed in dynamic showed severe evolutive modifications: in the beginning there was hypoploidia in 5-8% of metaphases, and afterwards severe structural changes were present in 45% of metaphases examined. These alterations were pronounced by irradiation of cell cultures. Conclusions: The evolution of the cases was not favorable, death appearing before the age of 14. The cause of death was severe immunodeficiency in three cases and malignancy – malign nonhodgkin lymphoma – in one case, the basis of these modifications being the chromosome instability syndrome. P33. BTK – GENE IN AGAMMAGLOBULINEMIA M. Cucuruz1, E. Boeriu1, Z. Ellenes2, M. Bataneant1, E. Ursu1, G. Brad1 1 IIIrd Clinic of Pediatrics, Emergency Children Hospital „Louis Turcanu” Timisoara 2 Children Hospital Oradea Objective of the study consists in analyzing Btk gene using molecular methods in 3 patiens, clinically and imunologicaly diagnosed with X-linked agammaglobulinemia, also their mothers were evaluated. Method: Screening for mutation was performed using SSCP, after PCR amplification of all Btk exons. The exons showing altered electrophoretic pattern on SSCP were sequenced to determine the nature of the mutation. Results: Two patients were found to have chain termination mutations in the kinase domain: a 4 bp detection at positions 527-528 resulting in frame shift and a premature termination codon at position 528 and a non sense mutation at codon 520 at the second patient. The third patient has a missense mutation c 29T<A in exon 2. All patient’s mothers were proved to be heterozygous for the mutation found in their sons. Conclusions: The mutations leading to premature stop codons at close sites of the Btk gene resulted in different disease severity in the patients, demonstrating a phenotypic heterogeneity in the XLA caused by junctionally similar Btk mutations. Mutation detection in the Btk gene provides the opportunity of definitive diagnosis in X-linked agammaglobulinemia, indispensable for adequate genetic counseling and carrier detection. 66 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P34. VESICO-URETERAL REFLUX IN PLURIMALFORMATIVE SYNDROMES UNFAVORABLE PROGNOSTIC FACTORS Camelia Daescu1,2, Ioana Maris1,2, Ioan Sabau1, Ioan Simedrea1,2, Oana Belei1,2, Tamara Marcovici1,2, Adela Emandi-Chirita2, Maria Puiu1,2 1 University of Medicine and Pharmacy „Victor Babes” Timisoara 2 Clinical Emergency Hospital for Children „Louis Turcanu” Timisoara Introduction: Disruption of embryogenesis in plurimalformative syndromes may be accompanied by reno-urinary anomalies, including vesico-ureteral reflux. Material and methods: Presentation of three cases with plurimalformative syndromes that associate vesico-ureteral reflux: Townes-Brocks syndrome, situs inversus and Cornelia de Lange, whom were evaluated in the First Pediatric Clinic in Timisoara. Results: The renal involvement in Townes-Brocks syndrome patient resulted nephrectomy of ectopic kidney affected by vesico-ureteral reflux grade IV. The patient with situs inversus, which has vesico-ureteral reflux grade IV associated with other malformations, now has unilateral ureterostoma and chronic renal failure due to hypodisplasia in the other kidney. The patient with Cornelia de Lange syndrome has vesico-ureteral reflux with recurrent urinary tract infections. Conclusions: It becomes important to actively search for vesico-ureteral reflux in the context of plurimalformative syndromes in the perspective that the reflux nephropathy is an unfavorable prognostic factor that may favor the evolution of these patients to renal failure. Keywords: vesico-ureteral reflux, syndrome, prognosis P35. IS PECTUS EXCAVATUM A GENETIC DISEASE? DavidVL1, Popoiu MC2, Anca Popoiu2, Puiu M2, Boia ES2 1 Emergency Children’s Hospital „Louis Turcanu” Timisoara, Romania 2 University of Medicine and Pharmacy „Victor Babes” Timisoara, Romania Background: Pectus Excavatum (PE) is the most common malformation of the anterior chest wall. The incidence of the disease is 1/ 1000 – 1200 new born. The cause of the disease remains yet unknown but there is an increasing evidences that genetic factor play an important role in the etiopathogeny of the disease. Several reports indicated that in 45% of the cases the disease has a hereditary pattern. The disease has an increased incidence in several genetic syndromes that involves structural defects of the hyaline cartilage: Marfan, Ehlers Danlos, and Noonan. Material and Methods: We reviewed the medical records of all the cases of PE admitted in our clinic during an 11 years period in order to identify the genetic factors that might be involved in the etiopathogeny of the disease. Results: A total number of 79 children with PE were referred to the Pediatric Surgery Department from January 2000 to August 2010. In the analyzed group PE affected at least one other member of the family in 17 patients (22%). Marfan syndrome affected two patients, Turner syndrome one patient, Noonan syndrome one patient, Down syndrome one patient and Piere Robin syndrome one patient. Conclusions: Genetic factors play an important role in the etiopathogeny of PE. In our series, genetic factors were present in almost 30% of the patients. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 67 P36. GENETIC COUNSELING IN DUCHENNE MUSCULAR DYSTROPHY, A MULTILEVEL APPROACH Amelia Dobrescu1, Maria Puiu2, Lavinia Sîrbu3, Elena Buteica1, Daniela Tache4, Dorina Iovanescu5 1 UMF Craiova, Department of Medical Genetics 2 UMF Timisoara, Department of Medical Genetics 3 Poiana Mare Psychiatric Hospital 4 UMF Craiova, Department of Biochemistry 5 Emergency Hospital of Craiova, Department of Neurology Duchenne Muscular Dystrophy (DMD), the most common muscular dystrophy in children, is a devastating condition that continues to affect many boys and their families. Recent advances in symptomatic management are able to improve patient`s status, minimize the side effects and even slow down the evolution of the disease (have provided improvements in function, ambulation, quality of life and life expectancy), but while there is no treatment for this pathology, it is essential to establish an accurate diagnosis and for reliable genetic counselling and prenatal diagnosis. Our study, that was performed among 20 families affected by such a pathology, presumed that the genetic advice for Duchenne Muscular Dystrophy must be a multilevel medical act and its purpose was to identify the specific characteristics of each case and the lacunas conditioned by the medical act or external agents (socio-economical, ethnical, religious). The approach plan of the genetic advice was structured on two levels: a) The management of counselling the DMD patient; b) The management of approaching the pathology within the family in which there is or was DMD The bond between these two levels is the establishment of compulsory molecular diagnosis; identifying the mutations specific to each case is, at the moment, a compulsory support for: 1) establishing a prenatal diagnosis; 2) clinical studies access, studies that are in progress on an international scale for particular therapies according to the mutation type (PTC 124, exon-skipping) We have created a decisional tree for both aims that should contain three aspects: medical education related to disease, coordination and genetic tests demonstration, psychological counselling. Conclusion: genetic counselling can offer several benefits for the families living with these diseases, especially when genetic counsellors work with families and their healthcare providers to determine the best testing strategy, the best therapeutic approach, the management of preventing the disease within the family and also the establishment of the necessary coordinates in order to obtain a safe pregnancy. A multilevel approach of genetic counselling in Duchenne Muscular Dystrophy, the collaboration between the genetic counsellor, families and healthcare professionals offer a continuity of the management strategy which can cover all aspects of the DMD disease and which has maximum benefits for all family members; patients are encouraged to inform themselves and, thus, to make the right choices, while specialists can act accordingly to risk conditions of each case. 68 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P37. CONSIDERATIONS OF REPRODUCTIVE COUNSELLING IN A MOSAIC 45, X/46, X, I(XQ) TURNER SYNDROME Amelia Dobrescu1, Maria Puiu2, Popa Cristina2, D. Iliescu3 1 UMF Craiova, Department of Medical Genetics 2 UMF Timisoara, Department of Medical Genetics 3 UMF Craiova, Department of Obstetric and Gynecology Turner syndrome (TS) occurs in one out of every 2,500 to 3,000 live female births and it is determined by quantitative and/or structure failure of X chromosomes with frequent presence of mosaicism. On chromosomal analysis, the percentage occurrence of the various karyotypes observed in TS are: 45, X (50%), 45, X / 46, XX (20%), 46, X, i(Xq)(15%), 46, X,r(X) or 46,X, del(X)(10%) and others (5%). Fertility and sexual development are often major concerns for patients with Turner syndrome. Usually women with Turner syndrome (TS) have a risk of premature ovarian failure; so fertility preservation may not be feasible for most patients with TS, but fertility preservation may be offered to young females with mosaic TS. Case and discussion The purpose of our study is to establish TS that could be potential candidates for fertility preservation and to determine their present reproductive and fertility status. The case was that of a 26 year old women diagnosed with Turner syndrome at the age of 15, having the basic symptoms: growth retardation and primary amenorrhea; the case did not have broad chest, neck webbing, low posterior hairline, without the presence of renal or cardiovascular symptoms. The patient took the hormonal treatment, the consequence of which was the return of mense and its effects related to height and breast growth were moderate. The patient was re-evaluated recently in order to establish the possibility of obtaining a pregnancy. The multidisciplinary approach has highlighted: • a 45, X/46, X, i(Xq) cytogenetic formula • a small uterus with almost normal endometrius • two moderate fibrotic ovaries • normal cardiovascular function Conclusion: The clinical status of the patient lacking in severe manifestations, supported by the cytogenetic formula that is characteristic to a moderate phenotype, along with a favourable evolution and the ultrasound results of the genital and cardiovascular system as well that encourage the possibility of carrying a pregnancy, are all arguments for proposing the in vitro fertilization during counselling as well as planning a counselling strategy that should define the steps of a medical act, explaining the boundaries and risks of the procedure and the continuous attendance of the patient. P38. NON-COMPACTION LEFT VENTRICLE, DIAGNOSTIC FEATURE FOR 1P36 MONOSOMY SYNDROME G. Doros1, V. Stan1, A. Popoiu1, M. Gafencu1, J. Puiu1, G. Miclaus2, B. Zoica1 1 University of Medicine and Pharmacy ”Victor Babes” Timisoara, Romania 2 Neuromed Clinic, Timisoara, Romania Aim: To present a 10 yo. boy, with distinct facial features and intellectual disability, treated from birth for recurrent seizures; he associated right small cerebral cavernoma and arachnoid cyst. After many years of neurologic follow up, he was sent for a cardiological examination, where a noncompaction left ventricle was found. This diagnose sugest a 1p36 monosomy syndrome. Material and methods: This patient was sent for a cardiologic examination due to a muscular hypotrophy. We performed clinical examination, ECG, Echocardiography and angio MRI. A genetic examination was done. Investigations were completed with FISH and tissue Doppler. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 69 Results: No cardiac murmur and normal ECG was registered. Non-compaction of the left ventricle (NCLV), with deep trabeculations and deep inter-trabecular spaces were detected in apical, mid ventricular, inferior and lateral wall; normal ejection fraction was found. Angio MRI confirmed the diagnose. The laboratory tests were normal, so a muscular pathology was excluded. Genetic examination mentioned a normal karyotype. In the context of particular face, with deep-set eyes, straight eyebrows, midface hypoplasia, flat nose and associated cardiomyopathy, mental delay, seizures and cerebral tumors, an extended FISH test was performed. The suspicion of monosomy 1p36 syndrome was confirmed. Discusions: Muscular hypotrophy in a patient with particular face and complex neurologic modifications conduct to a cardiac exploration, because a muscular disease was suspected. Left ventricular non-compaction was detected. This is a rare cardiomyopathy, present in muscular diasease and in other genetic syndromes. In the clinical context, a monosomy 1p36 syndrome was suspected and confirmed by FISH. Conclusions: NCLV when present, the genetic involvement has to be searched. Associated with particular facial features and neurological pathology, NCLV can indicate 1p36 monosomy syndrome. Genetic examination and FISH has to be done to confirm the syndrome. Follow up for LVNC is mandatory. P39. EVALUATION OF TROMBOPHILIC FACTORS AND MTHFR GENE POLYMORPHIMS IN RECURRENT PREGNANCY LOSS Cristina Dragomir, Adriana Stan, DT Stefanescu, L Savu Genetic Lab, Bucharest, Romania The Factor V Leiden – G1691A and prothrombin - G20210A mutations are the most common autosomal dominant inherited variants associated with recurrent pregnancy loss. About 30% of women with history of pregnancy loss or stillbirth turn out to be thrombophilic. Beside these trombophilic factors, the C677T and A1298C mutations of the MTHFR gene ”methylentetrahydrofolat reductase”, a gene that is normally involved in homocysteine levels regulation, are considered another genetic cause that can lead to pregnancy complications. Elevated levels of homocysteine have been associated with placental disease, preeclampsia and pregnancy loss. 21% of women with high levels of homocysteine experience recurrent miscarriage. Referring all these aspects we evaluate the relation between these mutations and pregnancy loss. The genetic status of the factors II and V of coagulation were evaluated by Real Time PCR – FRET technology and Melting Curve Analysis. The two variant of the MTHFR gene, C677T and A1298C, were identified by PCR-RFLP using Hinf I and Mbo II restriction enzymes. Initially we tested the patients with unexplained pregnancies loss for Factor V and Factor II mutations. Of the 260 patients tested we found that 77 (29.6%) were heterozygous for Factor V Leiden allele, 4 (1.5%) were homozygous for the factor V Leiden and 10 (3.8%) were heterozygous for the factor II gene mutation. 172 patients (66.15%) were found negative for two mutations. Last year we introduced in our laboratory the detection of the C677T and A1298C polymorphisms for the MTFR gene. We tested for these 35 patients with recurrent pregnancy loss. The patients, previously investigated for factor V and factor II status had no mutation at the proaccelerin (factor V) and prothrombin (factor II) genes level. 21 of 35 patients were affected: 10 patients compounds heterozygous (677CT/1298ACgenotype), 6 patients homozygous for C677T polymorphism (677TT genotype) and 6 patients homozygous for A1298C polymorphism (1298CC genotype). 12 of 35 patients presented one of the two mutations in heterozygous form: 6 patients with 677CT genotype and 6 patients with 1298AC genotype. The heterozygous genotype is not involved in pregnancies loss. Only one patient was normally for these polymorphisms (677CC/1298AA genotype). We consider that pregnancy loss at 35% of 260 patients investigated for factor V and Factor II status may be caused by G1691A or G20210A mutations. Also, the mutations detected in either compound heterozygous or homozygous forms at the MTHFR gene level in 62,85% of the 35 tested patients, without factor V Leiden or prothrombin gene mutation, suggests that these polymorphisms could be the cause of the pregnancies losses. 70 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P40. ASPECTS OF MOLECULAR GENETICS IN PROSTATE CANCER Dumache Raluca1, Miclea Florin2, David Dana1, Kaycsa Adriana1, Negru Serban3, Ionescu Daniela4, Puiu Maria5 1 Department of Biochemistry, University of Medicine and Pharmacy ‘’Victor Babes’’, Timisoara, Romania 2 Department of Urology, University of Medicine and Pharmacy ‘’Victor Babes’’, Timisoara, Romania 3 Department of Medical Oncology, University of Medicine and Pharmacy ‘’Victor Babes’’, Timisoara, Romania 4 Department of Toxicology, University of Medicine and Pharmacy ‘’Victor Babes’’, Timisoara, Romania 5 Department of Medical Genetics, University of Medicine and Pharmacy ‘’Victor Babes’’, Timisoara, Romania After lung cancer, prostate adenocarcinoma represents the second leading cause of male cancer deaths in the United States and Europe. Many studies have been developed trying to understand the complex molecular mechanisms involved in oncogenesis and progression of prostate cancer. The molecular pathogenesis of prostate cancer involves many contributing factors, such as: alterations in signal transduction pathways, angiogenesis, adhesion molecules expression and cell cycle control. The discovery of new molecular biomarkers for prostate cancer represents another relevant advance resulting from molecular genetics studies of prostate tumors. Serum, urine and tissue biomarkers have been discovered, resulting in useful novelties to diagnostic and prognostic approaches to follow-up prostate cancer. Also, gene therapy comes as an important approach for therapeutic intervention in prostate cancer. Key words: prostate cancer (PCa), biomarkers, gene therapy P41. PROBLEMS IN ETIOLOGY DIAGNOSIS AND INTERVENTION IN COGNITIVE REGRESSION IN CHILDREN Dumitriu Simona, Medic Specialist Psihiatrie Pediactrica, Dr. Ageu Luminita, Medic Rezident Psihiatrie Pediatrica, Clinica de Psihiatrie si Neurologie pentru Copii si Adolescenti, Timisoara Objective: Our work aims at a review of clinical and laboratory data and management in adrenoleukodystrophy X linked, starting from a clinical case. Material and methods: It is presented a child of 9 years old, living in rural areas and he is in third class of urban, who came in our Clinic for evaluation in school difficulties: attention deficit, difficulties centering on the task, impulsivity, hyperactivity, dyspraxia, dysgraphia, relationship difficulties, episodes of anxiety, impulsivity, school refusal, conduct repeated stereotype. Mother and teacher describe the occurrence of behavioral changes approximately one month after the beginning of the school year. Evaluation of initial symptoms were suggestive of a school adjustment disorder. Laboratory investigations included EEG, brain MRI, evaluation of adrenal function and very long chain fatty acids of carbon. Results: Historical data obtained later clinical and laboratory test results and psychological diagnosis Adrenoleukodystrophy claimed as X-Linked. Cognitive disharmony with regression at performance QIP = 65WISC. Conclusions: Complex genetic consultation and evaluation on development stages of children from families where are neurodegenerative diseases may offer an early etiological treatment. Key words: school difficulties, adrenoleukodystrophy, regression Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 71 P42. A RARE CASE OF CONGENITAL ARTHROGRYPOSIS: THE PENA-SHOKEIR I PHENOTYPE Corina Duncescu1, Elena Pop1, Ramona Giurescu1,2, Adela Chirită1, Monica Mărăzan1,2, Ioana Micle1,2 1 st 1 Pediatric Clinic, „Louis Ţurcanu” Emergency Hospital for Children, Timişoara, Romania 2 „Victor Babeş” University of Medicine and Pharmacy, Timişoara, Romania Introduction: Arthrogryposis multiplex congenita (AMC) comprises a very heterogeneous group of disorders that present with multiple congenital joint contractures. AMC with polyhydramnios, short umbilical cord, pulmonary hypoplasia, camptodactyly, facial anomalies and death during the first months of life is called Pena-Shokeir I phenotype. Affecting an estimated 1 in 12,000 newborns, there are about 100 reported cases. Objective: We report a case with multiple malformations and the difficulties concerning the diagnosis. Material and method: The newly born B.V., with a characteristic phenotype, multiple congenital joint contractures, hypertonic syndrome and respiratory distress was transferred in our clinic for proper diagnosis. The approach was complex: we performed a complete history and clinical assessment, laboratory and imaging studies, various consults. Results: The radiographs and both pediatric orthopedic surgeon and neurologist confirmed the AMC diagnosis. Furthermore, the thoracic radiograph revealed pulmonary hypoplasia. Corroborating the several facial anomalies (hypertelorism, small, markedly recessed jaw, low-set ears), the history data and the international experience, we included our case in the Pena-Shokeir I phenotype. The patient had frequent episodes of apnea and cyanosis, which finally resulted in his death. Conclusions: To pinpoint a particular syndrome in a case with AMC is a challenge, which requires excluding the more frequent causes. Each newborn with multiple malformations must benefit from the joined experience of a team consisting of: an obstetrician, a neonatologist, a radiologist, a neurologist, a pediatric orthopedic surgeon and a clinical geneticist. Establishing the correct diagnosis leads to proper genetic counseling, which, for the parents, could be the only comfort. Keywords: arthrogryposis multiplex congenita, Pena-Shokeir I phenotype P43. PERSISTENT SEVERE HYPOGLYCEMIA SINCE INFANCY: CASE REPORT Corina Duncescu1, Monica Mărăzan1,2, Adela Chirită1, Elena Pop1, Ramona Giurescu1,2, Ioana Micle1,2 1 st 1 Pediatric Clinic, „Louis Ţurcanu” Emergency Hospital for Children, Timişoara, Romania 2 „Victor Babeş” University of Medicine and Pharmacy, Timişoara, Romania Introduction: In children, persistent hypoglycemia has a major negative impact on structural and functional brain development. Aim: We present the difficulties in establishing the diagnosis and treatment of a patient with persistent severe idiopathic hypoglycemia and secondary seizures since infancy. Material and methods: The child was admitted in our clinic at age three in order to establish the etiology and proper treatment of the hypoglycemia. The approach was extensive. We assessed growth, as well as the biological and hormonal profile, until the presentation in our clinic. In addition to anthropometric measurements, we obtained repeated metabolic and hormonal serum tests, a growth hormone (GH) stimulation test with arginine, bone age X-ray. Interpreting the results was difficult considering the long time treatment with prednisone (2 years and 6 months), started at the onset of the disease. Results: We have limited the differential diagnosis to two pathological entities: persistent idiopathic hyperinsulinemia and GH deficiency. Finally we stopped prednisone therapy and started GH substitution. The outcome was favorable in terms of symptoms, anthropometric, biological and hormonal profile Conclusions: The management of a child with persistent severe hypoglycemia is difficult and requires an extensive approach. Keywords: persistent hypoglycemia, GH deficiency 72 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P44. GENETIC EVALUATION OF SEX DEVELOPMENT DISORDERS – CASES REPORT Simona Farcas¹, Valerica Belengeanu¹, Nicoleta Andreescu¹, Monica Stoian¹, Dorina Stoicanescu¹, Anca Muresan², Dana Amzar3, Marius Craina4 ¹Department of Medical Genetics, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania 2 Department of Morphopathology, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania 3 Department of Endocrinology, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania 4 Department of Obstetrics and Gynecology, „Bega” Hospital, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania We set as objective to study the spectrum of chromosomal anomalies in three cases as related to the phenotypic variability of patients with anomalies of disorder of sexual development. First case is a 20 years old female with primary amenorrhea, who had infantile but otherwise normal external genitalia. High levels of serum FSH, LH and a very low plasma estradiol level indicated gonadal failure. A small tumor on right gonad site was found using laparoscopy and histological analysis from gonadal biopsy revealed characteristic aspect of gonadoblastoma. A 46,XY karyotype was established analysing 30 metaphases by conventional cytogenetics. The patient was diagnosed with Swyer syndrome and gonadectomy was performed. Second case. Female 18 years old with secondary amenorrhea, stigmata of Turner’s syndrome, Tanner stage II. Ultrasound showed streak gonads. The patient presented elevated levels of FSH, LH and a low plasma estradiol level. The chromosomal investigation showed gonosomal mosaicism 45,X(40%)/46,XY(60%). The XY clone was found in 65% of the cells using FISH analysis. Molecular analysis for 20 polymorphic markers and SRY region of Y chromosome revealed normal pattern. The patient was reffered for gonadectomy. Third case. Newborn evaluated for ambiguity of the phenotypic sex and for establishing the gender. The patient presented labioscrotum, micropenis and hypospadias. The cytogenetic analysis showed the presence of chromosome Y in all evaluated cells. The SRY gene was found on the Y chromosome using metaphase FISH analysis. The gonads were discovered by laparascopy, as ultrasonography failed to reveal their presence neither in the scrotum nor along the inguinal canals and histopathological investigation revealed immature testes. Normal values of testosterone and FSH for male gender were found. The surgical intervention and testosterone therapy were recommended. Detailed molecular cytogenetic characterization and the clinical follow-up of the patients are very useful in defining the phenotypic range of these chromosomal patterns. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 73 P45. A CASE OF SEPTO-OPTIC DYSPLASIA Simona Farcas¹, Cristina Popa¹, Nicoleta Andreescu¹, Monica Stoian¹, Alina Belengeanu², Marioara Boia³, Elena Bernad4 ¹Department of Medical Genetics, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania ²Department of Cellular and Molecular Biology, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania ³Department of Neonatology, „Louis Turcanu” Children Hospital, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania 4 Department of Obstetrics and Gynecology, „Bega” Hospital, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania Septo-optic dysplasia (SOD) was first recognized by De Morsier, in 1956, as a new syndrome that included optic nerve hypoplasia to septum pellucidum agenesis. De Morsier syndrome, as it was later named, is a rare developmental disorder that determines impairment in infancy manifested by optic nerve hypoplasia, midline central nervous system malformations and pituitary dysfunction. We present a one-year-old girl evaluated for acquisition of motor and language developmental milestones delay. Upon clinical examination we noticed: dystonia, telecantus, epicantus, broad nasal bridge, high arched palate, large and posterior rotated ears, short neck, pubic pilosity, breast hypertrophy and labioscrotal folds. The infant manifested seizures, renal failure and systolic bruit was present. The ophthalmologic evaluation showed bilateral partial atrophy of the optic nerve, hypermetropia and astigmatism. IMR revealed agenesis of corpus callosum and ventricular asymmetry with moderate enlargement of lateral ventricles and mega-cistern magna. EEG showed hypsarrhythmia. Endocrine dysfunction was revealed by laboratory tests. Adrenal congenital hyperplasia was taken into consideration as differential diagnosis because of the genital anomalies, but was excluded after hormone testing. All the clinical data and paraclinical investigations, sustain as possible diagnosis De Morsier syndrome. P46. TREACHER COLLINS SYNDROME- CASE REPORT Valeria Filip1, Cristina Skrypnyk2, Elena Popescu3, Radu Galis1. 1 Obstetrics and Gynecology Clinical Hospital, Neonatology Department, Faculty of Medicine and Pharmacy, University of Oradea. 2 Clinical Municipal Hospital, Genetics Department, Faculty of Medicine and Pharmacy, University of Oradea, 3 Obstetrics and Gynecology Clinical Hospital, Anatomopathology Department Introduction: Treacher Collins syndrome, also referred to as mandibulo–facial dysostosis, a rare genetic disorder found in 1 in 10,000 births. The typical physical features include micrognathia, underdeveloped zygoma, downward slanting eyes, malformed or absent ears, conductive hearing loss. The disorder is inherited in an autosomal-dominant pattern and caused by TCOF1 gene mutation at the locus 5q32q33.1. Materials and methods: The reported case is a female newborn with birth weight 2100 gr and Apgar score 3/4 from a GI, PI pregnancy of a young, healthy and non-consanguineous couple with negative family history of congenital anomalies. The clinical evaluation revealed the craniofacial dysmorphysm: microcephay, hypertelorism, midface hypoplasia, micrognathia, left ear microotia with absence of the upper portion of the helices, atresia of the external meati and absent right ear. The Xray examination revealed hypoplasia of the zygomatic bones and significant airway distortion due to choanal atresia and abnormal thachea. Results: The tracheal intubation was a difficult task, the disease outcome unfavourable and the child died in the second day of life. The anatomopathological examination shown the right choanal atresia, signifiant narrow of the trachea diameter and lung hypoplasia. 74 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Conclusion: The association between tracheal hypoplasia, choanal atresia and malformed spectrum of Treacher Collins syndrome is a rare one and sustain the case severity. The authors sintetised the literature on this syndrome and pointed the management for the cases with neonatal severe forms. P47. OCULO-AURICULO-VERTEBRAL SPECTRUM – UN ENTITY EASY TO DIAGNOSE Cerasela Munteanu1, Marius Galiţă1, Laura Ionescu1, Mihail Voloşciuc2 Oculo-auriculo-vertebral spectrum (OAV) is a first and second branchial arch embryopathy , wide and extremely complex, usually unilateral. Clinical diagnosis is considered for patients with auricular, mandibular, ocular and vertebral anomalies; other anomalies may also occur. For clinical practice we divide the spectrum: the lower part of OAV (first and second branchial arch anomalies), Goldenhar syndrome (classic clinical expression), the upper part of OAV (defects of embryo median field, caudal regression or mesodermal defects). We presented the case of BN patient, a 7 years 2 months old boy. Family history was negative. Delivery and pregnancy have a normal evolution. At the clinical examination we can observe craniofacial dysmorphia, right complete cleft lip and palate, macrostomia (lateral facial cleft), hypoplasia of ascending mandibular ramus, right pre-auricular pit, bilateral epibulbar dermoids, undescended testes, mild mental retardation. The clinical features are extremely characteristic and lead us to the diagnosis of oculo-auriculo-vertebral spectrum (Goldenhar syndrome). In conclusion cleft lip and/or cleft palate and lateral facial clefts point out from new-born period. The diagnosis (of which further depends the correct medical management of patient) is possible only if the physicians recognize the clinical features. P48. CLEIDO – CRANIAL DYSPLASIA – CLINICAL STUDY Madalina Grigoras 1 , Alexandru Vlad2, Cristina Rusu 3, 4 1 „Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania – student 2 Radiology Unit, „Sf. Maria” Children’s Hospital, Iasi, Romania 3 Medical Genetics Center, „Sf. Maria” Children’s Hospital, Iasi, Romania 4 Medical Genetics Department, „Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania Cleidocranial dysplasia is a skeletal dysplasia characterized by delayed closure of the cranial sutures, hypoplastic or aplastic clavicles, and multiple dental abnormalities. Manifestations may vary among individuals in the same family. Diagnosis is based on typical clinical and radiologic features. The disorder has autosomal dominant inheritance, the gene involved being RUNX2. However, many of the cases are sporadic, due to new mutations. Patients are monitored by a multidisciplinary team (geneticist, dentist, orthopedics, ENT). Differential diagnosis is done with Crane-Heise syndrome, Mandibulo- acral dysplasia, Picnodisostosis, Yunis- Varon syndrome, CDAGS syndrome, Hypophosphatasia, Foramina parietalis with cleido- cranial dysplasia and congenital hypothyroidism. We present a typical case of cleido- cranial dysplasia to illustrate this rare genetic disorder, to present a comprehensive literature review and to discuss the complex management of the disorder. Patient L.A.M., female, 11 years 8 months old, has been evaluated in Iasi Medical Genetics Center due to increased joint mobility. Family history is negative. The patient is the only child of a young couple, apparently healthy, not related. Pregnancy has been uneventful. The child was born naturally, at term, cranial presentation, Wt G 3,200 g. Staturo- ponderal and intellectual development have been normal. However, motor development has been delayed. She has been diagnosed with left hip displasia at 1 year of age and followed orthopedic treatment. Later on, multiple dental abnormalities have been detected. Anterior fontanel closed at 8 years of age. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 75 Physical examination done at 11 years 8 months revealed: stature- ponderal development in the lower normal range, relative macrocephaly, typical dysmorphic face, dental anomalies, narrow shoulders with excessive mobility, winged scapulae, normal intellectual development. Radiologic investigations confirmed hypoplastic clavicles and dental anomalies. Based on clinical and radiologic data, we have established the diagnosis of cleido- cranial dysplasia. Management is multidisciplinary. In conclusion, we present a typical case of cleido- cranial dysplasia to illustrate this rare genetic disorder, to present a comprehensive literature review and to discuss the complex management of the disorder. P49. PRENATAL DIAGNOSIS AFTER ESTABLISHED CARRIER STATUS OF BALANCED STRUCTURAL CHROMOSOME ABNORMALITY Cristina Gug1,2, T. Cioată1, D. Grigoraş1, D. Chiriac1, G. Budău1, N. Hrubaru1, C. Mureşan1, A. Creţu3, V. Karadja4, V. Gorduza5, G. Furău6, 1 University of Medicine and Pharmacy „Victor Babes”, Timişoara, România, 2 Genetics Medical Center „Dr. Cristina Gug”, Timişoara, România, 3 Obstetrics-Gynecology „Dumitru Popescu” Hospital, Timişoara, România, 4 Obstetrics-Gynecology Private Practice, Timişoara, Romania, 5 University of Medicine and Pharmacy „Gr.T. Popa” Iaşi, România, 6 West University „Vasile Goldis”, Arad, România. About 5% of couples investigated in our laboratory for recurrent miscarriage are carriers of a structural chromosome abnormality. We offered all of them prenatal diagnosis for future pregnancies. We collected data from 53 couples with chromosomal rearrangement investigated for karyotyping after two or more miscarriages. We found 24 couples with a balanced translocations. Seven of these couples performed prenatal diagnosis. Parental karyotype showed the following translocations: 46,XX,t(7;10)(p22;p12.1), 46,XY,t(1;5)(q23;p12), 46,XX,t(17;20)(q12;q11), 46,XX,t(3;15)(q12;q13), 46,XX,t(4;10)(q22.2;q22.2-ter), 46,XY t(15p;19p), 46,XX,t(7;9)(p14q24). Fetal karyotype showed 3 normal fetuses, 4 carriers of a balanced translocation and one carrier of non-balanced translocation 46,XY,dup(10q22.2-ter), consequently this last pregnancy was terminated. One couple performed twice amniocentesis and fetal karyotype showed in both cases carriers of the balanced translocation t(7;10)(p22;p12.1), as was their mother. We encountered 6 couples with Robertsonian translocations: 3 cases with trob(13;14) and 3 cases with trob(13;22). At one case of each category, prenatal diagnosis was performed and pregnancy evolved normaly. One of these fetuses was normal and one was a carrier with balanced translocation trob(13;22), just like the mother. Twentythree couples were identified with chromosomal inversions and 10 of them had prenatal diagnosis performed, as following: 1 case, a female, with inv(7), 1 case, a female, with inv(10) and 8 couples with inv(9), 6 males and 2 females. In all cases amniocentesis showed fetuses carriers of invesions and pregnancies continued. Detection of a structural chromosome abnormality in couples with recurrent miscarriage has a striking influence on their decision to undergo prenatal diagnosis. Genetic counseling helps these couples to understand the recurrence risk and consequently to evaluate their options. Keyword: carrier of balanced chromosome abnormality, prenatal diagnosis; karyotype 76 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P50. PRENATAL DIAGNOSIS OF IPEX SYNDROME AND IDENTIFICATION OF 2 NEW FOXP3 MUTATIONS Radu Harbuz1, James Lespinasse2, Stéphanie Boulet 3, Christine Francanet 4, Isabelle Crevaux 5, Pierre-Simon Jouk1, Joël Lunardi1, Marius Bembea6 , Pierre F Ray 1 1 UF de Biochimie et Génétique Moléculaire, CHU de Grenoble, France 2 Génétique Chromosomique, CH de Chambéry, Chambéry, France 3 Gynécologie Obstétrique, CH de Chambéry, Chambéry, France 4 Génétique Médicale, CHU de Clermont-Ferrand, Clermont-Ferrand, France 5 UF de Biologie Moléculaire, CHU de Clermont-Ferrand, Clermont-Ferrand, France 6 Clinical Municipal Hospital «Dr.Gavril Curteanu» Oradea, Romania The syndrome of immunodysregulation, polyendocrinopathy, enteropathy, X linked (IPEX) is a rare disorder usually leading to the death of affected boys in early infancy. Mutations in the FOXP3 gene are found in approximately 60% of the patients while the remaining cases remain unexplained. We present here the genetic investigation and prenatal care of two women suspected to be IPEX carriers. A single nucleotide variant was identified in each patient after sequencing FOXP3 coding sequences and exon boundaries. Both variants: c.816+7G>C and c.816+4A>G, were localized near intron 7 donor site and were suspected to yield exon 7 skipping. This abnormal splicing could be verified by reverse transcription polymerase chain reaction on RNA extracted from both of our patient’s leukocytes. Prenatal diagnosis was carried out for one patient who bore a male fetus. The analysis demonstrated that it did not carry the identified mutation. The baby was delivered at term and is in good health. The other female pregnancy was terminated following the detection of an unrelated chromosomal abnormality. IPEX cardinal signs are quite evocative and include severe diarrhea, eczema and diabetes. However, since the disease is rare, clinical diagnosis is often considered with delay. Both patients reported here were already pregnant at the beginning of the genetic investigation and one of them had previously interrupted a male pregnancy for lack of diagnosis. It is thus important for all clinicians to bear in mind the possibility of IPEX and to rapidly propose a molecular diagnosis to all patients at risk. Key words: IPEX syndrome, FOXP3 gene, prenatal diagnosis P51. PSYCHOLOGICAL TREATMENT IN GENETIC DISEASES Hogea Lavinia University of Medicine and Pharmacy „Victor Babes” Timisoara, Department of Psychology This paper aims to highlight the role and importance of psychological treatment in different genetic disorders. It is importance to consider the impact of mental, emotional and behavioral genetic diseases that can have on patients and their families. Counseling can help patients better manage the difficulties of disease and contributes to improvement of the quality of life. Due to the diversity genetic diseases and their complexity psychotherapeutic methods should be selected specifically for them to prove their effectiveness. The study valorises the data collected over time, through careful research, by health psychology, clinical psychology, psychiatry, psychopathology and psychotherapy. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 77 P52. PARACETAMOL USAGE DURING EARLY PREGNANCY CAN CAUSE CONGENITAL ABNORMALITIES? Daniela Iacob1, RE Iacob2, C Ilie1, Marioara Boia1, Aniko Manea1, Maria Julieta Puiu3 1 University of Medicine and Pharmacy „Victor Babes” Timisoara – Deparment of Neonatology, Romania 2 County Emergencies Clinical Hospital of Arad – Department of Pediatric Surgery, Romania 3 University of Medicine and Pharmacy „Victor Babes” Timisoara – Deparment of Genetics, Romania Introduction: Congenital abnormalities and malformations, birth infirmity are terms used to describe structural, functional and metabolic disorders at the birth time. Many studies show the negative effects of some drugs over the embryo, there are insufficient data that will present a correlation between occasionally and short usage of this drugs and the risk of abnormalities. The aim of this paper was to identify the role of paracetamol in malformations appearance. Material and methods: For this study have been used 20 pregnant females of white mouse. The mice were divided into two groups; first group received paracetamol, the second group was the control group. First group of females has received for three days successively the paracetamol, corresponding to the days 5-7 of pregnancy. At the birth the kidneys and the liver from the cubs were microscopically analyzed. Results and discussions: The disorders in the liver and kidneys have a reversible character and are in different level of severity. The necrosis and tissular fibrosis were not presented. Conclusions: The administration of paracetamol during gestation had no influence over the pregnancy. The cubs from the group with paracetamol have similar development with the cubs from the control group. Microscopically, the changes in the aspect of liver and kidneys were erratic, in remission. P53. PREMATURE NEW BORN WITH HEART CONGENITAL MALFORMATIONS Daniela Iacob1, RE Iacob2, C Ilie1, Marioara Boia1, Aniko Manea1, Mirabela Dima1 1 University of Medicine and Pharmacy „Victor Babes” Timisoara – Deparment of Neonatology, Romania 2 County Emergencies Clinical Hospital of Arad – Department of Pediatric Surgery, Romania Introduction: Heart congenital malformations represent an important problem in pediatry because of the growth incidence and of the medical and social implications. Purpose: This study wants to establish the incidency of heart congenital malformations when compared to the other congenital malformations and their frequency according to some factors: risk, social background, sex and prematurity. Method: The study is based on clinical, paraclinical examinations and imagistic explorations which were performed on premature new born hospitalized in the Neonatology and Health Care Clinic Timisoara between 2007 and 2009. Results: Of the 72 studied with congenital malformations, 33 (45,8%) had heart congenital malformations. Of these 3 (9%) were cyanotic lesions (transpositions of the great arteries) and 30 (91%) – acyanotic lesions. Regarding the social background 18 (54,5%) were from urban areas and 15 (45,5%) from the rural areas. The repartition of the cases according to sex showed that 16 (48,4%) were female and 17 (51,6%) were male. According to the prematurity: 12 (36,3%) were 1st grade prematures, 12 (42,4%) – second grade prematures, 3 (9%) – 3rd grade prematures and 4 (12%) – 4th grade prematures. Conclusions: 1. the incidency of heart congenital malformations is high, representing 45,8% of the total congenital malformations. 2. the frequency of the acyanotic lesions is superior to the cyanotic lesions. 3. there is a slight predominance of the cases which come from the urban areas comparatively to those from the rural areas, possibly because of higher pollution. 4. distribution according to sex is approximately equal. 78 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P54. CHROMOSOMAL ABERRATIONS DETECTED BY PRENATAL CYTOGENETIC ANALYSIS OF 3000 CASES Cristina Ionescu, Luminita Roibu, Lorand Savu Genetic Lab, Bucuresti, Romania More than 50% of the spontaneous abortions are caused by chromosomal aberrations, and up to 96% of these are numerical abnormalities. Cytogenetic studies performed on live born showed that the frequency of chromosomal aberrations is between 1:150 and 1:200. Also, about 20 – 30% of the cases of infantile deaths are caused by genetic anomalies. Some anomalies appear de novo or they are the result of a balanced choromosomal rearangement present in one of the parents. The aim of this study was to investigate the different types of chromosomal aberrations and their relativ frequency in our laboratory in order to highlight the importance of prenatal cytogenetic test. This study included 3000 prenatal karyotypes carried out in our laboratory in over 10 years. The age of the pregnant women was between 21 and 45 years, 98% of them being caucasian. The study included karyotypes performed by cultivation of amniocytes, trophoblastic cells and lymphocytes obtained by cordocentesis. Standard GTG banding was used. At least 15 – 20 cells were analysed, and at least 40 cells were counted when mosaicism was suspected. 93,46% from a total of 3000 fetal karyotypes were normal, and 4,78% presented a chromosomal abnormality. Sex chromosome aberrations were present in 9,17% of the abnormal karyotypes. Autosomal aneuploidies represented the highest percent of the chromosomal aberrations, respectively, 51,67. Translocations were the second most frequent, with a total of 15%. Lower rates were found in inversions (3,34%), duplications (5,83%), deletions (4,17%) and marker chromosomes (3,33%). The frequency of the chromosomal aberrations increased with the mother’s age, when a trisomy was involved, although this anomaly was noted in women aged between 25 and 30. Poliploidies, sex chromosome aberrations and single cell aberrations were, almost exclusively, found in women under 30 years old. P55. A PRELIMINARY EVALUATION OF MANGANESE SUPEROXIDE DISMUTASE (MNSOD) GENETIC POLYMORPHISMS, DIETARY ANTIOXIDANTS, AND RISK OF BREAST CANCER Daniela Ionescu 1*, Raluca Dumache 2 , Maria Puiu 3, Cristina Dehelean 1, 1 Toxicology Department, University of Medicine and Pharmacy Timisoara 2 Biochemistry Department, University of Medicine and Pharmacy Timisoara 3 Department of Genetics, University of Medicine and Pharmacy Timisoara Oxidative stress, resulting from the imbalance between prooxidant and antioxidant states, damages DNA, proteins, cell membranes, and mitochondria and seems to play a role in human breast carcinogenesis. Dietary sources of antioxidants (chemical) and endogenous antioxidants (enzymatic), including the polymorphic manganese superoxide dismutase (Mn-SOD), can act to reduce the load of oxidative stress. We hypothesized that the valine-to-alanine substitution that seems to alter transport of the enzyme into the mitochondrion, changing its efficacy in fighting oxidative stress, was associated with breast cancer risk and that a diet rich in sources of antioxidants could ameliorate the effects on risk. Data were collected in a study of diet and breast cancer in Municipal Hospital between 2005 to 2008. Caucasian women with incident, primary, histologically confirmed breast cancer were frequency-matched on age and county of residence to community controls. Blood specimens were collected and processed from a subset of participants in the study (114 cases and 109 controls). Using a RFLP (Restriction fragment length polymorphism) that distinguishes a valine (V) to alanine (A) change in the 29 position in the signal sequence of the protein for MnSOD, we characterized MnSOD genotypes in relation to breast cancer risk. We also evaluated the effect of the polymorphism on risk among low and high consumers of fruits and vegetables. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 79 Premenopausal women who were homozygous for the A allele had a 4-fold increase in breast cancer risk in comparison to those with 1 or 2 V alleles (odds ratio, 4.3; 95% confidence interval, 1.7–10.8). Risk was most pronounced among women below the median consumption of fruits and vegetables and of dietary ascorbic acid and a-tocopherol, with little increased risk for those with diets rich in these foods. Relationships were weaker among postmenopausal women, although the MnSOD AA genotype was associated with an almost 2-fold increase in risk (odds ratio, 1.8; confidence interval, 0.9– 3.6). No appreciable modification of risk by diet was detected for these older women. These data support the hypothesis that MnSOD and oxidative stress play a significant role in breast cancer risk, particularly in premenopausal women. The finding that risk was greatest among women who consumed lower amounts of dietary antioxidants and was minimal among high consumers, indicates that a diet rich in sources of antioxidants may minimize the deleterious effects of the MnSOD polymorphism, thereby supporting public health recommendations for the consumption of diets rich in fruits and vegetables as a preventive measure against cancer. P56. IMPACT OF ACQUIRED POSTPARTUM HEMOPHILIA ON THE NEWBORNS A. Isac, L.Pop, M.Puiu, H. Ionita, D. Savescu, A. Balan University of Medicine and Pharmacy Timisoara Introduction: Acquired hemophilia A is an uncommon but potentially life-threatening hemorrhagic disorder caused by the development of autoantibodies directed against coagulation factor VIII (F VIII). The clinical picture is dominated by severe hemorrhage in the majority of patients with an inhibitor - related mortality rate up to 22 %. Acquired hemophilia may develop in association with various diseases and also in women following childbirth. Usually, factor VIII inhibitors appear after the first pregnancy, in rare cases, the autoantibodies are formed during pregnancy through transplacental passage of fetal blood they bind with factor VIII, therefore, the fetus or newborn, may develop severe bleeding syndrome, which can result in death. Patients and method:bIn the last five years, the Children’s Emergency Hospital „Louis Turcanu Timisoara, we recorded two cases of postpartum acquired hemophilia: a case where factor VIII inhibitors appeared after the second pregnancy and another case after the first pregnancy. We studied the dynamics of anticoagulant status, including determination of serum titer of factor VIII and factor VIII inhibitors, both in newborns and their mothers. Results and discussions: In the first case, the high titer of autoantibodies was confirmed a couple months after the second pregnancy. Factor VIII inhibitors were also present in children without significant clinical events, the autoantibodies probably occurred during the third trimester of pregnancy and passed through placenta. In the second case, inhibitors were present at 6 months after first pregnancy. Infant screening revealed normal serum levels of factor VIII and the absence of autoantibodies. Conclusion: In most cases, antibodies disappear spontaneously within a few months, but there are situations in which deaths were recorded in newborns due to factor VIII inhibitors, which emphasizes that prompt recognition and early therapy of this coagulopathy is crucial and can save patients’ lives. P57. CONGENITAL MANDIBULAR ANOMALIES – CLINICAL CONSIDERATIONS Oana Iuhas, Claudia Jurca, Radu Harbuz, Kinga Kozma, Marius Bembea Spitalul Clinic Municipal „Dr. G. Curteanu” – Oradea Background. Mandibular anomalies are frequently encountered among craniofacial dysmorphisms. They can be classified into congenital and acquired. Congenital mandibular anomalies are, usually, associated in plurimalformative syndromes. Objectives and methods. This study presents a retrospective analysis (1983-2010) of all the children with syndromic and nonsyndromic congenital mandibular anomalies investigated at the Clinic Municipal Hospital „Dr. G. Curteanu” – Oradea. 80 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 The aim of study is to determine the incidence and to identify the main clinical and therapeutical aspects. Patients and results: We evaluated 3660 children with congenital anomalies during the last 26 years; of these, 76 (2%) presented mandibular anomalies: 29 cases of Robin sequence, 21 cases of microretrognathism associated in plurimalformative syndrome, 8 cases of Treacher Collins syndrome, 3 cases of Patau syndrome, 1 case of oculo-auriculo-vertebral spectrum (Goldenhar syndrome), 1 case of velo-cardio-facial spectrum (di George syndrome), 1 case of Silver Russell syndrome. The remaining 9 patients were identified as having congenital mandibular isolated anomalies not associated with other anomalies. Conclusion: congenital mandibular anomalies represent 2% of all congenital anomalies but probably the incidence is underestimated. The majority are associated in plurimalformative syndromes. The isolated anomalies are, probably, produced by unusual mechanical forces. Key words: mandibular anomalies, cranio-facial dysmorphisms, deformations P58. MULTIPLEX RT-PCR AS A USEFUL AND COST EFFICIENT METHOD FOR FUSION GENE TRANSCRIPTS DETECTION IN ACUTE LEUKEMIA. D. Jardan2, C. Jardan2, R. Talmaci2, D. Coriu1,2. 1 Institut Clinic Fundeni 2 UMF „Carol Davila” Acute leukemia is a group of heterogeneous diseases which often present chromosomal translocations as a sole cytogenetic aberration. Expression of this fusion genes is often pathognomonic and detection of their transcripts is an important mean of risk stratification of acute leukemia. In this study we present a multiplex RT-PCR assay used for identification of 8 most common fusion genes in acute leukemia (E2A-PBX1, TEL-AML1, AML1-ETO, PML-RARa, CBFb-MYH11, MLL-AF4, BCR-ABL, SIL-TAL). For comparison of the method performance we tested this assay in parallel with a commercial kit (HemaVision Full kit). A cohort of 119 patients was studied (67 – AML si 52 – ALL) of which 29 (12 -ALL si 17 – AML) presented one of the fusion genes detectable by this method. There were detected these fusion genes - BCR-ABL – 4, PML-RARa – 9, MLL-AF4 – 5, CBFb-MYH11 – 2, AMLETO – 6, TEL-AML – 2, SIL-TAL – 1. Results: this method allowed risk stratification for 24% of acute leukemia patients. For these patients it was possible to improve disease management by adapting treatment options to specific genetic defect. In combination with the detection of other genetic malformations performed by our laboratory (MLL-AF9, MLL-PTD, FLT3-ITD, FLT3-TKD and tetranucleotid insertions in NPM1) this method allows risk stratification for the majority of AML patients. P59. INTERPHASIC FISH ON PERIPHERAL BLOOD IS A SENSIBLE METHOD FOR EVALUATION OF MINIMAL RESIDUAL DISEASE IN CML C. Jardan2, D. Jardan2, R. Talmaci2, D. Coriu1,2. 1 Institut Clinic Fundeni 2 UMF „Carol Davila” Chronic myeloid leukemia (CML) is the first malignancy for which a specific chromosomal abnormality was described – Philadelphia chromosome. Detection and monitoring of minimal residual disease in CML by detection of Philadelphia chromosome is an important part of response evaluation of patients in treatment with tirozinkinase inhibitors, because it allows early detection of suboptimal response or resistance to treatment. Fluorenscent in situ hybridisation (FISH) is a molecular cytogenetics technique which allows minimal residual disease (MRD) monitoring on interphase nuclei. MRD monitoring on peripheral blood has the advantage of generating results in short time (even in 24h) and is less invasive than Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 81 bone marrow biopsy. In comparison with bone marrow culture it has the advantage of being a direct method which abolish artifacts introduced by the culture. On the over side – it has disadvantage of being able to identify only specific chromosomal malformations – philadelphia chromosome in this case, all other important aberrations important in disease progression being omitted. For use of this method in MRD there is a need for large standardization studies, present clinical standard being conventional cytogenetics. In this study we analyzed FISH techniques as a rapid and uninvasive alternative method for MRD monitoring in CML patents undergoing tirozine-kinase inhibitor treatment. Results: in comparison to conventional cytogenetics – interphasic FISH on peripheral blood is a robust and efficient direct method for MRD monitoring in CML. P60. CHIMERISM ANALYSIS – ELECTIVE METHOD FOR THE OUTCOME PREDICTION OF ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION C. Jinca1, S. Arghirescu1, M. Puiu1, A. Oprisoni1, L. Balint-Gib1, V. Ordodi2, M. Serban1 1 University of Medicine and Pharmacy „V. Babes”, Emergency Children’s Hospital „L. Turcanu”, Center for Bone Marrow Transplantation 2 University of Medicine and Pharmacy „V. Babes”, Laboratory for Transplant Immunology Purpose of the study. The method used for the investigation of the presence of donor cells in the host after allogeneic hematopoietic stem cell transplantation (HSCT) is represented by the chimerism analysis. In this retrospective study, the objective was to evaluate the predictive value of chimerism analysis for the post-transplant outcome. Material and methods. The study was conducted on a lot of 20 patients who underwent allogeneic HSCT in the Centre for Bone Marrow Transplantation Timişoara. Chimerism was prospectively analyzed on days: + 30, +90, +180, + 270, +365 post-transplant. In 7 patients the investigation was done by fluorescent in situ hybridization whereas the rest were analyzed by means of real-time PCR. One patient died prior to the first chimerism assessment. Results. In 8 patients, the evolution towards a complete donor chimerism occured within the first six months post-transplant. In 3 patients, the last performed analysis revealed a progressive varied chimerism at least for one lineage, pattern which turned out to be highly predictive for the unfavorable outcome for two of them. Granulocytes, monocytes and CD 19 lymphocytes were 96 – 100 % donor cells already on day + 30 in the majority of patients. Discussion and conclusions. The two methods used for the assessment of chimerism offer predictive information for the outcome. Our patients with progressive mixed chimerism presented an unfavorable clinical outcome as compared to those with a full donor chimerism who were in complete remission at the time of evaluation. Key words. Hematopoietic stem cell transplantation, chimerism, polymerase chain reaction, fluorescent in situ hybridization. P61. CHEDIAK-HIGASHI SYNDROME-CLINICAL AND EVOLUTION ASPECTS E. Boeriu 1,2, M. Cucuruz1,2, M. Lelik1,2, G. Brad1,2,. I. Ursache1,2, A. Botiz1, S. Tamas1, M. Serban1,2 1 Children’s Emergency Hospital „Louis Turcanu, Timisoara 2 3rd Pediatric Clinics, Hemato-Oncology Department University of Medicine and Pharmacy „V. Babes „Timisoara Chediak-Higashi syndrome, diagnosed and followed by us, was defined by the presence of oculocutaneous albinism (hair, skin, eyes), the common bacterial infections associated, the presence of positive azurophillic peroxidase granular found in the leukocytes from the peripheral blood and 82 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 bone marrow and positive family history. Our diagnosis consisted in the identifications of the typical clinical, biological, immunological and evolutionary features of this disorder. Clinical, we identified the typical symptoms characteristic for severe Chediak-Higashi syndrome (significant visceromegaly, early appearance of immunodeficiency syndrome manifested as abscesses, impaired of central nervous system). Biological events can be assessed as moderate (triglycerides = 2.45-3mmol/L, fibrinogen=1.191.65g/L, ferritin=341-490ng/ml). The macrophage activation syndrome found in the bone marrow and manifested as severe citophage pathological aspects was responsible for the presence of severe tricitopenia. In these conditions, immunological, we noted a satisfactory humoral immune system. The lymphocytes B remained constant over 1500/mm3, while the serum Ig levels were normal or even higher than the normal values (IgG = 15.06 to 18.5 g / L). NK cells CD3-CD16 + CD56 + were within normal limits, sometimes slightly increased (689-822/mm3). The case had an unfavorable evolution under the treatment according to „Treatment Protocol of the Second International Study Hemophagocytic Limphohistiocytosis 2004”. In conclusion, we noted the complexity and diversity of the constitutional repercussions of the phagocytic deficiencies responsible for macrophage activation. It involves complex disorder in macrophage system / immune. P62. COMBINED EFFECT OF DIET AND THE RS1799883 POLYMORHISM OF THE INTESTINAL FATTY ACID BINDING PROTEIN 2 GENE ON THE RISK OF DEVELOPING THE METABOLIC SYNDROME Katalin Csep1, Gyongyi Dudutz1, Claudia Banescu1, Anamaria Butila Todoran1, Laszlo Koranyi2 1 University of Medicine and Pharmacy Tg. Mures – Romania 2 Drug Research Center Balatonfured – Hungary Introduction: Fatty acid-binding proteins (FABPs) belong to a multigene family coding for proteins participating in the uptake, intracellular metabolism and transport of long-chain fatty acids. The human intestinal fatty acid binding protein 2 (FABP2) is an abundant cytosolic protein in the epithelial cells of the small intestine, involved in mediating fat absorption by the binding and the intracellular trafficking of long-chain fatty acids. Because of these effects, the gene is considered a candidate for the metabolic syndrome. The polymorphism at codon 54 due to the A>T mutation is present in approximately 1/3 of the population, and the resulting Ala54Thr change determines a two-fold increase of affinity for long-chain fatty acids, leading to an increased fat oxidation and insulin resistance. Based on these considerations, we proposed to appreciate the risk determined by the combined effect of the predisposing allele and an unhealthy diet. Material and methods: We have carried out a case control study on 109 unrelated middle-aged patients and 73 healty persons from the Tg Mures urban area. Metabolic syndrome was asessed according to the IDF (International Diabetes Federation) criteria published in 2005, insulin resistance was measured by the HOMA and QUICKI indices based on fasting glucose and insulin, genotyping was carried out by PCR-RFLP, and diet was assesed by a simplified semi-quantitative food frequency questionnaire (FFQ). For risk assessment, data was analyzed by Fischer’s exact and chi2 tests using a 2x4 contingency table. Results: We found that the TT genotype as compared to the AA genotype associates with an increased risk for disease development (OR = 3.25, p = 0.01, CI 95%: 1.26-8.4). A diet characterized by a reduced intake of calories, sugar and fat associates with a significant decrease of the disease risk (OR = 0.27, p = 0.0013, CI 95%: 0.1173-0.6295). The combined effect of an unhealthy diet characterized by an increased fat and calorie intake and the rs1799883 polymorphism (presence of the predisposing Thr54 allele in the AT or TT genotype combination vs. its absence in the AA genotype) was found to associate with an increased risk for Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 83 developing the metabolic syndrome (ch2 = 16.7, df: 3, p = 0.001). Conclusion: An increased calorie and fat intake in the presence of the rs1799883 polymorphism of the IFBAP2 candidate gene associates with an increased risk for developing the metabolic syndrome; a heathy diet, however, is an efficient measure for decreasing the risk in the presence of inherited predisposing factors. P63. RARE FAMILIAL CASE OF LISSENCEPHALY Kinga Kozma1, Marius Bembea1, Claudia Jurca1, Radu Harbuz2, Cristina Skrypnyk1, Oana Iuhas1, Marius Ivascu2 1 Universitatea din Oradea, Facultatea de Medicina, Disciplina de Genetica, Oradea 2 Spitalul Clinic Municipal „Dr. Gavril Curteanu”, Oradea The term lissencephaly describes a „smooth” brain, result of the absence of normal gyri of cerebral cortex. The incidence is very rare, is estimated at approximately 1:100.000 live births. This is one of the most severe neuronal migration disorders. Several types of lissencephaly have been described in attempt to correlate the type of malformation with etiology. Currently five types are described: (1) classic lissencephaly (autosomal dominant and X-linked), (2) cobblestone lissencefaly (autosomal recessive) (3) lissencephaly with agenesis of the corpus callosum (X-linked); (4) lissencephaly with cerebellar hypoplasia (autosomal recessive) (5) microlissencephaly (not applicable). We present two cases of morph pathological confirmed lissencephaly of two died brothers, with consanguinity of the parents (uncle-niece once removed); both patients are male, birth rank IV and V, first three children were clinically normal female. Both patients had strikingly similar clinical symptoms: early onset neonatal seizures, severe neuro-psychical retardation, hypotonia, and fatal evolution marked by death in the first month of life. In the absence of possibility to identify the mutation, the authors present clinical and differential diagnosis with other congenital disorders and they are tryng to include on clinical criteria in one of those five categories described. Keywords: lissencephaly, cerebral cortex, neonatal seizures. P64. DIAGNOSIS AND MANAGEMENT OF OSTEOGENESIS IMPERFECTA M. Macovei, R. Cojocariu, M. Panzaru, R.Popescu, L.Caba, L.Butnariu, E. Braha, M.Volosciuc, C. Rusu Department of Medical Genetics, „Gr.T.Popa” University of Medicine and Pharmacy, Iasi Osteogenesis imperfecta is a term used for defining a heterogeneous group of hereditary diseases caused by qualitative and / or quantitative anomalies of type 1 collagen molecules which predispose to skeletal deformities and bone fractures as a consequence of minor trauma. There are more than 200 genetic mutations affecting type 1 collagen genes; two major types of mutations being involved in the etiology of osteogenesis imperfecta: mutations determining a decrease in type 1 collagen production and respectively, mutations determining synthesis of structurally abnormal collagen. Patients’ phenotype is variable related to the affected type of pro-collagen 1 chain and also to the mutation type found at each locus. There are 7 types of osteogenesis imperfecta, the majority having an autosomal dominant pattern of inheritance. The hallmark of the clinical exam is represented by bone fractures (multiple fractures, at minor trauma) associated with bone deformities, kyphoscoliosis, dental anomalies, articular hyperlaxity, bruising tendency. In order to illustrate the main elements of diagnosis and managements will report to 2 clinical cases: 84 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 The first case, A.I. (17 years old at the evaluation time), female, diagnosed with type-1 osteogenesis imperfecta on the following criteria: positive family history (autosomal dominant pattern of inheritance), multiple bone fractures, dorsal kyphosis, blue sclera, dentinogenesis imperfecta, radiologic appearance of spine and feet. The particularity of this clinical case is represented by 4th toe brachydactyly (feet, bilateral) along with normal audiogram (at the patient, her mother and her sister). The second case, C.A. (3 years old at the evaluation time), female, institutionalized child, diagnosed with type-3 osteogenesis imperfecta on the following criteria: severe bone fragility (multiple bone fractures at birth and after), major bone deformities, triangular face, blue sclera. In order to diagnose osteogenesis imperfecta, there are used a series of investigations from imagistics (postnatal radiological exam, prenatal ultrasound, quantitative CT scan to measure bone mineral density) to the anatomopathologic and molecular diagnosis. The management of this genetic disease involves a fracture prevention strategy, orthopedic and medical treatment (calcium, vitamin D, bisphosphonates). New methods of treatment are in experimental studies, among which we enumerate mesenchymal cell therapy, ribozyme therapy, as well as gene therapy. P65. DOWN SYNDROME-CLINICAL AND IMAGISTIC CORRELATION A.Manea, C. Ilie, M. Boia, J. Puiu, D. Iacob, M. Dima University of Medicine and Pharmacy Timisoara, Romania Introduction: Down syndrome is the most common and the most known chromosomal disorder. This syndrome is characterized by mental retardation, facial dimorphism and other phenotype distinctive traits. There cytogenic are three problems to be considered in appearance of Down syndrome: trisomy 21, mosaicism and chromosomal translocation. Objectives: In this study authors aimed to realize a correlation between anamnestic, clinical and paraclinical data of newborns with Down syndrome. Material and method: The study was developed based on clinical, cytogenetic and imagistic examination at a number of 10 newborns with Down syndrome, hospitalized in the Clinic of Neonatology and Prematures, Children Emergency Hospital „L.Ţurcanu” Timişoara, during three years. Results: From 10 case studied: 6 cases were newborns with various grades of prematurity , the other 4 cases were term newborns. Regarding maternal age in the moment of conception the situation was the following: maternal age between 20-25 years- 1 case (10%), between 25-30 years1 case (10%), between 30-35 years-3 cases (30%), between 35-40 years-4 cases (40%), between 4045 years- 1 case (10%). Genetical consult confirms existing somatic changes and chromosomal analysis shows trisomy 21 in 7 cases and mosaicism in 3 cases. Increased incidence of cardiac malformations requested cariologic consult in every case. Echocardiography showed ventricular septal defect in 4 (40%) cases, atrial septal defect in 3 (30%) cases and persistancy of arterial ductus in 3 cases (30%). Also, in one case there was an association between the last two disorders. Conclusion: Down syndrome is the most chromosomal disorder in neonatal period. Diagnosis is easy to set both in antenatal and perinatal period, based on particular phenotipical feature followed by case confirmation with cariotype performing. Key words: Down, chromosomal analysis, cardiac malformation Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 85 P66. DOWN SYNDROME, DIAGNOSIS AND PREVENTION Marchian Sanda Victor Papilian Medical School, Sibiu Chromosomal diseases, especially Down syndrome, in Romania continue to be an important cause of morbidity. In our country are registered 30 000 persons born with the Down syndrome. This condition, with major physical and mental disabilities, creates a special psycho-emotional and material problem to the patient and his family. While European countries report a decrease in the number of the newborns with this condition, in Romania the rate of this disease is still at high levels. The neonatologists are drawing attention that in the last year, the number of the newborns with congenital diseases, including the Down syndrome, has increased. Purpose of the study: this work is part of an interdisciplinary study in which the purpose is: to identify inside the Clinic of Obstetrics and Gynecology Hospital, Sibiu, the cases of newborns suffering from chromosomal syndromes; to confirm through cytogenetic examination (sample G and Fish heel), these cases in order to establish their rate in the amount of newborns registered each year, for a better understanding of the link between the clinical signs and the positive diagnostic; Material and method: Genetic consultation and lab examination was performed on 18 cases with suspicion of Down syndrome, examination by connecting to G-soft Metasystem in the Cytogenetic Laboratory of the Faculty beginning January 2009. Results: 10 cases were confirmed with Down syndrome and mosaicism with normal cell lines with trisomy 21. The consultations also aimed to genetically assess the possible antenatal risk factors incriminated in the induction of disease and characteristic signs.Conclusion:This study will continue to monitor all births in our Clinical Hospital and evaluate the projection incidence of this syndrome among births recorded here. Until now a small number of cases have been studied – due to the short time this laboratory was set up – but we believe that the number of confirmed cases, allows us to say that the incidence of this syndrome remains high. Although prevention method practiced in the Romanian Medical Universities could lead to reduction of births of children with Down syndrome. We want to draw attention that prevention through amniocentesis should become a medical act promptly recommended by doctors, when needed and also affordable for all the couples wishing to conceive. P67. DISCUSSION OF A CASE STICKLER SYNDROME Otilia Marginean 1, Ioan Simedrea1, Maria Puiu1, Belei Oana1, Bochean Camelia2, Tamara Marcovici1, Camelia Daescu1, Daniela Chiru1 1 st I Pediatric Clinic of Clinical children ‘s Emergency Hospital”Louis Turcanu” Timisoara Romania 2 Specialty Ambulatory of Clinical children ‘s Emergency Hospital”Louis Turcanu” Timisoara Romania Introduction : Stickler syndrome is a connective tissue disease that may include ocular findings ; hearing loss ; midfacial underdevelopment and cleft palate and mild spondyloepiphyseal dysplasia and/or precociuos arthritis. Aim:The authors present the case of a 7 month infant with ocular findings and specific facial phenotype that was admitted for diagnosis establishment. Presentation of case: T.V seven months old, is hospitalized in October 2009 for an intercurrent respiratory. Particularly striking phenotype with facial dimorphism, microcephaly, congenital cataracts, horizontal nystagmus, and psycho-motor retardation. In his family two maternal uncles with acquired amblyopia of unknown etiology. In his personal history, are not significant dates. Clinical examination showed an infant with normal anthropometric dates according to age, the undeveloped middle part of face, micrognathia, horizontal nystagmus. No baby sitting sits. The child give polysyllabic and shows the chaotic movements of the eye, not follow objects. TORCH serology was negative. Ophthalmologic examinations: microphthalmia. Cornee transparency reduced in size in horizontal and vertical axis. 86 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Crystalline opacity, subcapsular previous whitish color. Ocular pressure: normal; papillary reflex, photomotor present. FAO: no shine . Neurological exam - muscular hypotonia. Horizontal nystagmus. Opsoclonii. Psychomotor retardation with stereotypy. EEG - theta rhythm - delta, with many artifacts. ORL- Audiogram - mixed deafness, transmission and neuro-senzory. Karyotype 46 XY 1qh +, 21s +, yq +. Heterochromatic elongation chromosome with secondary constriction on chromosome 1, satellite at 21 and heterochromatic of the Y chromosome. Conclusions: 1. Stickler syndrome is under diagnosis 2. For diagnosis is necessary history, clinical exams and karyotype. 3. Genetic counseling is appropriate in this case. P68. EARLY TREATMENT MANAGEMENT IN HYPODONTIA Popa Malina, Dinu Stefania, Luca Magda, Lazar Cristina, Bratu Cristina, Szuhanek Camelia, Balan Raluca, Ogodescu Emilia, Ogodescu Alexandru Department of Pedodontics-Orthodontics, University of Medicine and Pharmacy „Victor Babes” Timisoara, School of Dentistry Introduction: Congenital missing teeth is one of the most common developmental problems in children and hypodontia is the term most often applied to this situation. The early treatment of nonskeletal orthodontic anomalies in early mixed dentition is intended to prevent the development of pronounced anomalies in the permanent dentition with the ultimate aim of reducing or even eliminating the need for later orthodontic treatment. Aim and objectives: The purpose of this paper is to describe the need for early examination and diagnosis of hypodontia in growing children and the necessity of the interceptive therapy in this malloclusion. Material and methods: Five patients presenting reduced hypodontia, ages between 6-10 years, have received orthodontical treatment. We have managed to reduce the complications such as malposition of the teeth next to the agenetic site, with consequent midline deviation or resorption of the alveolar bone. Early detection of teeth involved by the reduction process offers optimal conditions for closing the spaces through controlled mesial migration of the teeth. Discussions: Early diagnosis of hypodontia may allow a more favorable prognosis and minimal functional, aesthetical and psychological complications. Keywords: hypodontia, early treatment P69. HLA ROLE IN CHRONIC AUTOIMMUNE THYROIDITIS PREDISPOZITION Alina Martinescu1, Irina Durbala2, Eduard Circo3 1 Medical Genetics Department 2 Cell and Molecular Biology Department, 3 Endocrinology Department, Faculty of Medicine, Ovidius’’ University, Constanta, Romania Introduction: Class II HLA genes (HLA-DRB1 and DQB1), are well established risk genes for autoimmune thyroiditis and other autoimmune diseases. In autoimmune thyroid diseases, however, both susceptible and protective alleles have been described, influencing the development of autoimmunity and progression to overt chronic autoimmune thyroiditis. This study determines the class II genes in patients with Graves’ disease and Hashimoto’s thyroiditis. Material and methods: The subjects included in this study comprise 70 patients with chronic autoimmune thyroiditis registered at the Endocrinology Clinic of the Emergency Clinical Hospital Constanţa and 84 healthy subjects. The method used for the assignment of alleles at HLA-DRB1 and DQB1 loci was molecular genotyping, primarily by the sequence specific oligonucleotide hybridization method, and when required, by the sequence specific primers method. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 87 Results: The analysis of the DRB1 alleles frequencies showed DRB1*0301 as a predisposing allele in HT patients (pc<0.04), but failed to demonstrate DR4 alleles as a high-risk alleles (p=0.30) and for DR11 (p=0.18). HLA-DRB1*16 was more frequent in patients than in controls (p=0.05). Conclusion: This study demonstrated the positive association of DRB1*0301 allele with Hashimoto’s thyroiditis and Graves’disease. We also found a borderline association of HLA-DR*16 (p=0.05). We failed to demonstrate association with the disease for other known susceptible and protective alleles, fact that is probably due, when such polymorphic loci were involved, to the relatively low number of cases analyzed. P70. 13P CHROMOSOME POLYMORPHISM IN A COUPLE WITH RECURRENT MISCARRIAGE – CASE REPORT Anca Mitroi1, Iuliana Dimofte2, Mariana Aschie2 1 Emergency Clinical County Hospital of Constanta 2 „Ovidius” University of Constanta, Faculty of Medicine Introduction. Variations in homologue human chromosome morphology at one or different persons, consist largely of heterochromatin are called chromosome polymorphisms (heteromorphisms). One type of chromosome polymorphism is represented by blocks of heterochromatin found in the short arms of acrocentric chromosomes. Materials and methods. This paper present a case report of a couple with recurrent miscarriage investigated through family anamnesis and karyotype examination in G-banding and Q-banding. Results. Family anamnesis revealed: non cosanguineous couple, with ages around 29 and 28 years old, clinical healthy. In man’s family one sister had a spontaneous abortion, and in woman’s family there is no reproduction failure or genetics disease. From investigation of reproduction we identified: one miscarriage in 13 weeks of gestational age and another pregnancy echo graphic diagnosed with occipital meningocele in 21 weeks of pregnancy and spontaneous aborted at 24 weeks of pregnancy, in which the necropsy revealed a plurimalformative syndrome. The woman karyotype revealed the presence of 13p polymorphism. Conclusions. 13p polymorphism, though considered normal variant, without any phenotype anomalies, is mentioned in the studies to have an increase frequency in the couples with recurrent miscarriage P71. MOLECULAR PROFILING OF ADAM12 IN BREASTCANCERS Diana Narita1, Andrei Anghel1, Edward Seclaman1, Razvan Ilina2, Natalia Cireap2 1 University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania, Biochemistry Department 2 University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania, Surgical Oncology Department Background. ADAMs (a desintegrin and metalloproteinase) family have been associated with the process of proteolytic „shedding” of membrane-associated proteins ectodomain and hence the rapid modulation of key cell signalling pathways in tissues microenvironment. A variety of cytokines, chemokines and growth factors that are initially produced as transmembrane proforms are activated by sheddase activities. Purpose. ADAM12 is highly expressed in rapidly growing tissues such as placenta and malignant tumors and it was found as one of the Candidate Cancer Genes in a comprehensive mutational analysis of human breast cancers. Among genes detected to be mutated with high frequencies in breast cancers, only 14 had higher mutation prevalence than ADAM12. In this context, we purposed us to determine the gene expression profiles of ADAM12 in breast cancer, in comparison with normal breast and benign breast tumors. 88 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Experimental Design. ADAM12 gene expression (ADAM12L and 12S spliced variants) was eva luated in tumor cells and stroma obtained by laser microdissection from 50 patients with breast cancers and compared with benign breast tumors and adjacent normal breast from 23 patients. Results. ADAM12 mRNA expression was significantly up-regulated in breast cancers, as determined by real-time RT-PCR, especially in the tumor cells. In the normal breast we found an absent or very low expression. For benign tumors, we found an increased expression, especially in atypical hyperplasia. ADAM12L was up-regulated in both tumor cells and stroma while ADAM12S was upregulated only in the tumor cells. Conclusion. From these preliminary results, we found that ADAM12 could be an interesting marker and eventually a therapeutic target for breast cancer. P72. ADAM17: SIGNALING SCISSORS IN THE TUMOR MICROENVIROMENT Diana Narita1, Andrei Anghel1, Edward Seclaman1, Razvan Ilina2, Natalia Cireap2 1 University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania, Biochemistry Department 2 University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania, Surgical Oncology Department Background: The shedding activities of ADAMs (a desintegrin and metalloproteinase) place them in pivotal positions in the extracellular regulation of cell signaling. They can potentially regulate many activities: immune regulation, angiogenesis, cell migration and proliferation. ADAM17 is a key regulator of EGFR signaling pathway, being a major sheddase for the ErbB receptor ligands, including the proforms of heparin-binding EGF-like growth factor, transforming growth factor-α, amphiregulin and epiregulin. Purpose: The aim of this study is to determine the ADAM17 expression profiles in breast cancers versus benign tumors and normal breast. We compared also the expression of ADAM17 with another member of ADAMs family, ADAM12. Experimental design: ADAM17 gene expression was evaluated using real-time RT-PCR in the tumor cells obtained by laser capture microdissection from 50 patients with breast cancers and compared with benign breast tumors and respectively adjacent normal breast from 23 patients. Results: Using laser capture microdissection in order to obtain from the heterogeneous tumor environment only the cells of interest, we extracted a quantity of RNA ranged between 2.7 – 12.8 ng/μl, depending on the number of the captured cells. ADAM17 gene expression was significantly lower than ADAM12 in breast cancers, but significantly up-regulated in cancers versus normal breast and benign tumors. Although there was an obvious increased expression of ADAM17 in cancers versus benign tumors, the difference was not statistically significant. Conclusions: Our study is only at the beginning and we need further correlations with tumor phenotypes, but from these preliminary results we assume that ADAM17 could be a marker of tumor progression. The study of ADAMs expression confers a better understanding of molecular mechanisms that are implied in the regulation of signaling pathways in breast cancers. P73. CLINICAL STUDY REGARDING THE LINK BETWEEN MTHFD POLYMORPHISM AND TRISOMY 21 Daniela Neagoş, Ruxandra Creţu, L.Camil Bohiltea Genetic Department, UMF Carol Davila, Bucharest Introduction: Methylenetetrahydrofolate (MTHFD1) is an enzyme that interconverts tetrahydro folate derivatives for purine, methionine and thymidylate synthesis. The folate (members of the B9 vitamins family) cycle is involved in two essential physiological processes: the synthesis of purines Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 89 and pyrimidines equired for DNA synthesis and repair; and the methylation associated with the methionine cycle. Lowered folate has also been associated with increased risks of trisomy 21 and folic acid supplementation is now commonly recommended during early pregnancy, pre- and postconception. DNA hypomethylation can lead to chromosomal abnormalities such as DNA breakage and genes polymorphisms with reduced expression. Impairments in folate metabolism associated with polymorphisms of metabolic enzymes may increase the risk in women for giving birth to infants with Down Syndrome (DS). The MTHFD gene is located on chromosome 14 (14q24). A common MTHFD1 1958G > A polymorphism (Arg653Gln) has a reduced enzymatic activity and stability and has been associated with increased risk of DS. Materials and methods: Our study include 72 women: 26 of them (younger than 40 years age), that gave birth to DS babies, confirmed as regular trisomy 21; 7 of them have a history of spontaneous miscarriages (26.92%); and 46 control mothers that gave birth only to healthy children and without any history of miscarriages or abnormal pregnancies. All women in our study reside in the same geographic area and have a similar social background. Genomic DNA was isolated from whole blood by means of the peqGOLD blood DNA mini kit following the manufacturer’s instructions. The presence of MTHFD1958 G>A mutation was analyzed by polymerase chain reaction (PCR-RFLP) and allele specific restriction digestion with Msp I. Results: Allele frequencies were calculated for each genotype, and the differences in allele frequencies between mothers of children with Down Syndrome and control mothers were determined using chi-square test. Expected genotype frequencies were calculated from the allele frequencies under the assumption of Hardy-Weinberg equilibrium. The interaction between genotypes was evaluated by calculating the odds ratios (OR) for mutant genotypes, as compared to wild types genotypes. Analyses were performed using the software SPSS. MTHFD 1958 A allele frequency was 40,22% (37/92 allele) in control mothers and 44,23 % (23/52 allele) in case mothers (χ2: 0,22, P: 0.63) . The heterozygous genotype frequency (GA) at position 1958 was higher in case mothers than in controls (65,38% versus 54,34 % respectively) with an odds ratio of 1,7 (95% CI 0,54 to 5,26). Interestingly, homozygous and heterozygous genotypes frequencies of MTHFD at position 1958 (GA or AA) were higher among case mothers than controls (76,92 % versus 67,39%) with an odds ratio of 1,61 (95% confidence interval [CI] 0.53–4.85), indicating that this polymorphism may have genetic impact in DS. Conclusions: We didn’t find a statistical significant association between women with MTHFD polymorphism and history of DS pregnancies; the second step in our study is the catamnestic evaluation of our patients with DS babies for two years. P74. REPORT OF CYTOGENETIC ANALYSIS FOR 68 CASES WITH ANOMALIES OF SEXUAL DEVELOPMENT Diana Ochiana1, Vasilica Plaiasu1, Gabriela Motei1, Amalia Costin2 1 Genetics Department, Institute for Mother and Child Care ”Prof.dr.Alfred Rusescu”, Bucharest, Romania 2 Genetics Department, District Hospital Ilfov ”Sf.Imparati Constantin si Elena”, Bucharest, Romania The cytogenetic diagnosis of sexual development anomalies is crucial for the clear determination of the genetic sex of the individual, but also for the identification of other subsequent genetic abnormalities. In the present study we analysed 68 cases with various sex anomalies; after applying conventional cytogenetic methods (GTG banding), out of all analysed samples, for 4 patients the cytogenetic result was 45, X, for 3 patients it was 47, XXY and for one patient it was 47,XXX. 90 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 In two cases an inconsistency was observed between phenotype and karyotype: two patients of apparent female gender had 46, XY karyotype. Out of 35 male individuals and 21 female individuals, approximately 19% showed complementary chromosomal alterations (structural changes, mosaics and heteromorphisms) that involve the X chromosome and autosomal chromosomes. To extend routine cytogenetic banding methods, we used FISH analysis to verify the presence of the SRY region for 7 cases. The early determination of gender anomalies is important for the medical and social management of identified cases with this type of disorders and for proper genetic counselling. P75. CONGENITAL HEART DEFECTS IN CHAR SYNDROME Monica Panzaru 1,2, Mihail Volosciuc2, Cristina Rusu 1,2, Elena Braha1,2, Lavinia Caba1, Lacramioara Butnariu1,2, Mihai Macovei1, Roxana Cojocariu1, Cristina Rusu 1,2 1 „Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania, 2 Medical Genetics Centre, Iasi, Romania Char syndrome (CS) is an autosomal dominant condition comprising patent ductus arteriosus (PDA), typical facial dysmorphism (flat midface, mild ptosis, a short philtrum with prominent philtral pillars with an upward pointing vermilion border resulting in a triangular mouth, and thickened everted lips) and hand anomalies (aplasia or hypoplasia of the middle phalanges of the fifth fingers). The prevalence of Char syndrome has not been determined but is thought to be quite low. Less common heart defects associated with Char syndrome are septal defects or complex congenital diseases. The prognosis depends on the potential presence of associated heart malformations. Management of PDA after the immediate newborn period is determined by the degree of shunting from the aorta to the pulmonary artery. Char syndrome has been mapped to a narrow region of chromosome 6p12-p21. Further studies have shown that Char syndrome is caused by defects in TFAP2B, encoding a neural crest-related transcription factor (also known as AP-2β). We present two cases in order to illustrate this rare entity and to discuss the variability of heart defects. C.C.A (0,8 y old male): brachycephaly, high forehead, mild hypertelorism, bilateral epicanthic folds, short nose, anteverted nostrils, a short philtrum, a triangular mouth, and thickened lips; ecocardiography: patent ductus arteriosus, patent foramen ovale. G.N.L. (2 y old female): broad forehead, thin eyebrows, right palpebral ptosis, right epicanthic fold, flat nasal bridge, short nose, anteverted nostrils, , a short philtrum, a triangular mouth, clynodactyly of the fifth fingers; ecocardiography: atrial septal defect - ostium secundum, tricuspid insufficiency; laboratory findings: Ig A and Ig G deficiency. In conclusion, we present two cases of CS in order to illustrate this rare genetic disorder but also to discuss the variable expression of heart defects, long term follow-up, management and genetic counseling. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 91 P76. MILD HYPERHOMOCYSTEINEMIA SECONDARY TO HOMOZYGOUS C677→T MUTATION IN THE METHYLENETETRAHYDROFOLATE REDUCTASE GENE, ASSOCIATED WITH PORTAL-VEIN THROMBOSIS AND PORTAL CAVERNOMA Petrescu Carmen1, Grigorescu-Sido Paula2, Bârsan Mircea3, Zoica Bogdana1, Scridon Traian3 1 Department of Hematology and Oncology, IIIrd Pediatric Clinic, University of Medicine „Victor Babeş” Timisoara, Romania 2 Department of Medical Genetic, Ist Pediatric Clinic, University of Medicine „Iuliu Haţeganu” Cluj, Romania 3 Department of Cardiovascular Surgery, Institute of Heart „Niculae Stăncioiu” Cluj, Romania Background. Hyperhomocysteinemia was associated with vein thrombosis but whether this relation is causal is still unclear. The C677→T mutation in the methylentetrahydrofolate reductase (MTHFR) gene mildly increases homocysteine levels being citated one of the congenital predisposing factors for vein thrombosis. Aims. The study presents a child with the homozygous type of the mutation presenting early portal vein thrombosis and portal cavernoma. Methods and results. A male patient with developmental delay, motor and gait abnormalities, mental retardation and myoclonus, admitted in our service at the age of 4 years and 11 months (December 2005) for important splenomegaly and hematological hypersplenism with the initially presumption of sphingolipidosis (excluded by enzymatic analysis), was than diagnosed with portal cavernoma; the suspicion of MTHFR deficiency suggested by the high level of plasma homocysteine (MEIA) was confirmed by the finding of homozygous C677 → T mutation in the MTHFR gene (PCR). The family history was negative for thrombosis (autosomal recessive inheritance). Acquired predisposing factors for venous thrombosis were found in the patient’s personal history (intrauterine and neonatal hypoxia), but other investigated congenital or inherited predisposing factors were negative (factor V Leiden, G20210A mutation in the prothrombin gene, antithrombin, protein S or protein C deficiency, aso). Two episodes of gastrointestinal hemorrhage (2006) (rupture of esophageal and gastric varices) were temporarily resolved by ligature and treatment with sandostatin); chronic administration of beta-adrenergic blocking agent was started; in the mean time, treatment with betaine and folic acid aiming to control the homocysteine level. After splenectomy and a left gastro-renal shunt (January 2007) the patient had three more episodes of gastrointestinal hemorrhage - the last one on November 2007. Since that time, under medical treatment and periodically prophylactic ligature of the esophageal varices, the evolution was satisfactory without other hemorrhagic episodes, and the homocysteine level remained normal. Conclusions. The association between the homozygous C677→T mutation in the MTHFR gene, folate deficiency, and the intrauterine and neonatal hypoxia, could probably explain the early thrombotic event in a patient without other predisposing factors for vein thrombosis. P77. RARE CHROMOSOMAL DELETIONS WITH BREAKPOINTS OVERLAPPING THOSE OF THE WILLIAMS-BEUREN SYNDROME V. Plaiasu1, D.Ochiana1, G.Motei1, A.Costin2, T.Ciomartan3, I.Anca3 1 IOMC ”Prof.dr.Alfred Rusescu”, Genetics Department, Bucharest, Romania 2 District Hospital Ilfov ”Sf.Imparati Constantin si Elena” 3 IOMC ”Prof.dr.Alfred Rusescu”, Pediatrics Department, Bucharest, Romania Williams-Beuren syndrome (WBS) is a rare multisystemic disorder, which is caused by a chromosomal microdeletion in the q11.23 region of one copy of chromosome 7. This microdeletion can not be detected by conventional karyotyping and is revealed by FISH analysis, which leads to diagnosis in 95% of the cases. The most common symptoms of WBS are mental disability, heart defects and unusual facial features. 92 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 We report two cases with large, atypical, visible deletions on the long arm of the chromosome 7, from two different unrelated families. The male proband revealed the WBS phenotype and in addition showed more severe mental retardation than is commonly observed with this syndrome and infantile spasms, indicating a larger than usual deletion affecting neighbouring genes as well. Additional findings in this patient included also horsekidney, hypotonia, complex cardiac malformation. Cytogenetic analysis of the parents was normal, indicating that the interstitial deletion had de novo occurrence. The second patient, the female proband, born very prematurely, showed a striking dysmorphic features, very characteristic for WBS. Parental karyotypes have not been performed, but the patient has four healthy siblings. We conclude that FISH analysis is not sufficient for genotypic characterization of a patient with Williams phenotype and for complete and correct genetic counseling. P78. CHROMOSOMAL CHANGES OF THE ACROCENTRICS IN ACUTE LEUKEMIA Popa Cristina1, Ionita Hortensia1,2, Surducan Dan1, Nicoleta Andreescu1, Alina Belengeanu1, Maria Puiu1, Margit Serban1,3 1 University of Medicine and Pharmacy„Victor Babes” Timisoara 2 City Universitary and Emergency Hospital Timisoara 3 Emergency Children Hospital „Louis Turcanu” Timisoara Acute leukemia is a clonal disturbance due to malign transformation of a myeloid or lymphoid progenitor cell.. Acute myelocytic leukemia (AML) and Acute lymphoblastic leukemia (ALL) is characterized by a variety of numerical and structural chromosome aberrations. Even if 21 and 22 trisomies are not particularities of any type of acute leukemia, they are frequently encountered. During the period between 2004 and 2010, in Cytogenetics Laboratory of University of Medicine and Pharmacy „Victor Babes” Timisoara, there were performed 110 cytogenetics tests of the patients with the suspicion of acute lymphoblastic leukemia and acute myeloblastic leukemia. The diagnostic cytogenetic analysis has been shown to be an important prognostic factor, capable of predicting remission duration and the therapeutic management. For the confirmation of previous data, CGH may provide useful information regarding the nature of genomic aberrations that take place in cases with complex karyotypes. Keywords: Small Acrocentrics ( 21 and 22), chromosomal abnormalities, acute leukemia P79. Cytogenetic issues in „CRI DU CHAT” syndrome The experience of Iasi medical genetics center R. Popescu1, C. Rusu1, M. Volosciuc2, L. Butnariu1, E. Braha1, M Panzaru1, L Caba1, M. Macovei1, R. Cojocariu1, M. Gramescu1, C.Bujoran2, I. C. Ivanov3, E. V. Gorduza1 1 „Gr. T. Popa” University of Medicine and Pharmacy Iasi -Department of Medical Genetics,, Romania . 2 Children’s Hospital Iasi - Medical Genetics Center, Iasi, Romania 3 Saint Spiridon Hospital Iasi – Immunology and Genetics Laboratory, Iasi, Romania Cri-du-chat syndrome is a genetic disease resulting from a deletion of variable size occurring on the short arm of chromosome 5. Main clinical features are: high-pitched monochromatic cry, microcephaly, broad nasal bridge, epicanthal folds, micrognathia, abnormal dermatoglyphics and severe psychomotor and mental retardation. Cardiac, neurological and renal abnormalities may be associated. We present 5 cases diagnosed with „Cri du Chat” syndrome between 2002-2010 in Iasi Medical Genetics Center, in order to illustrate some rare variants and to discuss the cytogenetic complexity Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 93 of this syndrome. In this period our Cytogenetics Lab has performed 2074 karyotypes Cytogenetic diagnosis was performed using peripheral lymphocytes with G banding and FISH analysis. Case 1: (3-year-old female): pregnancy - uneventful; birth - term, Apgar score 5, Wt 2,700 g; clinical features - failure to thrive, microcephaly, typical cri-du-chat face and cry, hyperextensible joints, hypertrichosis, severe mental retardation; karyotype - 46,XX,del(5p). Normal echocardiography and ophtalmologic examination. Case 2 (11 months-old female): pregnancy - polyhydramnios; birth - term, intrauterine growth retardation, Apgar score 8; clinical findings - growth retardation and microcephaly, dysmorphic face, typical cry, moderate mental retardation. Echocardiography - subaortic ventricular septal defect, tricuspid insufficiency; karyotype- 46,XX,r(5)(p21;q35)/45,XX,-5/47,XX,r(5)(p21;q35),r(5) (p21;q35). Case III: (6-year-old female): pregnancy - uneventful; birth- term, intrauterine growth retardation; clinical findings - microcephaly, dysmorphic face, severe mental retardation; echocardiography and renal ultrasound - normal, psychological examination - IQ 29. Karyotype: 45,XX,-22,t(5;22)(5p15.1;22q.1)/46XX,t(5;22)(5p15.1;22q.1)der(22)(pter→q11.1) [32]/[2]. FISH analysis confirmed the translocation Case IV (11 months-old female): pregnancy - uneventful; birth - term, Apgar score 8, intrauterine growth retardation; clinical findings - growth retardation and microcephaly, dysmorphic face, typical cry, hypotonia, ectopic anus, moderate mental retardation, echocardiography - atrial septal defect. Karyotype: 45,XX,-21, t(5,21)(p10). FISH analysis confirmed the translocation. Case V (5 months-old female): pregnancy - imminent abortion; birth - term, Apgar score 6 (cardiopulmonary resuscitation); clinical findings - growth retardation and microcephaly, dysmorphic face, typical cry, right simian palmar creases, bilateral metatarsus adductus, moderate phsycomotor retardation. Karyotype: 46,XX,del(5)(p13-pter); Our study confirms the cytogenetic complexity of „Cri du Chat” syndrome. So we find 2 deletions on the short arm of chromosome 5 (in one case it wasn’t possible to identify precisely the breakpoint), and surprising (reported to literature data which indicate the presence of deletion in 85% of the cases), we find 3 complex cytogenetic abnormalities: one mosaic with a ring of chromosome 5; one mosaic with a translocation t(5;22) and one unbalanced translocation t(5;21). P80. ETHICAL CONSIDERATIONS REGARDING GENETIC MODIFIED ORGANISMS Pusta Dana1, Mariela Militaru2, I. Paşca1, Rodica Sobolu1 1 USAMV Cluj-Napoca 2 UMF Iuliu Haţieganu Cluj-Napoca Frequently it is said that the science is „per se” meaning that science is made just for the science sick, this being the reason why the science cannot be neither „good” nor „bad” this being the reason why it is considered being moral and ethical neutral. This is true just for the concept of science as a process, but the development of the scientific process and especially the applications resulting from this process are those risings the questions especially about ethics. Also science is made by researchers, which surely are not neutral, both from ethical and moral points of view. Considering these is almost impossible to separate the science, in its essence, by its practical applications. In the followings rows there will be presented some ethical considerations regarding the utilisation of the genetic modified organisms in medicine, in biotechnologies and also some other ethical considerations regarding the existence and utilisation of the transgenic organisms. In medicine there are just a few oppositions regarding the producing by biotechnologies of some medicines of some therapies. A possible area regarding the medical application which rise ethical problems are the information regarding the human genome development. The genetic screening, and the possibility of genetic discrimination, is that which rise ethical problems in present. 94 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 The industrial biotechnologies field is a difficult field because of its diverse applications. The first ethical problem is that linked to the commercial interests. Many persons see the biotechnological applications as a result of the commercial urges. Also ethical questions may be raised regarding the technological processes of obtaining the respective products and also regarding their utilisation. In this case, the public opinion plays an important role; this can determine the success or the disappearance of one product from the market. The transgenic organisms, by themselves, raise many ethical problems. The transgenic plants were ignored till the second part of ’90 when there appeared many public reactions regarding the transgenic food. The reactions were in two directions, on one hand the human health and on the other hand the environment protection. The transgenic animals didn’t raise so many ethical problems compared to the plants; first of all because they cannot be easily dissipate into the nature. Also the transgenic animals are generally used in medical context, field in which the benefits are clear. The most frequent ethical problems regarding the transgenic animals are those regarding their welfare, but the breeding standard of these animals is very high and attentive monitoring in all the countries using transgenic animals. Also, the acceptance of the transgenic mouse as a model for the human disease evolution and of the transgenic animals as bioreactors seemed to rise less ethical questions compared to the transgenic plants. The possibility of the xeno-transplantation offers a hope but also raises many questions, especially in some religious groups which do not agree with these. In other words, the new opportunities which have come out as a result of obtaining the genetic modified organisms not just solve some problems but also determine the apparition of the new ones. P81. ULTRASOUND SOFT MARKERS IN SECOND TRIMESTER AND FETAL ANEUPLOIDY Viorica Radoi1,2, Dana Mierla1, Luminita Neagu1, Andreea Mustata1, .Silviu Predoi1, L.C.Bohiltea2 1 Life Memorial Hospital Bucharest, Maternal- Fetal Medicine Department 2 UMF Carol Davila Bucharest Introduction: Soft markers are minor ultrasound abnormalities, considered variants of normal, which do not constitute a structural defect. The most commonly studied soft markers of aneuploidy include nuchal pad edema, echogenic bowel, echogenic intracardiac focus, choroid plexus cyst, mild fetal pyelectasis, ventriculomegaly. Ultrasound soft markers are also common among karyotypically normal fetuses, it may not be clear when genetic amniocentesis should be offered. The risk adjustment secondary to presence of markers, and the issue of which markers are most significant remain controversial. Material and method: This study analyses the prevalence of numerical fetal karyotype changes among the genetic amniocentesis performed in 78 pregnant women referred Life Memorial Hospital for ultrasonographic soft markers of aneuploidy in second trimester. Result: 11 of these were karyotypically abnormal. 4 of abnormal result were among patients with choroid plexus cysts, 3 - with echogenic intracardiac focus- left ventricle ; 2 – with echogenic bowel, 1 – with ventriculomegaly and 1- with nuchal pad edema. Conclusions: Ultrasonography cannot be used to diagnose aneuploidy.The management of the soft markers is different for each of them, but the detection of any abnormal ultrasound finding should prompt an immediate detailed evaluation by an experienced sonographer. If there is more than one abnormal finding on ultrasound or the patient is over 35 years of age an amniocentesis should be recommended to rule out aneuploidy. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 95 P82. CORRELATIVE STUDY OF HLA ANTIGENS (A, B AND C LOCI) IN SOLID BRAIN TUMORS A.Rodewald1, A.Koenig2, Angelika Kroll1, Georgeta Cardos3 1 Institute of Human Biology-Anthropology of the University of Hamburg-Germany 2 Medical University UKE-Hamburg; present Neurological Hospital Karlsruhe-Germany 3 National Institute of Pathology „Victor Babes” Bucharest-Romania The Human Leukocyte Antigens (HLA) or the Major Histocompatibility Complex (MHC) is very important in immunological reactions, being the principal antigenic barrier to organ transplantation. Genes codifying HLA antigens are localized on human chromosome 6, region p21-22 (6 HLA loci) and are known as susceptibility genes in different diseases with genetic determinism. In the last few decades, several studies showed statistical association between different types of HLA antigens and both some diseases with genetic background (such as rheumatological, infectious, autoimmune or endocrine diseases) and tumors and cancer types. The aim of our study was to find (if any) correlation between susceptibility to brain tumors and certain HLA antigens or haplotypes in patients. HLA antigens were investigated in 98 German patients of Caucasian origin with solid brain tumors (39 meningiomas, 49 gliomas and 10 patients with brain metastases: neuriomas, oligodendrogliomas and glioblastomas) and RR index (RR=”relative risk”) was counted. HLA antigens were determined in A (18 antigens), B (30 antigens) and C (5 antigens) loci. The control sample consisted of 2163 of German healthy people. Biostatistical analysis revealed high significant positive correlations, patients with solid brain tumors showing more high frequencies of Cw1, Cw2, B21 and B37 antigens than healthy people. Significant positive corelation was also found for Cw3, Cw4,Cw5, A2,A30/31,A32 , B16 and B18 antigens. Our results were found to be in accordance with another similar study on 65 patients with brain tumors, made by University of Medicine in Halle, Germany, sugessting that certain HLA antigens and/or hapotypes are significant associated with a high risc of appearance and development of several brain tumors. P83. EHLERS – DANLOS SYNDROME – DISCUSSIONS CONCERNING DIAGNOSIS AND MANAGEMENT Alexandra Rusu1, Mihail Volosciuc2, Lacramioara Butnariu2, 3, Cristina Rusu2, 3 1 „Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania – student 2 Medical Genetics Center, „Sf. Maria” Children’s Hospital, Iasi, Romania 3 Medical Genetics Department, „Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania Ehlers Danlos syndromes are a genetically, biochemically and clinically diverse group of heritable connective tissue conditions. Once classified as 11 separate disorders, a new nomenclature has significantly reduced this number. The Ehlers – Danlos syndromes have a combined estimated prevalence of 1/ 5.000. Features common to all forms include joint laxity and dermal findings. We present 3 cases diagnosed with Ehlers danlos syndrome in Iasi Medical Genetics Center to illustrate different forms of the disorder and to discuss in detail literature data concerning clinical picture, pathogenic mechanism and especially management. Case 1 – female, 34 years old, presents joint laxity, marked skin abnormalities (soft and lax skin, bruises at minimal trauma, „cigarette paper” and pigmented scars), moderate mental retardation and seizures. Personal history reveals that birth was produced before term, the child being resuscitated. Family history is negative. The patient has been diagnosed with Ehlers Danlos classical type (I). 96 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Cases 2 and 3 are sibs diagnosed with Ehlers Danlos kyphoscoliosis type ( VI ). Both present blue sclerae, soft and lax skin, muscle hypotonia, marked kyphoscoliosis and joint laxity. Osteopenia determined in time pathologic fractures. In conclusion, we present 3 cases of Ehlers Danlos syndrome to illustrate different forms of the disorder and to discuss in detail literature data concerning clinical picture, pathogenic mechanism and especially management. P84. CLINICAL VARIABILITY IN VELO- CARDIO- FACIAL SYNDROME Cristina Rusu1,2, Mihail Volosciuc1, Monica Panzaru1,2, Constantin Iordache3, Florentina Cucer 4, Vlad Gorduza 2 1 Medical Genetics Center, „Sf. Maria” Children’s Hospital, Iasi, Romania 2 Medical Genetics Department, „Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania 3 st 1 Pediatrics Clinic (Cardiology), „Sf. Maria” Children’s Hospital, Iasi, Romania 4 th 4 Pediatrics Clinic (Nephrology), „Sf. Maria” Children’s Hospital, Iasi, Romania DiGeorge syndrome associates immune defect (duet o hypoplastic thymus), cono- truncal heart defects and mild dysmorphic face (long nose with hypoplastic nostrils, high/ cleft palate/ velopalatine insufficiency, small ears). Velo – cardio – facial syndrome associates heart defects (frequently cono- truncal), cleft palate/ velo- palatine insufficiency and learning difficulties. By international consensus it has been established that both syndromes represent in fact one and the same entity, clinical picture being very variable. The disorder is due to a microdeletion of the 22q11 region, most of the cases having autosomal dominant inheritance and only rarely cases being sporadic. We present 3 cases diagnosed with velo- cardio- facial syndrome in Iasi Medical Genetics Center to illustrate very variable expressivity of this disorder and to underline the importance of both multidisciplinary evaluation and detailed history. Case 1 – male, 3 years 11 months old, presents mild dysmorphic face, nasal speech, hipermobile joints, hypospadias, mild developmental delay and frequent respiratory infections. Investigations revealed left renal agenesis and tortuous blood vessels, but no heart defects. Normal karyotype. FISH test confirms 22q11 deletion. Family history is negative. Case 2 – male, 5 months old, presents mild dysmorphic face, postaxial polydactyly in both hands (clinical feature not cited in the literature), mild developmental delay. His mother presents dysmorphic face, nasal voice and mild mental retardation. Investigations revealed a minor heart defect in the child. Both mother and child had normal karyotype. FISH test confirmed 22q11 deletion in both mother and child. Family history is positive on maternal side for heart defects and on paternal side for polydactyly. Case 3 – female, 17 years old, presents mild dysmorphic face, nasal speech, slender fingers and insulin- dependent diabetes. Investigations revealed a heart defect. Normal karyotype. FISH test confirms 22q11 deletion. Family history is negative. In conclusion, we appreciate that clinical picture in velo- cardio- facial syndrome could be very variable and sometimes basic diagnostic criteria could be missing. For this reason, careful multidisciplinary evaluation of both the patient and parents is crucial to deterct minor forms of the disorder. Moreover, in cases with suspicion of diagnosis (especially those with positive family history), FISH test is essential for the diagnosis. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 97 P85. KABUKI SYNDROME – IASI MEDICAL GENETICS CENTER’S EXPERIENCE Cristina Rusu1,2, Mihail Volosciuc1, Adriana Sireteanu2, Elena Braha1,2, Lacramioara Butnariu1,2, Constantin Iordache3, Florentina Cucer4 1 Medical Genetics Center, „Sf. Maria” Children’s Hospital, Iasi, Romania 2 Medical Genetics Department, „Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania 3 st 1 Pediatrics Clinic (Cardiology), „Sf. Maria” Children’s Hospital, Iasi, Romania 4 th 4 Pediatrics Clinic (Nephrology), „Sf. Maria” Children’s Hospital, Iasi, Romania Kabuki syndrome ( Kabuki makeup syndrome, Niikawa-Kuroki syndrome) is a multiple-malformation syndrome characterized by distinctive facial features (long palpebral fissures, eversion of the lower lateral eyelid, arched eyebrows with sparse or dispersed lateral one third, depressed nasal tip, and large, prominent ears), mental retardation, and postnatal growth deficiency. The etiology is unknown, however, it is suspected to be an autosomal dominant disorder. Malformations involving almost every system have been described but the most common are heart (40-50%) and renal defects (25%). We present the clinical study of 22 patients diagnosed with Kabuki syndrome in Iasi Medical Genetics Center, to show the variable aspect of the disorder, to evaluate the most common and suggestive features for the diagnosis, as well as to demonstrate the increased frequency of renal defects in Romanian patients and to underline the importance of early diagnosis for a correct management. Clinical diagnosis was based on diagnostic criteria mentioned in the literature. Only 2 cases have been familial, but in 3 other cases one of the parents presented mild Kabuki syndrome features. Typical dysmorphic face and fetal pads were present in all cases. Developmental delay/ mental retardation was present in 21/22 cases, moderate/ severe mental retardation being recorded in 6 cases. Postnatal short stature was present only in 6/22 children, but some of the patients were quite young and have to be followed. Heart defects affected 10/22 cases, whereas renal defects affected 9/22 cases. The entire panel of cardio-vascular and reno-urinary abnormalities will be presented. The most severe defects associated have been renal (agenesis, nephrotic syndrome, severe vesico-ureteral defects), thus influencing the prognosis of the child. The most severe 3 cases will be presented in detail. In conclusion, we appreciate that renal defects and kidney function should be checked in every patient with the suspicion of Kabuki syndrome, early diagnosis being very useful for appropriate treatment and complication prevention. P86. OCCURENCE OF HIGH TITER INHIBITORS IN A PATIENT WITH MILD HEMOPHILIA A – ANALYSIS OF RISK FACTORS M. Serban1, L. Pop1, D. Mihailov1, E. Ursu1, M. Puiu1, D. Savescu2 1 University of Medicine and Pharmacy ``Victor Babes`` Timisoara, Romania 2 Children`s Emergency Hospital ``Louis Turcanu``, Timisoara, Romania Background Inhibitor development is the most serious complication of modern hemophilia care with an incidence of 25-30% in patients with severe hemophilia A and a prevalence of ~6% in the total hemophiliac population. The cumulative incidence of inhibitors in patients with mild hemophilia A has been reported to be 3 to 13% by the age of 33 in some populations. Case report A 42-year-old male with mild hemophilia A (FVIII:C=13%) developed aproximatelly 40 days after an orthopedic surgery high titer inhibitors (51.2 Bethesda units) with a drop of his residual FVIII level to 0.4%. He experienced spontaneous severe, life-threatening bleedings: large subcutaneous, cutaneous bleeding, urogenital bleeding and a massive right iliopsoas muscle hematoma (~2.000 cc) with secondary neuropathy of the right thigh. Potential risk factors for inhibitor development in our patient would include the recent orthopedic surgery with intensive exposure to plasma derived FVIII (Beriate P), use of continuous infusion for the first 4 days postsurgery, 98 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 and the use of different factor VIII concentrates for haemorrhagic episodes experienced until the age of 42, and during a previous orthopedic surgery performed approximately one year before the appearance of inhibitors. The patient has no evidence of family history for inhibitors. Analysis of exon 11 of the F8 gene showed that the causative mutation of mild hemophilia in our patient has been identified as a c.1648C>T* substitution. This base change predicts the replacement of the native Arginine residue at codon 550 with a Cysteine (p.Arg550Cys*). This mutation has multiple entries on the Hemophilia A mutation database and is associated with mild phenotype. Analysis of 3 non F8 genetic factors (TNFα-308G>A, CTLA-4-318C>T, and IL-10 345 bp VNTR) associated with the formation of FVIII inhibitors showed heterozygosity for the high risk TNFα-308A variant, and a lack of `high risk` allele at the other gene loci. The treatment of his severe bleedings, appeared after development of high titer inhibitors, consisted of repeated administration of rFVIIa, Corticotherapy (Prednisone 1 mg/kg/day), DDAVP, and transfusions of packed red blood cells. Avoidance of FVIII concentrates and the above mentioned treatment resulted in undetectable inhibitors levels 7 weeks after therapy start, with complete restoration of his previous FVIII level. Unfortunately, due to the extremely high costs, we were not able to perform ITI. For fear of anamnesis, the patient will require rFVIIa or aPCC upfront for bleeds and for any further surgery. Conclusion The development of inhibitors in mild hemophilia A is generally considered uncommon and less frequent than in severe haemophilia A, but the increased use of FVIII products have made this complication more frequent than previously thought. The inhibitors occur later in life than in severe haemophilia A and their development is generally associated with a change in bleeding pattern. There is a distinct risk of inhibitor development in mild hemophilia A. Genetic factors are strongly involved. Intensive replacement therapy appears to be a risk factor for the development of inhibitors in mild hemophilia A, but other non-genetic risk factors may also be present. The optimal treatment regimen for the eradication of inhibitors in mild hemophilia A and for the treatment of bleeding episodes have yet not been established. The return to baseline FVIII level, the resurgence of a mild bleeding state, and the disappearance of inhibitors should be the goals in treating mild hemophilia A with inhibitors. P87. PARTIAL MONOSOMY 9P22-PTER - CLINICAL AND CYTOGENETIC STUDY A. Sireteanu1, M. Voloşciuc2, E. Braha1, V. Gorduza1, C. Rusu1 1 Department of Medical Genetics, ”Gr. T. Popa” University of Medicine and Pharmacy, Iasi, Romania 2 Medical Genetics Center, „Sf. Maria” Children’s Hospital, Iasi, Romania The distal region of the short arm of chromosome 9 is of interest to scientists concerned with sexual development and the phenotype of patients with 9p deletion syndrome, characterized by trigonocephaly, dysmorphic facial features, congenital heart defects, and abnormal genitalia. In most cases, the breakpoint is located at band 9p22 and the deletion is de novo. Specific genes responsible for various aspects of the phenotype have not been identified. We present 3 patients aged 4 months-4 years diagnosed with partial monosomy 9p in order to compare the phenotype of the patients with the cases reported in the literature and to highlight some particularities. Physical examinations revealed facial dysmorphism, genital abnormalities and mental retardation in all patients. The family history was negative. Investigations showed congenital heart abnormalities in 2 patients and partial optic nerve atrophy, which has not been reported before, in one patient. Cytogenetic analysis showed a deletion of the short arm of chromosome 9 with the breakpoint being located at band 9p22 in all cases. The chromosomal analysis for parents is normal in one case, the other two being in progress. In conclusion, we present three cases with monosomy 9p to discuss the main clinical signs and the problems posed by genetic counseling and individualized management. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 99 P88. FETAL GENDER DETERMINATION BY ANALYZING THE FREE FETAL DNA CIRCULATING IN MATERNAL BLOOD - POSTER Adriana Stan1, Cristina Dragomir1, Emilia Severin2, Lorand Savu1 1 Genetic Lab, Bucuresti, Romania 2 Facultatea de medicina si farmacie „Carol Davila”, Bucuresti, Romania Prenatal diagnosis in Romania is exclusively based on standard invasive techniques for fetal cells sampling such as chorionic villi sampling, amniocentesis and cordocentesis. These sampling techniques present a 1-5% risk to induce pregnancy loss or amniotic fluid and blood leakage, infections, fetal distress (respiratory distress, benign bone deformity) and also maternal anxiety. Considering the development strategy of prenatal diagnosis based on minimizing the risks, rapidity and certainty, this field evolution become more dynamic; for the last 10 years the main objective in research centers in Asia, Europe and USA was the noninvasive approach. The noninvasive approach of prenatal diagnosis was based on the free fetal DNA and fetal cells circulating in maternal plasma; using this procedure the risks for which are submissive both the fetus and the mother, are therefore eliminated. Considering the fact that there are no other prenatal diagnosis methods for the first trimester of pregnancy other then chorionic villi sampling and early amniocentesis this new approach is the only viable option. The aim of this study was to determine the fetal gender by analyzing the free fetal DNA in maternal plasma, because the fetal gender is important in cases at risk for X-linked disease, both dominant and recessive. Knowing the fetal gender in an early stage of pregnancy will reduce the number of invasive procedure for prenatal diagnosis tests; will be selected the male fetuses at risk for recessive X-linked disease and female fetuses at risk for dominant X-linked disease when the father is affected. This new approach is preferred in „in vitro” fertilization centers where the noninvasive methods are correlated with preimplantation genetic diagnosis (PGD). The working protocol included the extraction of total plasmatic DNA from 10ml of peripheral maternal blood withdrawn in vacutainer with EDTA (QIAamp DSP Virus kit, QIAGEN) in accordance with the SAFE Network of Excellence protocol (Special Non-invasive Advances in Fetal and Neonatal) optimized by Legler et al.,2008. DNA sample is analyzed in same day by monoplex and multiplex classical PCR and Real-Time PCR techniques, using primes selected from scientific literature; primers were designed for chromosome’s Y genes - DYS14 si SRY. We used in the PCR reaction the housekeeping gene beta globin as extraction control. We conclude that this noninvasive method for fetal gender determination can be used as a screening procedure in high risk pregnancies for X-linked diseases thanks to the possibility to detect free fetal DNA in maternal blood. P89. DISMORFISM PHENOTYPE WITH MINOR AND PARTIAL AGENESIS OF BODY CALOS Dana Metea Stefanescu1, Liviu Pop1, Cristina Dragomir1, Simona Farcas2 1 „Department II Pediatrics”, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania. 2 „Department of Health Hydrogenated”, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania. Introduction: Total or partial agenesis of the body calos is a malformation frequency nervous system. Incidence is difficult to estimate because many asymptomatic forms discovered by chance. Diagnosed in the neonatal few observations reported in the literature, asymptomatic forms are exceptional. Materials and Methods: Report the case of a newborn, female, admitted to Pediatric Clinic II Timisoara, particularly for cranial facial phenotype. Study methods used are: family investigation, history of physiological, clinical examination on equipment and systems, common and 100 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 complementary paraclinical ( etiological investigation to exclude a chronic intrauterine infection, electroencephalogram, fundus, karyotype, ultrasound transfontanelar). Results and Discussions: Clinical examinations showed associating with cranial facial dismorfisme suggestive (hipertelorism, microphthalmia, ear implanted below) with axial hipotonia. Transfontanelar ultrasonography was the one who chepped the diagnosis of partial agenesis of body calos. Conclusions: 1. Abnormal growth of the fetal biparietal diameter a neonatal hypotonia or dismorfic cranial facial syndrome indicate the likelihood of agenesis of the body calos and can association with brain malformation or extracerebrale. 2. Malformations prognosis depends not on whether they fully or partially associated malformations but. 3.Imaging method of choise as required by endovaginala ultrasound in antenatal, transfontanelar ultrasonography in neonatal, infant magnetica nuclear resonance after 4 mouths of age. Key words: agenesis of body calos, particular phenotype, cranio- facial dismorfism. P90. DELINEATION OF CHROMOSOMAL ANOMALIES IN METAPHASE AND INTERPHASE CELLS BY FISH AND ARRAY CGH TECHNIQUES Monica Stoian¹, Valerica Belengeanu¹, Eli Ormerod2, Nicoleta Andreescu¹, Simona Farcas¹, Sorin Iurian3 ¹ Department of Medical Genetics, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania 2 Department of Medical Genetics, Oslo University Hospital, Oslo, Norway 3 Pediatric Clinic, Pediatric Clinic Hospital Sibiu, Romania Detection of balanced or unbalanced chromosomal rearrangements and the cryptic mosaicism is most efficiently achieved by a combination of classical cytogenetic methods with FISH techniques as well as array CGH. We have chosen to focus on four cases in whom the cytogenetic abnormalities were principally described by G-banded karyotyping for which FISH analyses and aCGH increased the chance of positive diagnosis for patients. First case, we report on the characterization of a de novo double translocation involving chromosomes 1 and 13 and chromosomes 5 and 11 in a male patient. Another patient had neurodevelopment delay and facial dysmorphism with additional material on distal long arm of chromosome 2 on conventional cytogenetic G-banded metaphases. FISH analysis using chromosome 2 telomere and WCP 4 probes and later array CGH analysis helped to delineate the chromosomal imbalance. The third patient is a 35 years old female investigated for primary amenorrhea, and because of the borderline abnormal clones revealed by chromosomal analysis, FISH was performed using CEP X probe, and revealed the karyotype: 46,XX(90,5%)/45,X(7,5%)/47,XXX(2%). The fourth patient is a male infant referred for Down syndrome suspicion. Surprisingly the conventional cytogenetics revealed 10 metaphases with 46,XX and one with 46,XY. FISH analysis using CEP X and SRY probes evaluating 235 nuclei and metaphases set the karyotype: 46,XY(87%)/ 47,XYY(6%)/47,XXY(5%)/46,XX(2%). Molecular cytogenetic analyses revolutionized the cytogenetic testing available for patients with learning disabilities who have ‘‘chromosomal’’ phenotypes with dysmorphic features and other anomalies. Molecular testing methods such as FISH, aCGH will continue to play an important role in identifying the exact chromosome complement. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 101 P91. PHENOTYPE AND CYTOGENETIC CHANGES IN A TRISOMY 8 IN MOSAICISM CASE FROM NEWBORN TO INFANCY Monica Stoian¹, Valerica Belengeanu¹, Nicoleta Andreescu¹, Simona Farcas¹, Cristina Popa1, Marioara Boia² ¹ Department of Medical Genetics, University of Medicine and Pharmacy ”Victor Babes”, Timisoara, Romania ² Department of Neonatology, „Louis Turcanu” Children Hospital, University of Medicine and Pharmacy „Victor Babes”, Timisoara, Romania Mosaic trisomy 8 is a relatively common chromosomal abnormality, which shows a great variability in clinical expression. The more discriminating findings for this condition are skeletal anomalies and specific plantar furrows. The present report describes phenotypic signs and cytogenetical characteristic in a trisomy 8 in mosaicism case diagnosed at birth and revaluated at the age of six weeks, age of 14 months and 5 years. At birth the proband was noted to have a particular facial appearance with telecanthus, upslanting palpebral fissures, notched nasal tip, cleft upper alveolar ridge, bifid tip of tongue, grooved uvula, low-set, dysplastic ears and short and wide neck. On both hands deep palmar creases and deep plantar grooves bilaterally were observed. The perinatal period was complicated with seizures and apneic episodes, and transfontanellar ultrasound revealed absence of corpus callosum which was confirmed by MRI. Chromosome analysis on peripheral blood lymphocyte culture revealed trisomy 8 in 12 metaphases of a total of 50 examined metaphases. FISH analysis on cultured blood lymphocytes with a centromeric probe of chromosome 8 (CEP 8-Spectrum Orange, AbbottVysis) showed 3 hybridization signals in 25 % of cells (120/ 492 nuclei). At six weeks revaluation she had soft dark hair on the forehead extending to the eyebrows and choanal atresia. We reassessed the patient at the age of 14 months and she had a slender body habitus with mild limitation of movement in the right ankle, and developmental milestones were grossly delayed. The radiography of the knees performed at 38 months revealed absence of patella. Our patient followed a specific rehabilitation program. She was revaluated at 5 years and she had tall stature, mild pectus excavatum, mils lumbar lordosis, limitation of motion in interphalangeal joints and disproportionately delayed language. Interphase FISH revaluation revealed a decrease of proportion of the trisomic cells (20%). We assess that our patient has a good prognosis regarding neurodevelopment evolution. P92. MEDICAL REHABILITATION FOR LIFE QUALITY IMPROVEMENT IN A CASE WITH ARTHROGRYPOSIS Dorina Stoicanescu1, Mariana Cevei2, Valerica Belengeanu1, Lucia Stoican3, Monica Stoian1, Nicoleta Andreescu1 1 University of Medicine and Pharmacy „Victor Babes”, Department of Medical Genetics, Timisoara, Romania 2 University of Oradea, Faculty of Medicine and Pharmacy, Oradea, Romania 3 University of Medicine and Pharmacy „Victor Babes”, Department of Anatomy, Timisoara, Romania Arthrogryposis is a is a rare non-progressive congenital disorder characterized by multiple joint contractures. Frequently, the contractures are accompanied by muscle weakness, which further limits movement. We present the case of a girl of 4.5 year-old with multiple congenital defects: arthrogryposis involving bilateral hip, knee and ankle joints, together with sacral agenesis with lombar dysmorphism. At the age of 2 years, surgery for the equinovarus deformities and for left genu flexum was performed. As there is no reversal of this condition, is essential that individual quality of life can be greatly improved. Each person will respond differently, and will have different needs, thus a combination of therapies is needed. 102 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Medical rehabilitation tries to maximize independent function. The main goals were increasing the muscle tonus of upper-limbs, increase the rate of motion of the joints, establishment of stability for ambulation, learning different schemes of movement according to her needs, obtaining of a functional independency. The patient followed a complex rehabilitation program with hydrokinetotherapy, electrotherapy, massage, occupational therapy, psychological counseling. The therapies were successful, after 10 weeks an improvement of the moving capacity and of the transfer in orthostatism with minimal external assistence, with the obvious increasing of the patient’s satisfaction, were noticed. In order to fulfil our objectives the child will be hospitalized every two months for functional evaluation and for continuing the physical therapies. P93. IMPROVED FUNCTIONAL PROGNOSIS AFTER MEDICAL REHABILITATION IN A CASE WITH CRI DU CHAT SYNDROME Dorina Stoicanescu1, Mariana Cevei2, Valerica Belengeanu1, Lucia Stoican3, Simona Farcas1, Cristina Popa1 1 University of Medicine and Pharmacy „Victor Babes”, Department of Medical Genetics, Timisoara, Romania 2 University of Oradea, Faculty of Medicine and Pharmacy, Oradea, Romania 3 University of Medicine and Pharmacy „Victor Babes”, Department of Anatomy, Timisoara, Romania Cri-du-chat syndrome is a relatively rare chromosomal disorder with an estimated incidence of 1 in 20000 to 50000 newborns, resulting from loss of varying lengths of the short arm of chromosome 5. We present a case report on the rehabilitation treatment of a patient with this condition, aged 18 months, who received multidisciplinary treatment and precocious stimulation. The baby was born from healthy, young parents, with a birth weight of 2250 g. At examination this patient presented cranio-facial dysmorphy, transverse flexion creases, important growth and psychomotor retardation, axial hypotonia and hypotrophy, bilateral varus equine, positive Babinski sign, lively deep tendinous reflexes, hypertrophic cardiomyopathy, unclosed sagittal and lambdoid sutures. The child had a happy aspect, was able to sit and to roll, but was not able to stay in the quadruped position. Rehabilitation program started at 15 months and consisted of hydrokinetotherapy in the pool for 15 minutes each day, physiotherapy, re-education of motor control stages, tonisation of paravertebral, quadriceps, gluteal muscles, paraffin therapy for the legs, massage. Occupational therapy, coordination exercises and speech therapy were also performed. The evolution of the child was very good, he was able to stand with assistance after the first rehabilitation period of three weeks. Improvements in management of patients with this disorder by a multidisciplinary team, with the application of early rehabilitation programs increase psychomotor development, improve autonomy and finally lead to a better social adaptation. P94. RARE VARIANT E19A2 BCR-ABL FUSION TRANSCRIPT IN TYPICAL CHRONIC MYELOID LEUKEMIA: CASE REPORT Andreea Tutulan-Cunita1, Sorina Mihaela Chirieac1, Gabriela Mocanu2, Catalina Luca3, Marieta Costache3, Agripina Lungeanu1, Aurora Arghir1 1 „Victor Babes” National Institute of Pathology, Bucharest, Romania 2 Coltea Clinical Hospital, Clinic of Hematology, Bucharest, Romania 3 University of Bucharest, Faculty of Biology, Department of Biochemistry, Bucharest, Romania The landmark of chronic myeloid leukemia (CML) is the Philadelphia (Ph) chromosome, that originates in the reciprocal translocation t(9;22)(q34;q11) and is the first chromosomal aberration linked to a specific neoplasm in humans. The main consequence of this rearrangement is the Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 103 apparition of the BCR/ABL hybrid gene, encoding for a constitutively active tyrosine kinase central to the malignant process. While the breaking points inside ABL gene fall within a ∼300 kb segment at the 5’ end of the gene, there are 3 breakpoint cluster regions inside BCR, therefore resulting a range of fusion transcripts occurring with different incidences. Here, we report clinic, cytogenetic and molecular data regarding a new case of CML with the rare e19a2 transcript. P95. SEVERE MICROGNATHIA, QUESTION MARK EARS – A RELATIVELY RECENT SYNDROME, AURICULO-CONDYLAR SYNDROME M. Volosciuc, Elena Braha, Aurica Rugină Medical University Iasi Auriculo-condylar syndrome (ACS) is an autosomal dominant disorder of first and second branchial arches, with complete penetrance and great intra- and inter-familial phenotypic variation; ACS is characterized by dysplastic ears (‘question mark ears’), micro-retrognathia, microstomia, prominent cheeks. The psyhological development is normal. The genetic defect for ACS is still unknown, the gene could be probably map in 1p21.1-q23.3 region. We describe a pacient CPN, a 6 months old girl, with a highly suggestive dysmorphic face (severe micro-retrognathia, round facial appearance, Prominent cheeks, dysplastic ears with abnormal lobule). The patient is the one child of a young and non-consanguineous couple. The mother revealed the same facial malformations as the child. We discussed a possible differential diagnosis. We underline Good knowledge importance of the clinical features and syndromes of first and second pharyngeal arches for a correct diagnosis, clinical assessment anf genetic counsellig. P96. THE KLIPPEL FEIL SYNDROME: DIFFICULTIES OF GENETIC COUNSELING Andra But1, Corina Duncescu1, Adela Emandi-Chirita1, Loredana Covle1, Maria Puiu1,2 Narcis Dobre 1 „Louis Turcanu” Emergency Hospital for Children, Timisoara 2 „Victor Babes” University of Medicina and Pharmacy, Timisoara Klippel-Feil syndrome (KFS) is a rare congenital disease. It is characterized by the abnormal union or fusion of at least two of the seven cervical vertebrae. Some of those affected may have a short neck, restrictive head and neck movements. In some people with KFS, the disease may be associated with other physical problems: scoliosis, hearing problems, facial or head malformations (craniofacial area) or modified structures of the heart (congenital heart problems). In some cases, neurologic complications may occur. We present a 7 year old boy with typical manifestations of the disease (short neck, low mobility of the cervical area, difficulties in tilting the head back, front or rotating it, pain in the neck area, including head, neck, shoulder blades, stiffness and muscle tension of the neck and shoulders). Radiological cervical fusion was found (C0-C1) and the spaces between the unfused segments of the cervical spine are extremely mobile explaining muscle tension, pain and other neurological problems. This increased mobility may entail risks of paralysis or even death in case of a minor shock (car accident, riding). Given the complex etiology of the syndrome (autosomal recessive transmission, autosomal dominant, with complete or incomplete penetration, X-linked recessive, disruptive, vascular or other type of trauma occurring during embryo development), the management of the case involved considering all the etiological variants of the syndrome. After Clarke et al. the SGM1 gene is responsible for some types of KFS. The lack of technical means prevented the exclusion of this case. In some KFS family types, Clarke et al. assumed a pericentric inversion of chromosome 8: inv (8) (q22.2q23.3). This was not our case, because the karyotype was normal. Family history was negative, thus the environmental causes we considered were excluded. 104 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 In conclusion, until now, the interdisciplinary approach didn’t allow an exact framing of the KFS type, making it impossible to grant appropriate genetic counseling. Our patient remains in observation, while we seek an international collaboration for establishing etiology. P97. HUNTER SYNDROME: THE HISTORY OF A DIAGNOSIS Maria Puiu1,2, Cristian Jinca1,2, Laura Pop1,2, Margit Serban1,2 1 „Louis Turcanu” Emergency Hospital for Children, Timisoara 2 „Victor Babes” University of Medicina and Pharmacy, Timisoara Mucopolysaccharidosis (MPS) are a group of lysosomal storage diseases, each caused by an inherited deficiency of an enzyme involved in the degradation of glycosaminoglycans. This group includes MPS type I- Sheie syndrome, type II - Hunter syndrome, type III - Sanfilippo syndrome, type IV- Morquio syndrome, type VI- Maroteaux-Lamy syndrome, type VII- Sly syndrome, each with several subtypes with a specific enzyme deficiency and distinctive clinical picture. Often, the clinical picture, especially the first clinical signs, are misleading and may suggest a particular form of MPS that will be infirmed by enzymatic tests. Correct etiological diagnosis raises many technical problems and is hardly accessible. In its absence, the existing therapeutic options may not be used and appropriate genetic counseling is impossible. In the case we present, San Filippo syndrome was suspected. The first laboratory studies were sent through Bioclinica Laboratories, with a financial effort made by the parents to the laboratory of Heidelberg University. The results supported the diagnosis of San Filippo syndrome, but they were accompanied by the request of a second blood and urine sample. The patient was transported by his family in Germany, where tests for MPS type I-III and Gaucher syndrome were performed, establishing the final diagnosis as Hunter syndrome. The conclusion is that for the high accuracy diagnosis of very rare diseases, for which we are not technically equiped, with the possibility of differentiating the diagnosis, accurate classification of different types of MPS is hazardous. P98. LOBSTEIN’S DISEASE – CASE REPORT Boia ES1, Marioara Boia1, Popoiu MC1, David VL2 1 University of Medicine and Pharmacy „Victor Babes” Timisoara, Romania 2 Emergency Children’s Hospital „Louis Turcanu” Timisoara, Romania Lobstein’s disease or Osteogenesis Imperfecta (OI) is a rare is a systemic heritable condition, with an estimated prevalence of 1/10,000. The cardinal manifestation of OI is severe bone fragility due to a primary defect of the type I collagen. In approximately 90% of individuals with OI, there is a mutation encoding the type I collagen. There are eight types of OI, type II is the most severe and type I and IV are the most common. We report the case of a male newborn child with OI. The boy had multiple bone fractures during the intrauterine life. The mother had no follow-up until the 9th month when the disease was diagnosed. Postnatal genetic consult identified a type II OI. This is a particular case because even if the prenatal diagnosis was practically absent and severe bone deformities were present at birth, the child survived trough the neonatal period. Usually, most of the children with this type of OI die during first few weeks after birth due to poor circulatory/pulmonary conditions resulting from ribs and vertebral deformations. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 105 P99. MUSCULOSKELETAL DISORDERS IN PATIENTS WITH PRADER WILLI SYNDROME Popoiu MC1, David VL2, Boia ES1, Maria Puiu1 1 University of Medicine and Pharmacy „Victor Babes” Timisoara, Romania 2 Emergency Children’s Hospital „Louis Turcanu” Timisoara, Romania Prader-Willi (PWS) syndrome is a genetic disorder with an incidence of 1 in 25000–30000, characterized by severe hypotonia and feeding difficulties in early infancy, followed later by excessive eating and gradual development of morbid obesity. The management of patients with PWS requires the attention of many professionals. Obesity is the major feature of PWS and exacerbates all the other health issues. Musculoskeletal disorders like scoliosis, hip dysplasia, and lower limb alignment, are universal features of PWS. In early childhood, hip dysplasia is a frequent finding in patients with PWS. The management of hip dysplasia includes orthopedic measures and in very rare cases surgery. Scoliosis is the main musculoskeletal issue, affecting up to 80% of the PWS patients. Scoliosis is a major concern for the orthopedist and choosing the proper treatment for these patients is challenging job. Nonsurgical treatment is the primarily option for PWS patients with scoliosis. Regular physical exercises corroborated with physiokinetotherapy besides improving the spinal curve increase the muscle tonus and help maintaining body weight, and can control the scoliosis in up to 90% of the patients. Spinal surgery has a higher rate of complications than general populations and the death rates have been found to be higher than in patients with Idiopathic Scoliosis. Osteoporosis is affecting most of the patients with PWS and leads in junction with the weight excess to severe fractures. Other musculoskeletal manifestations include limb malalignment like genu varum deformity, foot and hand abnormalities like flat foot. In conclusion, there is a high prevalence of musculoskeletal disorders in PWS, especially spinal deformity and osteopenia related fractures often complicated by obesity, osteopenia, growth hormone treatment, defiant behaviors, and diminished pain sensitivity charachteristic for the PWS. For these reasons, the pediatric orthopedist should play an important part in the management of the patients with PWS and periodic systemic clinical and radiographic evaluations should be conducted. P100. PRADER WILLI SYNDROME – FROM RESEARCH PROJECT TO MULTICENTRIC APROACH M. Puiu1, D. Dan2, M. Serban1, M. Gafencu1, G. Anton3, N. Cucu4 1 Universitatea de Medicina si Farmacie „V. Babes”, Timisoara 2 Asociatia Prader Willi din Romania 3 Universitatea Bucuresti 4 Institutul de Virusologie, Bucuresti The purpose of this research is to establish the new hypotheses that are responsible for PraderWilli/Angelman syndromes. Our study has as objectives the implementation of new molecular methods for genetic/epigenetic investigation and establishment of national centers with high expertise in approaching the two syndromes, the rare genetic diseases that will develop educational reference and release centers. We also aim to evolve efficient partnership with patients associations through specific modalities like dialogue. The power of these associations will propel the research, will inform the patients and will respond to civil society questions. The study will establish international collaboration and partnerships with researchers having similar scientific interest, establish partnerships with PWS-Organizations, IPSWO, research groups from each country aiming financial support on programs that intend to stimulate collaboration between specialists, researchers and nongovernmental organizations. Finally, we aim to develop a multidisciplinary partnership, to build a common platform of activities for new innovative solutions in respect to rare disease needs. These new bridges of real and effective collaboration will ascertain on the national level the setting up of a solid network comprising institutions with high expertise in this domain, well connected to other national or international research networks. The results of the research will be published in well-known journals with high impact factors for enlargement of Romanian research visibility in 106 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 international domain of rare diseases. In conclusion our project aims to implement in Romania the European model of the network for rare diseases research, model adapted with success in management of these diseases. P101. SOLITARY MEDIAN MAXILLARY CENTRAL INCISOR SYNDROME – CASE REPORT Dinu Stefania, Popa Malina, Bratu Cristina, Jianu Rodica, Szuhanek Camelia, Lazar Cristina, Ogodescu Emilia Department of Paedodontics-Orthodontics, School of Dentistry,”Victor Babes” University of Timisoara, Romania Introduction: Solitary median maxillary central incisor syndrome is a unique developmental abnormality involving midline structures of the head including the cranial bones, the maxilla and maxillary dentition, the nasal airways and sometimes the brain (holoprosencephaly) together with other midline structures of the body. Materials and methods: This report describes a 6 year old girl with a unique primary maxillary incisor enveloped in the half line of the maxillary.In the first visit we performed the case history, the clinical and radiological examinations. Results: The examinations revealed a small head, hypotelorism, a small narrow nose with an arched midline of the upper lip, primary and permanent solitary median central incisor tooth, oval palate and absence of the normal midline labial frenulum. Discussions: Examination and testing may reveal a lot of characteristic abnormalities of the head and the face. Orthodontic treatment should be prompt and accurate to prevent the esthetic and functional problems. Key Words: solitary median maxillary central incisor syndrome, short stature, holoprosencephaly P102. TREATMENT MANAGEMENT OF OLIGODONTIA Dinu Stefania1, Bratu Elisabeta1, Glavan Florica1,Schiller Eleonora1, Popa Malina1, Ogodescu Alexandru1, Dinu Cristian Dorin2, Luca Magda1,Balan Raluca1 1 Department of Paedodontics-Orthodontics, School of Dentistry, Timisoara, Romania 2 * Private Practice , Timisoara, Romania Introduction: Oligodontia is defined as a congenital absence of 6 or more teeth, excluding the third molars.Dental and orthodontical treatment depends on the severity of case and generally requires a multidisciplinary approach. Purpose: The present study describes an interdisciplinary approach to the treatment of patients with oligodontia in permanent dentition.It also presents the importance of the clinical examinations and optimal options in the management of the dental rehabilitation of young patients with oligodontia. MATERIALS and METHODS: Patients with oligodontia requires an accurate clinical examination. The orthopantomogram and the study casts of both jaws need to be performed to chose the adequate way of treatment. Results: Dental treatment depends on many aspects as: severity of the case, patient age, hygiene status, treatment expectations and socioeconomic background. Our main objectives are to restore the masticatory function and to improve esthetics. Conclusions: The diagnosis and treatment of oligodontia should be prompt and accurate to prevent the esthetic and functional problems in the dentition. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 107 P103. GENETIC DETERMINISM OF CHRONOLOGIC AGING SKIN Anca Dragomirescu University of Medicine Timisoara, Faculty of Pharmacy, Department of Dermatopharmacy and Cosmetology The process of skin aging is having two main components: photo-aging (aging caused by environmental factors) and intrinsic aging (aging caused by natural and chronological factors). This last component has a deep genetic determinism, in which are involved pathways interconnecting the action of genes in inflammation, the extracellular matrix and protease activity (MMP-s proteinasis). The massive release of those enzymes is the critically sequence in collagen tridimensional network destruction. Additionally, melanine is the most important factor of UV skin rejection. More than 130 genes contribute to the control of the human skin colour, involved in the following process: - embriogenesis and surviver of melanocyte systhem, - melanosoms byogenesis, - malanogenesis process, - the transfer of melanosoms into near-by cells, - ratio eumelanine / pheomelanine constant, all live long, - melanosoms turnover. There are also epidermal factors involved in aging skin. So, expression of genes involved in lipid biosynthesis and epidermal differentiation, genes involved in the biosynthesis of cholesterol, fatty acids and sphingolipids must be take in consideration. P104. KLINEFELTER SYNDROM – CASE REPORT Rodica Urtila1, Valeria Belengeanu2, Eulalia Boceanu1, Sonia Tanasescu1, Patricia Urtila3 1 Clinic II Pediatrics Timisoara 2 Medical Genetics, University of Medicine and Pharmacy „V.Babes” Timisoara 3 Clinic II Pediatrics Timisoara Klinefelter Syndrom (KS) is a sex chromosome abnormality affecting approximately 1 in 800 males. KS is most often identified during late puberty or early adulthood, the most common indications for karyotyping are hypogonadism and infertility. We discuss a case of a 1year 2month old boy, presented at a pediatrician for a respiratory intercurrence and that we notice his particular phenotype. The karyotype of this child is 47XXY - Klinefelter Syndrom. The result was surprising, due to the fact that the patients with KS didn’t present any phenotypic abnormalities, the most overwhelming of cases being diagnosed at puberty. In this case the early diagnosis due to join the discussion and present relatively obvious phenotypic changes, intellectual development difficulties and psychosocial adjustment problems. The early diagnosis permit a substitutive treatment with testosterone which is likely to remedy the difficulties mentioned. Key words: Klinefelter Syndrom, phenothype, child 108 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P105. SURGICAL APPROACH OF A CHILD WITH CONGENITAL ANOMALIES OF THE KIDNEY AND URINARY TRACT Vasilie D. 1, 2, Puiu M. 2, Poclid I. 1, Cristina1, Mejdi R. 1 1 Pediatric Surgery Clinic, „Louis Turcanu” Emergency Hospital for Children, Timisoara 2 „Victor Babes” University of Medicine and Pharmacy, Timisoara Anomalies of the kidney and urinary tract represent 20-25% of congenital defects. The most common are the obstructive ones: pieloureteral or vesicoureteral junction stenosis (PUJS, VUJS), followed closely by posterior urethra valves (PUV). Today they are mostly found when performing an antenatal ultrasound screening. PUV is the main cause of subvesical obstruction and has an incidence of 1/5.000-1/8.000 children. Aproxomativ one third of the children with PUV will develop kidney failure during childhood or adolescence. Although very rare, PUV can cause a secondary VUJS by thickening the bladder wall. We present the case of a prematurely born infant who was diagnosed during the neonatal period with acute renal failure caused by a congenital subvesical obstruction associated with bilateral ureteral obstruction. In this case, due to a relatively late diagnosis, unilateral nephrectomy was performed and the prognosis depends heavily on the functionality of the remaining kidney. Negative prognostic factors are: a serum creatinine level under 0.8 mg% at 1 year of age, bilateral vesicoureteral reflux, urinary incontinence at the age of 5 years. Patients with PUV require long-term urologic and nephrologic follow-up, for monitoring renal function and the appearance of postobstructive bladder malfunction. Our patient’s prognosis is reserved especially because of a single, hidronephrotic kidney. In conclusion, in order to detect these malformations a multidisciplinary approach for an early diagnosis is required. This will lead to an improvement of the quality of life of these patients, but also to providing proper genetic counseling to address optimally a potentially high risk pregnancy. P106. SPECIAL NEEDS CHILDREN’S DAY Ionela Alina Moaca, Iulia Jurca-Simina, Florin Jurca-Simina Iosif Reascu, Norbert Pop, Stefan Berci, Iulia Popa, Irimia Cristina, Oana Rosca, Graziella Ecob, Adrian Juverdeanu, Carmen Dumitranoiu, Mihai Gafencu, Maria Puiu, Narcis Dobre Save the Children, Timis branch. Introduction: As a social being, man is dependent on other people. This addiction means help and possibility of communication. Every child is born with certain skills, with a potential, sometimes with talent, sometimes genius, but some are born with special needs. Those with special needs require much more attention in terms of their desires and a special treatment, too. Special Needs Children’s Day is an event full of color, joy and life, is an event for children, and we, Save the Children organization volunteers, with the Timis County Council and Children’s Emergency Hospital „Louis Turcanu” from Timisoara, tried to create a festive atmosphere for all children and especially for children with special needs. Objectives: Joy and smiles on children faces, public awareness on child rights to a decent life and a happy childhood, despite the problems caused by disease, encouraging the idea of volunteering and training team of willing volunteers to help these children. Material: Puzzles, call for children, Lego’s, short books, plasticine, balloons, diploma for each child, all purchased from funds made available of the project sponsor (County Council Timis). Methods: Recruitment of two groups of volunteers to handle these children, to be able for creative and entertaining activities with children, awarding each child, depending on talent and participation at the event. Results: The event was attended by children aged between 1 and 18 years who lives dominated Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 109 by chronic or rare diseases (hemophilia, leukemia, lymphoma, neoplasm, thalassemia, digestive, renal disease, etc.). Volunteers applied various activities with children, drew, cut, color, plasticine built, but most important, they made the children feel that is their day, and have not been forgotten just because they are hospitalized or because they are sick. One of volunteers, dressed in clown, delighted children with songs, interactive games, poetry, bestowing many gifts (diploma, puzzles, books, Lego, etc.), not in a certain order but depending on the activity of each child. Conclusion: According to the poll, the event had an important impact on participants, both children, volunteers and parents who attended and thanked us for the great opportunity offered to their children to forget the illness and treatment. The main attraction, volunteer-clown has managed to improve mood for all through the game, music, poems, ensuring that this day will remain in children memory that a beautiful celebration of their day . P107. THE IMPORTANCE OF EARLY DIAGNOSIS IN MARFAN SYNDROME Anca Popoiu1, Gabriela Doros1, Bogdana Zoica1, Popoiu MC1, David VL2, Boia ES1, M Puiu1,2 1 University of Medicine and Pharmacy „Victor Babes” Timisoara 2 Children Hospital „Louis Turcanu” Timisoara Marfan syndrome is a genetic disorder of the connective tissue, the body-wide system that provides strength and flexibility to bones, ligaments, muscles, blood vessel walls, and heart valves. It occurs in one out of 5,000 people worldwide and can develop even when there is no family history of the disorder.) Affected individuals often are tall and slender with elongated fingers and toes, skin that is stretchy, and an arm span that exceeds their body height. Many have unusually flexible joints, long and narrow faces, abnormal curvature of the spine (scoliosis), and a sunken or protruding chest. About half of all patients have vision problems; most have some degree of nearsightedness. Marfan’s patients may develop cataracts in mid-adulthood and are at higher than normal risk for glaucoma (increased pressure within the eye). Our studies have highlighted the importance of early diagnosis and Marfan syndrome. Early diagnosis allows the correct approach to the manifestations of the disease and avoid complications, sometimes fatal, disease. Marfan patients are also at higher than normal risk for respiratory problems, including sudden collapse of the lungs. As adults, patients are at increased risk of early emphysema, even if they don’t smoke. Despite all of this, most patients with Marfan syndrome who get proper medical management now have a normal life expectancy. The most troublesome aspect of Marfan is involvement of the heart and major blood vessels. Weakness of the aorta, the main artery that carries blood from the heart to the rest of the body, can result in a tear in the lining that allows blood to leak into the aortic wall, compressing its interior and blood flow through it. This is a surgical emergency that can result in sudden death if it is not quickly diagnosed and treated. Because strenuous exercise can stress the aorta, patients should avoid it. This means no contact sports or heavy lifting, although less vigorous activities such as golf, biking and swimming may be possible with physician guidance. Affected children and teens need to be checked yearly for signs of scoliosis and should also have annual eye exams by an ophthalmologist. Any nearsightedness can be corrected with glasses or contact lenses. Later in life, early detection and treatment of cataracts and glaucoma usually can head off or lessen vision problems. When detected early, Marfan complications usually can be managed by a team of specialists including a cardiologist to watch out for heart problems. The team should be headed by a physician who is familiar with all aspects of the syndrome. 110 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 P108. NEED SURGERY ON GENETIC DISEASES Jurca- Simina Florin Ioan, Jurca- Simina Iulia Elena, David Vlad, Narcis Dobre, Puiu Maria University of Medicine and Pharmacy “Victor Babes” Timisoara VitroBioChem Aim.To determine the effectiveness and the need of surgery ingeneticsyndromes (Lobstein’s disease, Turner syndrome, achondroplasia, and Marfan syndrome). Material and Method. The study included 9 children with ages between 7-17 years old, (2 females and 7 males) diagnosed with genetic disorders in Louis Turcanu Emergency Hospital for Children - Department of Pediatric Surgery and Orthopedics, Timisoara, Romania, during the period of 2 years. Results.1 patient required surgery due to Lobstein,s disease complications (fracture of the third external right clavicle) osteosynthesis with kirscanerpin was performed. 2 patients underwent interventions due to Marfan syndrome associated malformation (Genuvalgum), where performed osteotomy of the tibia. - 1 patient required an ovariectomy and anexectomydue to tumors (about 10 cm) in the left ovary. 1 patient required surgery due to infectious complications occurred (adenoidectomy and tympanostomy). 1 patient suffered a thoracoplasty intervention through Nusseprocedure which aimed to have negative repercussions upon lungs due topectusexcavatum. 1 patient remains under the supervision of cardiologists and thoracic surgeons with multiple cardiac dysfunction Conclusion.This study proves the need for surgery in these patients in order to improve life quality. P109. LIMITS OF STANDARD KITS FOR CYSTIC FIBROSIS GENETIC TESTING IN ROMANIA L.Pop1, I.M. Ciuca1, L.Tamas2, Z.Popa3, Gh. Budau4, I.Popa1 1 Pediatric II Department, UMF V. Babes, Timisoara, Romania. 2 Biochemistry Department, Molecular Genetic Unit, UMF V. Babes , Timisoara 3 National Cystic Fibrosis Centre, Clinical County Hospital, Timisoara, Romania 4 Obstetrics and Gynecology Clinic , UMF V. Babes, Timisoara Introduction: Genetic diagnosis is now available in Romania. Starting 2004 we use standard Elucigene CF 29 kit, who identify the most frequent 29 european mutations (1717-1G>A, G542X, W1282X, N1303K, ΔF508, 3849+10kbC>T, 621+1G>T, R553X, G551D, R117H, R1162X, R334W, A455E, 2183AA>G, 3659delC, 1078delT, ΔI507, R347P, S1251N, E60X, D1152H, 3120+1G>A, 2789+5G>A, 1898+1G>A, 711+1G>T, G85E, 2184delA, I148T, R560T). Aim study: evaluation of kit`s eficacy in relation to mutations identified in Romania. Methods: We evaluated 40 CF patients(pts) with typical forms, pozitive sweat test and at list one identified allele. Results: Mutation were, in order of frequency: ΔF508, G542X, N1303K, 621 + 1 G>T, I148T; they’re cumulative percentage corresponded to 17, 2% from Elucigene kit’s mutations. Genotypes found: 21 patients ∆F508 homozygous ,10 pts ∆F508/ x compound genotype , 5 pts ∆F508/G542X genotype, 1 ∆F508/ N1303K, and 3 pts with compound genotype: non ∆F508 (I148T, N1303K,G542X)/x. Parent’s genetic tests excluded the possibility of homozygous genotype for this alleles. Thus in 13 pts (32,5%) we could not identify the completed genotype. Conclusions: Concordance of kits mutation with identified mutation in CF Romanian patients is very low. Use of Elucigene CF 29 kit in Romania is limited, probably because of important genetic heterogeneity, and limits significantly heterozygous diagnosis. Development of complementary or wide kits, besides the existing ones is in discussion. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 111 P110. THE DIAGNOSTIC VALUE OF SERUM CARBOHYDRATE ANTIGEN LEVELS 19-9 IN DIABETIC PATIENTS Claudia Borza1, Rodica Mateescu2, Germaine Savoiu3, Corina Serban1, Lelia Susan 4 1 Pathophysiology Department, University of Medicine and Pharmacy Victor Babes Timisoara 2 Physiology Department, University of Medicine and Pharmacy Victor Babes Timisoara 3 Anatomy, Physiology and Pathophysiology, University of Medicine and Pharmacy Victor Babes Timisoara 4 th IV Medical Clinic of University of Medicine and Pharmacy Victor Babes Timisoara Aim: The association between diabetes and pancreatic cancer remains controversial. The study aimed to establish the diagnostic value of the serum carbohydrate antigen 19-9 (CA 19-9) levels in type 2 diabetic patients in comparison with healthy subjects. Materials and methods: The study comprised 32 patients with type 2 diabetes mellitus (10 males and 22 females) and 34 healthy subjects (11 males and 23 females). We recorded in every patient the duration of diabetes, the presence of the complications of the diabetes, the lipid profile, fasting glucose levels and HbA1c levels. The concentration of CA 19-9 was measured by an electrochemiluminescence assay (ECLIA) on a Roche E170 analyzer. In all the patients we performed abdominal echography in order to eliminate the cases of pancreatic cancer. Results: Serum CA 19-9 levels were significantly higher for diabetic patients (45 ± 23.8 U/ml) comparative with the control subjects (10.2 ± 6.8 U/ml) (p < 0.001). In patients with type 2 diabetes, CA 19-9 did not correlate with BMI, duration of diabetes or number of complications. CA 19-9 levels were positive correlated with HbA1c levels (r=0.15, p<0.05). Conclusions: Diabetic patients have increased values of serum CA 19-9. CA 19-9 that is generally used for the diagnosis of pancreatic cancer can be also used as a marker of pancreatic tissue damage that might be caused by diabetes. Diabetic patients should be carefully followed up for an early diagnosis of a possible pancreatic cancer. P111. SERUM PROSTATE-SPECIFIC ANTIGEN (PSA) LEVELS IN MEN WITH TYPE 2 DIABETES Claudia Borza1, Rodica Mateescu2, Germaine Savoiu3, Corina Serban1, Lelia Susan 4 1 Pathophysiology Department, University of Medicine and Pharmacy Victor Babes Timisoara 2 Physiology Department, University of Medicine and Pharmacy Victor Babes Timisoara 3 Anatomy, Physiology and Pathophysiology, University of Medicine and Pharmacy Victor Babes Timisoara 4 th IV Medical Clinic of University of Medicine and Pharmacy Victor Babes Timisoara Objective: Prostate Specific Antigen (PSA) is a valuable prostatic cancer marker that is used in present for screening, diagnosis and monitoring of the patients with prostate cancer. The objective of the study was to compare the levels of prostate specific antigen (PSA) in men with type 2 diabetes and in healthy men and to detect a possible relationship between PSA levels and age of the subjects. Material and methods: In the study were included 62 subjects: a control group of 30 men, and a diabetic group of 32 men. We divided the diabetic patients after the duration of diabetes (8 patients were with recent diagnosed diabetes, 22 with duration of diabetes 6-10 years and 2 of them with more than 10 years of diabetes). We examined the serum level of PSA (normal range < 4.0 ng/ml) with the most sensitive immunoassay method, Electro-Chemi-Luminescence-Immunoassay (ECLIA), on a Roche E170 analyzer. Results: The mean PSA levels were significantly lower in diabetics than in healthy subjects (p <0.001). PSA levels were not associated with the duration of diabetes. The average levels of PSA increased with age (40–49 years: 0.69 ng/ml; 50–59 years: 0.89 ng/ml; 60–69 years: 1.08 ng/ml; 70–79 years: 1.69 ng/ml; over 80 years: 2.22 ng/ml). 112 Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Conclusions: The study showed that the serum PSA levels were lower in diabetics than in healthy. We observed a dependent relationship between PSA levels and age of the patients. Other studies must be done in the future to investigate if the diabetes status and the duration of diabetes should be considered when interpreting a PSA test result. P112. rare Diseases Week in Timisoara Oana Rosca, Ionela Alina Moaca, Iulia Jurca-Simina, Florin Jurca-Simina, Iosif Reascu Norbert Pop, Stefan Berci, Iulia Popa, Irimia Cristina, Graziella Ecob, Adrian Juverdeanu, Carmen Dumitranoiu, Narcis Dobre, Mihai Gafencu, Maria Puiu Save the Children” organization, Timis office VitroBioChem The importance lies in the fact that rare diseases are little known even by the medical system and consequently do not enjoy as a diagnostic and care. The campaign launched on the International Day of rare diseases of February 28 was an opportunity to celebrate community diversity of rare diseases in Romania and abroad and make their presence known among us. Promoting this project began by teasing campaign performance and continued through work programs on rare diseases. From February 1 to 15 Prader Willi Association of Romania has run a campaign to launch teasing message „You know that there are rare people?”; . During the teasing message was supported and promoted by staff from the following cities: Timisoara, Oradea, Cluj, Bucharest, Brasov, Iasi, Sibiu, Carei Simleul Silvana, Deva, Hunedoara, Lupeni, Anina, Lugoj, Bacău, Baia Mare, Piatra Neamţ, Turnu Severin. The theme and slogan: „Rare diseases - a public health priority” „Healthy and sick, Partners for Life!” Period: February 1 to 28 of the Week February 22 to 28 accounted for Rare Diseases Objectives: increase awareness, opportunities propulsion social integration of persons with rare diseases, rare diseases spreading, raising awareness and local authorities. Materials used: • Lessons pupils organized by teachers and civic culture, information on rare diseases in schools • Conference at Children’s Hospital L. Turcanu on rare diseases • Street Campaign - tent material, leaflets distributed by volunteers (Medical students) • March promotion; • Roundtable medical television personalities, political, cultural involvement in the Banat and rare diseases. Method: The message was communicated on the Internet, through posters placed in public places, schools, medical institutions, people distributed flyers on the street, tents located with information materials distributed by medical volunteers. The campaign was initiated by Maria Puiu, vice president ANBRaRo, Michael Gafencu, President Save the Children Timisoara branch and Alina Cristescu, bearing the image of Rare Disease Campaign in Timisoara. Manifestations of Timisoara had as partners: UMF Timisoara, Timis Save the Children, DSP Timis, Timisoara City Hall, Timis County Council, Children’s Hospital L Turcanu, RATT, media personalities. Results and conclusions: we enjoyed the support of many people in this project and the feedback which we received from people who we informed, gives us confidence that rare diseases will remain rare in the absence of informare.People were very interested in this subject and we have requested more information. The campaign also has received many media appearances, including the presence of Mrs. Dorica Dan, president of the National Alliance for proper rare disease in the show Day’s Theme of the Romanian Television, who explained the need for a national strategy for rare diseases emphasizing that information is key to improving quality of life for patients with rare diseases. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 113 Romanian Prader Willi Association and International Prader Willi Organization are organizing the 2nd Eastern European Conference on PWS in cooperation with UMF Timisoara and Local Authorities Salaj „Management of the daily life” and the main themes will be: clinic, genetic and epigenetic aspects in PWS; GH Therapy; weight management and behavior therapy. Zalau, 29-30 OCTOBER 2010 ZALAU Placed in north-western Romania, at the crossing between Eastern Carpathians and the Apuseni Mountains, Salaj county was known as Sylvania County (i.e. the County of Woods). With an area of 3.850 sq. km., its neighbors are Satu-Mare and Maramures counties in the north, Bihor county in the west and south-west and Cluj county in south-east. Placed in the centre of the county it is the administrative residence of the Salaj county. It is situated close to the border with the former Roman Empire, more precisely 8 km from the Roman camp of Porolissum- the strongest fortress in north-western Roman Dacia. During the medieval times the town represented the passage between Central Europe and Transilvania, through the well-known „salt road”. Today the town is connected to the European road on the axis Cluj- Satu-Mare- Petea Vama, DN1F-E81. Apart from its economic importance, it is a powerful cultural educational and touristic centre. It is not long since, apart from the well-known schools and high schools, two more colleges have been authorized to carry on their activities in the town. Being a municipal town of the county, Zalau is also its cultural centre. The cultural events take place in the Union Culture House, the Municipal Cultural House, The „Ionita Scipione Badescu“ LIbrary, the History and Arts Museum, „Ioan Sima” Gallery as well as in the cinema hall of the town. Zalau also possesses a Documentary Library created in 1646 within the Reformed College. Today the library owns more than 40.000 books of which a good part are very rare or even unique. Among the writers one could mention the ancient Greeks- Homer, Platon, Aristotel, Plutarch, Dyionisus of Halicarnas, Ptolemeu and Plinius Secundus or the writers Erasmus of Rotterdam, Calvin, Descartes as well as other writers of the time. 114 Latest research has revealed 7 Dutch sole exemplars, documents belonging to the 17th and 18th centuries (The „Ionita Scipione Badescu” County Library). History The media of the time recorded the inauguration - on the 23 th August 1950- of a „Central Library” owning 7000 Roman and Hungarian books. Among other personalities, Rodica Fanateanu, Elena Fanateanu, Karl Kun and many others were present. The collections comprise very old books like Petrarca’s „De remediis” (1649). In 1952 it became „District Library”, coordinating all the Salaj county libraries. In 1957 took the name of the Salaj scholar Ionita Scipione Badescu. It became „County Library” in 1968. Throughout the times it functioned in the Municipal Cultural House building, in the former Museum location, in the building of the today’s Bookstores Centre and since 1971 in the building on the Iuliu Maniu Square no. 13. It possesses at the moment more than 160.000 books. Cultural institutions Among the most representatives the following institutions are to be mentioned: The Salaj County Cultural, Cults and National Cultural Heritage Control, The Zalau County Arts and History Museum, The “Ionita Scipione Badescu” County Library, The Popular School of Arts and Trades, The Salaj Centre for the Valorization and Promotion of the Traditional Culture, The Zalau Cultural House of Unions. We are waiting you to learn together about PWS and to enjoy the beauties of this Transylvanian town. We are looking forward to welcome you again in Romania! Dorica Dan – president RPWA Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 Day 1 Day 2 12.00 - 13.00 REGISTRATION 13.00 - 14.30 13.00 13.15 - 14.30 OPENING OF THE CONFERENCE Wellcome and introduction Particular aspects of PWS including news from IPWSO conference in Taiwan, May 2010 Genetics, psychology and psychiatry and brain studies Endocrinology (hormones), general health incl. nutrition Questions and answers 30 min 30 min 15 min 14.30-15.00 15.00 – 16.15 15 min 15 min 15 min 10 min 10 min 10 min 16.15 16.30 - 17.45 18.00 17.45 - 18.00 COFFEE BREAK Changes over life time: what do we know and what can we do? Best practice Preparing for school, education of teachers Preparing for adulthood Work and PWS Psychological problems and psychiatry in adults Nutrition management: infantschildren- adults Physical activities 09.00 – 09.30 REPORT OF THE REZULTS OF THE 1ST DAY 09.30 - 10.00 GENERAL HEALTH IN PWS (children and adults) 10.00 - 10.30 COFFEE BREAK 10.00 - 12.00 EUROPEAN RESEARCH NETWORKS EXPERIENCES ACROSS EUROPE – ways to work together 12.00 - 13.00 DEVELOPING SERVICES FOR PEOPLE WITH PWS - examples 13.00 - 14.00 LUNCH 15.30 - 16.00 Assessment of accessibility to and quality of health and social services for patients with PWS in Europe 17.30 - 18.30 Future collaborative research projects Cooperation with Eastern European countries CONCLUSIONS 18.30 - 19.30 DINNER Short break, coffee available Experiences to give to others Recommendations: focus on examples of good practice, and mention mistakes not to be repeated! What is possible in our country? What resources do we need and how can we change the attitudes? Norway (FRAMBU), Italy (BIRD), Greece, Romania, Denmark, France, Germany; Italy, Bulgaria, others; 7-10 min each DINNER BIRD: Experiences from our group stay: Positive aspects and difficulties. ABSTRACTS Authors are invited to write and submit abstracts for oral and poster presentations according to the following instructions: Instructions for writing abstracts • abstracts must be submitted in English • abstracts should not exceed 250 words • font should be Times New Roman, size 10 pts • title of the abstract should be written in capital letters (size 12 pts, Times New Roman) • authors: start a new line and write the last name and first name initial for all authors without titles; underline the name of the presenting author • start a new line and write the name of the institution, city and country for all authors • then write down the e-mail address of the author presenting the paper • abstracts should consist preferably of: aim of the study, methods, results and conclusions • your abstract will be published exactly as submitted (linguistic accuracy is the responsibility of the author) • accepted abstracts and full-text presentation will be published in the supplement of Journal of Paediatrics Instructions for submitting abstracts • online submission: [email protected] • abstracts submitted by fax will not be accepted Submission Deadline extended to September, 30, 2010 Notification of Acceptance of Abstracts will be sent electronically to the corresponding author by October 5, 2010 Instructions for Speakers and Poster Presenters Oral presentations Invited speakers: 20 minutes plus 5 minutes discussion Presentation time: 10 minutes plus discussion Technical equipment LCD projector will be available for oral presentation. Speakers are kindly asked to submit their computer presentations one day or at least two hours before their lecture in the preparation room and give them to the technician. Preparation room will be open daily from 8:00 am to 7:00 pm. Poster presentations Posters must be written in English. Poster dimensions: max. 80 cm (width) x 120 cm (height) Posters will be displayed in the congress area on poster panels. The authors should bring posters in person. Supplies for poster mounting will be provided by the Congressional Service. Romanian Journal of Rare Diseases 2010 | Supplement 1/2010 115
