Impresum
PUBLISHER: ISABS – International Society for Applied Biological Sciences
CIRCULATION: 600 copies
Zagreb, September 2007
Copyright 2007
Video and/or audio-taping of any session is not permitted without prior approval from the speakers and from the scientific committee of the 5th ISABS Conference in Forensic Genetics and Molecular Anthropology
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 1 Table of Contents
Table of Contents
Welcome note …………………………………………………. 3 Conference Organizer ……………………………………….. 4 About ISABS …………………………………………………... 5 ISABS Committees …………………………………………… 6 Scientific Program Information …………………………….. 9 General Information ………………………………………….. 10 Speakers ………………………………………………………... 11 Scientific Program ……………………………………………. 26 Abstracts – Oral Presentations ……………………………. 28 Invited Lectures ………………………………………………… 29 Selected Lectures ………………………………………………. 62 YIA ……………………………………………………………….. 70 Sponsor’s Lectures …………………………………………….. 77 Abstracts – Poster Presentations ………………………….. 83 Forensic Genetics ……………………………………………… 84 Molecular Anthropology ……………………………………….. 120 Advances in Genomic Methods ………………………………. 171 About Invited Speakers . …………………………………….. 174 Sponsor’s Information ……………………………………….. 185 Author index …………………………………………………… 196 Keyword index ………………………………………………… 202
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 2 September 3-7, 2007, Split, Croatia Welcome note
Welcome note
Dear Colleagues
Welcome to the 5th ISABS Conference in Forensic and Anthropological Genetics! This year the International Society for Applied Biological Sciences (ISABS) has taken over the organization with the intent to remain the driving force for our meetings in the future. In addition, this meeting solidifies the idea of linking forensic genetics and anthropological genetics based on the similarity of laboratory methods and intertwining interpretive concepts.
We hope this meeting continues as a forum for the exchange of pertinent information, ideas and technical developments in an informal and beautiful setting. Previous meetings (Split, 1997; Dubrovnik, 2001 and 2005; Zagreb, 2003) received good reviews for the timeleness of presentations and international flair. A decade after our inauguaral meeting in Split, we are returning to this seventeen- century old city; we are certain it will inspire you by its blend of tradition and vibrancy of the Mediterranean.
Enjoy!
Moses Schanfield Dragan Primorac Stanimir Vuk-Pavlović
Conference/Program Directors
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 3 Conference Organizer
CONFERENCE ORGANIZER
Organizers
International Society of Applied Biological Sciences e-mail: [email protected] http://www.isabs.hr
Program Directors
Moses Schanfield, George Washington University, Washington, DC, USA Dragan Primorac, University of Split, Split and Josip Juraj Strossmayer University, Osijek, Croatia Stanimir Vuk-Pavlović, Mayo Clinic College of Medicine, Rochester, MN, USA
Program Committee
Damir Marjanović (Rudjer Boskovic Institute, Zagreb, Croatia) Pavao Rudan (University of Zagreb, Zagreb, Croatia)
Organizing Committee Šimun Anđelinović Damir Marjanović Ivana Erceg Ivkošić, Chair Petar Projić Ante Ivkošić Ivana Šamija Dalibor Marijanović Vedrana Škaro – coordinator Inga Marijanović Assistance to the Organizing Committee Ivana Furčić Duje Rako – webmaster Andreja Leskovac Zrinka Romić Igor Matić Jelena Todorović Maja Radnić Ante Vulić
Conference Service
IDEJA kongres Vukovarska 17/III Dubrovnik Tel: +385 20 356 488 Fax: +385 20 356 489 E-mail: [email protected] URL: www.ideja-kongres.hr
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 4 September 3-7, 2007, Split, Croatia About ISABS
About ISABS
Dear Participants, Friends and Colleagues, our future Members, International Society for Applied Biological Sciences (ISABS) is a non-profit organization established to promote, enhance and improve research, development and education in molecular biology, genomics, proteomics, forensic and anthropological genetics, and biotechnology. Mission of the ISABS is to serve the medical, forensic and humanitarian communities applying biological sciences to the solution of medical, forensic and humanitarian problems through interdisciplinary expert collaboration in the disciplines of applied biological sciences by facilitating international contact, educational opportunities and engendering personal trust. Although the first authorized gathering of the ISABS was held in 2004, the society has actually started with its activities in 1997 by organizing the international meetings in forensic and clinical genetics. The previous meetings held in Split (1997), Dubrovnik (2001 and 2005) and Zagreb (2003) were highly successful with lectures, poster sessions and workshops provided by the outstanding leaders in the fields related to the conference program. With total of about 1600 participants from more than 60 countries worldwide these events were truly international. Since 2003 meetings have been organized in collaboration with Mayo Clinic (Rochester, Minnesota, USA) and University of Zagreb through the approved program Mayo Clinic College of Medicine-University of Zagreb International Program in Advanced Medical Education. All these years Croatian Medical Journal (CMJ) was official journal of our conferences and we had thematic issues of CMJ. Now we are proud to announce that CMJ, one of the major scientific journals in Croatia and respectable journal in whole scientific world, is now official journal of the ISABS. In order to make ISABS recognizable as the organizer of our meetings for the first time the ISABS name appears as the part of the conference title at this fifth anniversary held again in Split, one of the most beautiful cities on the Adriatic coast. We are happy to share this unique scientific and social experience with you and we hope that you will recognize our efforts to achieve objectives of our Society: to promote for education and research in the applied biological sciences; to encourage the study, improve the practice, elevate the standards and advance the cause of the applied biological sciences; to promote interdisciplinary communications; and to plan, organize and administer meetings, reports and other projects for the stimulation and advancement of these and related purposes. I wish you very successful gathering at the 5th ISABS Conference in Forensic Genetics and Molecular Anthropology hoping that all of us will have benefit from this distinctive event. Please do not hesitate to join us and become ISABS member.
Best wishes,
Ivana Erceg Ivkosic, M.D. ISABS president
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 5 ISABS Committees
ISABS Committees
ISABS registration number: 21003655 Date of registration: August 27, 2004
President: Ivana Erceg-Ivkošić (General Hospital "Sveti Duh", Zagreb, Croatia) Vice President: Ante Ivkošić (General Hospital "Sveti Duh", Zagreb, Croatia) Secretary: Dalibor Marijanović (Business Innovation Center of Croatia- BICRO, Zagreb, Croatia)
ISABS Conferences Founding Members:
Dragan Primorac (University of Split, Split and University of Osijek, Osijek, Croatia) Moses Schanfield (George Washington University, Washington, DC, USA) Stanimir Vuk-Pavlović (Mayo Clinic College of Medicine, Rochester, MN, USA)
Scientific Committee:
Forensic Genetics:
Antonio Alonso (National Institute of Toxicology and Forensic Sciences, Madrid, Spain) Zoran Budimlija (Office of Chief Medical Examiner, New York, NY, USA) Cecelia Crouse (Palm Beach Sheriff's Office, West Palm Beach, FL, USA) Jürgen Henke (Institut für Blutgruppenforschung, Köln, Germany) Mitchell Holland (Forensic DNA Consultants, Manassas, VA, USA) Henry Lee (University of New Haven, West Haven and Connecticut Forensic Science Laboratory, Meriden, CT, USA) José Lorente (Department of Legal Medicine, University of Granada, Granada, Spain) Marilyn Menotti-Raymond (National Cancer Institute, Frederick, MD, USA) Antti Sajantila (Department of Forensic Medicine, University of Helsinki, Helsinki, Finland) Molecular and Cellular Medicine: Henry Erlich (Roche Molecular Systems, Inc., Alameda, CA, USA) Francis Glorieux (Genetics Unit, Shriners Hospital for Children and McGill University, Montreal, QC, Canada) Robert Huber - Nobel Laureate 1988 (Max Planck Institute for Biochemistry, Martinsried, Germany) Pier Franco Pignatti (Institute of Biology and Genetics and Faculty of Medicine and Surgery, University of Verona, Italy) David I. Smith (Mayo Clinic College of Medicine, Rochester, MN, USA)
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 6 September 3-7, 2007, Split, Croatia ISABS Committees
Molecular Anthropology:
Pavao Rudan (Institute for Anthropological Research, Zagreb, Croatia) Peter Underhill (Stanford University Medical Center, Stanford, CA, USA)
Course Committee:
Frederick Bieber (Harvard Medical School and Birgham and Women's Hospital, Boston, MA, USA) Damir Marjanović (Institute for Genetic Engineering and Biotechnology Sarajevo, Sarajevo, B&H and Ruđer Bošković Institute, Zagreb, Croatia) Timothy Palmbach (Connecticut Forensic Science Laboratory, Meriden, CT, USA) Thomas Parsons (International Commission on Missing Persons, Sarajevo, B&H)
ISABS Young Investigator Programme Committee:
Šimun Anđelinović (Clinical Hospital Split, Split, Croatia) Ivana Erceg-Ivkošić (General Hospital "Sveti Duh", Zagreb, Croatia) Edwin Huffine (Bode Technology Group, Lorton, VA, USA) Inga Marijanović (Faculty of Science, University of Zagreb, Zagreb, Croatia)
Science, Society & Ethical Committee:
José Lorente (Department of Legal Medicine, University of Granada, Granada, Spain)
Fellowship Committee:
Katja Drobnič (Forensic Science Centre, Ministry of Interior, Ljubljana, Slovenia) Alemka Markotić (Department for Research and Imunology, Cellular Immunology Unit, Institute of Immunology, Zagreb, Croatia) Daniel Vanek (Forensic DNA Service, Prague, Czech Republic)
Publications, Electronic Information & Communications Committee:
Ante Ivkošić (General Hospital "Sveti Duh", Zagreb, Croatia) Dalibor Marijanović (Business Innovation Center of Croatia- BICRO, Zagreb, Croatia)
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 7 ISABS Committees
Membership & Publication Committee:
Petar Projić (Ruđer Bošković Institute, Zagreb, Croatia) Ivana Šamija (Clinical Hospital Center Zagreb, Zagreb, Croatia) Vedrana Škaro (Ruđer Bošković Institute, Zagreb, Croatia)
Students Committee:
Ante Vulić (Medical School, University of Zagreb, Zagreb, Croatia)
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 8 September 3-7, 2007, Split, Croatia Scientific Program Information
Scientific Program Information
Certificate of Attendance Confirmations of attendance will be issued at the registration desk.
Young Investigator Awards Members of the Program Committee selected abstracts among entries submitted by investigators under the age of 40. Author(s) of each selected abstract will receive the Young Investigator Award Certificate and EUR 500 and will present their work from the podium at the times scheduled in conference program.
Credits The 5th ISABS Conference in Forensic Genetics and Molecular Anthropology has been approved for 20 (participants) or 25 (lecturers) points by the Croatian Medical Chamber. Credits are intended for medical doctors, members of Croatian medical Chamber, in order to extend their medical doctor's license.
Sponsor Exhibition Set up: Sunday, September 2, 2007 18:00 to 20:00 Monday, September 3, 2007 noon to 18:00 Tuesday, September 4, 2007 08:30 to 20:00 Thursday, September 6, 2007 08:30 to 20:00 Dismantling: Friday, September 7, 2007 18:00 to 20:00
Language The official language of the conference is English (no simultaneous translation)
Poster Setup Sunday, September 2, 2007, 18:00 – 20:00 Monday, September 3, 2007, 09:00 – 11:00 Poster board numbers can be found in the author’s index. The staff at the registration and info desk will help you in finding both the number and location of the board.
Poster Discussion Tuesday September 4, 2007, 12:30-15:00 Posters presentation (even numbers) Thursday September 6, 2007, 11:30-14:30 Posters presentation (odd numbers)
If you or a co-author will not be able to be at your board at this time, please leave a note on your poster stating date and time when you will be present.
Wine and cheese meeting with speakers Tuesday September 4, 2007, 19:00 - 20:30 Thursday September 6, 2007, 18:15 - 19:45
In order to make speakers more available to the participants we invite you all to participate in speaker/participant program. A speaker will be assigned a table (tables will be scattered around the various venues) so that the participants can come and talk and move around to various speakers.
Poster Removal Friday, September 7, 2007, 12:30 - 13:30
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 9 Scientific Program Information
Program Changes The organizers cannot assume liability for any changes in the program due to external or unforeseen circumstances.
Registration Desk Opening Hours Sunday, September 2, 2007 18:00 – 20:00 Monday, September 3, 2007 07:30 – 20:00 Tuesday, September 4, 2007 07:30 – 16:00 Wednesday, September 5, 2007 07:30 – 16:00 Thursday, September 6, 2007 07:30 – 16:00 Friday, September 7, 2007 07:30 – 16:00
PowerPoint Preview Room PowerPoint preview room will be available to all speakers. We encourage all lecturers to present data as a PowerPoint presentation.
Recipients of the 2001 Young Investigator Awards Forensic IdentityTesting: Frontiers in Molecular and Cellular Medicine: • Lucia Cifuentes Ovalle, Chile • Rima Dada, India • Katja Drobnič, Slovenia • Anna Gareeva, Russia • Nguyen Hoai Giang, Vietnam • Tomasz Kupiec, Poland
Recipients of the 2003 Young Investigator Awards • Robert J. Shelton, CO, USA (Forensic Genetics) • Chiara Magri, Italy (Molecular and Cellular Medicine)
Recipients of the 2005 Young Investigator Awards • Caroline Round, United Kingdom (Forensic Genetics) • Tracy Johnson, USA (Forensic Genetics) • Vedrana Montana, USA (Molecular and Cellular Medicine) • Mirela Baus Lončar, Germany (Molecular and Cellular Medicine)
Recipients of the 2007 Young Investigator Awards • Grzegorz Kaczmarczyk, Poland (Forensic Genetics) • Agnieszka Krzyżańska, Poland (Forensic Genetics) • Kaye Ballantyne, Australia (Molecular Anthropology) • Tomislav Domazet-Lošo, Croatia (Molecular Anthropology) • Coralie Frassati, Switzerland (Molecular Anthropology) • Taeko Kashima, Japan (Molecular Anthropology)
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 10 September 3-7, 2007, Split, Croatia General Information
General Information
Badges Badges will be provided to participants, accompanying persons, exhibitors and press at registration and will be required for admission to all conference facilities and scientific and social events during the Meeting. The badges will be checked by security guards at the conference venue. Any individual who is not wearing an official meeting badge will be directed to the registration desk to register or, if already registered, to purchase a replacement badge. Handling fee for replacement badges is €10.
Bank Services The official currency in Croatia is the Croatian kuna (HRK). The official exchange rates: 1 EUR = 7.32 HRK* (August 27, 2007) 1 USD = 5.36 HRK* (August 27, 2007) * Please note that these exchange rates are variable
Busines hours for post offices and banks Banks and post offices are normally opened from 8:00 a.m. to 7:00 p.m., Monday through Friday and from 8:00 a.m. to 12:00 p.m. on Saturday.
Cash Machines ATMs accepting all major bank cards and credit cards are located at numerous sites in Split.
Electricity Supply 220-240 V, 50 Hz
Insurance Participants need to make their own arrangements pertinent to health and travel. By registering for the 5th ISABS Conference in Forensic Genetics and Molecular Anthropology, participants agree that neither the organizers and its agents nor the sponsors and exhibitors nor the Hotel Le Meridien Lav, Split assume any liability whatsoever.
Message Center A Message Center is available at the registration desk.
Public Transportation in Split Buses are the main means of public transportation in Split. Almost all hotels have easy access to buses.
Restaurants Most restaurants in Split are opened from 8:00 a.m. to 11:00 p.m. Service charges are included in the price, unless explicitly mentioned otherwise, but an additional tip of 5 to 10 percent is expected. Some restaurants may have a cover charge.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 11 General Information
Service Center Photocopying, typing, production of overhead transparencies and computer printouts are available at the Le Méridien Lav Service Center for a fee.
Shopping Store hours in Split are usually 8:00 a.m. – 9:00 p.m. Monday to Saturday. Some are open on Sundays. Most shops accept all major credit cards.
Smoking Policy The 5th ISABS Conference in Forensic Genetics and Molecular Anthropology is a non-smoking event. Smoking is prohibited everywhere on conference grounds and at all functions associated with the conference.
Special requirements Registrants with special requirements for physical communication and dietary requirements should contact Ideja kongres d.o.o., official congress service agency in advance.
Information Conference staff will be pleased to help you with any conference and travel information you may need.
Taxi Numerous Taxi stands are located throughout Split city centre and in front of the hotels. All receptionists will be glad to help you.
Hotel Information At the Hotel Le Méridien Lav, Split a wide choice of accommodation offers luxury and modern comfort. The hotel is perfect choice for both leisure and business travelers. This five star luxury hotel is located at Podstrana on the Adriatic coast, approx 6 km south of historic Diocletian’s Palace, the core of the Split and approx 30 mins from Split International Airport (SPU). The hotel is a fully integrated resort destination, unique within the Croatian market. It is the only internationally- affiliated hotel within the area of Split (Croatia’s second commercial city and one of the largest ports on the Mediterranean) and it has the largest hotel convention facilities on the Adriatic. The Hotel enjoys a spectacular coastal position overlooking the sea, Split, and surrounding islands.
The highly acclaimed Le Méridien hospitality is reflected in the understated elegance of the 382 well appointed and large (min 32 sq.m.) Rooms and Suites, in 4 interlinked buildings, each of which boasts its own balcony or terrace and over 80% enjoy magnificent views over the sea towards Split and its many islands.
The leisure facilities include a world class Spa and Wellness Centre, with a long list of treatments and massages, the latest Fitness equipment and a circular indoor pool—all of which give an added impetus to the experience of rejuvenation. Other activities include retail therapy at the Shopping Arcade and Promenade, four 5th ISABS Conference in Forensic Genetics and Molecular Anthropology 12 September 3-7, 2007, Split, Croatia General Information tennis courts, water sports and scuba diving as well as an extensive animation/activities programme. The children have not been forgotten either as Le Méridien’s Penguin Club provides creative activities which will arouse the senses and inventiveness of the Resort’s younger guests.
There are three Restaurants and four Bars in the Hotel as well as five Restaurants and four Cafés and Bars overlooking the fascinating yachts in the Resort’s very own Marina.
Other facilities: Underground Car Park for 350 cars, Medical Centre, Private Marina for 60 yachts, Non Smoking Rooms, Wheelchair friendly, Transfers from and to airport by Motor Launch, Shuttle Bus Service to Split, Night Club, Casino.
Congress tour Wednesday, September 5, 2007 A boat trip to Islands of Brač and Hvar Price per person: EUR 50 (Transfer, local drink aboard, lunch) Menu: fish, meet, vegetables, vine and juice. Duration: half day This boat excursion will take you to the nearby islands of Brac and Hvar, to their most attractive towns. A sweeping view of small island places, accompanied with pleasant notes of Dalmatian songs, grilled fish and homemade vine reminds the passengers of the ancient pirate tradition. In the desired place, you have time for sight-seeing, sunbathing and swimming.
Please note that congress tour is subject to alteration. Please check at registration desk.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 13 Speakers
SPEAKERS
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 14 September 3-7, 2007, Split, Croatia Speakers
Plenary lecture
Svante Pääbo (Max-Planck-Institute of Evolutionary Anthropology Leipzig, Germany )
Forensic Genetics
Theodore Anderson (Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD, USA)
Frederick Bieber (Harvard Medical School and Birgham and Women's Hospital, Boston, MA, USA)
Zoran Budimlija (NYC Office of Chief Medical Examiner, New York, NY, USA)
Bruce Budowle (FBI Laboratory, Quantico, VA, USA)
Casandra Calloway (Roche Molecular Systems, Alameda, CA, USA)
Angela van Daal (Faculty of Health Sciences & Medicine, Bond University, Gold Coast, Queensland, Australia)
Katja Drobnič (Forensic Science Centre, Ministry of Interior, Ljubljana, Slovenia)
Nils Gerke (Eppenforf AG, Hamburg, Germany)
Andreas Hellmann (Bundeskriminalamt, Kriminaltechnisches Institut, KT32, Wiesbaden, Germany)
Mitchell Holland (Forensic Science, Pennsylvania State University, University Park, PA, USA)
Edwin Huffine (Bode Technology Group, Lorton, VA, USA)
Rebecca Just (Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD, USA)
Grzegorz Kaczmarczyk (Department of Molecular Biology and Clinical Genetics, Internal Medicine, Krakow, Poland)
Agnieszka Krzyżańska (Molecular Technique Unit, Department of Forensic Medicine, Wroclaw Medical University, Wrocław, Poland)
Henry Lee (University of New Haven, West Haven and Connecticut Forensic Science Laboratory, Meriden, CT, USA)
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 15 Speakers
José A. Lorente (Department of Legal Medicine, University of Granada, Granada, Spain)
Damir Marjanović (Institute for Genetic Engineering and Biotechnology Sarajevo, Sarajevo, B&H and Ruđer Bošković Institute, Zagreb, Croatia)
Marilyn Menotti-Raymond (National Cancer Institute, Frederick, MD, USA)
Rebecca Mikulasovich (NYC Office of Chief Medical Examiner, New York, NY, USA)
Tahnee M. Nelson (San Francisco Police Department, San Francisco, CA, USA) Pieter van Oers (Applied Biosystems, Foster City, USA)
Thomas Parsons (International Commission on Missing Persons, Sarajevo, B&H)
Lutz Roewer (InstituteOf Legal Medicine, Charité University of Medicine, Berlin, Germany)
Antti Sajantila (Department of Forensic Medicine, University of Helsinki, Helsinki, Finland)
Brion C. Smith (Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD, USA)
Poonam Taneja (Molecular Devices, MDS Analytical Technologies Sunnyvale, CA, USA)
Daniel Vanek (Forensic DNA Service, Prague, Czech Republic)
Molecular Anthropology
Kaye Ballantyne (Victoria Police Forensic Services Department, Macleod, VIC, Australia)
Ranjan Deka (University of Cincinnati College of Medicine, Cincinnati, OH, USA)
Tomislav Domazet-Lošo (Ruđer Bošković Institute, Zagreb, Croatia)
Henry Erlich (Roche Molecular Systems, Alameda, CA, USA)
Coralie Frassati (Department of Anthropology and Ecology, University of Geneva, Switzerland)
Taeko Kashima (Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, Japan)
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 16 September 3-7, 2007, Split, Croatia Speakers
Eliza Khusnutdinova (Institute of Biochemistry and Genetics, Ufa, Bashkortostan, Russian Federation)
Laszlo Mark (Institute of Biochemistry and Medical Chemistry, University of Pecs, Pecs, Hungary)
Irena Martinović Klarić (Institute for Anthropological Research, Zagreb, Croatia)
Erika Nagle (Department of Medical Biology and Genetics, Riga Stradiņš University, Riga, Latvia)
Giuseppe Passarino (Department of Cell Biology, University of Calabria, Rende, Italy)
Mario Pirastu (Institute of Population Genetics, National Research Council, C.N.R., Alghero and Shardna Life Sciences Cagliari, Italy)
Pavao Rudan (Institute for Anthropological Research, Zagreb, Croatia)
Irina Udina (N.I. General Genetics Institute RAS, Moscow, Russian Federation)
Peter Underhill (Stanford University Medical Center, Stanford, CA, USA)
Richard Villems (Institute of Molecular and Cell Biology, University of Tartu and Estonian Biocentre, Tartu, Estonia)
Advances in Genomic Methods
Jean-Pierre Kocher (Mayo Clinic College of Medicine, Rochester, MN, USA)
David Smith (Mayo Clinic College of Medicine, Rochester, MN, USA)
George Vasmatzis (Mayo Clinic College of Medicine, Rochester, MN, USA)
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 17 Scientific Program
5TH ISABS CONFERENCE IN FORENSIC GENETICS AND MOLECULAR ANTHROPOLOGY
Hotel Le MERIDIEN LAV Split Croatia September 3-7, 2007
SCIENTIFIC PROGRAM
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 18 September 3-7, 2007, Split, Croatia Scientific Program
Please note that the programme and speakers are subject to alteration.
Sunday, September 2, 2007
18:00 -20:00 Registration and Poster Set-Up (Conference floor, lobby and foyer)
Monday, September 3, 2007
Introductory Plenary session (Part One) "Grand Dalmacija" Conference Room
9:00 - 11:00 Registration and Poster Set-Up (Conference floor, lobby and foyer)
9:30 Henry Lee (Connecticut Forensic Science Laboratory, Meriden, CT, USA) Homicide Investigation - Anthropological & Genetic Analysis of the Crime Scene
10:15 Pavao Rudan (Institute for Anthropological Research, Zagreb, Croatia) Reality and perception in molecular anthropological studies of population structure
11:00 Coffee Break
11:15 David I. Smith (Mayo Clinic College of Medicine, Rochester, MN, USA) The dawn of the $1,000 genome
12:00 Opening of Sponsor Exhibits and Poster Presentation (Conference floor, lobby and foyer)
13:00 - 15:00 Lunch
Introductory Plenary session (Part Two) "Grand Dalmacija" Conference Room
15:00 Introduction from Chair
15:05 Bruce Budowle (FBI Laboratory, Quantico, VA, USA) Microbial forensic investigation of biocrime and bioterrorism
15:35 José Lorente (University of Granada and Genio Andalucia, Spain) Missing kids identification: The Prokids International Program
16:05 Coffee Break
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 19 Scientific Program
16:25 Marilyn Menotti-Raymond (National Cancer Institute Frederick, MD,USA) The domestic cat: Domestication, breed development and a population genetic database for computation of match probabilities
16:55 Peter Underhill (Stanford University, Stanford, CA, USA) Y chromosome phylogenetics and phylogeography
17:20 Adjourn
19:00 Annual Meeting of the ISABS ("Grand Dalmacija" Conference Room)
20:30 Welcome Reception ("Vila Dalmacija"; "Trio Gušt" band)
Tuesday September 4, 2007
Forensic Genetics I "Grand Dalmacija I" Conference Room
8:30 Introduction from Chair
8:35 Zoran Budimlija (Office of Chief Medical Examiner, New York, NY, USA) Forensic biology techniques in the separation of mixed tissues: from routine to the extremes
9:05 Frederick Bieber (Harvard Medical School, Cambridge, MA, USA) Close encounters: Partial DNA profiles and family searching in forensic investigations
9:40 Coffee Break
10:00 Inauguration of Conference ("Grand Dalmacija I" Conference Room)
11:00 Tahnee Nelson (San Francisco Police Department, San Francisco, CA, USA) Development of a multiplex single base extension assay for mtDNA haplogroup typing
11:30 Grzegorz Kaczmarczyk (Department of Molecular Biology and Clinical Genetics, Internal Medicine, Krakow, Poland) DNA identification of leather relics in the memorial and museum Auschwitz-Birkenau
11:50 Daniel Vanek (Forensic DNA Service, Prague, Czech Republic) DNA identification of Premyslid royal dynasty remains by AMPFLSTR Minifer PCR
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 20 September 3-7, 2007, Split, Croatia Scientific Program
12:20 Discussion
12:30-15:00 Posters presentation (even numbers) (Conference floor, lobby and foyer)
13:00-15:00 Lunch
Forensic Genetics II "Grand Dalmacija I" Conference Room
15:00 Introduction from Chair
15:05 Theodore Anderson (Armed Forces Institute of Pathology, Washington, DC, USA) Validation of demineralization and low-copy-number autosomal and Y- chromosomal short tandem repeat typing and its application to the identification of aged skeletal remains associated with previous United States military conflicts
15:45 Rebecca Just (Armed Forces Institute of Pathology, Rockville, MD, USA) The application of new non-CODIS MiniSTRS to resolve commingled skeletal remains from Korean War
16:25 Coffee Break
16:40 Katja Drobnič (Forensic Science Centre, Ministry of the Interior, Ljubljana, Slovenia) Validation of new SRY marker for use in forensic casework
17:20 Mitchell Holland (Pennsylvania State University, University Park, PA, USA) GeneMarker® HID: An effective new software tool for analysis of STR data
17:50 Discussion
Molecular Anthropology I "Grand Dalmacija II" Conference Room
8:30 Introduction from Chair
8:35 Mario Pirastu (Institute of Population Genetics, National Research Council, C.N.R., Alghero and Shardna LifeSciences, Cagliari, Italy) Ogliastra Genetic Park: a link of history and genetic makeup of an ancient population
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 21 Scientific Program
9:05 Katja Drobnič (Forensic Science Centre, Ministry of the Interior, Ljubljana, Slovenia) Interaction of personnel at the crime scene involving a buried body: The role of forensic archaeologist
9:40 Coffee Break
10:00 Inauguration of Conference ("Grand Dalmacija I" Conference Room)
11:00 Giuseppe Passarino (Department of Cell Biology, University of Calabria, Rende, Italy) Evolution of human populations and the study of the genetic basis of human longevity
11:40 Coralie Frassati (Department of Anthropology and Ecology, University of Geneva, Switzerland) The polymorphism of KIR genes in worldwide human populations
12:00 Taeko Kashima (Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, Japan) TP53 codon 72 polymorphism in 12 populations of insular Southeast Asia and Oceania
12:20 Discussion
12:30-15:00 Posters presentation (even numbers) (Conference floor, lobby and foyer)
13:00-15:00 Lunch
Molecular Anthropology II "Grand Dalmacija II" Conference Room
15:00 Introduction from Chair
15:05 George Vasmatzis (Mayo Clinic College of Medicine, Rochester, MN, USA) Gene expression profiling to identify genes predictive of cancer patient outcome
15:45 Ranjan Deka (University of Cincinnati College of Medicine, Cincinnati, OH, USA) Linkage Disequilibrium in an Isolated Population from Polynesia: Prospects for Mapping Complex Traits
16:25 Coffee Break
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 22 September 3-7, 2007, Split, Croatia Scientific Program
16:40 Kaye Ballantyne (Victoria Police Forensic Services Department, Macleod, VIC, Australia) Increased amplification sensitivity and success with locked nucleic acids
17:00 Tomislav Domazet-Lošo (Ruđer Bošković Institute, Zagreb, Croatia) Landmarks of animal adaptive history uncovered by genomic phylostratigraphy in Drosophila
17:20 Discussion
19:00 Wine and cheese meeting with speakers ("Grand Dalmacija" terrace)
21:00 Hotel Le Meriden Lav Split poolside program ("Bravo" band)
Wednesday September 5, 2007
8:30 Half-day excursion (Brač/Hvar)
EMBO Lecture "Grand Dalmacija" Conference Room
18:00 Introduction from Chair
18:05 Svante Pääbo (Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany) Neandertal Genomics
19:00 Conference Dinner and Young Investigator Awards Ceremony ("Grand Dalmacija" Conference Room)
Thursday September 6, 2007
Joint Plenary Session "Grand Dalmacija I" Conference Room
8:30 Introduction from Chair
8:35 Bruce Budowle (FBI Laboratory, Quantico, VA, USA Statistical issues surrounding national DNA databases
9:15 Lutz Roewer (Institute of Legal Medicine, Charité University of Medicine, Berlin, Germany) Structured population databases in forensic genetic practice 5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 23 Scientific Program
9:55 Coffee Break
10:15 Ranjan Deka (University of Cincinnati College of Medicine, Cincinnati, OH, USA) Common Variants in 19q13 are associated with Prostate Cancer in European Men
10:55 Edwin Huffine (Bode Technology Group, Lorton, VA, USA) The developing role of forensics in deterring violence and genocide
11:25 Richard Villems (University of Tartu and Estonian Biocentre, Tartu, Estonia) Lessons on mtDNA phylogeny learnt from complete sequences
12:05 Discussion
12:10-14:30 Poster Presentation (odd numbers) (Conference floor, lobby and foyer)
Forensic Genetics III "Grand Dalmacija I" Conference Room
15:00 Introduction from Chair
15:05 Rebecca Mikulasovich (NYC Office of Chief Medical Examiner, New York, NY, USA) A Retrospective of High Sensitivity DNA Testing for Criminal Cases
15:45 Thomas Parsons (International Commission on Missing Persons, Sarajevo, Bosnia & Herzegovina) ICMP: Integrating forensic sciences in high throughput identification of missing persons from mass graves
16:25 Coffee Break
16:40 Damir Marjanovic (Institute for Genetic Engineering and Biotechnology, Sarajevo, B&H and Ruđer Bošković Institute, Zagreb, Croatia) DNA identification of skeletal remains from the second World War mass graves uncovered in Slovenia - First results
17:20 Poonam Taneja (Molecular Devices, MDS Analytical Technologies Sunnyvale, CA, USA) New Developments and Recent Advances in Laser Capture Microdissection
18:00 Discussion
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 24 September 3-7, 2007, Split, Croatia Scientific Program
Molecular Anthropology III "Grand Dalmacija II" Conference Room
15:00 Introduction from Chair
15:05 David I. Smith (Mayo Clinic College of Medicine, Rochester, MN, USA ) Gene expression profiling to analyze the transcriptional output of the entire genome: The emergence of non-coding transcripts
15:45 Jean-Pierre Kocher (Mayo Clinic College of Medicine, Rochester, MN, USA) Data management solution and analytical infrastructure to support basic clinical and translational research
16:25 Coffee Break
16:40 Laszlo Mark (Institute of Biochemistry and Medical Chemistry, University of Pecs, Pecs, Hungary) Evolutionary paleoproteomics: analysis of pathological proteins by mass spectrometry
17:05 Eliza Khusnutdinova (Institute of Biochemistry and Genetics, Ufa, Bashkortostan, Russian Federation) Y-chromosome diversity in southern Urals: A geographic border between Europe and Asia
17:25 Erika Nagle (Department of Medical Biology and Genetics, Riga Stradiņš University, Riga, Latvia) Craniofacial parameters as phenotypic markers for the susceptible to clefts genotypes
17:45 Irina Udina (N.I. General Genetics Institute RAS, Moscow, Russian Federation) Differentiation of indigenous Asian and American populations revealed by autosomal SNPSTR systems
18:05 Discussion
18:15 Wine and cheese meeting with speakers (Conference floor, lobby and foyer) Friday, September 7, 2007
Forensic Genetics IV "Grand Dalmacija I" Conference Room
8:30 Introduction from Chair
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 25 Scientific Program
8:35 Agnieszka Krzyżańska (Department of Forensic Medicine, Wroclaw Medical University, Wrocław, Poland) Non-destructive method of DNA extraction from teeth and bones
8:55 Cassandra Calloway (Roche Molecular Systems, Alameda, CA, USA) Improving the discrimination of mtDNA typing using a multiplex PCR and linear array probe panel: HV and beyond
9:35 Coffee Break
10:00 Brion Smith (Armed Forces Institute of Pathology, Washington, DC, USA) The human tooth as a source of forensic DNA
10:40 Antti Sajantila (Department of Forensic Medicine, University of Helsinki, Finland) Integrated forensic analysis of FinnishWorld War II soldiers
11:20 Andreas Hellmann (Bundeskriminalamt, Kriminaltechnisches Institut, KT32, Wiesbaden, Germany) DNA analysis of trace evidence from animal and plant origin - experiences in forensic casework
11:40 Angela van Daal (Faculty of Health Sciences & Medicine, Bond University, Gold Coast, Queensland, Australia) The genetic basis of human pigment variation
12:00 Discussion
12:00 – 12:30 Closing Ceremony ("Grand Dalmacija I" Conference Room)
12:00 - 13:30 Take down posters
12:00 - 14:30 Lunch
Molecular Anthropology IV "Grand Dalmacija II" Conference Room
8:30 Intro from Chair
8:35 Irena Martinović Klarić (Institute for Anthropological Research, Zagreb, Croatia) Roma in Croatia: Inferences on paternal genetic heritage of the Bayash
9:15 Henry Erlich (Roche Molecular Systems, Alameda, CA, USA) HLA allelic diversity and the phylogenetic analysis of human population
09:55 Coffee Break
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 26 September 3-7, 2007, Split, Croatia Scientific Program
10:20 Nils Gerke (Eppenforf AG, Hamburg, Germany) Real-time PCR in forensics
10:40 Pieter van Oers (Applied Biosystems,Nieuwerkerk aan den IJssel, Netherlands) Quantitative and qualitative assessment of total human and human male DNA in forensic type biological samples using a multiplexed real-time PCR system
11:00 Discussion
12:00 – 12:30 Closing Ceremony ("Grand Dalmacija I" Conference Room)
12:00 - 13:00 Take down posters
12:00 - 14:30 Lunch
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 27 ORAL PRESENTATIONS
ABSTRACTS – ORAL PRESENTATIONS
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 28 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS
INVITED LECTURES
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 29 ORAL PRESENTATIONS Plenary Lecture Plenary Lecture
Abstract number: ABS-001-ISABS-2007
NEANDERTAL GENOMICS
Pääbo S1
1Max-Planck Institute for Evolutionary Anthropology, Leipzig, Germany [email protected]
Neandertals, who diverged from modern humans about half a million years ago and became extinct around 30,000 years ago, are the closest relatives of current humans. A Neandertal genome sequence would allow nucleotide sequence differences between the human and the chimpanzee genomes to be assigned to fully modern humans during and after their descent from the Neandertal-modern human ancestor. The Neandertal genome, which falls partially within the coalescence of human genes, would also allow novel approaches to detect positive selection in modern humans. We are applying large-scale parallel pyrosequencing on the GS-FLX platform to DNA extracts of Neandertal bones with the ultimate goal of achieving 1-fold coverage of the entire genome. Strategies to estimate the authenticity and reliability of the DNA sequences will be reviewed as well as preliminary analyses of approximately 1% of the Neandertal genome.
Keywords: Neandertal, Ancient DNA, Human origins, Population history, DNA damage
Suggested reading: Green, R.E., J. Krause, S.E. Ptak, A.W. Briggs, M.T. Ronan, J.F. Simons, L. Du, M. Egholm, J.M. Rothberg, M. Paunovic and S. Pääbo: Analysis of one million base pairs of Neanderthal DNA. Nature 444, 330-36 (2006). Pääbo, S., H. Poinar, S. Serre, V. Jaenicke-Després, J. Hebler, N. Rohland, M. Kuch, J. Krause, L. Vigilant and M. Hofreiter: Genetic analyses from ancient DNA. Annual Review of Genetics, 38, 645-79 (2004). Enard, W. and S. Pääbo: Comparative primate genomics. Annual Review of Genomics and Human Genetics 5, 351-78 (2004).
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 30 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Forensic Genetics
Abstract number: ABS-8234-ISABS-2007
VALIDATION OF DEMINERALIZATION AND LOW COPY NUMBER AUTOSOMAL AND Y-CHROMOSOMAL SHORT TANDEM REPEAT TYPING AND ITS APPLICATION TO THE IDENTIFICATION OF AGED SKELETAL REMAINS ASSOCIATED WITH PREVIOUS UNITED STATES MILITARY CONFLICTS
Edson S1, Loreille O1, Maynard K1, Irwin J1, Desnoyers A1, Oliver R1, Barritt-Ross S1, Anderson T1, Coble M1
1Armed Forces DNA Identification Laboratory, Rockville, Maryland, USA [email protected]
Mitochondrial (mt) DNA analysis of degraded skeletal remains has been a highly effective tool with which to support the association of these remains with family members. However, mtDNA analysis is typically only a putative means of identification, as even a complete mtDNA haplotype is not unique to an individual. For this reason, improvements to existing technologies are necessary. Original DNA extraction protocols required using 1.0-2.5g of pulverized bone powder in an extraction buffer and proteinase K. Modifications to this method have decreased the amount of bone needed to 0.25g and resulted in greatly-increased DNA yield through complete dissolution of the osseous material. Short tandem repeat (STR) analysis has not been widely used for ‘aged’ skeletal remains largely because the nuclear DNA from such bones is often degraded and present in low quantity (<<0.25ng). Therefore, low copy number (LCN) STR procedures (using elevated concentrations of polymerase and increased cycle numbers) are being validated to generate STR profiles (using the Promega PowerPlex® 16 and Applied Biosystems AmpFlSTR® Yfiler® STR kits and the Applied Biosystems 3130xl Genetic Analyzer) from such samples. However, these STR profiles exhibit elevated levels of stutter, drop-in and drop-out which complicate data analysis. Thus, LCN STR allele calling guidelines, based on the analysis of replicate amplifications, are being established. The views expressed are those of the author(s) and do not reflect the official policy of the Department of the Army, the Department of Defense or the United States Government.
Keywords: Skeletal Remains, Demineralization, Low Copy Number, Autosomal STR Analysis, Y-Chromosome STR Analysis
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 31 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-4732-ISABS-2007
CLOSE ENCOUNTERS: PARTIAL DNA PROFILES AND FAMILY SEARCHING IN FORENSIC INVESTIGATIONS
Bieber FR1
1Departments of Pathology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA USA [email protected]
This presentation will provide a discussion of using formal genetic kinship analysis of forensic DNA data banks to identify additional investigative leads to resolve serious crimes. When comparing crime scene DNA samples to those from known offenders the ideal result is a precise multiple locus match. Partial DNA profile matches, however, can also offer opportunities to indirectly identify those who may be closely related to the actual perpetrator. That is, purposeful familial searches of DNA data banks can often identify a possible suspect who is not himself in the forensic database, but is closely related to someone who is. Such search successes can also occur unexpectedly when close, but not quite identical, DNA matches are observed between crime scene samples and the profiles of known convicted offenders. Addition of Y-chromosome or mtDNA typing of possible leads will anchor the partial match results to kindred and thereby help eliminate false leads developed from kinship searching methods. Worldwide more than 100 applications of familial searching of offender databases in criminal investigations have been reported around the world, principally in the U.K., with success in more than a dozen. To date, proactive family searching methods have not yet been widely embraced and are, to this point, under-utilized in most countries. This is surprising, given the potential power of such analysis to identify suspects indirectly and the fact that legislation in most countries does not bar use of the databases in this manner. Legal, ethical and policy implications will be fully discussed.
Keywords: family searching, DNA database, kinship analysis, partial DNA profile, forensic DNA
Suggested Reading: 1. Bieber FR, et al. Science 312 : 1315-1316, 2006, 2. Bieber, FR. J. Law, Med. Ethics 134(2): 222-233, 2006, 3. Bieber, FR. Leg Med, 7(4):230-243, 2005, 4. Biesecker LG, et al. Science 310 (5751) : 1122 – 1123, 2005, 5. DNA and the Criminal Justice System, MIT Press, 2004.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 32 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-9171-ISABS-2007
FORENSIC BIOLOGY TECHNIQUES IN THE SEPARATION OF MIXED TISSUES: FROM ROUTINE TO THE EXTREMES
Budimlija ZM1, Popiolek D2, West AB2, Fogt F3, Caragine T1, Mikulasovich R1, Lu C1, Prinz M1
1New York City Office of Chief Medical Examiner, Department of Forensic Biology, New York, New York, U.S.A.; 2New York University School of Medicine, Department of Pathology, New York, New York, U.S.A.; 3University of Pennsylvania School of Medicine, Department of Pathology, Philadelphia, Pennsylvania, U.S.A. [email protected]
Specimen mix-up and tissue cross contamination are well known problems in the field of surgical and anatomic pathology, having serious diagnostic, therapeutic and medicolegal implications. The aim of this study is to present a diagnostic algorithm designed to resolve such issues. Twenty cases of different complexity were included in the study of the genetic identity testing of the following ieter n: possible floater tissue contamination, identity of biopsy tissue questioned by patients, possibility of malignant tissue introduction by transplanted organs, laboratory biopsies mix-up. Following protocol was established: case triage – tissue separation (scraping or laser microdissecting microscopy) – DNA extraction (phenol-chlorophorm-isoamylalcohol or low copy number DNA extracting protocol) – DNA quantitation (real-time PCR) – DNA profiling (regular STR protocols – mitochondrial DNA linear array screening protocol – high sensitivity STR protocols). Using the proposed sample flow and decision matrix, all cases in question could be positively solved. All steps of the protocol were found to be reliable, reproducible and specific.
Keywords: forensic biology, techniques, tissue contamination, QA/QC, pathology
Suggested Reading: 1. Popiolek DA, Prinz MK, West AB, Nazzaruolo BL, Estacio SM, Budimlija ZM. Multiplex DNA short tan,Alonso A, Alves C, Suárez-Mier MP, Alberrán C, Pereira L, Fernández de Simón L, Martín P, García ,Gill P. Application of low copy number DNA profiling. Croat Med J. 2001 Jun;42(3):229-32. 2. Worsham MJ, Wolman SR, Zarbo RJ. Molecular approaches to identification of tissue contamination in surgical pathology sections. J Mol Diagn. 2001 Feb;3(1):11-5. 3. Hunt JL, Swalsky P, Sasatomi E, Niehouse L, Bakker A, Finkelstein SD. A microdissection and molecular genotyping assay to confirm the identity of tissue floaters in paraffin-embedded tissue blocks. Arch Pathol Lab Med. 2003 Feb;127(2):213-7.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 33 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-4787-ISABS-2007
MICROBIAL FORENSIC INVESTIGATION OF BIOCRIME AND BIOTERRORISM
Budowle B1
1FBI Laboratory, Quantico, USA [email protected]
Because of the availability of pathogenic microorganisms and the relatively low cost of preparing and disseminating bioweapons, there is a continuing threat of biocrime and bioterrorism. The risk and threat of bioterrorism and biocrime have become a large concern and challenge governments and society to enhance biosecurity. Law enforcement plays an important role in the assessment and investigative activities involved in an event of bioterrorism or biocrime. Key to a successful biosecurity program is increased awareness and early detection of threats facilitated by an integrated network of responsibilities and capabilities from government, academic, private, and public assets. To support an investigation the microbial forensic sciences will be employed to analyze and characterize forensic evidence with the goal of attribution or crime scene reconstruction. Microbial forensics focuses on the characterization, analysis and interpretation of evidence for attributional purposes from a bioterrorism act, biocrime, hoax or inadvertent agent release. A forensic investigation will attempt to determine the etiology and identity of the causal agent, often in a similar fashion as in an epidemiologic investigation. However, higher resolution characterization is needed. The microbial forensic tools include genetic and non-genetic based assays, phylogeny, ecology, etc. of the target microorganism. In addition, chemical and physical assays may help determine the process used to prepare, store, or disseminate the bioweapon.
Keywords: Bioterrorism, Microbial Forensics, Epidemiology, Genetics, Attribution
Suggested reading: 1. Budowle B, Murch RS, Chakraborty R. Microbial forensics: the next forensic challenge. Int J Leg Med., 2. Carus SW. Bioterrorism and biocrimes: the illicit use of biological agents since 1900. Fredonia Book, 3. Morse SA, Budowle B. Microbial forensics- application to bioterrorism preparedness and response. Infect Dis Clin North Am. 4. Van Ert MN, et al. J Clin Microbiol. 2007 Jan;45(1):47-53. 5. Budowle B, et al. Appl Environ Microbiol. 2006 Oct;72(10):6431-8.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 34 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-5627-ISABS-2007
STATISTICAL ISSUES SURROUNDING NATIONAL DNA DATABASES
Budowle B1
1FBI Laboratory, Quantico, USA. [email protected]
DNA databases have proven extremely useful for developing investigative leads for a variety of crimes where there is no known suspect, for linking multiple cases committed by the same individual, and for identifying human remains attributed to a missing person. These databases have been designed solely to develop investigative leads. However, some issues have been raised regarding the use or the analysis of the DNA profiles contained within the data bases and particularly the CODIS data. These are 1) using the large number of reference offender DNA profiles to invalidate the current assumptions for assessing the statistical significance of an evidence profile; 2) using the frequency of observing a DNA profile in the offender database in lieu of the more appropriate random match probability (or likelihood ratio) for assessing the rarity of a DNA evidence profile in a particular case; and 3) using the current searching algorithms in an attempt to identify perceived highly probable kinship relationships between an evidence sample and a offender DNA profile. If not properly understood, the information contained within CODIS and/or the interpretation of candidate matches obtained through CODIS searches may be misinterpreted or inappropriately applied. These issues will be addressed as they apply to the CODIS and its operational constraints.
Keywords: DNA Database, CODIS, Statistics, Validity, Interpretation
Suggested Reading: 1. P.D. Martin, National DNA databases: practice and practicability. A forum for discussion. Prog. Fore, 2. B.S. Weir, Matching and partially-matching DNA profiles. J. Forens. Sci. 49 (2004) 1009-1014. 3. W. Feller. An Introduction to Probability Theory and its Applications, Vol. 1, Third Edition. 4. John W,R. Chakraborty and L. Jin, A unified approach to study hypervariable polymorphisms: Statistical cons. 5. Budowle, B., Planz, J.V., Chakraborty, R., Callaghan, T.F., and Eisenberg, A.J.: Clarification of Statistical Issues Related to the Operation of CODIS.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 35 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-8367-ISABS-2007
VALIDATION OF NEW SRY MARKER FOR USE IN FORENSIC CASEWORK
Kastelic V1, Drobnič K1
1Forensic Science Centre, Ministry of the Interior, Ljubljana, Slovenia [email protected]
Abstract Accurate gender determination is crucial for forensic investigations, which is now in commercially available kits established on amelogenin sex test. Many studies identify that this test is not reliable enough due to the failure to amplify the amelogenin Y-homolog from a phenotypically normal male. In our previous work we have reported successful co-amplification of a new sex marker residing on SRY gene resulting in 96 based pair long PCR product only in male samples, utilizing AmpFlSTR SGM Plus and Power Plex 16 identification kits, respectively. The aim of this study is to get more information about mutation rates within binding sequences of a new set of primers and to determine their specificity and sensitivity. To validate reproducibility of amplification of SRY gene with the mentioned pair of primers, we tested 115 unrelated male Slovenians. We did not observe any null allele. Male DNA in different concentrations was used for determination sensitivity of new set of primers. Reproducible and reliable results were obtained from as low as 100 pg of template of male DNA, indicating high sensitivity of the assay. Studies with females revealed a high specificity of the new SRY marker because no PCR product was detected even at concentration of 10.0 ng/mL of female DNA.
Keywords: validation, sex determination, amelogenin gene, sry gene, mutation
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 36 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-184-ISABS-2007
INTERACTION OF PERSONNEL AT THE CRIME SCENE INVOLVING A BURIED BODY: THE ROLE OF FORENSIC ARCHAEOLOGIST
Drobnič K1
1Forensic Science Centre, Ministry of the Interior, Ljubljana, Slovenia. [email protected]
The paper addresses the pivotal role that forensic archaeologists should play in the exhumation of human remains, as the archaeological methods could be a deciding factor in the reconstruction of the events surrounding a death, although there are countries that have yet to grasp its importance. As the excavation of human remains is a complex process that requires integrated involvement of many forensic personnel such as crime scene officers, archaeologist, anthropologists and pathologists. The roles and responsibilities of each of them are discussed. While between them, there is an overlapping area of expertise, a crime scene manager is the person who controls and directs their activities at the crime scene. I claim also that proper collection and preservation of evidence are paramount to efficient and successful identification of the dead and that people involved in a criminal justice system should be systematically informed about the development in the forensic science. Before any final decision about the identity is made all evidence, not only from DNA analysis but also from anthropological and other “classical” forensic data, as well as conditions of disappearance must be compared and should be consistent.
Keywords: buried body, exhumation, archaeology, crime scene officer, evidence collection
Suggested Reading: 1. Skinner M, Alempijevic D and Djuric-Srejic M. Forensic Sci Int 2003;134: 82-83. 2. Skinner M, Sterenberg J. Forensic Sci Inter 2005;151: 221-232. 3. Menez LL, Forensic Sci Int 2005;152:311-315. 4. Cattaneo C, Forensic Sci Int 2007;165:185-193. 5. Steadman DLW and Haglund WDJ, Forensic Sci 2005;50:23-29.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 37 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-1123-ISABS-2007
GeneMarker® HID: AN EFFECTIVE NEW SOFTWARE TOOL FOR THE ANALYSIS OF STR DATA
Holland M1, Liu CSJ2, Hulce D2, Wan N2
1Penn State University, University Park, PA; 2SoftGenetics®, LLC, State College, PA, USA [email protected]
The analysis of STR data generated from forensic evidence requires specially designed software. Essential features of the software include: 1) user-friendly interface and functionalities, 2) user-defined parameters, 3) accurate processing of raw data, 4) quality assessment of analyzed data, 5) mechanism for flagging data, 6) effective management of data files, 7) rapid analysis times, and 8) adaptable reporting features. Through collaboration with the Forensic Science program at Penn State University, a forensic version of the software package GeneMarker® was developed by SoftGenetics®, a company in State College, Pennsylvania, for the analysis of STR data from forensic samples. GeneMarker® HID meets or exceeds the criteria listed above and is an attractive alternative to GeneMapper® ID. GeneMarker® HID uses the raw data files generated on any Applied Biosystems instrument (e.g., 310, 3130). The analyzed data is presented in a user-friendly manner and provides functionalities that meet or exceed forensic expectations. Validation studies were performed by doing side-by- side analysis of STR profiles from single source, mixed, and LCN samples, comparing the data analyzed with GeneMarker® HID with the same data analyzed using GeneMapper® ID. This presentation will walk participants through the features of GeneMarker® HID and illustrate the effectiveness of the software as a tool for the analysis of STR data in forensic casework, paternity testing, analysis of data from customized STR kits, and for virtually any other associated STR-related fragment analysis. SoftGenetics® has been recognized by Applied BioSystems as a valued 3rd party provider of analytical software for their electrophoresis systems.
Keywords: STR, Analysis, Software, GeneMarker, Forensics
Suggested reading: 1. K.E. Roberts, J.J. McElroy, W.P.K. Wong, E. Yen, A. Widlitz, R.J. Knowles, J.H. Morse. BMPR2 Mutations in Pulmonary Arterial Hypertension with Congenital Heart Disease. European Respiratory Journal 2004; 24: 371-374 2. Rajagopalan H, Jallepalli PV, Rago C, Velculescu VE, Kinzler KW, Vogelstein B, Lengauer C. Inactivation of hCDC4 can cause chromosomal instability. Nature. 2004 Mar 4;428(6978):77-81. 3. Robert R. Freimuth, Ming Xiao, Sharon Marsh, Matthew Milton, Nicholas Addleman, Derek J. Van Booven, Howard L. McLeod and Pui-Yan Kwok. Polymorphism discovery in 51 chemotherapy pathway genes. Human Molecular Genetics, 2005, Vol 14, No. 23.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 38 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-6593-ISABS-2007
THE DEVELOPING ROLE OF FORENSICS IN DETERRING VIOLENCE AND GENOCIDE
Huffine E1, Crews J1, Davoren J1
1Bode Technology Group, Lorton, VA, USA [email protected]
The development of DNA-testing capabilities within the former Yugoslavia to identify the missing began the process of returning names to thousands of the missing. The most striking example was the successful application of a DNA-led identification system for the victims of Srebrenica. To date, almost 4,100 individuals related to the Fall of Srebrenica in July of 1995 have been identified, almost all of whom were Bosnians. Without DNA testing, very few of these victims could be identified. The overwhelming weight of evidence and number of identifications, made possible by forensic DNA testing, was a critical factor in helping to compel a national entity to admit their role in genocide. Forensic DNA testing was instrumental in the admission of genocide and in the indictment of hundreds of alleged perpetrators. Forensic DNA testing provided critical evidence that was used to hold individuals and nations accountable for crimes against humanity and genocide. The rapid advances in the understanding and application of not only the science of forensic DNA testing, but also its legal-judicial implications, permits such testing to be used in a new context; to help deter violence, systematic rape, and genocide.
Keywords: DNA, Forensics, Genocide, Bode, Violence
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 39 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-7780-ISABS-2007
THE APPLICATION OF NON-CODIS MiniSTRs TO RESOLVE COMMINGLED SKELETAL REMAINS FROM THE KOREAN WAR
Just R1, Sturk K1, Barritt S1, Irwin J1, Coble M1
1Armed Forces DNA Identification Laboratory, Rockvile, MD, USA. [email protected]
Highly degraded human remains pose a challenge to many forensic laboratories due to the difficulty of producing short tandem repeat (STR) profiles using standard nuclear DNA typing protocols. Some laboratories perform mitochondrial DNA (mtDNA) sequence analysis on such highly degraded samples. However, mtDNA is not a unique identifier, and in cases involving commingled remains from a large number of individuals there will frequently be multiple non-related individuals whose mtDNA profiles are identical. To address these limitations, we have multiplexed 8 high heterozygosity non-CODIS STR markers (D2S441, D2S1776, D3S4529, D6S474, D9S2157, D10S1248, ATA63, and D22S1045), utilizing small amplicons (~70-170 basepairs) in four dye channels. The markers were used to type and reassociate samples matching common mtDNA types from a large set of commingled remains repatriated from North Korea. Additionally, the markers were used to add additional loci when likelihood ratios (LRs) calculated from standard STR and mtDNA typing were low. These data demonstrate that sorting and reassociation of such samples can be accomplished through the use of these non-CODIS nuclear markers when multiple skeletal elements cannot be distinguished using mtDNA. The data additionally show that these markers can be used to complement low copy number (LCN) nDNA results and add weight to the statistics generated with CODIS markers, mtDNA control region profiles, and select coding region mtDNA single nucleotide polymorphisms (SNPs).
Keywords: STR, miniSTR, non-CODIS, multiplex, likelihood ratio
Suggested Reading: 1. Coble and Butler (2005) Characterization of New MiniSTR Loci. J. Forensic Sci. 50(1): 43-53. 2. Hill, Coble, and Butler (2007) Characterization of 26 miniSTR loci. J Forensic Sci. in press. 3. Parsons et al. (2007) Application of novel \”mini-amplicon\” STR multiplexes. FSI Genetics 1:175-179. 4. Dixon et al. (2006) Analysis of artificially degraded DNA. Forensic Sci. Int. 164: 33-44. 5. Gill et al. (2006) The evolution of DNA databases. Forensic Sci. Int. 156: 242-244.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 40 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-8264-ISABS-2007
HOMICIDE INVESTIGATION: ANTHROPOLOGICAL & GENETIC ANALYSIS OF CRIME SCENE
Lee H1
1Forensic Science, University of New Haven; Forensic Laboratory, Connecticut State Police; Forensic Research and Training Centre [email protected]
Homicide Investigation Anthropological & Genetic Analysis of Crime Scene Dr. Henry C. Lee Chair Professor, Forensic Science, University of New Haven Chief Emeritus, Forensic Laboratory, Connecticut State Police Director, Forensic Research and Training Centre Despite countless historical cases from which to learn and improve upon, we continue to watch serious crimes go unsolved or end without justice served because of errors associated with the crime scene investigation. Virtually no jurisdiction in the United States or abroad has shed this plague. It seems ironic that while forensic science has experienced significant advances and improvements, the utilization of forensic science is still at the mercy of the crime scene. If the crime scene is not properly managed, all of the technology and advances in DNA we currently possess will be unable to overcome these shortcomings, and true justice will continue to be unattainable in far too many cases. A high-quality crime scene process need not be elaborate or sophisticated; rather, the simple adherence to fundamental principles and procedures will suffice. The procedure of anthropological and genetic analysis of crime scene will be presented. The essential crime scene functions involve the recognition, documentation, collection, and preservation of all relevant physical evidence: pattern, conditional, transient, transfer, and associative in nature will be addressed. If these elements are achieved, then there is a much higher probability that the case will be resolved through an anthropological and genetic reconstruction of the events and an accurate interpretation of the available information and data. The goal of this lecture is to see the day when it is no longer common for a trial or investigation to conclude abruptly or implode because of mistakes made at the crime scene. Unlike a great novel or epic movie, Crime scene investigators and forensic scientists have only one opportunity to conduct and original crime scene search. History is a valuable, but also expensive lesson. Case examples will used to illustrate for forensic scientists and crime scene investigator to learn from history and avoid making similar mistakes in crime scene investigation.
Keywords: Homicide, Crime Scene, Genetic Analysis, Anthropology, Investigation
Suggested reading: 1. Dr. Henry C. Lee\’s Crime Scene Handbook
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 41 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-2129-ISABS-2007
MISSING KIDS IDENTIFICATION: THE PROKIDS INTERNATIONAL PROGRAM
Lorente JA1, Alvarez JC1, Martinez-Espin E1, Martinez-Gonzalez LJ1, Entrala C2, Fernandez-Rosado F2, Villanueva E1, Luna A3
1Dept. Legal Medicine, University of Granada, Granada, Spain; 2LORGEN GP, BIC-P.T.Ciencias Salud, Armilla, Granada; 3Genetic Laboratory, PGR, Mexico City, Mexico. [email protected]
Missing person’s identification is a collaborative effort where DNA analysis is usually of the greatest help, especially in those cases where samples are degraded or the identification is based in badly preserved bones or partial skeletons. Nevertheless, DNA is also useful to identify \”other kind\” of missing persons that are not dead: missing kids. According to UN and some NGO reports, there are over 1 million kids reported as missing; these are kids that have been taken from their families and are being exploited or trafficked. We have developed the PROKIDS Program to generate two independent databases that are automatically compared each other. The first database or reference database (KD) is composed of DNA (autosomal and mitochondrial data) from mothers (other family members could be also included) of missing kids; the second database or questioned database (QD) is composed of DNA (autosomal and mitochondrial data) from children that have been found without the family, and/or exploited and/or known as human traffics victims. The University of Granada has already signed agreements with the governments of Mexico and Guatemala (other countries to come) to collect, analyze and generate the above mentioned databases. A global and coordinated effort is necessary to help to solve this problem.
Keywords: DNA, missing persons, missing kids, mitochondrial DNA, autosomal DNA
Suggested Reading: 1. Lorente JA et al. Croat Med J (2001) 42: 267-270 2. Lorente JA et al. Int J Legal Med (2001) 116: 187-190 3. Lorente JA et al. Science (2000) 290 : 2257-2258
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 42 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-7358-ISABS-2007
DNA IDENTIFICATION OF SKELETAL REMAINS FROM THE SECOND WORLD WAR MASS GRAVES UNCOVERED IN SLOVENIA – FIRST RESULTS
Marjanovic D1,2, Durmic-Pasic A1, Bakal N1, Haveric S1, Kalamujic B1, Kovacevic L1, Ramic J1, Pojskic N1, Skaro V2, Projic P2, Bajrovic K1, Hadziselimovic R1, Drobnic K3, Huffine E4, Davoren J4, Primorac D5,6
1Institute for Genetic Engineering and Biotechnology, Sarajevo, Bosnia and Herzegovina, 2“Rudjer Boskovic” Institute, Department of Molecular Medicine, Forensic Genetics Group, Zagreb, Croatia, 3Forensic Laboratory and Research Center, Ministry of Interior, Ljubljana, Slovenia, 4 Bode Technology Group Inc, Springfild, VA, USA, 5 Medical School, University of Split, Split, Croatia, 6 Medical School, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia [email protected]
To present joint effort of three institutions in the identification of human remains from the World War II found in two mass graves in Škofja Loka area, Slovenia. DNA from bone and teeth samples from 27 remains, found in two small and closely located mass graves from Skofja Loka area (Slovenia), was isolated using either standard phenol/chloroform alcohol extraction or optimized Qiagen DNA extraction procedure. Some recovered samples required employment of additional DNA purification methods, such as N-buthanol treatment. QuantifilerTM Human DNA Quantification Kit was used for DNA quantification. PowerPlex 16 kit was used to simultaneously amplify 15 STR loci. Electrophoresis of the amplification products was performed on ABI PRISM 310 genetic analyzer. Matching probabilities were estimated using the DNA View program. Out of all processed samples (bones or teeth), taken from the remains, 15 remains were fully profiled at all 15 STR loci. The other twelve profiles were partial. The least successful profile included 13 loci. Also, 69 referent samples (buccal swabs) from potential living relatives were collected and profiled. Comparison of victims’ profile against referent samples database resulted in 4 strong matches. In addition, five other profiles were matched to certain referent samples with lower probability. Our results show that in over six decades since the end of the Second World War, DNA analysis is the solution to the identification of the remains from that period. Additional analysis of Y-STRs and mtDNA markers should be performed in the second phase of the identification project.
Keywords: DNA identification, mass graves, skeletal remains, Slovenia, Second World War
Suggested Reading: 1. Andjelinovic S. et al. Croat Med J 2005;46(4):530-9. 2. Alonso A. Et al. Croat Med J 2001 ;42 :260-266. 3. Fisher DL. Et al J Forensic Sci 1993 ;38(1) :60-8. 4. Dixon LA. Et al. Forens. Sci. Int 2006; 164(1):33-44. 5. Butler JM. London: Elsevier Academic Press; 2005.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 43 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-3039-ISABS-2007
THE DOMESTIC CAT: DOMESTICATION, BREED DEVELOPMENT AND A POPULATION GENETIC DATABASE FOR COMPUTATION OF MATCH PROBABILITIES
Menotti-Raymond M1, David V1, O’Brien SJ1
1Laboratory of Genomic Diversity, National Cancer Institute at Frederick, Frederick, Maryland, USA [email protected]
A panel of ten tetranucleotide simple tandem repeat (STR) loci, developed for the genetic individualization of domestic cat samples, has been used to generate a population genetic database of domestic cat breeds. The multiplex was co- amplified in a sample collection of 1240 domestic cats representing 38 cat breeds and a small sample set of outbred domestic cats. The power for genetic individualization of domestic cat samples of the multiplex is moderate to high within cat breeds, with a probability of match (Pm) range of 2.4E-06 – 1.3E-14, and within outbred cats, which represent approximately 97% of household cats, a Pm of 7.8E-13 was observed. We have utilized this dataset utilizing the STRUCTURE algorithm and an additional dataset of 284 single nucleotide polymorphisms (SNPs) to examine population substructure and breed relationships. Results of these studies and a recent completed work on cat domestication will be presented.
Keywords: Domestic cat, forensic typing system, STR, genetic database, cat breeds
Suggested Reading: 1. Menotti-Raymond, M., David, V.A., and O’Brien, S.J. Pet cat hair implicates murder suspect. Nature 1997 Apr 24;386(6627):774. 2. Menotti-Raymond, M., David, V.A., Stephens, J.C., Lyons, L.A., and O’Brien, S.J., Genetic Individualization of Domestic Cats Using Feline STR Loci for Forensic Applications. J Forensic Sci. Nov;42(6):1039-51 1997 3. Menotti-Raymond, M., David, V., Wachter, L., Yuhki, N., and O’Brien, S.J. Quantitative polymerase chain reaction-based assay for estimating DNA yield extracted from domestic cat specimens, Croat Med J. 2003 Jun;44(3):327-31 4. Menotti-Raymond, M., David, V.A., Wachter, L.L., Butler, J.M., and O’Brien, S.J. An STR forensic typing system for genetic individualization of domestic cat (Felis catus) samples, J Forensic Sci. 2005 Sep;50(5):1061-70. 5. Murphy, W.J., Davis, B., David, V.A., Agarwala, R., Schäffer, A.A., Pearks-Wilkerson, A.J., Neelam, B., O’Brien, S.J., and Menotti-Raymond, M. A 1.5 megabase resolution radiation hybrid map of the cat genome and comparative analysis with the canine and human genomes. Genomics, 2007. in press
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 44 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-1586-ISABS-2007
A RETROSPECTIVE OF HIGH SENSITIVITY DNA TESTING FOR CRIMINAL CASES
Mikulasovich R1, Caragine T1, Baum H1, Prinz M1
1Department of Forensic Biology, Office of Chief Medical Examiner, New York, NY, USA. [email protected]
The Office of the Chief Medical Examiner’s Department of Forensic Biology has validated a method for recovery of Low Copy Number quantities of DNA from challenging samples. Previously, the bulk of DNA analysis was reserved for the more promising blood and sexual assault kit evidence collected during the investigation of homicides and sexual assaults. Items considered for Low Copy Number DNA testing may include any touched item from a crime scene. This effectively expands the type of case in which DNA analysis would prove a useful investigative tool. Today, crimes of arson, property theft, and assault may be investigated with DNA analysis as an option. For example, DNA has been recovered from detonation devices, fingerprint smudges identified at the scene of burglaries, and presumably degraded samples from cold cases. In order to successfully implement this testing all concerned parties including evidence crime scene specialists, forensic biologists and attorneys should be educated regarding legal considerations. This study covers the analytical methods practiced in the department and the interpretation guidelines applied to the data collected as well as a review of a select number of cases in which Low Copy Number DNA evidence has been presented.
Keywords: Low Copy Number DNA Testing, High Sensitivity DNA Testing, Criminal Cases, Validation, Testimony
Suggested Reading: 1. Gill P. Croat Med J 2001; 42:229-232, 2. Tamariz J. J Forensic Sci. 2006 Jul;51(4):790-4., 3. Schiffner LA. Croat Med J. 2005 Aug ;46(4) :578-86. 4. Gill P. Forensic Sci Int. 2000 Jul 24;112(1):17-40. 5. Gill P. Forensic Sci Int. 2006 Jul 13;160(2-3):90-101
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 45 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-5462-ISABS-2007
DEVELOPMENT OF A MULTIPLEX SINGLE BASE EXTENSION ASSAY FOR mtDNA HAPLOGROUP TYPING
Nelson TM1,3, Just RS2, Loreille O2, Schanfield M3, Podini D3
1San Francisco Police Department, Forensic Services Division, San Francisco, CA, USA; 2The Armed Forces DNA Identification Laboratory, Rockville, MD, USA; 3The George Washington University Department of Forensic Sciences, Washington, DC, USA [email protected]
Previous forensic human identification and anthropological studies have demonstrated the utility of coding region and/or complete mtDNA genome sequencing for the differentiation of common mtDNA haplogroups following standard control region (CR) sequencing methods. Since current mtDNA sequencing entails the analysis of multiple, overlapping amplicons to account for large stretches of the mtDNA molecule, we sought to develop a single, multiplex amplification strategy to rapidly distinguish major mtDNA haplogroups A, B, C, D, E, F, G, H, I, L1/L2, L3, M, N and X. This method targets twelve known polymorphic locations using the SnaPshot single base extension assay, also referred to as minisequencing, in order to classify individuals into one of the listed haplogroups as well as provide inference of maternal ancestral origin. The single base extension assay described in this study has the ability to type various haplogroups using both pristine samples and highly degraded forensic specimens. A survey of 146 specimens demonstrated a marked increase in haplogroup or macro- haplogroup assignment (87%) relative to conventional CR sequence analysis (78%), which is considerably more labour intensive and costly. This single base extension assay was also successful in analyzing decades old, embalmed skeletal remains dating back to WWII, thereby demonstrating its effectiveness as a rapid, high throughput method for mtDNA haplogroup typing in human identification efforts (e.g. identification and re-association of remains) and anthropological studies.
Keywords: mtDNA, haplogroups, SNP, minisequencing, ancestry
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 46 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-8166-ISABS-2007
ICMP: INTEGRATING FORENSIC SCIENCES IN HIGH THROUGHPUT IDENTIFICATION OF MISSING PERSONS FROM MASS GRAVES
Parsons T¹
¹ International Commission on Missing Persons, Sarajevo, Bosnia and Herzegovina [email protected]
This presentation will provide an overview of integrated forensics sciences used by the International Commission on Missing Persons to identify missing persons, primary from mass graves in the former Yugoslavia. To date, over 12,000 individuals have been identified using a system that is \”led\” by high throughput DNA typing and matching of skeletal remains to family reference samples. However, the success of this system is not based on DNA alone, but rather on fully integrated processes involving: 1) state-of-the art forensic archaeological excavation that documents remains and associated evidence, and recovers the remains in anatomical association to the best extent possible, 2) physical anthropological analysis of age, sex, stature, distinctive characteristics and antemortem: postmortem comparison, 3) pathology examination, 4) personal effects, 5) DNA comparisons. Central to all these processes are custom informatics systems that track and link data, and perform DNA matching and statistical analysis. The particularly complex problem presented by secondary mass graves, with fragmented and commingled bodies scattered among multiple secondary sites will be discussed, as well as practical and theoretical considerations relating to the combination of DNA and non-DNA evidence in determining identity.
Keywords: Mass graves, DNA Identification, STRs, forensic archeology, forensic anthropology
Suggested Reading: 1. Parsons, T.J., Huel, R., Davoren, J., Katzmarzyk, C., Milos, A., Selmanovic, A., Smajlovic, L., Coble, M.D., Rizvic, A. Application of novel “mini-amplicon” STR multiplexes to high volume casework on degraded skeletal remains. FSI Genetics 2007 1:175-179.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 47 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-349-ISABS-2007
STRUCTURED POPULATION DATABASES IN FORENSIC GENETIC PRACTICE
Roewer L1
1Dept. Forensic Genetics, Institute of Legal Medicine, Charité – University of Medicine, Berlin, Germany. [email protected]
Over the last years, the analysis of Y chromosomal short tandem repeat (STR) loci has become a powerful tool for resolving unbalanced stain mixtures in forensic casework as well as for a broader clientele interested in kinship and genealogical testing. The need of information regarding the spatial distribution of Y-STR haplotypes in order to obtain reliable frequency estimates has produced an enormous amount of regional population studies, most of which were submitted to the online Y-STR Haplotype Reference Database (YHRD). The database is the largest repository of its kind in the world (currently > 51.000 DNA profiles from > 450 different populations) and aims to support the decision-making process of forensic analysts. YHRD considers reduction of the available number of polymorphisms on the Y chromosome to a uniform data string of 11 highly variable Y-STR loci as an efficient way to rapidly scan population samples and to make their Y chromosome make-up comparable. Typing of the YHRD 11-locus core set is facilitated by commercial products, namely diagnostic PCR kits, and endorsed by scientific and practitioner’s societies as ISFG or SWGDAM. YHRD is structured by the assignment of each population sample to a system of hierarchically arranged metapopulations (a group of discrete populations interacting via gene flow and migration) which facilitates the statistical evaluation of haplotypes matches due to a significant enlargement of sample sizes. Due to the rapid growth and diversification of the database the definition of substructures within continents or language groups is now feasible solely on the basis of the very dense genetic data as exemplified for the European dataset. Large sample numbers within certain metapopulations also allowed the development of extrapolation methods to estimate the frequency of unobserved or rare haplotypes (“haplotype frequency surveying method” based on the availability of a dense dataset of close neighbours surrounding the haplotype in question). Essential for the YHRD project as well as for similar databases (e.g. EMPOP for mtDNA sequences) is its collaborative character relying on the engagement of individual laboratories to make their data accessible and to share high standards regarding data quality. Due to the availability of Y- SNP haplogroup data the YHRD will be developed further to supply forensic experts with information on the biogeographical ancestry in no-suspect cases. The utility of the database in forensic and genealogical casework will be illustrated.
Keywords: Population database, Y chromosome, Substructure, Haplotype, STR
Suggested Reading: 1. Willuweit S, Roewer L (2007) Y chromosome haplotype reference database (YHRD): Update. FSIGen 1(2) 83-87 2. Parson W, Dür A (2007) EMPOP – A forensic mtDNA database. FSIGen 1(2) 88-92 3. Kayser M et al (2007) Relating two deep-rooted pedigrees…FSIGen 1(2) 125-8 4. Jobling, Hurles, Tyler-Smith (2004) Human Evolutionary Genetics. Garland Science, New York, Abingdon 5. Butler J, Forensic STR Typing, Elsevier
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 48 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-7949-ISABS-2007
INTEGRATED FORENSIC ANALYSIS OF FINNISH WORLD WAR II SOLDIERS
Sajantila A1, Hedman M1, Palo J1
1Department of Forensic Medicine, University of Helsinki, Finland [email protected]
In the course of the Second World War (WWII) Finland fought three separate wars: the Winter War (1939-1940) and Continuation War (1941- 1944) against the Soviet Union and the so called Lapland war against Germany in 1944-1945. Altogether 93 500 Finnish soldiers lost their lives, and 15 500 soldiers went missing in action or were left to the Soviet territory. There has been a growing public interest in repatriation of the Finnish soldiers MIA since the previous battlegrounds became accessible after the collapse of Soviet Union in 1991. The repatriations were enabled by an agreement between Finland and Russian Federation signed in 1992. This marked the start of ongoing field searches, repatriation, identification and reburial of Finnish soldiers. At present (2007), the skeletal remains of roughly 1000 soldiers have been found and repatriated, and 210 of these have been identified.
Keywords: forensic genetics, forensic anthropology, war victims, DNA, forensic pathology
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 49 ORAL PRESENTATIONS Invited Lectures – Forensic Genetics Abstract number: ABS-9471-ISABS-2007
THE HUMAN TOOTH AS A SOURCE OF FORENSIC DNA
Smith B1, Loreille O1, Lee D1, Barritt-Ross S1
1Armed Forces DNA Identification Laboratory, Rockville, Maryland, United States. [email protected]
Teeth have long represented a fascinating centerpiece for forensic scientists. To the odontologist, teeth are the basis for human individualization, particularly when radiographic documentation of past clinical treatment is available for comparison. To the anthropologist, teeth provide cultural clues and evidence of race and age. Now, the forensic biologist has come to rely increasingly on the tooth as the most enduring remnant of the human skeleton. The human tooth represents a protective crucible for DNA evidence that withstands harsh environmental challenges which would otherwise compromise hair, bone and soft tissues as viable sources for DNA evidence. Some laboratories are quite comfortable extracting DNA from blood, muscle or even bone, but are reluctant to work with teeth because of the unique anatomy of the sample. This presentation will elaborate upon the casework and research-based experiences of the Armed Forces DNA Identification Laboratory (AFDIL) in working with dental specimens over the last 15 years. The various technical approaches to sectioning or grinding teeth will be compared and casework examples will be presented to include the analysis of teeth recovered from a variety of forensic scenarios including tropical and salt water environments. The views expressed are those of the author(s) and do not reflect the official policy of the Department of the Army, the Department of Defense or the United States Government.
Keywords: Teeth, DNA, Forensic, Dental, AFDIL
Suggested Reading: 1. Shiroma et al; J Forensic Sci, July 2004, Vol. 49, No.4, 2. Gaytmenn and Sweet; J Forensic Sci, May 2003, Vol. 48, No. 3, 3. Sweet and Hildebrand; J Forensic Sci, 1998, Vol. 43, No. 6, 4. Smith et al; J Forensic Sci, September 1993, Vol. 38, No. 5, 5. Schwartz et al; J Forensic Sci, 1991, Vol. 36, No. 4
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 50 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Molecular Anthropology Molecular Anthropology
Abstract number: ABS-6438-ISABS-2007
LINKAGE DISEQUILIBRIUM IN AN ISOLATED POPULATION FROM POLYNESIA: PROSPECTS FOR MAPPING COMPLEX TRAITS
Deka R1
1University of Cincinnati Medical Center, Cincinnati, Ohio, USA [email protected]
We have witnessed phenomenal progress in human genetics that has paved the way for genome-wide association studies. The HapMap project that provided data on genetic variation on four major human populations, has played a pivotal role in this development. A key question, however, is whether the catalog of common genetic variation in the continental populations would capture the patterns of genetic diversity in world-wide populations, particularly the isolated populations with recent evolutionary histories, who are likely to offer significant advantage in mapping complex traits? To address this issue, we have studied a Polynesian population, the Samoans, who have a relatively recent founding history and compared the patterns of genetic variation with the HapMap populations – the African, the European, the Chinese, and the Japanese. We have previously shown that the extent of genetic variation at microsatellite loci in Samoans is significantly reduced. Single Nucleotide Polymorphism (SNP) data on chromosome 21 (~8,000 SNPs spanning 12.1Mb of DNA with a density of ~2.0kb per SNP) show that long- range linkage disequilibrium (LD > 50kb) is significantly elevated in the Samoans compared to the other populations. Extension of this work to a genome-wide level (>250,000 SNPs at a resolution of ~11.5kb per SNP) provides comparable results – Samoans have extended levels of LD. Patterns of LD sharing among populations reveal portability of tagging SNPs across populations, particularly among those of recent shared ancestry. These observations highlight the benefits of population isolates in genetic association studies.
Keywords: Linkage Disequilibrium, Samoa, HapMap, Single Nucleotide Polymorphism, Complex Trait
Suggested Reading: 1. Dr. Pavao Rudan, Dr. Stanimir Vuk-Pavlovic, Dr. Bruce Budowle, Dr. Peter Underhill, Dr. Irena Martinovic Klaric
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 51 ORAL PRESENTATIONS Invited Lectures – Molecular Anthropology Abstract number: ABS-3985-ISABS-2007
ROMA IN CROATIA: INFERENCES ON PATERNAL GENETIC HERITAGE OF THE BAYASH
Martinović Klarić I1, Peričić Salihović M1, Barać Lauc L1, Pokupčić K2, Cukrov S2, Janićijević B1
1Institute for Anthropological Research, Zagreb, Croatia; 2Forensic Centre “Ivan Vučetić”, General Police Directorate, Ministry of the Interior of the Republic of Croatia, Zagreb, Croatia. [email protected]
The Bayash consist of numerous and small Romani groups speaking different dialects of the Romanian language and living dispersedly in Croatia, Hungary, Bosnia and Herzegovina, Serbia, Romania, Bulgaria, and to the lesser extent in Macedonia, Greece, Ukraine, Slovakia and Slovenia. The etnonyms Bayash or Boyash are used in Croatia and Hungary, Banyash, Karavlasi or Romanian Gypsy in Serbia and Băeşi in Romania. Diverse Bayash groups are also often called by professionyms based on their traditional occupations (Lingurari – spoon makers, Fusari – spindle makers Koritari – trough makers, Rudari – miners). Larger Bayash groups migrated to Croatia most likely during the 19th century, after abolition of slavery in Romania. This study is based on the field work in eastern (Baranja) and northwestern (Međimurje) Croatia where 156 Bayash adult men were recruited. As the exact place of origin and the time of arrival of the Bayash in Croatia are not precisely documented in written historical records or kept in the collective Bayash memory, we attempt to address the ambiguities regarding their origin, migration history and admixture based on the analysis of the battery of 22 diagnostic SNP/indel and 17 microsatellite (DYS19, DYS385, DYS389I, DYS398II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, GATA H4) Y chromosome markers.
Keywords: Roma, Bayash, Croatia, Y chromosome, high-resolution analysis
Suggested Reading: 1. Pericic et al. MBE 22 :1964-1975, 2005. 2. Barac et al. EJHG 11 :535-542, 2003. 3. Gresham et al. AJHG 69 :1314-1331, 2001. 4. Morar et al. AJHG 75:596-609, 2004. 5. Kalaydjieva et al. EJHG 9:97-104, 2001.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 52 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Molecular Anthropology Abstract number: ABS-1082-ISABS-2007
EVOLUTION OF HUMAN POPULATIONS AND THE STUDY OF THE GENETIC BASIS OF HUMAN LONGEVITY
Passarino G1
1Department of Cell Biology, University of Calabria, Italy. [email protected]
Individual variability of human lifespan is influenced by interplay of environmental, genetic and stochastic factors. The disentangling of such factors is complicated by cultural and genetic heterogeneity of human populations. Human societies undergo continuous changes with the improvement of the environment (for instance cleaner water and better food) and medical assistance. This leads to the increase of average and of maximum life span that are now far beyond where they were a few decades ago. Although the improvement of environmental conditions is occurring all over western societies, it has a different pace in different areas, and this reflects on the conditions of aging and of longevity. In addition, it has been shown that genetic factors are able to predispose to longevity in certain populations but not in others, either because some variants are population specific or because the interaction of that variant with the environment is specific for the geographic area. On the whole it is emerging that most of the factors influencing longevity are heterogeneous and population specific due to specific interactions between environment and genetic background. This may clearly affect the fraction of phenotypic variance due to genetic/environmental variance. Thus, it seem important a close monitoring in different populations of the trajectories of lifespan and of their correlation with different factors that are likely to affect longevity in that population.
Keywords: Aging, longevity, association studies, human demography, male- female ratio
Suggested Reading: 1. Christensen K et al (2006) Nat Rev Genet. 7(6) :436-448, 2. vB Hjelmborg J et al. (2006) Hum Genet. 119(3) :312-321 3. Passarino G et al. (2006) Hum Hered. 62(4):213-220 4. Passarino G et al. (2007) Biogerontology 8(3):283-290 5. Passarino G. Et al. (2002) Exp Gerontol. 37(10-11):1283-1239.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 53 ORAL PRESENTATIONS Invited Lectures – Molecular Anthropology Abstract number: ABS-xxxx-ISABS-2007
OGLIASTRA GENETIC PARK: A LINK OF HISTORY AND GENETIC MAKEUP OF AN ANCIENT POPULATION
Pirastu M1
1Institute of Population Genetics, National Research Council, C.N.R., Alghero and Shardna Life Sciences Cagliari, Italy [email protected]
Abstract not provided
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 54 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Molecular Anthropology Abstract number: ABS-2923-ISABS-2007
REALITY AND PERCEPTION IN MOLECULAR ANTHROPOLOGICAL STUDIES OF POPULATION STRUCTURE
Rudan P1
1Institute for Anthropological Research, Zagreb, Croatia [email protected]
The evaluation of influences of hereditary (intrinsic) and environmental (extrinsic) factors are reflected differently through phenotype of various human populations. This is why today the questions on the formation of population structure of a group are still topical. The overview of definitions of population structure given by demographers, geneticists and anthropologists show how present evaluations are still based on analyses of different sets of biological features, among which the molecular analyses are, after all, the most competent, as far as genetic structure is concerned. However, the anthropological definition considers also the much needed notion of culture – all what is transferred from one generation to another by non biological means. On the several examples of Croatian populations, as convenient models for studying population structure, the shaping of human populations is presented as a reflection of historical processes which are laboratories for their formation. Diversity of current findings is explained by migrational waves, different intensity of reproductive isolation and the reflection of historical layers. They are especially evident through a certain degree of congruity between linguistic and genetic distinctiveness. The reflection of the centuries of reproductive isolation (which is clearly evident on village inhabitants of eastern Adriatic), offers a possibility to apply the holistic analytic approach in the population structure studies.
Keywords: Anthropology, Population structure, Microevolution, Historical Migrations, Genetic epidemiology
Suggested Reading: 1. Angel, J.L. 1954. Human biology, health and history in Greece from first settlement until now. Yrbk. 2. Baker, P.T. 1966. Human biological variations as an adaptive response to the environment. Eugen. Qua 3. Cavalli-Sforza, L.L. and W.F. Bodmer 1971. The Genetics of Human Populations. W.H. Freeman and Co. 4. Harrison, G.A. and A.J. Boyce 1972. The Structure of Human Populations. Clarendon Press. Oxford 5. Hiernaux, J. 1963. Heredity and environment: Their influence on human morphology. A comparison of two independent lines of study. Am J Phys Anthropol. 1963 Dec;21:575- 89.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 55 ORAL PRESENTATIONS Invited Lectures – Molecular Anthropology Abstract number: ABS-5932-ISABS-2007
Y-CHROMOSOME PHYLOGENETICS AND PHYLOGEOGRAPHY
Underhill PA1
1Department of Genetics, Stanford University School of Medicine, Stanford, California [email protected]
Human history has experienced numerous episodes of population size fluctuations, fissions/ fusions and migrations that create considerable complexity in the contemporary gene pool. One approach is to disentangle the sequestered genetic memory to help understand migrations, population origins, substructure and histories. Non-recombining haploid systems like the Y-chromosome provide an accessible way forward. While there is no a priori reason for a 1:1 correlation between the evolution of a DNA molecule and other non-genetic evidence, the patterns of Y-chromosome substructure in contemporary populations provides alternative evidence for re-evaluating non-genetic views of pre-historical affinity and diversification. Since most of the haploid Y- chromosome has the special property of not recombining the sequential accumulation of genetic differentiation can be deciphered allowing the construction of an unequivocal genealogy reflecting the geographic relationships of various lineages. Pronounced non- random correlations between Y- chromosome varieties and geography often manifest as clines in frequency and/or accumulated diversity. These phylogeographic patterns of genetic diversification provide a molecular perspective to migratory routes, genetic barriers and polarity of net gene flow. Thus the phylogeographic pattern of Y chromosome differentiation offers stimulating clues concerning the origins of contemporary population affinities and substructure and provides an independent perspective to investigate ambiguities concerning resemblances and origins of populations based upon material culture, linguistic and other genetic knowledge. Illustrative examples will be presented.
Keywords: Y chromosome, cladistics, phylogeography, genetic diversity, population structure
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 56 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Molecular Anthropology Abstract number: ABS-7139-ISABS-2007
LESSONS ON mtDNA PHYLOGENY LEARNT FROM COMPLETE SEQUENCES
Villems R1
1Estonian Biocentre, Tartu, Estonia [email protected]
Gender-specific uniparental non-recombining haploid genetic systems – mitochondrial DNA and the Y chromosome (its non-recombining portion, NRY) – allow us to reconstruct well-resolved phylogenetic trees. Matrilineally inherited mtDNA, though tiny compared to autosomal chromosomes as well as to NRY, has been exploited in population genetics already for more than two decades, yet only recently structural information available from its little more than 16500 nucleotides, has been fully explored. The first lesson learned in this new and in a way final structural resolution was, somewhat reassuringly, that the established earlier robust skeleton of the global phylogenetic tree was fundamentally correct. Nevertheless, as it soon turned out, complete genomic resolution allowed to formulate new questions, hitherto inaccessible. Perhaps most importantly, it became a tool for the quest for global spread of autochthonous, region-specific basal maternal lineages, likely arisen soon after the African exodus. A nearly continuous line of such lineages alongside the southern flank of Eurasia, extending from India to Australia and, as its independent branch, to South China, lend strong credence to the southern pioneer human settlement phase of Eurasia and beyond. As about the second lesson, one may highlight greatly improved \”nested cladistic\” phylogeography for many otherwise seemingly uniformly spread haplogroups, allowing to draw conclusions about the source populations, expansion trajectories, etc. Last, but not the least, work in progress allows to identify maternal founders of specific populations at high resolution.
Keywords: phylogeny, phylogeography, mtDNA genomes, out-of-Africa, nested cladistics
Suggested Reading: 1. Torroni et al. (2006) Trends Genet 22:339-356 2. Underhill & Kivisild (2007) Annu Rev Genet (in press) 3. Macaulay et al. (2005) Science 308 :1034-1036 4. Palanichami et al. (2004) AJHG 75:966-978 5. Loogväli et al. (2004) Mol Biol Evol 21:2012-2021
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 57 ORAL PRESENTATIONS Invited Lectures – Advances in Genomic Methods Advances in Genomic Methods
Abstract number: ABS-1612-ISABS-2007
DATA MANAGEMENT SOLUTION AND ANALYTICAL INFRASTRUCTURE TO SUPPORT BASIC, CLINICAL AND TRANSLATIONAL RESEARCH
Kocher JP1
1Mayo Clinic, Rochester, Minnesota, United States [email protected]
The Bioinformatics Core at Mayo Clinic is contributing to the development of an integrated IT infrastructure to support research projects as well as the management and analysis of research data. This effort leverages two major initiatives at Mayo Clinic: 1) the development of the Enterprise Data Trust (EDT) data warehouse (in collaboration with IBM) and 2) the implementation of a Laboratory Information Management System for research. The integration of these components will enable IRB controlled access to patients’ clinical information and research data obtained from patients’ biospecimens. In addition, the Bioinformatics Core is actively engaged in developing a third component to enable the fast design, implementation and execution of analytical procedures. This development utilizes ontology’s and common data elements to describe both the analytical functions and the data they consume or produce in order to facilitate an environment for rapid analytic implementation and adoption. The system will use the InforSense Knowledge Discovery Environment (KDE) to provide a graphical user interface for the creation of analytical pipelines. The resulting integrated infrastructure will support the different phases of a research project, namely the identification and selection of patients for a study, the tracking, archival and retrieval of biological data derived from biospecimens and the analysis of these data. This presentation provides an overview of our current work and discusses some of the challenges faced.
Keywords: Data Management, Data Analysis, Infrastructure Development, Centralized Repository System, Project Support
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 58 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Advances in Genomic Methods Abstract number: ABS-5393-ISABS-2007
GENE EXPRESSION PROFILING TO ANALYZE THE TRANSCRIPTIONAL OUTPUT OF THE ENTIRE GENOME: THE EMERGENCE OF NON-CODING TRANSCRIPTS
Perez D, Pritchett J, Ducharme-Smith A, Halling M, Smith D
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA [email protected]
The sequencing of the human genome revealed that less than 2% of the entire human genome actually codes for protein. Much of the rest of the genome was previously thought of as junk. However, newer technologies have been developed including whole genome tiling arrays (which contain tiled oligonucleotides across the entire non-redundant portion of the human genome) which can now be utilized to characterize all genome wide transcripts, not only those that are associated with traditional genes and encode protein. Utilizing these arrays researchers have discovered an entire new world of transcription and have begun to realize that there are many more non-coding transcripts than genes. Some of the non-coding transcripts have already been found to play very important regulatory roles within cells and others are targets of alteration during cancer development. We have utilized these arrays to uncover an entirely new group of extremely large non- coding transcripts which are both abundantly expressed and high evolutionarily conserved. We have begun to examining these in cancer and have now found that many of them have altered expression in different cancers and some are specifically mutated in certain cancers. The discovery of new classes of regulatory transcripts makes one realize that the genome is much more complex than previously anticipated.
Keywords: tiling arrays, whole genome, non-coding transcripts, conserved, cancer markers
Suggested Reading: 1. Structured RNA in the ENCODE selected regions of the genome. Genome Res. 2007; 17: 852-864., 2. Science 2007; 1484-1488., 3. Science 2005; 308: 1149-1154., 4. Curr Mol Methods 2007; 7: 103-120.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 59 ORAL PRESENTATIONS Invited Lectures – Advances in Genomic Methods Abstract number: ABS-8348-ISABS-2007
THE DAWN OF THE $1,000 GENOME
Smith D1
1Mayo Clinic, Rochester, Minnesota, USA, Department of Laboratory Medicine and Pathology [email protected]
The technologies that were developed to sequence complex genomes, including humans, have changed the way that basic research and its translation into clinical practice can and will be conducted. In just a few years technologies advanced to the point that a complete sequence for the 3 billion base pair human genome could be generated at a cost of several hundred million dollars. In less than 7 years after that the technology has advanced further and this resulted in the complete nucleotide sequence for James Watson at a cost of only $300,000. Each year faster and cheaper technologies are developed for rapid DNA sequencing thus in less than 3 years the cost of sequencing your entire genome will cost less than $1,000 and the cost will continue be even cheaper over time. This will change both research and clinical practice. It will be possible to not only measure your DNA sequence at birth but to monitor that sequence through ones lifetime for the early detection of disease. This will also herald in the era of personalized medicine where detailed sequence information can best inform clinical strategies for each individual patient.
Keywords: Genome, sequencing, Next Generation, personalized medicine, high throughput technologies
Suggested Reading: 1. Gene Sequencing- The race for the $1,000 genome. Science 311: 1544-1546.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 60 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Invited Lectures – Advances in Genomic Methods Abstract number: ABS-4780-ISABS-2007
GENE EXPRESSION PROFILING TO IDENTIFY GENES PREDICTIVE OF CANCER PATIENT OUTCOME
Vasmatzis G1
1Department of Laboratory Medicine and Pathology, Division of Experimental Pathology, Mayo Clinic, Rochester, Minnesota, USA [email protected]
Improved prediction of prostate cancer systemic progression could immediately impact the management of high risk patients. This paper describes a process for the identification of genes that can report on the aggressiveness of prostate tumors and thereby add to the information provided by current pathological analysis. The analysis presented is based on the premise that tumors are characterized by aberrant gene expression that is heterogeneous among tumors. The first step was to identify genes expressed at abnormally high levels in only some tumors. These genes were subsequently stratified by their association with aggressive phenotypes and were filtered to exclude genes with redundant expression patterns. Genes with the most predictive value for prostate cancer aggressiveness were selected for validation in a case/control study. Cases (systemic progression within 5 y) and controls (no systemic progression at 7 y follow-up) were matched for all clinical and pathological criteria from time of prostatectomy. Both cases and controls, therefore, could have nodal invasion or seminal vesicle involvement at the time of initial treatment. In the case-control analysis TOP2A, RRM2, and SSTR1 were strongly associated with poor outcome. These genes may add information not available from current clinical measures and improve the prognosis of prostate cancer. A model incorporating topoisomerase 2a (TOP2A) gene expression, cadherin 10 (CDH10) gene expression, the ploidy status, and fusion status based on ERG ETV1 and ETV4 expression resulted in a 85% sensitivity and 67% specificity for the identification of men with systemic progression and death from high grade prostate cancer.
Keywords: Bioinformatics, Prognosis, Cancer, proliferation, prostate
Suggested Reading: 1. Kosari F, Parker AS, Kube DM, Lohse CM, Leibovich BC, Blute ML, Cheville JC, Vasmatzis G. Clear cell renal cell carcinoma: gene expression analyses identify a potential signature for tumor aggressiveness. Clin Cancer Res. 2005 Jul 15;11(14):5128-39. 2. Parker AS, Kosari F, Lohse CM, Thompson RH, Kwon ED, Murphy L, Riehle DL, Blute ML, Leibovich BC, Vasmatzis G, Cheville JC. High expression levels of ieter n protein independently predict a poor outcome for patients who undergo surgery for clear cell renal cell carcinoma. Cancer 2006 Jul 1; 107(1):37-45. 3. Vasmatzis G, Klee EW, Kube DM, Therneau TM, Kosari F. Quantitating tissue specificity of human genes to facilitate biomarker discovery Bioinformatics.[Epub ahead of print] 2007 Mar 26. 4. Kube DM, Savci-Heijink DC, Lamblin AF, Vasmatzis G, Cheville JC, Connelly DP, Klee GG, Kosari F. Optimization of laser capture microdissection and RNA amplification for gene expression profiling of prostate cancer. BMC Molecular Biology 2007; 8(25). 5. Tomlins SA, Rhodes DR, Perner S, et al. Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science. 2005 Oct 28;310(5748):644-8.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 61 ORAL PRESENTATIONS Selected Lectures
SELECTED LECTURES
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 62 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Selected Lectures – Forensic Genetics Forensic Genetics
Presentation number: FG 1 Abstract number: ABS-1297-ISABS-2007
DNA IDENTIFICATION OF PREMYSLID ROYAL DYNASTY REMAINS BY AMPFLSTR® MINIFILER™ PCR
Vanek D1, Hajkova J 1, Saskova L 1, Pospisek M2,5, Frolík J3, Bravermanová M4
1Forensic DNA Service, Prague 7, Czech Republic; 2Centre of Biomedical Informatics, Institute of Computer Sciences AS CR, Prague 8, Czech Republic; 3Archaelogical Institute, Academy of Science, Prague 1, Czech Republic; 4Prague Castle Administration, Prague 1, Czech Republic; 5Biologicals, Ricany, Czech Republic [email protected]
The aim of the presented study was to test the suitability of mini-STR system AmpFlSTR® MiniFiler™ for genotyping of degraded and ancient DNA samples. The set of ancient bone samples tested comprised the aged specimen (bronze age – 12th century) from archaeological sites in Czech Republic and the results proved the usefulness of robust mini-STR for DNA typing of compromised samples. DNA extraction and quantitation procedures as well as counter- contamination issues will also be discussed. DNA identification of Premyslid family skeletal remains on Prague castle is a multidisciplinary project covering fields of archaeogenetics, anthropology, archaeology and genealogy and seems to be the biggest archeo – DNA identification effort.
Keywords: Mini-STR, ancient DNA, inhibitors, archeogenetics, degraded DNA
Suggested reading: 1. J Forensic Sci. 2005 Jan;50(1):43-53. 2. J Forensic Sci. 2004 Jul;49(4):733-40. 3. J Forensic Sci. 2003 Sep;48(5):1054-64. 4. Biotechniques. 2007 Mar;42(3):343-52. 5. Hum Genet 1999; 104:164-166.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 63 ORAL PRESENTATIONS Selected Lectures – Forensic Genetics Presentation number: FG 2 Abstract number: ABS-7860-ISABS-2007
DNA ANALYSIS OF TRACE EVIDENCE FROM ANIMAL AND PLANT ORIGIN – EXPERIENCES IN FORENSIC CASEWORK
Hellmann A1, Morzfeld J1, Schleenbecker U1
1Bundeskriminalamt, Kriminaltechnisches Institut, KT32, Wiesbaden, Germany. [email protected]
Until now biological traces of non-human origin have been analysed by traditional descriptive and comparative techniques based on the knowledge of morphological patterns. Morphological aspects provide a firm basis for species classification but are normally not sufficient to identify an individual out of a population. With the introduction of STR analysis of human traces using polymerase chain reaction, experts are increasingly interested in applying the power of DNA investigations to other species as well. For this purpose several species-specific STR markers for dog, oak, and cannabis were established and basic loci data and population statistics were implemented. Different cases of successful applications of STR analysis on animal and plant traces in forensic case work will be illustrated in this study: - A single oak tree leaf recovered from a suspect’s car trunk was analysed 6 years after a homicide in 1998. The leaf matched in all loci to a single tree directly beneath the place where the dead body was found. This finding led to the conviction of the suspect. – Several dog hairs were collected from a submachine gun used in a murder case. The suspect predicated that he found the gun and simply hid it from his wife. The hairs matched in all loci to one of his two Rottweiler dogs. He was sentenced to lifelong imprisonment. – Furthermore, different plantlets of an illegal cannabis plantation were subjected to DNA analysis. The findings should state whether plantlets in culture derived from only one single origin plant by clonal reproduction.
Keywords: non-human DNA, short tandem repeats, identity testing, forensic evidence, case work
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 64 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Selected Lectures – Forensic Genetics Presentation number: FG 3 Abstract number: ABS-7737-ISABS-2007
THE GENETIC BASIS OF HUMAN PIGMENT VARIATION van Daal A1
1Faculty of Health Sciences & Medicine, Bond University, Gold Coast, Queensland, Australia [email protected]
Current methods for forensic DNA analysis require identification of a suspect for comparison purposes. The ability to determine the physical characteristics of the source of a crime scene sample would provide important probative information. There are potentially useful human physical biometric indicators, namely height, colouring and facial features, that are amenable to molecular analysis since they are all highly heritable. The considerable understanding of the genetics of mouse coat colour determination has assisted the understanding of normal human pigment variation. Over 100 pigmentation genes have been identified in the mouse. SNPs in a handful of these genes have been associated with various human hair, skin and eye colour phenotypes. A population database of approximately 700 samples with corresponding phenotypic information relating to pigmentation, height, weight and facial morphology has been collected. These samples have been used to screen 58 SNPs in 26 different candidate human pigmentation genes. Associations with pigmentation phenotypes have been identified in greater than half of these genes. Associations with different gene SNP combinations was also investigated.
Keywords: pigmentation, phenotype, variation, SNP, polymorphism
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 65 ORAL PRESENTATIONS Selected Lectures – Molecular Anthropology Molecular Anthropology
Presentation number: MA 1 Abstract number: ABS-8086-ISABS-2007
EVOLUTIONARY PALEOPROTEOMICS: ANALYSIS OF PATHOLOGICAL PROTEINS BY MASS SPECTROMETRY
Mark L1
1Institute of Biochemistry and Medical Chemistry, University of Pecs, Pecs, Hungary [email protected]
The Mycobacterium genus comprises several human pathogens such as Mycobacterium leprae and Mycobacterium tuberculosis. It is estimated that more than one-third of the worldwide human population is Mycobacterium tuberculosis infected, and 2 million die of the disease each year. Moreover it is expected that over the next 25 years, 40 million people will die from tuberculosis. The Mycobacterium tuberculosis complex (MTC) is an ancient pathogen, its emergence dates several thousands of years. It has widely accepted view that diagnosis of tuberculosis from archaeological human skeletal remains is not an easy task. By this time numerous studies have detected the presence of the MTC in archaeological bone findings by gross osteological examination and biomolecule analyses. The survive of pathological biomarkers in ancient skeletal remains is a major prerequisite for any molecular analysis and is thus essential for the pathological examination of archaeological bones. In this study, ancient mycolic acids and mycobacterial proteins were successfully extracted and identified by matrix-assisted laser desorption ionization tandem time of flight mass spectrometry (MALDI TOF/TOF MS) for the first time from archaeological human bones. The identification and sequencing of ancient mycobacterial proteins has the potential to expand our understanding of ancient epidemiology and evolution of these pathogens.
Keywords: paleoproteomics, evolution, paleoanthropology, paleopathology, tuberculosis
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 66 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Selected Lectures – Molecular Anthropology Presentation number: MA 2 Abstract number: ABS-8289-ISABS-2007
Y-CHROMOSOME DIVERSITY IN SOUTHERN URALS: A GEOGRAPHIC BORDER BETWEEN EUROPE AND ASIA
Khusnutdinova E1, Lobov A1, Yunusbayev B1, Yusupov R3, Bermisheva M1, Kutuev I1, Khusainova R1, Villems R2
1Institute of Biochemistry and Genetics, Ufa, Russian Federation; 2Estonian Biocenter, Tartu, Estonia; 3Institute of Language and Literature, Ufa, Russian Federation. [email protected]
Turkic-speaking Bashkirs are dispersed throughout the southern Ural region. They are considered by historians as descendants of Turkic- speaking nomadic communities that arrived in southern Urals at Early Medieval ages and assimilated indigenous population. We performed phylogenetic analysis of Y-chromosome lineages in a sample of 587 Bashkirs drawn from different parts of the southern Ural region and neighbouring areas: Abzelilovskiy (N=152), Sterlibashevskiy (N=54), Baimakskiy (N=95), and Burzyanskiy (N=82) districts of Bashkortostan republic, Orenburg (N=79), Perm (N=72), Samara and Saratov (N=51) Oblasts of Russia. Obtained samples of Y-chromosomes were analyzed using 24 biallelic markers of the Y chromosome non-recombining region. A total of 17 haplogroups were identified among which R1b3-M269, R1a1- SRY 1532, and N3-M46 lineages were predominant. Since N3-M46 lineage is prevalent among neighbouring Finno- Ugric populations and is rarely found in Central Asia, where numerous Turkic- speaking confederations dominated over a long period ancestors of Finno-Ugric groups are probable source population that contributed N3 lineage into Bashkirs. Y- Chromosome lineages specific to Central and East Asian populations (C3c- M48, O-M175) were absent or found with very low overall frequency (less than 10%). The only exception is R1b2 lineage which was found with very high frequency among Transural Bashkirs. Overall prevalence of typical West Eurasian (R1a-SRY 1532 and R1b3- M269) and North Eurasian (N3) lineages imply that Turkic-speaking newcomers were either admixed or genetic input associated with their arrival was limited.
Keywords: population, phylogeography, Y-chromosome, diversity, Southern Urals
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 67 ORAL PRESENTATIONS Selected Lectures – Molecular Anthropology Presentation number: MA 3 Abstract number: ABS-436-ISABS-2007
CRANIOFACIAL PARAMETERS AS PHENOTYPIC MARKERS FOR THE SUSCEPTIBLE TO CLEFTS GENOTYPES
Nagle E1, Teibe U2, Kažoka D3, Balode I4
1Department of Medical Biology and Genetics, Riga Stradiņš University; 2Department of Medical Physics; 3Institute of Anatomy and Anthropology; 4Children`s Clinical University, Riga, Latvia. [email protected] and [email protected]
The purpose of this study was to determine if a set of morphological parameters in the craniofacial region could define phenotypic markers that could be used in risk assessment of nonsyndromic facial clefts. In this cross-sectional study the subjects were parents of children with clefts born in Latvia, and clinically healthy Latvia residents (control). Craniofacial measurements were carried out with the GPM Anthropological instruments, Siber-Hegner & Co. A total 20 linear measurements from 201 individual were taken between well defined landmarks of the head and face. Descriptive and inferential statistics have been used for interpretation of the results. Statistically significant differences for 18 measurements between the cleft parents and controls (p<0.05) were obtained. Four craniofacial indices (cephalic index, facial index, intercanthal index, and upper face index) were calculated also. Only upper face index did not show significant differences between cleft parents and controls. Results of this study support hypothesis that craniofacial morphology in parents of cleft children is different from that observed in control individuals. Data about morphological features of the parents who have nonsyndromic cleft child could be valuable as phenotypic markers in identifying susceptible genotype to have a child with a cleft.
Keywords: craniofacial parameters, nonsyndromic clefts, linear measurements, craniofacial indices, phenotypic markers
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 68 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Selected Lectures – Molecular Anthropology Presentation number: MA 4 Abstract number: ABS-4917-ISABS-2007
DIFFERENTIATION OF INDIGENOUS ASIAN AND AMERICAN POPULATIONS REVEALED BY AUTOSOMAL SNPSTR SYSTEMS
Udina I1, Zhivotovsky L1, Knight A2, Feldman M3, Mountain J2
1 N.I. General Genetics Institute RAS, Moscow, Russia; 2Department of Anthropology, Stanford University, California, USA, 3Department of Biology, Stanford University, California, USA [email protected]
Genetic roots of indigenous American populations were intensively traced in Siberia by mitochondrial DNA and Y chromosome NRY markers. DNA autosomal markers, which include main proportion of genetic information, were fairly studied in the context of the American population origin. Therefore, we investigate DNA variation in 19 samples from human populations of northern Asia and the Americas, using the simultaneous analysis of tightly linked pairs of autosomal SNPs and STRs (SNPSTRs). Data from 13 populations sampled in southern Asia, Europe and Africa are included to establish geographic context. We demonstrate that SNPSTRs can aid in the historical and geographic analysis by using SNPs for the spatial distribution of DNA variation, and calibrating evolutionary time using the associated STR. According to SNPSTR data analysis of three autosomal SNPSTR systems (5SR1, 8SR2 and 22SR1, rapid long-distance gene flow from northern Asian populations to the Americas, as well as towards southern Asia, might have been frequent in this historically recent period. In terms of genetic distances, the American Indian populations are differentiated from the rest of the world, while the Greenland Inuit fall in a distinctive cluster formed by northern Asian populations except for those that appear to include a European component. Both admixture and random genetic drift have played a significant role in population diversification in small northern Asian and American indigenous populations. We suggest that the peopling of the Americas involved movement of people from a geographically broad range in Siberia and the Far East through the Bering Land Bridge.
Keywords: Human populations, Differentiation, Autosomal SNPSTR systems, Asia, Americas
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 69 ORAL PRESENTATIONS Young Investigator Award
YIA
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 70 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Young Investigator Award – Forensic Genetics Forensic Genetics
Presentation number: FG 4 Abstract number: ABS-5386-ISABS-2007
DNA IDENTIFICATION OF LEATHER RELICS IN THE MEMORIAL AND MUSEUM AUSCHWITZ-BIRKENAU
Sanak M1, Opolska-Bogusz B2, Kaczmarczyk G2, Piniewska D 2, Bartosik I3, MA Cywinski P3
1Department of Molecular Biology and Clinical Genetics, Internal Medicine, Krakow, Poland; 2Jagiellonian University Medical College Department of Forensic Medicine, Krakow, Poland; 3Memorial and Museum Auschwitz-Birkenau, Oswiecim, Poland [email protected]
Since twenty years molecular methods have permitted DNA analysis of archive materials. Revaluation of old criminal cases and identification of missing people was done successfully with these methods. The aim of this study was to verify if six exhibits from Memorial and Museum Auschwitz-Birkenau were made of human skin. Genomic DNA from small leather fragments was isolated by ion-exchange columns (A&A Biotechnology). Autosomal DNA STR loci were genotyped using a commercial kit (AmpFlIdentifiler, Applied Biosystems, USA) according to the manufacturer recommendations. In addition, amplification of hypervariable region- 1 (HV1) of mitochondrial DNA was made by the nested PCR reaction. Separation of amplifications products was completed using ABI Prism 377 (Applied Biosystems). Gel image was analyzed with Gene Scan Analysis 3.7NT and Genotyper 3.7NT software. One of the relics, a fragment of a parchment appearance, revealed a full STR profile of the male. Only a few STR alleles and amelogenin products characterizing human sex were obtained from the other samples. Sequencing of VH1 mitochondrial DNA region was completed for each of the samples within nucleotides 16183 – 16389, according to Anderson’s numbering. Three samples out of six had mitochondrial HV1 region sequence identical to the reference one. Three remaining samples differed from the reference sample by a single C>T transitions. DNA STR and mitochondrial analysis proved that all studied items were relics made of human skin. Thus, it documented Nazis crimes against humanity during the Second World War.
Keywords: DNA, identification, archive materials, skin, STR
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 71 ORAL PRESENTATIONS Young Investigator Award – Forensic Genetics Presentation number: FG 5 Abstract number: ABS-1638-ISABS-2007
NON-DESTRUCTIVE METHOD OF DNA EXTRACTION FROM TEETH AND BONES
Krzyżańska A1, Dobosz T1
1Molecular Technique Unit, Department of Forensic Medicine, Wroclaw Medical University, Wrocław, Poland [email protected]
DNA analysis can be used to widen our knowledge of extinct populations, world and human history. Archaeological excavations and museum specimens are convenient sources for various samples, but in almost all extraction methods, a piece of bone or tooth is powdered before extraction thus inflicting damage on them. The samples are usually irreplaceable and unique so we must choose to research or to keep them completely intact. Facing these facts, we require fieldwork to obtain a non-destructive method of DNA extraction from bone and tooth samples. We used entire human teeth and long bones as subjects. Samples were from fourteen to one thousand years old. The system of DNA extraction is based on rinsing the dental root canal and dental cavity or spongy part of bones, which are accessible after the decalcification process, by using a ieter n io pump and special buffers. The samples, clean and apparently untouched after rinsing in water and parafine oil, can be stored in museum collections for long periods of time. We used Quantifiler® Human DNA Real Time Quantification Kit to quantify the amount of amplifiable human DNA present in a sample and afterwards amplified five STR loci nuclear DNA and/or sequenced hypervariable region of mtDNA. We obtained human DNA from all tested teeth. The concentrations of DNA attained were comparable to other, more destructive methods. The detection of mtDNA from the 1000-year-old samples is particularly pleasing.
Keywords: ancient DNA, degraded DNA, DNA extraction, teeth, bones
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 72 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Young Investigator Award – Molecular Anthropology Molecular Anthropology
Presentation number: MA 5 Abstract number: ABS-8352-ISABS-2007
INCREASED AMPLIFICATION SENSITIVITY AND SUCCESS WITH LOCKED NUCLEIC ACIDS
Ballantyne KN1,2, van Oorschot RAH1, Mitchell RJ1,2
1Victoria Police Forensic Services Department, Macleod, VIC, Australia 2 Genetics Department, La Trobe University, Bundoora, VIC, Australia [email protected]
PCR primers used for amplification of forensically relevant STR loci are frequently designed to achieve maximum sensitivity, ieter n the target size, and to increase the multiplexing ability. However, these demands can lead to compromises in the efficiency of amplification, with some primer pairs amplifying less effectively at the chosen conditions than others. In particular, multiplexing often requires compromising the amplification parameters to ensure all loci are amplified at similar levels. This is in contrast to the desire to ieter n the amplification potential for ‘difficult’ forensic samples. An emerging technology which may negate the need for compromise is locked nucleic acids (LNAs). These nucleic acid analogs can be incorporated into oligonucleotide primers, and can significantly increase the binding specificity and strength during the PCR. They have previously been shown to increase amplification success of real-time PCR probes and primers, sequencing primers, and SNP mismatch discrimination probes. In a forensic context, results from our experiments show that incorporating LNA bases into existing STR primers can significantly decrease minimum template requirements, and obtain cleaner profiles. Furthermore, our empirical testing has shown that LNA primers have wider optimum reagent and temperature levels than traditional DNA primers, and, as such, can be incorporated into multiplexes without decreasing their efficiency. Multiplex ieter n ion time can be decreased, and amplification success increased with the use of LNAs.
Keywords: Locked Nucleic Acids, Trace DNA, Multiplexing, STR, Primer design
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 73 ORAL PRESENTATIONS Young Investigator Award – Molecular Anthropology Presentation number: MA 6 Abstract number: ABS-739-ISABS-2007
LANDMARKS OF ANIMAL ADAPTIVE HISTORY UNCOVERED BY GENOMIC PHYLOSTRATIGRAPHY IN DROSOPHILA
Domazet-Lošo T1, Brajković J1, Tautz D2
1Ruđer Bošković Institute, Zagreb, Croatia; 2Institute for Genetics, Cologne, Germany. [email protected]
Macroevolutionary trends are traditionally studied by analysis of fossils, comparative morphology or evo-devo approaches. With the availability of genome sequences and associated data from a large diversity of taxa it becomes possible to add a novel level of analysis: genomic phylostratigraphy. In this approach, one searches for genes that appear to have arisen at specific time points in evolution and have then be retained in the respective clade. Expression data from extant members of the clade are then used to assess the role of these genes, which allows inferring their contribution to the evolution of the clade. We exemplify the approach using a phylogenetic framework and embryo expression data from Drosophila to show that grouping of genes based on their phyletic origin uncovers footprints of important adaptive events in animal evolutionary history. Specifically, we find support that the germ layers evolved in the ectoderm- endoderm- mesoderm stepwise progression, and that aptations, achieved by action of novel genes, were biased towards ectoderm through most of the metazoan evolution. A remarkable exception is the lineage leading to the last common ancestor of bilaterian animals where the novel genes had balanced contribution to the germ layers. In the same phylostratum we detect unprecedented rate of fixation of novel genes, suggesting that the internal trigger preceded Cambrian explosion. These results have profound implications on the understanding of animal macroevolutionary processes. However, genomic phylostratigraphy can be readily applied, if suitable data are available, to any evolutionary lineage.
Keywords: genomics, phylostratigraphy, macroevolution, adaptation, metazoa
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 74 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Young Investigator Award – Molecular Anthropology Presentation number: MA 7 Abstract number: ABS-7671-ISABS-2007
THE POLYMORPHISM OF KIR GENES IN WORLDWIDE HUMAN POPULATIONS
Frassati C1, Picard C2, Chiaroni J2, Sanchez-Mazas A1
1Laboratory of Anthropology, Genetics and Peopling history (AGP), Dpt. Of Anthropology and Ecology, University of Geneva; 2EFS Alpes-Méditerranée, Marseille, France. [email protected]
Killer-cell Immunoglobulin-like Receptors (KIR) of natural killer (NK) cells recognize HLA class I molecules and are responsible for destroying infected and malignant cells. Due to this functional link, KIR genes are expected to display a high level of polymorphism. Indeed, their HLA ligands are encoded by highly polymorphic genes, the evolution of which has been submitted in part to natural selection (heterozygous advantage). However, HLA genetic variation among human populations is significantly correlated to geography, as observed for neutral markers, indicating that it has also been shaped by human migrations history. A key task is thus to investigate KIR allelic diversity in various human populations and analyze its patterns in relation to HLA. In this study, we investigate KIR genetic relationships of 23 human populations, among which 5 represent original data, within and among the main broad geographic areas of the world. Fifteen KIR genes and 2 pseudogenes were first genotyped by PCR-SSP in 5 populations from Congo (107 Bantu and 79 Pygmies), Syria (n=124), Comoros Islands (n=54) and France (n=38). Different approaches (e.g. MDS, hierarchical analysis of variance) were then used to explore the genetic variation of these genes in the whole set of 23 populations. The results reveal a high level of KIR allelic diversity in all samples studied. Like for HLA, KIR genetic relationships are geographically structured. Despite their functional properties, KIR genes thus represent a reliable set of molecular markers to be used for the reconstruction of human peopling history.
Keywords: molecular anthropology, human genetic diversity, KIR polymorphism, HLA polymorphism, human migrations history
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 75 ORAL PRESENTATIONS Young Investigator Award – Molecular Anthropology Presentation number: MA 8 Abstract number: ABS-966-ISABS-2007
TP53 CODON 72 POLYMORPHISM IN12 POPULATIONS OF INSULAR SOUTHEAST ASIA AND OCEANIA
Kashima T1, Makino K1, Soemantri A2 Ishida T1
1Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, Japan; 2Department of Child Health, Faculty of Medicine, Diponegoro University, Semarang, Indonesia [email protected] p53 tumor suppressor protein is a multifunctional protein, which plays central roles in the genomic stability of a cell. A well known functional polymorphism in the TP53 codon 72 (p53Arg/p53Pro) is ubiquitously distributed in human populations with various allelic frequencies; however, many Asian-Pacific populations were left unexamined. To add more information on this polymorphism, we have surveyed this polymorphism among 12 populations in Southeast Asia-Pacific region. A total of 733 subjects from 12 populations in insular Southeast Asia and Oceania were the subject of this study. These populations were classified either an Austronesian speaking group or Papuan speaking group. A PCR-RFLP method was employed for the genotyping. Distribution of the genotype in each population was in Hardy-Weinberg equilibrium except for the Timorese (p<0.05). The p53Arg frequencies ranged from 0.06 in the Seramese to 0.62 in the Kahayan with an average frequency of 0.38. No significant correlation between the p53Arg frequency and latitude was observed in the 12 populations tested (p>0.05), whereas a significant correlation was obtained for the relationship between frequency and longitude among nine Austronesian or the whole 12 populations tested (p<0.01). A longitudinal cline of the p53Arg frequencies may reflect history of the Austronesian’s migration and local admixture with indigenous Papuan speakers who had probably harboured low p53Arg frequencies. To overview the worldwide distribution and to clarify the bases for the ubiquitous presence of the TP53 codon 72 polymorphism, more extensive screenings for the polymorphism in Asian populations are awaited.
Keywords: p53 codon 72 polymorphism, Austronesian, Papuan, Southeast Asia, Oceania
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 76 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Sponsor's Lectures
SPONSOR’S LECTURES
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 77 ORAL PRESENTATIONS Sponsor's Lectures Abstract number: ABS-9930-ISABS-2007
IMPROVING DISCRIMINATION OF mtDNA TYPING BY MULTIPLEX PCR AND A LINEAR ARRAY PROBE PANEL: HV AND BEYOND
Calloway C1,2, Stuart S1,2, and Erlich HA1,2
1Roche Molecular Systems, Alameda, California, USA; 2Children’s Hospital, Oakland Research Institute, Oakland, California, USA. [email protected]
Mitochondrial DNA is an informative and useful target for the forensic genetic analysis of limited and/or degraded samples. However, there are some inherent limitations to targeting only the hypervariable regions I/II independent of the method of analysis. The power of discrimination is limited for all population groups as a result of a few common HVI/II sequences. Therefore, additional sequence polymorphisms outside the HVI/II regions need to be targeted to increase the power of discrimination of mtDNA analysis. To address this need, we have developed a multiplex PCR and immobilized probe assay which targets 48 polymorphic sites (24 HVI/II and 24 VR/CR) using 12 primer pairs and 83 immobilized probes. Primers and probes targeting polymorphic sites that subdivide common HV types as well as rapidly evolving polymorphic sites that subdivide multiple haplogroups were added to increase the discrimination power of the LINEAR ARRAY mtDNA HVI/HVII Region-Sequence Typing Kit. In addition, probes that target sequences within HVI/HVII regions to reduce the number of ‘0’ and weak signal types were added to improve the robustness of the assay. We have greatly increased the discrimination power beyond that of the HVI/II assay for all populations with the additional 52 probes and 10 primer pairs. Moreover, the expanded array is more informative than sequencing the HVI/II regions for the Caucasian population (h= 0.996 compared to 0.995). We present here population data to illustrate the increased discrimination power as well as the ease of use of this assay.
Keywords: mtDNA, linear array, coding region, HVI/HVII, immobilized SSO probes
Suggested Reading: 1. Divne A.M, et al. J For Sci 2005; 50(1):548-554.,3. 2. Date Chong M, et al. For Sci Int 2005 ; 54 :137-148. , 3. Kline M, et al. J For Sci 2005; 50(2): 377-385., 4. Calloway C. and Erlich H. For Mag 2007; 4(3):32-37., 5. Roberts K and Calloway C. J For Sci 2007; 52(1):40-47.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 78 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Sponsor's Lectures Abstract number: ABS-498-ISABS-2007
HLA ALLELIC DIVERSITY AND PHYLOGENETIC ANALYSIS OF HUMAN POPULATIONS
Erlich HA1,2, Mack SJ 2
1Roche Molecular Systems, Inc. USA; 2Children’s Hospital Oakland Research Institute, USA [email protected]
The HLA class I (A, B, and C) and class II loci (DR, DQ, and DP) are the most polymorphic genes in the human genome and, consequently have served as highly informative genetic markers for analyzing the genetic relationships among contemporary human populations. As part of the 13th, 14th, and 15th International Histocompatibility Workshops, HLA-A, - B, -C, -DRB1, -DQB1, and –DPB1 genotypes were determined for more than 100 populations using our immobilized probe linear array HLA typing system. These data, as well as other datasets, have been deposited in and are available from The NCBI’s dbMHC database (www.ncbi.nlm.nih.gov/projects/mhc/). Phylogenetic analyses of the allele and haplotype frequencies for these populations, and historical inferences made from these analyses will be discussed. We have used HLA population data to address some specific anthropological questions such as the colonization of the Pacific, the peopling of the Americas, and the origins of the Saami. HLA population data, along with mtDNA analysis, has also been used to identify the geographic/ethnic origin for the remains of a murder victim in Southern California.
Keywords: Phylogeny, Populations, HLA, Genetics, Anthropology
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 79 ORAL PRESENTATIONS Sponsor's Lectures Abstract number: ABS-7726-ISABS-2007
REAL-TIME PCR IN FORENSICS
Gerke N1
1Eppendorf AG, Germany [email protected]
Nowadays, real-time PCR is an established and powerful method for quantitating human DNA in forensic samples. In addition to commercially available Kits there are several methods described in respective articles of forensic journals. Each application has its strengths and weaknesses. Furthermore, there are maybe pitfalls and challenges when optimizing and establishing such an application in a forensic DNA laboratory. Besides, the potentials of more advanced real-time PCR applications, like low volume PCR, low copy number PCR, mRNA detection and SNP-Genotyping are discussed with regard to their usefulness in forensic DNA analysis.
Keywords: DNA quantitation, real-time PCR, optimization, low volume, low copy number
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 80 September 3-7, 2007, Split, Croatia ORAL PRESENTATIONS Sponsor's Lectures Abstract number: ABS-6410-ISABS-2007
QUANTITATIVE AND QUALITATIVE ASSESSMENT OF TOTAL HUMAN AND HUMAN MALE DNA IN FORENSIC TYPE BIOLOGICAL SAMPLES USING A MULTIPLEXED REAL-TIME PCR SYSTEM
Barbisin M1, Fang R1, O’Shea C1, Calandro L1, Furtado M1, van Oers P2 and Shewale J1
1Applied Biosystems, 850 Lincoln Centre Drive, Foster City CA 94404, USA; 2Applied Biosystems, Hoogeveenenweg 100, 2913LV, Nieuwerkerk aan den Ijssel, Netherlands [email protected]
Forensic DNA samples often contain mixtures of DNA from male and female contributors and may be exposed to environmental insults. It is therefore desirable to determine the relative quantities of male and female DNA and detect the presence of PCR inhibitors at an early stage in the analysis. To facilitate this process we have developed a multiplex TaqMan® assay to 1. Quantitate total human DNA and human male DNA simultaneously 2. Determine the ratio of human male and female DNA 3. Detect PCR inhibitors 4. Allow selection of appropriate STR amplification kit 5. Predict success with downstream STR amplification The multiplex assay is designed for the 7500 Real Time PCR System using the ribonuclease P RNA component H1 (RPPH1) human target and the sex determining region Y (SRY) male-specific target together with a synthetic oligonucleotide sequence co-amplified as an internal PCR control (IPC). Results of experiments performed to demonstrate the correlation between the quantification results and the nature of the subsequent STR profiles will be discussed to show how the multiplex assay provides guidance for selection of the appropriate AmpFℓSTR® system. This approach will reduce the number of samples requiring re-processing thereby decreasing the turn around time.
Keywords: Mixtures, Quantities, AmpFℓSTR® system, 7500 Real Time PCR System, Multiplex TaqMan® assay
Suggested Reading: 1. Nucleic Acids Res. 25:2657–2660. 2. Journal of Forensic Science, July, 2005, Vol 50, No.4. 3. Nucleic Acids Res. 25:3718–3723. 4. Martens, H. and Naes, T. 1989. Multivariate Calibration 5. Lakowicz, J.R. 1983. Energy Transfer
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 81 ORAL PRESENTATIONS Sponsor's Lectures Abstract number: ABS-2055-ISABS-2007
NEW DEVELOPMENTS AND RECENT ADVANCES IN LASER CAPTURE MICRODISSECTION
Taneja P1
1Molecular Devices (now part of MDS Analytical Technologies), USA [email protected]
This presentation will provide an overview of microdissection systems that combine the gentle laser capture microdissection (LCM) and rapid UV laser cutting in integrated platforms to enable isolation of pure sample populations with speed, sensitivity, reliability, and precision. LCM has wide applications for research and forensic applications. LCM is the safe and validated process for selection, isolation and analysis of trace biological crime scene samples. It can improve the quality of existing molecular methods as individual cells captured by this technology can be used to perform a broad spectrum of forensic and other life science applications. Analysis of DNA from microdissected cells has included polymerase chain reaction (PCR) amplification of alleles to observe the loss of heterozygosity, DNA fingerprinting, and detection of mutations by single- strand conformation polymorphism. We will provide examples of the use of LCM technology for forensics medicine for DNA isolation and profiling from cells of interest from tissue samples, stains, or smears (vaginal, sperm and buccal swabs). The new technologies have been expanded for alternate applications and increased sample flexibility. Sample formats include contact or non-contact microdissection: frozen or formalin fixed tissue; thick or thin sections; stained, unstained or fluorescently stained; hydrated or dehydrated; cell cultures; and forensic smears. We will focus the discussions on the one- source solution for isolation, amplification, labeling and analysis of RNA from both frozen and formalin fixed tissue samples to obtain the profiling of native expression levels of thousands of genes, in a selected cell population.
Keywords: Laser capture microdissection, Trace evidence, DNA Analysis, Forensic Genetics, Crime scene.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 82 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS
ABSTRACTS – POSTER PRESENTATIONS
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 83 POSTER PRESENTATIONS – Forensic Genetics
POSTER PRESENTATIONS Forensic Genetics
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 84 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics
Presentation number: FG 6 Abstract number: ABS-1163-ISABS-2007
RAPD-PCR ANALYSIS OF LABORATORY BRED TICKS, HYALOMMA ANATOLICUM ANATOLICUM AND HYALOMMA MARGINATUM (ACARI: IXODIDAE)
Abdigoudarzi M1, Dinparast-djadid N2
1Razi Vaccine and Serum Research Ins., Karadje, Iran; 2Pasteur Ins. of Iran, Tehran, Iran [email protected]
Eighteen RAPD primers from AB1 and OPA1 series, M13 and a Pz primer were used to amplify polymorphic DNA of Hyalomma anatolicum anatolicum and Hyalomma marginatum for the first time. Three different DNA extraction methods were used for DNA extraction from live Laboratory- bred ticks. Sixteen primers out of eighteen and the Pz primer could be successfully used for PCR and genomic DNA have been amplified for two different ticks. Different patterns of bands have been seen reproducible and specific for Hyalomma genus and H. an. an. and H. m. species. In addition specific reproducible bands were measured and planned for their specific amplification, cloning and sequencing. RAPD-PCR for its simplicity, rapidity and ability to reveal variation at the DNA level is an additional molecular tool for the study of ticks in their ecological living environment.
Keywords: RAPD-PCR, ticks, hyalomma anatolicum anatolicum, acari, ixodidae
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 85 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 7 Abstract number: ABS-2510-ISABS-2007
MULTIPLE DISPLACEMENT AMPLIFICATION: A PROMISING TECHNIQUE FOR IMPROVING TRACE DNA ANALYSIS
Ballantyne KN1,2, van Oorschot RAH1, Mitchell RJ1,2
1 Victoria Police Forensic Services Department, Macleod, VIC, Australia 2 Genetics Department, La Trobe University, Bundoora, VIC, Australia [email protected]
Low DNA quantities are commonly encountered in forensic genetics, as trace DNA, but also in other fields, such as microbiology, medical genetics, anthropology, and population genetics. A solution currently being applied to overcome small DNA sample sizes is multiple displacement amplification (MDA), a form of whole genome amplification. We examined a commercial MDA kit for forensic use, and found that, when used as the manufacturer recommends, it can increase profiling success from both trace and degraded DNA. However, amplification bias is a significant problem, frequently causing allele and locus dropout, and complicating mixture profiling. Therefore, we have developed an optimised MDA procedure to increase amplification success and decrease bias. By adding a molecular crowder, such as PEG400, and performing a modified denaturation protocol, samples are amplified more consistently and accurately than with standard MDA. This modified MDA protocol has been applied to both pristine and casework samples, and more than doubled the amplification success with difficult samples. Although more testing and optimisation is required, this modified MDA protocol appears highly promising for use with trace DNA samples.
Keywords: Whole Genome Amplification, Multiple Displacement Amplification, Trace DNA, Molecular Crowding, Amplification Bias
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 86 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 8 Abstract number: ABS-7360-ISABS-2007
VARIANT ALLELES, TRIALLELIC PATTERNS AND POINT MUTATIONS OBSERVED IN NUCLEAR STR TYPING FROM POPULATIONS OF THE FORMER YUGOSLAVIA
Bašić L1, Huel R1, Madacki-Todorović K1, Smajlović L1, Eminović I1, Miloš A1, Parsons T1
1International Commission on Missing Persons, Sarajevo, Bosnia and Herzegovina [email protected]
The International Commission on Missing Persons (ICMP) was founded in 1996 to address the issue of persons missing as a result of the conflicts that occurred during the breakup of the former Yugoslavia during the 1990’s. ICMP employs a “population based, DNA-led” identification system in assisting with identification of missing persons in the region of former Yugoslavia. On a regional scale, DNA profiles from reference samples of living relatives of missing persons are continuously compared in batch mode to DNA profiles obtained from mortal remains of victims. We aim to present a compendium of off-ladder alleles and other genotyping irregularities relating to rare/unexpected population genetic variation, observed in a large STR database from the former Yugoslavia. This information is useful for the scientific community to help practitioners recognize and deal with these testing irregularities when they are encountered.
Keywords: off-ladder allele, variant allele, triallelic, point mutation, STR
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 87 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 9 Abstract number: ABS-323-ISABS-2007
NUCLEASE BASED PURIFICATION OF MALE DNA FROM SEXUAL ASSAULT CASES
Garvin AM1
1Bureco, AG, Reinach, Switzerland [email protected]
Large amounts of the victim’s epithelial cells contaminate the sperm present on swabs taken from rape victims. The standard method for obtaining pure sperm is to digest the epithelial cells with Proteinase K and separate the victim’s solubilised DNA from the sperm by pelleting and washing, steps that are labour intensive, difficult to automate, and highly dependent on the technician’s skill level. I have found that the alternative approach of destroying the victim’s DNA with a nuclease appears to be faster, easier, and better than the standard method. Following ProK digestion of a swab cutting, an aliquot is removed for the victim’s fraction and a nuclease is added and then incubated for 1 hour. A nuclease inhibitor and reducing agent are then added to lyse the sperm and release the suspect’s DNA into solution. The nuclease degrades more than 99% of the victim’s DNA but does not affect the sperm DNA sequestered in the sperm heads. No sperm are lost during the purification, so yields are good. Data will be presented showing that a vaginal swab cutting with 500 sperm yields a profile that is predominantly male. Since only pipetting steps are required, the process can be easily automated.
Keywords: Sperm DNA, Sexual assault, purification, backlog, automation
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 88 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 10 Abstract number: ABS-490-ISABS-2007
AUTOMATED DNA EXTRACTION BY EPPENDORF epMOTION 5075 LH WORKSTATION
Hajkova J1, Saskova L1, Wehrhahn D2, Vanek D1
1Forensic DNA Service, Prague, Czech Republic; 2Eppendorf AG, Hamburg, Germany [email protected]
Forensic DNA laboratories are experiencing rapidly growing demand to process large number of evidence samples. In an effort to meet the rising requirements for DNA analysis, liquid handling workstations are utilized for the automation of the liquid handling needs. The epMotion 5075 LH (Eppendorf AG) is flexible and extremely accurate robotic platform capable to extract DNA from forensic casework and reference samples. This study demonstrates that the epMotion 5075 LH workstation used in combination with either the ChargeSwitch (Invitrogen) or DNA IQ (Promega Corporation) extraction chemistries, is versatile enough to accommodate the whole spectra of samples encountered by a crime laboratory. The performance of the epMotion LH 5075 with regard to DNA yields and potential cross-contamination for a different sample types was also evaluated.
Keywords: automation, DNA extraction, Eppendorf epMotion, DNA IQ, RT-qPCR
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 89 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 11 Abstract number: ABS-9815-ISABS-2007
SENSITIVE DETECTION OF GC-RICH REGIONS BY PCR USING DEAMINATED DNA: APPLICATION TO SUDDEN INFANT DEATH SYNDROME IN ASSOCIATION WITH ONDINE’S CURSE
Hasegawa I, Sato F, Kimura R, Osawa M
Department of Forensic Medicine, Tokai University School of Medicine, Isehara, Kanagawa, Japan [email protected]
High GC content inhibits conventional PCR reactions. This report presents the circumvention using a bisulfite treatment of DNA, which was developed to reduce the GC content. We applied this modification to detect congenital central hypoventilation syndrome (CCHS), also known as Ondine’s curse, which is characterized by hypoventilation during sleep with an onset in infants. Characteristics of the clinical features suggest that undetected CCHS is potentially involved in cases of sudden infant death syndrome (SIDS). Recent studies indicate that the expansion of a polyalanine repeat in the PHOX2B gene is relevant to the pathogenesis of the disorder. However, the repeat region including the vicinity contains high GC, which is refractory to amplification. The bisulphite- converted DNA permitted direct PCR amplification using primers specific to the deaminated sequence of the coding strand, in which dropouts of expanded alleles were completely prevented. It yielded a product of 123 bp for the common 20- residue repetitive tract with converted T from original C by sequencing. The majority (90%) of clinically diagnosed CCHS patients carried heterozygous expansions of 25- to 33-residues at the polyalanine tract of PHOX2B. In contrast, analysis revealed no expansions in SIDS victims and healthy subjects. These results suggest that the major pathogenesis of SIDS is distinct from that of CCHS.
Keywords: GC content, bisulphite, SIDS, deamination, PCR amplification
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 90 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 12 Abstract number: ABS-8474-ISABS-2007
OUR EXPERIENCE OF FATHERLESS PATERNITY TESTS
Korabecna M1, Horinek A2
1Faculty of Medicine, Charles University, Pilsen, Czech Republic; 21st Faculty of Medicine, Charles University, Prague, Czech Republic. [email protected]
In cases, in which the putative father of a person is not available the paternity testing is dependent on the DNA analysis of his relatives. We try to achieve the best accuracy in each tested family. Herein we demonstrate the results of seven fatherless cases. DNA samples were isolated from buccal swabs using QIAamp DNA Blood Mini Kit (Quiagen). For STR analysis, PowerPlex16 (Promega) and AmpFlSTR Yfiler PCR (AppliedBiosystems) were used. The results of STR analysis on autosomal loci were evaluated using the programme Familias (Egeland et al, Forensic Science International 110 (1), 2000). Calculations were based on population data. In all cases, the satisfactory results were obtained. The probabilities calculated for the most probable pedigree provided strong evidence for or against paternity in each tested family. Our combination of STR analysis on autosomal and Y- chromosomal loci together with application of the programme Familias seems to be sufficient for the successful solution of fatherless cases.
Keywords: fatherless paternity cases, STR analysis, Y-chromosomal STR, probability calculation, Familias
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 91 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 13 Abstract number: ABS-1972-ISABS-2007
ANALYSIS OF TWO DIFFERENT METHODS IN DNA EXTRACTION FROM SALIVA SPECIMEN
Kovacevic L1, Makarevic E1, Bakal N1, Marjanovic D1,2
1Institute for Genetic Engineering and Biotechnology, Sarajevo, Bosnia and Herzegovina; 2Group for Forensic Genetics, Department of Molecular Medicine, “Rudjer Boskovic” Institute, Zagreb, Croatia [email protected]
Saliva is recognized as suitable source of genomic DNA for various genetic studies. That is the sample, which could be relatively easily collected in a painless and non-invasive manner. Also, when is collected, it could be successfully stored through extended period of time. The aim of this work was to observe possibility of application of two different approaches in DNA extraction from saliva specimens for DNA forensic analysis purposes. One is usage of commercial Qiagen protocol for DNA extraction from buccal swab, semen and blood, on saliva specimens collected with passive oral lavage in micro tubes and another-one is application of the Oragen TM DNA Self – Collection Kit. Of laboratory protocol for manual extraction of DNA from 0,5 ml Oragen TM /saliva solution. The samples of saliva were collected from the volunteers. Promega PowerPlex 16 kit was employed for simultaneous amplification of 15 STR loci in PE GeneAmp PCR Thermocycler 9700. ABI PRISM 377 Sequencer was used in analysis of the amplified fragments. Given results confirmed the successfulness of application of standard Qiagen protocol for DNA extraction from saliva without need for its additional modification in order to get an optimal amount of DNA for further phases of forensic DNA analysis. Furthermore, it was pointed that Oragen approach could be also used in the same purpose, but with supplementary optimisation of isolation protocol.
Keywords: collecting sample, saliva specimens, DNA extraction, different methods, DNA analysis
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 92 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 14 Abstract number: ABS-3892-ISABS-2007
CHARACTERIZATION OF NEW miniSTR KIT TO AID ANALYSIS OF DEGRADED DNA IN PCR MULTIPLEX
Krzyżańska A1, Jonkisz A1, Lebioda A1, Ciciała A1, Sarzyńska M1, Dmochowska G1, Dobosz T1
1Molecular Technique Unit, Department of Forensic Medicine, Wroclaw Medical University, Wrocław, Poland [email protected]
Forensic typing of highly degraded DNA based on short tandem repeat commercial kits have shown loss of signals which are assigned to larger- sized STR products. This loss of signal may be the results of PCR inhibitors present in the sample or highly fragmented DNA. Analysis of received results from PCR multiplex proves that in case when the concentration of DNA is low, polymerase preferential amplify small-size loci. The aim of our study was to create a new, reduced size miniSTR set to PCR multiplex. Primer sequences were selected to closely flank the repetitive region and reduce the amplicon length of four STRs down to 150bp for the longer alleles of the polymorphism. We designed a PCR multiplex kit containing shortened loci: CD4, D1S1677, D2S441, D4S2364. DNA isolated from blood or evidence samples, by chelex or Qiagen QIamp Mini Kit, was amplified with the new primer set and visualised in capillary electrophoresis on an ABI Prism 310. Allelic ladders for miniSTRs were created by using a combination of samples that represented the range of alleles observed in the population. Finally, homozygotes for each allele in each locus were sequenced to verify results. A comparison of the new PCR set and commercial kits show that, if the DNA was highly degraded, we received results only from the new miniplex. These new loci can be useful in typing closely related individuals where there is a need for additional genetic information.
Keywords: DNA typing, degraded DNA, STR, miniSTR, PCR multiplex
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 93 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 15 Abstract number: ABS-3112-ISABS-2007
CHARACTERIZATION OF 6 NON CODIS MINI STR LOCI ON TELOGEN HAIR, FINGERPRINT AND ANCIENT BONE SAMPLES
Nagy G1, Könczöl F1, Kricskovics A1 Márk L2, Bajnóczky I1
1 Institute of Forensic Medicine, Medical School, University of Pécs, Pécs, Hungary, 2 Depertment of Biochemistry and Medical Chemistry, Medical School, University of Pécs, Pécs, Hungary [email protected]
A number of studies have demonstrated that successful analysis of degraded DNA specimens from mass disasters or forensic evidence improves with smaller sized PCR products. NIST research group lead by Michael D. Coble, and John M. Butler, designed and tested new STR loci, unlinked from the CODIS markers, which can generate amplicons less than 125 bp in size and would therefore be helpful in testing degraded DNA samples. New STR loci D1S1677, D2S441, D4S2364, D10S1248, D14S1434, and D22S1045, arranged into two miniSTR triplexes labelled NoC1 and NoC2. We tested these NonCodis Mini STR triplexes on 34 fingerprint and telogen hair samples from the same individuals and on 26 ancient bone samples from a “Gorzsa” burial ground. DNA isolated from 6700 year old bone samples by various methods (AFDIL’s demineralization protocol, Invitrogen’s ChargeSwitch® Forensic and Plant kit, Promega DNAIQ® system, Qiagen EZ1 and Forensic Card, Phenol-Chloroform-Isoamyl Alcohol method). The triplexes were reliable and sensitive to at least 50-100 pg of DNA template under controlled laboratory conditions and pristine DNA samples. The new triplexe miniSTR assays were success for recovering useful information from fingerprint, telogen hair and ancient bone samples, while no profile or partial profiles were observed with the majority of the samples using present STR multiplexe kits (PP16 and IdentiFiler). We also made allele frequency database on two Hungarian populations. All loci show a moderate degree of polymorphism among 200 Roma and Caucasian samples tested.
Keywords: NoC1 NoC2, Ancient bone sample, Telogen Hair, Fingerprint, Allele frequencies
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 94 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 16 Abstract number: ABS-8637-ISABS-2007
CODIS SYSTEM STR LOCI POPULATION DATA AND MUTATION RATE FOR MOSCOW REGION OF RUSSIAN FEDERATION
Nikulin MV1, Shchagina OA1, Vasserman NN1, Kazakova SA1, Polyakov AV1
1Research Centre for Medical Genetics, Moscow, Russian Federation [email protected]
Paternity testing became a widespread type of analyzes in recent years. More than 570 cases of paternity disputes were processed in our laboratory. For these studies thirteen loci of CODIS system were analyzed using Promega PowerPlex®16 or Applied Biosystems AmpFlSTR® Identifiler® kits. The obtained PCR products were analyzed by Applied Biosystems 3130 Genetic Analyzer. During these studies, 462 male and 436 female meioses were observed for each CODIS locus. In total, 18 cases of mutations were detected: 12 cases in males, 5 cases in females and one unresolved case. Paternal mutations were detected in loci D8S1179, D13S317, TH01, vWA – each in one case, FGA, D5S818, D18S51, D7S820 – each in two cases. Maternal mutations were detected in loci TH01, D21S11, vWA – each in one case, D18S51 – in two cases. The unresolved mutation was in locus D5S818. Maternal mutation in locus TH01 led to change of allele length for two repeats. Other observed mutations led to one-step allelic change. Two incidents of simultaneous mutations were detected among the aforementioned cases: two paternal mutations in loci D5S818, D18S51 and two maternal mutations in loci D18S51, D21S11. These data confirms the conclusion that mismatching in at least three loci allows paternity exclusion. The profiles of 642 unrelated individuals were used to draw allele frequencies distributions in these 15 loci for population of Moscow region of Russian Federation. The obtained distributions demonstrate high level of resemblance to data presented by Promega Corporation for Caucasian-Americans (http://www.promega.com/techserv/apps/hmnid/referenceinformation/pops tat/custstat_Allelefreq.htm).
Keywords: CODIS, STR, mutation, frequencies distribution, paternity testing
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 95 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 17 Abstract number: ABS-5823-ISABS-2007
OCCURENCE OF OFF-LADDER ALLELIC VARIANT IN PATERNITY TESTING - CASE STUDY
Projić P1, Škaro V1, Šamija I 2, Primorac D3,4, Marjanović D 1,5
1Ruđer Bošković Institute, Department of Molecular Medicine, Forensic Genetics Group, Zagreb, Croatia; 2Institute of Gynecologic Cytology, University Department of Gynecology and Obstetrics, Zagreb University Hospital Center, Zagreb, Croatia; 3Medical School, University of Split and Medical School, Split, Croatia; 4Josip Juraj Strossmayer University of Osijek, Osijek, Croatia; 5Institute for Genetic Engineering and Biotechnology Sarajevo, Sarajevo, B&H [email protected]
The latest scientific breakthroughs in the field of molecular biology, as well as its joint treatment with newest population-genetic findings and technological improvement, made disputed paternity testing by applying DNA expertise, more suitable for wider public usage. Very important part in this process is to perform simple and precise way of final results presentation. During the official education for performing paternity caseworks, an inconsistency affecting segregation of allele was encountered in three paternity cases, detected as an off-ladder allelic variant (located between D16S539 and D2S1338 loci). Buccal swab from voluntary donors was used as the DNA source. The Qiagen DnaeasyTM Tissue Kit was employed for DNA extraction. The DNA samples were amplified at 15 STR loci using an AmpFlSTR® Identifiler® (ABI, Foster City, CA). The PCR amplification has been done in PE Gene Amp PCR System Thermal Cycler (ABI, Foster City, CA). The amplified products were separated by capillary electrophoresis using an ABI Prism 3130 Genetic Analyzer (ABI, Foster City, CA). Raw data were analyzed drawing on the ABI PRISM® Data Collection Software v3.0 and GeneMapperTM ID Software v3.1 to obtain numerical allele designations of the profiles. A rare, off- ladder allelic variant has been noticed on approximate position 298,30 bp in D2S1338 locus. In each of three paternity cases, the novel D2S1338 allele segregated from one of the parent to, at least, one of the children. If this allelic variant has been accidentally ignored, results for this locus would be misinterpreted. These cases indicated the possible existence of an additional D2S1338 allele, a previously unreported in Croatian population studies. Before the final conclusion about this item, we are planning, in the nearest future, to perform suitable additional monoplex analysis.
Keywords: DNA typing, paternity test, STR loci, rare allelic variant, Croatian population
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 96 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 18 Abstract number: ABS-9012-ISABS-2007
POLYMORPHISM ANALYSIS AT 16 STR LOCI IN DELHI'S (INDIA) RANDOM POPULATION AND THEIR FORENSIC SIGNIFICANCE
Raina A1, Yadav B1, Dogra TD1
1DNA Fingerprinting Laboratory, Dept. of Forensic Medicine, All India Institute of Medical Sciences, New Delhi, India [email protected]
Indian population has diverse ethnic, linguistic and geographical groups. Presently the complete genetic data is not available for all the Indian populations. As such data is required for the medico-legal purposes, it necessitates the study of the genetic polymorphism. Therefore, this study has been carried out on 100 samples for Delhi’s random population using STR markers at 15 different autosomal loci. The study is based on the estimation of allelic frequencies for 15 STR loci and also includes the calculation of observed and expected heterozygosity, Power of Discrimination, Degree of Paternity Exclusion, Polymorphism Information Content. The results observed indicate that all the 14 STR loci are highly polymorphic in this population except TPOX locus (Allelic diversity 0.6569). Thus suggesting that this set of STR markers can be used for forensic purposes.
Keywords: Delhi, STR markers, India, DNA, Forensic
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 97 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 19 Abstract number: ABS-060-ISABS-2007
RAPID GENOMIC DNA EXTRACTION (RGDE) IS A NEW TECHNIC FOR DEVELOPMENT FORENSIC GENETIC
Saremi MA 1 , 2, Saremi M 3 , Tavallaei M 4
1 Imam Hussein University, Faculty of Science, Tehran, Iran; 2 Kawsar Human Genetic Research Center, Tehran, Iran; 3 Reference Laboratories of Iran - Research Center, Tehran, Iran; 4 Baqeyatollah University – Molecular Biology Research Center, Tehran, Iran [email protected]
Nowadays expanded windows have been opened to the world especially in the area of Biotechnology and Biology science by performance of big projects such as human genome project. Production of great Bio Banks and DNA Banks in national & international levels is one of development pivots, which is very important in medical, agricultural, economical & forensic genetic fields. In this regard, one of the primary and important steps for all is DNA extraction with high quality & quantity in minimum time from cells. By using this method, genomic DNA with high quality & quantity can be acquired in the shortest time which has been presented in the world up to now. With this novel rapid method can isolation of high molecular weight genomic DNA from blood, sperm tissues, bone, forensic case work and etc. In this rapid method using of cell lysis buffer, nuclei lysis buffer (an equal volume), chloroform and absolute ethanol also. Quantity Control DNA extracted to perform by spectrophotometry and showed results that average for DNA yield (μg)= 5.86±2.53 and A260/A280 radio=1.91±0.16 if volume of blood used (μl)=500 (n=20). Quality Control DNA extracted to perform by agarose Gel 0.8% and showed results we have DNA with high quality. In this method genomic DNA can be extracted in the fastest time and preparation of extremely high molecular weight DNA by very simple materials and without any need to special equipment & apparatus. We could registration this method as an invention.
Keywords: Rapid DNA Extraction, Newest Technic, Development Forensic Genetic, Shortest time, High quality & quantity
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 98 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 20 Abstract number: ABS-8269-ISABS-2007
MONITORING, CONTROLLING AND RECORDING THE CRITICAL ENVIRONMENTAL CONDITION IN DNA LABORATORIES USING THE 4N6 XC- TEST SYSTEM
Saskova L1, Hajkova J1, Pospisek M2,3, Vanek D1
1Forensic DNA Service, Prague 7, Czech Republic; 2Biologicals, Ricany, Czech Republic; 3Centre of Biomedical Informatics, Institute of Computer Sciences AS CR, Prague 8, Czech Republic [email protected]
According to ISO 17025 requirements for testing and calibration laboratories and other ILAC recommendations and regulations shall DNA laboratories ensure that the environmental conditions do not invalidate the results or adversely affect the required quality of DNA typing. The aim of this presentation is to describe the process of monitoring, controlling and recording the critical environmental condition – background contaminating DNA- using the novel 4n6 XC-TEST system. Laboratory monitoring using the XC-TEST system is described in Standard Operating Procedure and comprise several steps: swabbing of critical objects and items in DNA laboratory, manual or automated DNA extraction, DNA quantitation using SYBR system and optional identification of contamination source (sample or person) using the capillary electrophoresis. The XC-TEST control also tests measures taken to prevent cross-contamination.
Keywords: STR, DNA contamination, laboratory environment, 17025, RT-qPCR
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 99 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 21 Abstract number: ABS-5491-ISABS-2007
SINGLE BASE PRIMER EXTENSION METHOD TO IDENTIFY 16 mtDNA SNP POLYMORPHISMS THAT INFER EUROPEAN ANCESTRY
Hung M1, Podini D1, Schanfield MS1
1George Washington University, Washington, DC, USA [email protected]
Our laboratory recently developed and validated a rapid and robust multiplex assay capable of typing the mtDNA haplogroups A, B, C, D, E, F, G, H, I, L1/L2, L3, M, and N using the SNaPshot® (Applied Biosystems, Foster City, CA) single base extension reaction kit. Though this global screening kit can identify most of the East Asian and African lineages it is weak at identifying Indo-European maternal lineages. We developed a second assay based on 16 SNPs that include the diagnostic polymorphic sites for the European haplogroups H, J, K, T, U, V, and W. 13 European samples typed by the global multiplex as N (5), L3 (4) and H (4) were respectively typed by the European multiplex as K*, T, U, V and W; J, J*, and K (2); and H (4). Where the * designates a published variant. These multiplexes allow a fast, robust, and low-cost screening tool could help eliminate unnecessary testing of reference samples from plane crashes or where remains from several individuals are found together and may be commingled. In anthropological studies, these assays can be applied as a screening tool to determine the haplogroups present in specific populations, as well as on archaeological samples for the better understanding of human history and evolution. These multiplexes are not intended to replace sequencing of HVI and HVII but rather serve as a rapid screening tool to allow for the determination of which samples need to be sequenced to provide maximum information and minimize costs.
Keywords: mtDNA, SNP, European haplogroups, Single base extension, PCR
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 100 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 22 Abstract number: ABS-6749-ISABS-2007
AUTOSOMAL STR DIVERSITY IN SOUTHEASTERN ANKARA: FORENSIC AND ANTHROPOLOGICAL IMPLICATIONS
Dogan Alakoc Y1, Gokcumen O2, Tug A2, Gultekin T3, Gulec E3, Schurr TG2
1Department of Forensic Medicine, Ankara University Medical Faculty, Ankara Turkey; 2Department of Anthropology, University of Pennsylvania, Philadelphia, U.SA. 3Department of Anthropology, Ankara University, Ankara, Turkey. [email protected]
Throughout history, there have been many biological, cultural and social interactions between populations living in Anatolia. However, some groups, at least based on their oral histories, have been isolated from others in this region. Our ethnographic work amongst these populations indicates that their traditional social structures have, indeed, restricted marriages outside of the group. Such isolated groups can give very interesting insights into the population history of the region. More importantly, the histories of these groups have important genetic implications for regional forensic databases in that they are crucial for accurately identifying regional genetic diversities. For this study, we collected 125 blood samples from a district in south-eastern Ankara, the capital of the Turkish Republic. To contextualize the biological analysis, we also collected detailed genealogical family histories and ethnographic information with questionnaires and interviews. Our ethnographic observations suggest that the population of this district, the known human history for which dates back to the 14th century, comprises four distinct groups, with different heritages, linguistic norms and social dynamics. In this paper, we comparatively analyze variation at 15 STR loci from these four groups to reveal the similarities and differences between them. This approach allows us to discuss, within an anthropological context, the extent and implications of the genetic diversity within local isolated groups for larger forensic genetic databases.
Keywords: Turkey, Alleles, Endogamy, Population Structure, Lineages
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 101 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 23 Abstract number: ABS-5285-ISABS-2007
APPLICATIONS OF MINISTRS IN AN AUSTRALIAN FORENSIC LABORATORY
Sears A1, Walsh SJ2, Goetz R1, Gunn P3, Roux C4
1Division of Analytical Laboratories, Lidcombe, NSW, Australia; 2Australian Federal Police, Forensic Biology Laboratory, ACT, Canberra, Australia; 3 FSSB, Forensic Services Group, NSW Police Service, Australia; 4 Centre for Forensic Science, University of Technology, Sydney, Australia. [email protected]
Identification of deceased individuals is a critical aspect in any coronial and criminal investigation. DNA provides an extra dimension to the identification process, especially for reconciliation of fragmented bodies. Increased prevalence worldwide of natural and terrorism-related disasters has prompted further advancements by scientists involved in human DNA identification. MiniSTRs offer clear advantages in the analysis of compromised samples and compliment existing STR testing systems currently used in Australian forensic laboratories. The modified primers target some routinely analyzed loci, allowing existing DNA databases to be utilized. In addition, non-CODIS miniSTRs can potentially contribute additional information, strengthening the likelihood of individual identification. This is of benefit in kinship reconstruction, especially where victims include multiple members of one family. In this research modified miniSTR primers (based on those developed by Butler et al., NIST) for both CODIS and non-CODIS loci and the recently released AmpFlSTR® miniFiler™ kit (Applied Biosystems) were validated and applied to degraded samples and human skeletal remains of varying ages and histories. MiniSTRs produce reliable results and increase the success rate of DNA testing on degraded samples and skeletal remains. The analytical problems encountered with degraded DNA samples can be reduced through miniSTR technology. This research has demonstrated the potential of miniSTRs to contribute an enhanced DNA analysis component for Disaster Victim Identification (DVI), missing persons as well as criminal and coronial identification cases in Australia.
Keywords: miniSTRs, DNA, identification, forensic, human
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 102 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 24 Abstract number: ABS-7021-ISABS-2007
INDIRECT APPROACH IN FINDING SOMEONE’S AUTOSOMAL GENETIC PROFILE – A FORENSIC TOOL IN INVESTIGATING HISTORICAL CASES
Stanciu F1,2, Potorac R1, Stoian V2
1Forensic Science Institute, General Inspectorate of Romanian Police, Bucharest, Romania; 2Department of Genetics and Biotechnologies, Faculty of Biology, University of Bucharest, Bucharest, Romania [email protected]
Starting with an intuitive schema and a rich historical information background, it has been possible to determine the origins and the most probable genetic profile of a male person who lived between 1954 and 1984. In lack of some biological sample from this person, an indirect approach was the most suitable way in determining his genetic autosomal profile. For this purpose it has been analyzed his genealogical tree - eight saliva samples taken from his relatives and 9 biological micro-traces taken from the surface of fifth personal paper documents - a school certificate, a driving license, a handwrote letter, a communist party notebook and a receipt. DNA Isolated from all biological samples had been amplified using Identifier and Yfiler kits. PCR products had been analyzed with ABI Prism 3100. For revealing the genealogical origins of the investigated male person, specific international DNA databases for Y and autosomal STR markers, such as YHRD, Ysearch, OmniPop and ENFSI WG STR Population Database, had been interrogated.
Keywords: most probable genetic profile, genealogical tree, personal documents, historical case, STR databases
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 103 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 25 Abstract number: ABS-5369-ISABS-2007
NUCLEAR DNA QUANTIFICATION OF BONE SAMPLES FROM DIFFERENT SOURCES: A COMPARISON BETWEEN REAL-TIME PCR, AGAROSE GEL ELECTROPHORESIS AND SPECTROPHOTOMETRY
Sutlović D1, Gugić D2, Definis-Gojanović M1, Anđelinović Š1
1Department of Pathology and Forensic Medicine, Split University Hospital and School of Medicine, Split, Croatia; 2Department of Pathology, University of Miami School of Medicine Jackson Medical Center, Miami, Florida, USA [email protected]
To comparate analytical methods to evaluate nuclear DNA concentrations of forensic and ancient bone samples. Three different methods were used: agarose gel electrophoresis, spectrophotometry and Quantitative Real-time PCR. QRT- PCR offers several advantages with respect to other compared methods. Twenty four DNA extracts from bone samples: eight DNA extracts from ancient bone samples (3 DNA samples retrieved from 10-50 years old bones and 5 DNA samples obtained from about 1300 years old bones recovered from an old church burial sight), eight DNA were extracted from mass graves bones and eight were extracted from fresh bones. The L containingμquantification assay was performed in total volume of 25 L of DNA extract, Quantifiler human primer mix and Quantifiler PCRμ2 reaction mix with thermal cycling conditions according to the manufacture’s protocols In the following trials total DNA was evaluated by gel electrophoresis (1.5% agarose) and ethidium bromide staining by UV illuminations (312 nm) of the gel . Absorbancies at 260 and 280 nm were measured by spectrophotometer. Highly sensitive methods for quantification of human DNA retrieved from forensic and ancient DNA samples are important because they ensure the consistency of low copy number DNA typing. RT- PCR offers several advantages with respect to other compared methods. In samples which contain high amounts of DNA all of the tested methods gave positive results with good agreement. All samples that possibly contain small amount of DNA or inhibitors therefore need to be quantitated by QRT-PCR as method of choice.
Keywords: bone samples, quantification, human DNA, forensic science, analytical methods
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 104 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 26 Abstract number: ABS-5182-ISABS-2007
MOTHER-BABY MIX-UPS, STOLEN BABY, ALLEGED “BABY DIED” CASES AND DNA ANALYSIS
Tug A1, Ozguven HD2
1Department of Forensic Medicine, Medical Faculty, Ankara University, Ankara, Turkey; 2Department of Pschychiatry, Medical Faculty, Ankara University, Ankara, Turkey [email protected]
Except in a very few cases, the birth of a baby is one of the happiest moments in parents’ lives. A suspicion right after birth or years later that they might not be the biological parents of the child, or the child that they were told to be dead might after all not be dead initiates a difficult-to-cope- with process. According to the Tanderberg Annual Report 1998 edition one out of eight babies born in American hospitals is sent home with the wrong parents. In many of these cases the mistake is recognized within a few days after hospital discharge and corrected. However, in big hospitals, where a reliable identification system cannot be established, this mistake is not recognized until after many years when the parents or the child get suspicious and start to investigate and the mother-child mix-up is uncovered. Furthermore, for families, whose child was stolen from hospital or whose child many years ago was told to be dead but might not be dead after fall, the investigation process is certainly long and traumatic. At the end, even if the child is found, the effects of the ensuing legal process and psychological trauma will not be overcome easily by both the parents and the children. In mother-baby mix-ups, stolen baby and alleged baby died cases help from the media is usually requested to reach out to wide masses and many parents or children having similar doubts apply for DNA analysis. Short Tandem Repeats (STR) or, where necessary, Y- STR and mitochondrial DNA analyses make it possible to determine genetic concordance between parties in a short period of time. If there are many reference samples and analysis is carried out in more than one centre, the use of the same methods is important in terms of the assessment of the results. Through such analyses mix-ups due to human error can be uncovered and individual or organized stolen baby incidents brought up to light.
Keywords: Forensic Sciences, DNA analysis, Mother-baby mix ups, Stolen baby cases, Alleged baby died cases
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 105 POSTER PRESENTATIONS – Forensic Genetics Usage of the Newest Technologies and Genetic Markers in Forensic Genetics Presentation number: FG 27 Abstract number: ABS-3574-ISABS-2007
COMPARISON OF THREE DIFFERENT SYSTEMS FOR NONINVASIVE DIAGNOSIS OF FETAL GENDER FROM MATERNAL PLASMA
Wagner J1, Džijan S1, Jukić-Pavan D2, Lauc G1
1DNA Laboratory, School of Medicine, J.J.Strossmayer University, Osijek, Croatia; 2Department of Gynecology, General Hospital Karlovac, Karlovac, Croatia [email protected]
For the last forty years, cytogenetic prenatal karyotyping is the golden standard for prenatal diagnosis. However, this commonly used invasive technique has some disadvantages, the most important one being a low, but definite risk both for fetus and mother. Since the discoveries of fetal cell free DNA in maternal circulation ten years ago, there were many attempts to develop safer noninvasive methods for prenatal diagnosis. In families with X-linked recessive genetic disorder it is important to determine fetal gender early. Several different systems have been used for this purpose, but they all suffer from the same disadvantage; the lack of positive control for the amplification of fetal DNA. Aiming to alleviate this disadvantage we used multiplexed STR system to amplify DNA from maternal plasma hoping that the presence of paternal alleles would serve as a positive control for the amplification of fetal DNA. Cell free DNA was isolated from 90 samples of maternal plasma (length of gestation 11-36 weeks) using QIAamp Blood DNA kit. Extracted cell free DNA was: a) amplified using AmpFl STR Identifiler kit (15 STR loci and gender specific amelogenin) or AmpFl STR Y-filer PCR Amplification Kit (16 STR loci on Y chromosome) and subsequently analyzed on capillary electrophoresis system ABI PRISM 310 Genetic Analyzer, or b) amplified and analyzed using Quantifiler Y Human Male DNA Quantification kit (SRY gene). Gender of fetuses was confirmed by cytogenetic analysis or phenotypically at birth. Unfortunately paternal STR alleles were amplified only sporadically, thus our attempt to develop positive control for the amplification of fetal DNA was not successful. However, we successfully determined presence of Y chromosome in all 49 pregnancies with male fetuses. AmpFl STR Y-filer was found to be the best system since it successfully detected Y chromosome in all cases. AmpFl STR Identifiler was correct in 45 cases (92%), while the results obtained by real time PCR were accurate in 46 cases (94%) of male fetuses. There were no falsely detected Y chromosomes in female fetuses.
Keywords: fetal cell free DNA, noninvasive prenatal testing, fetal gender, Y- chromosome, maternal plasma
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 106 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence, Recognition, Collection, Preservation Forensic Evidence Recognition, Collection, Preservation
Presentation number: FG 28 Abstract number: ABS-696-ISABS-2007
AGE CHANGES OF THE HUMAN FOOT TUBULAR BONES ROENTGENOMORPHOMETRICAL INDICES OF THE RUSSIA MIDDLE VOLGA REGION INHABITANTS
1 1 1 1 Akhmetova G , Ryakhovsky M , Khayrullin R , Mitchenko I
1Department of Human Morphology, Ulyanovsk state University, Ulyanovsk, Russia [email protected]
The purpose of the present research consists of an establishment of quantitative age anatomic variability of the human foot tubular bones roentgenomorphometrical parameters of the Russia Middle Volga region inhabitants. As the objects of investigation X-ray photographs of foot tubular bones of 42 men and 11 women aged 17-87 years were used. During the investigation the measuring of X-ray octeometrical indices was made with the standard vernier calliper. For the 5th metatarsi bone the additional parameter was introduced. After the final measuring all the linear parameters were converted into indices. The obtained data was subjected to one-dimensional dispersive analysis. As a result six indices with significant Fishers criterion were chosen, which trustworthy reflect expressiveness of age changes of bones. The present quantitative age change of tubular bones of human foot is the result of the first phase of fulfilment of proclaimed tasks and shows the expressiveness, the tendency of processes of ageing trustworthy detected for metatarsus bones and proximal phalanxes of II and V radiuses of human foot. The results of the investigation must be applied in forensic medicine personality identification, orthopaedics, biometrics, anthropology and other biological sciences.
Keywords: bone, tubular, age, vernier, calliper
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 107 POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence, Recognition, Collection, Preservation Presentation number: FG 29 Abstract number: ABS-9282-ISABS-2007
MITOCHONDRIAL AND NUCLEAR DNA ANALYSIS FOR RECONSTRUCTION OF HISTORICAL GENEALOGY OF PRINCES OF MOLDAVIA FROM THE SECOND HALF OF THE XVI-TH CENTURY
Cardos G1,2, Batrana A3, Batrana L3, Miritoiu N4, Rodewald A1
1Institute of Human Biology, University of Hamburg, Germany; 2Biology Faculty, University of Bucharest, Romania; 3National Museum of History, Bucharest, Romania; 4Institute of Anthropology, Bucharest, Romania. [email protected]
The genealogy of the princes of Moldavia from the last half of the XIVth century is a much-debated subject in Romanian historiography. In order to shed light on their genetic kinship we studied at molecular level the skeletal remains of 5 Moldavian princes and 2 other male of the royal family, buried in the “Saint Nicholas” Church from Radauti, Romania. Therefore, we analyzed mitochondrial (HVR I region) and nuclear DNA (VWA31A, TPOX, DYS392 and DYS393 Microsatellites) polymorphisms. The Amelogenin gene was used for identifying the genetic sex of the old individuals. The ancient DNA was extracted from teeth of those 7 old individuals by a phenol-chloroform-based method. After amplifying by PCR, the mtDNA as well as the nuclear DYS392 and DYS393 polymorphic markers were sequenced by the Sanger method. The nuclear DNA sequences were separated and identified on polyacrylamide gel, Ag- stained. Our results revealed the genetic kinship between the 7 old individuals, showing that there is no continuity along the entire male line. Two individuals are close related to each other along the paternal lineage and differ from the other 5 individuals also closed related among them as concerns the paternal line. There is a maternal linkage between the two groups. Thus, we could argue by genetic evidence that two princely dynasties succeeded on the Moldavian scepter in the last half of the XIVth century, infirming the historiography version which had considered the existence of only one dynasty and succession exclusively along the paternal line.
Keywords: aDNA, genealogy, Moldavia, DNA polymorphism, Amelogenin
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 108 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence, Recognition, Collection, Preservation Presentation number: FG 30 Abstract number: ABS-7967-ISABS-2007
A HIGH DEGREE OF CORRESPONDENCE OF THE ANTHROPOLOGICAL METHOD AND DNA ANALYSIS WHILE ESTABLISHING FATHERHOOD, THE EXPERIENCES OF DEPARTMENT OF FORENSIC MEDICINE OF THE CLINICAL CENTRE OF MONTENEGRO, PODGORICA
Čukić D1
1Department of Forensic Medicine, the Clinical Centre of Montenegro, Podgorica; Medical School, Podgorica; Medical and Stomatological School, Foča [email protected]
The work is of historiographic character, from the period of using the DNA analysis for identification of fatherhood and personality. But, through retrospective analysis I want to point out that the anthropological methods, as then the only ones known and available, were not useless. Ten cases were analyzed in Department of Forensic Medicine of the Clinical Centre of Montenegro, Podgorica, where fatherhood was being established, first by the anthropological methods, and then after a certain, shorter or longer period –also by the DNA analysis. In all 10 cases, the DNA analysis uncontroversially established, that is to say, confirmed fatherhood. By previous anthropological methods, it was concluded that there was the highest degree of probability that ’’...XXperson is father of XX child ...).Though Montenegro has none of the DNA laboratories, the anthropological methods are- what is expected- abandoned. For now, Department of Forensic Medicine is the ’’service’’ for the establishing of fatherhood etc. by the DNA analysis. In this moment, our Department is directing the clients toward the DNA laboratories. In this connection, the clients decide if they go to the recommended DNA laboratory or they use our Department as the ’’service’’ (taking and sending of the material, results’ submitting etc.). I hope that Montenegro will get the DNA laboratory soon, whether it will be located in Department of Forensic Medicine or in the Police (what would be effective for investigations but not for civil lawsuits and’ free analyses’’). If the DNA analysis were located in Department of Forensic Medicine, the subject would be useful in the widest degree.
Keywords: antropology, DNA, fatherhood, comparison, DNA analysis
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 109 POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence, Recognition, Collection, Preservation Presentation number: FG 31 Abstract number: ABS-4990-ISABS-2007
EVALUATION OF THE RELIABILITY OF THE DNA TYPING PROCESS IN THE IDENTIFICATION OF WAR VICTIMS IN CROATIA
Džijan S1, Ćurić G1, Pavlinić D1, Hercog R1, Lauc G1
1DNA Laboratory, School of Medicine, J.J. Strossmayer University, Osijek, Croatia [email protected]
Aiming to estimate frequency of various types of errors that may have occurred during the process of identification of victims of war in Croatia, we have identified and compared genotypes of 911 parents – child pairs in the database of nearly 3500 relatives of missing people. Genotypes of 891 pairs (97.8%) were matching perfectly, while 20 pairs did not match in one or more of the analyzed loci. Aiming to identify reasons for the observed lack of genetic matches between putative parents and children, we reanalyzed DNA in these 40 blood samples. We found that out of 1822 analyzed genotypes one was completely wrong as a result of clear human error. Three additional genotypes had one wrong allele, again because of a human error. Four additional genotypes had a single wrong allele due to either PCR of electrophoresis errors. Other mismatches were a consequence of the lack of biological link between the presumptive parents – child pairs.
Keywords: DNA typing, human identification, quality control, war victims, genetic match
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 110 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence, Recognition, Collection, Preservation Presentation number: FG 32 Abstract number: ABS-4826-ISABS-2007
VALIDATION STUDIES OF THE NOVEL FORENSIC SAMPLE COLLECTION SYSTEM – 4N6 DNA SWAB
Hajkova J1, Saskova L1, Giambra A2, Vanek D1
1Forensic DNA Service, Prague 7, Czech Republic; 2Copan Innovation Group, Italy [email protected]
The aim of this presentation is to inform the forensic community about the results of validation studies performed on novel sample collection system designed specially for forensic use. Sample collection capacity, assay sensitivity, DNase- free, RNase-free, Human DNA-free and PCR-inhibitor free status and suitability for automated DNA extraction are core features that have been tested and compared with “traditional” cotton, dacron or rayon swabs. 4N6 DNA swabs can be used not only for reference sampling (mouth swabs) but due to its sampling capacity and especially the efficiency of sample release from the swab matrix are extremely suitable for laboratory sampling and especially crime scene sampling. The best description of the 4N6 DNA Swabs is “What You Swab is What You Get”, as the swab with sample is placed in DNA LoBind tube where the subsequent DNA extraction takes place and almost 100% release rate of sample from the swab matrix ensures that the final recovery rate is exceptionally high if compared to the traditional sampling methods for DNA analysis.
Keywords: sampling kits, swabs, forensic, DNA analysis, automation
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 111 POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence, Recognition, Collection, Preservation Presentation number: FG 33 Abstract number: ABS-1811-ISABS-2007
PROTEIN-BASED IDENTIFICATION OF EPITHELIAL CELL TYPES IN FORENSIC SAMPLES
Simons, JL1, Vintiner, SK1
1ESR (The Institute of Environmental and Scientific Research), Auckland, New Zealand. [email protected]
Although the value of DNA profiles found at a crime scene is indisputable, there is increasing importance in identifying the cellular source of the DNA as well as the identity of the person to whom the profile belongs. Knowledge regarding the cellular source from which the DNA profile originates increases the evidential value of the sample. Our research involves investigation of protein candidates in order to find an epithelial cell type-specific protein that will enable differentiation of vaginal, buccal and skin cells in forensic samples. We have used several methods including histochemical stains, western analysis and immunohistochemistry to investigate candidate proteins known to be present in various types of epithelial cells1. Our most current results from these studies will be presented here.
Keywords: Epithelial Cells, Protein specificity, Immunohistochemistry, Dane\'s stain, Cytokeratins
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 112 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence from Non-Human DNA Forensic Evidence from Non-Human DNA
Presentation number: FG 34 Abstract number: ABS-9163-ISABS-2007
AN INVESTIGATION OF THE UTILITY OF MOLECULAR GENETIC ANALYSIS BY USING FORMALIN PRESERVED ANIMAL TISSUE
Kazic A2,3, Hammond JBW2, Merrett NR1, Crimmen O1
1Department of Zoology, The Natural History Museum (NHM), London, U.K.; 2North East Surrey College of Technology (NESCOT), Ewell, Epsom, Surrey, U.K.; 3Laboratory for Molecular Genetics of Natural Resources, Institute for Genetic Engineering and Biotechnology (INGEB), Sarajevo, Bosnia-Herzegovina. [email protected]; [email protected]
The damaging effect of formalin on DNA and the inhibition of PCR are serious problems in molecular studies. The possibility of using 20-yrs-old formalin-fixed, Steedman’s preserved museum specimens of fish with unstudied genomes (collection of the Natural History Museum, London, UK) in molecular analyses was investigated. A number of DNA extraction protocols and different tissue pre- washing/drying regimes were tested. These gave different levels of success, but a guanidinium-based protocol developed in this study gave the best results. RAPD- PCR methodology was used as a test of the efficiency of DNA extractions/amplifications and for developing specific PCR primers to amplify PCR product sizes 300 bp – 350 bp. Two genomic sequences of Nezumia aequalis and N. micronychodon were determined from cloned RAPD-PCR fragments of DNA extracted from formalin-fixed specimens and validated by direct sequencing of PCR products (derived from DNA extracted from formalin/Steedman’s, DMSO and ethanol preserved tissue). Sequences were reproducible regardless of the way of preservation. The sequences were submitted to the GenBank database under accession numbers AY826774 – AY826792. Attempts to amplify mitochondrial DNA sequences with the six mitochondrial genes were mostly unsuccessful. Sporadic amplifications were obtained with primers of 16S and COIII genes. The DNA extracted from preserved specimens possesses unique characteristics that make molecular investigations very difficult. However, this study has demonstrated that an appropriate strategy and molecular approach could lead to the successful use of formalin-fixed archival collections and in providing reliable sequence information even on organisms with unstudied genomes.
Keywords: formalin-fixed fish, museum collection, guanidinium-based protocol, RAPD approach, sequences
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 113 POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence from Non-Human DNA Presentation number: FG 35 Abstract number: ABS-8294-ISABS-2007
MEDICAL APPROACH OF PATIENTS WITH DYSGENESIS OF GONADS
Malikova R1
1Department of Obstetrics and Gynecology, Pediatric Faculty, Samarkand State Medical Institute, Uzbekistan [email protected]
In spite of its seldom occurance, the problem of dysgenesis of gonads is considered to be important all over the world. One of the forms of dysgenesis of gonads is dysgenesis of ovaries. The main symptoms are: amenorrhea, under- developed secondary sexual symptoms, short height in Shereshevsky-Terner syndrome. When biological age does not correspond to the calendar one the girls suffer from psychological and social problems, so they feel themselves defective and often lead a secluded life. So the pations with this pathology first must be prepared psychologically and after that they should be persuaded to undergo substitutional hormonal therapy. 50 patients with different forms of dysgenesis of gonads have been studied at the Department of Children’s and Teenager’s Gynecology in Samarkand Regional Multiprofile Children’s Hospital during 4 years. We have revealed clean (46, XY) dysgenesis of gonads or Swyer syndrome in 2 (4%); mixed (45, XO/46XY) in 2 (4%); typical (45, XO) or Shereshevsky- Terner in 10 (20%); mosaic form of Shereshevsky-Terner in 14 (28%); clean (46, XX) in 23 (46%). The patients in whose cariotype Y-chromosome presents, gonadectomy with following substitutional hormonal therapy was performed. The rest patients were explained the significance of hormonal therapy immediately after making diagnosis and the treatment was carried out. The treatment was effective. 20 (40%) patients had the development of lactiferous glands and appearing of menstrual-like reaction, and 15 (30%) – enlargement of the uteri in size. Pointed changes save the girls from feeling of being defective and contribute to their social adaptation. But this is not a single reason of necessity of this kind of therapy. Use of preparations of sexual hormones suppresses the discharge of gonadotropins by hypophysis and decrease the risk of development of malignant degeneration of gonads. Thus the patients with dysgenesis of gonads need in previous psychological preparation and further substitutional hormonal therapy. However an adequate approach and maintenance of ethic norms of behaviour in management of patients with dysgenesis of gonads are acquired.
Keywords: dysgenesis, gonads, hormonal, Y-chromosome, substitutional
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 114 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics Forensic Evidence from Non-Human DNA Presentation number: FG 36 Abstract number: ABS-4278-ISABS-2007
FORENSIC BOTANY: POTENTIAL USEFULNESS OF MICROSATELLITE- BASED GENOTYPING OF CROATIAN OLIVE (OLEA EUROPAEA L.) IN FORENSIC CASEWORK
Štambuk S1, Sutlović D2, Bakarić P3, Petričević S1, Primorac D4,5, Anđelinović Š2
1SMS - Food Development Center, Klis, Croatia; 2Clinical Hospital Split, Laboratory for Forensic and Clinical Genetics, Split, Croatia, 3Marka Marojice 47, Dubrovnik, Croatia; 4Medical School, University of Split, Split, Croatia, 5Medical School, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia [email protected]
Growing body of evidence points to the useful application of forensic botany in resolving legal issues. Genotyping by applying microsatellite based markers is the most common DNA profiling method used in forensic analysis of human DNA, but it is also extensively used in plant population genetics, plant specimen fingerprinting and plant varietal identification studies. Olive (Olea europaea L.) genotyping applying microsatellite based markers was carried out in order to assess the potential application of olive as legal case evidence. This potential depends on the degree of variability within the Croatian olive genomic pool and on the effectiveness of the chosen set of microsatellite based markers in revealing olive divergence. The total of 44 autochthonous Croatian olive specimens were subject to genotyping applying 16 previously described and developed microsatellite based markers. By previous morphological analyses those 44 specimens were classified into 30 cultivars with the exception of an additional, previously unassigned specimen. Genotyping of 44 specimens provided the possibility of distinguishing a total of 44 different genotype profiles over 16 microsatellite based loci. Average expected heterozigosity amounted to 0.758, which points to significant diversity of Croatian olives. Croatian olive genotyping performed by the set of 16 microsatellite based markers showed strong varietal discrimination up to the single tree and subsequently its considerable application potential in detection of crime, accident and suicide circumstances, in case olive is presented as evidence.
Keywords: Forensic botany, microsatellite based genotyping, Olea europaea L., average expected heterozygosity, SSR
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 115 POSTER PRESENTATIONS – Forensic Genetics DNA Identification of Victims of Mass Disaster – the Latest Approach DNA Identification of Victims of Mass Disasters – the Latest Approach
Presentation number: FG 37 Abstract number: ABS-863-ISABS-2007
DIFFERENT APPROACHES FOR DNA ANALYSIS OF OLD BONE SAMPLES – PRELIMINARY RESULTS
Ahmetaš L1, Bego T1, Hodžić B1, Kovačević L1, Marjanović D1,2
1Institute for Genetic Engineering and Biotechnology, Sarajevo, Bosnia and Herzegovina; 2Ruđer Bošković Institute, Zagreb, Croatia [email protected]
The process of identifying mortal remains from different mass fatalities has advanced significantly in last decade. However, obtaining DNA profiles from skeletal remains is among the most challenging types of DNA forensic testing. The development of STR profiles from bone or tooth samples is not without its limitation. While working with few decades old bones and teeth forensic scientists are usually confronted by many problems (insufficient quantity of DNA, high level of DNA degradation, presence of PCR inhibitors etc.). Each laboratory may encounter significant difficulties when developing their own procedures for this kind of testing. This presentation is based on preliminary and brief comparison of different approaches in three crucial stages of DNA analysis of skeletal remains: bone powdering procedure, DNA isolation and amplification protocols. We have briefly analyzed two different approaches of bone powdering (blender vs. Dremel tool), DNA isolation procedures (phenol-chlorophorm vs. Optimized Qiagen procedure) and PCR protocols for PowerPlex16 kit (regular vs. extended). Earlier processed and profiled 60 years old bone samples were used in this analysis. QuantifilerTM Human DNA Quantification Kit was used for DNA quantification and electrophoresis of the amplification products was performed on ABI PRISM 310 genetic analyzer. Obtained results indicate advantages but also some disadvantages of all examined methods.
Keywords: DNA analysis, old bone samples, bonepowdering, DNA extraction, PCR protocol
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 116 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics DNA Identification of Victims of Mass Disaster – the Latest Approach Presentation number: FG 38 Abstract number: ABS-9419-ISABS-2007
POPULATION SUBSTRUCTURE CAN SIGNIFICANTLY AFFECT RELIABILITY OF A DNA-LEAD PROCESS OF IDENTIFICATION OF MASS FATALITY VICTIMS
Ćurić G1, Kračun SK2, Biruš I1, Džijan S1, Lauc G1,2
1DNA Laboratory, School of Medicine, J.J.Strossmayer University, Osijek, Croatia; 2Department of Biochemistry and Molecular Biology, Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia [email protected]
Aiming to evaluate effects of population substructure on the reliability of a DNA match in the process of human identification, we used model of “in silico” constructed populations with and without substructure. Effects of population substructure were evaluated at the level of locus heterozygosity, Hardy-Weinberg equilibrium, and mini-haplotype distribution. Inbreeding in a subpopulation of 100 individuals through 10 generations did not significantly alter level of heterozygosity, and Hardy- Weinberg equilibrium. However, analysis of mini- haplotype distribution revealed significant homogenization in separated subpopulations. Average observed mini-haplotype frequency (fo) increased to threefold from expected values (fe), and the number of mini-haplotypes with fo/fe above 10 increased over six-fold, suggesting that effects of population substructure on calculated likelihood ratios might be larger than previously estimated. In most criminal cases this would not represent a problem, but for identifications in large-scale mass fatality events, population substructure might considerably increase the risk of false identification.
Keywords: DNA typing, human identification, population substructure, mini- haplotypes, inbreeding
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 117 POSTER PRESENTATIONS – Forensic Genetics DNA Identification of Victims of Mass Disaster – the Latest Approach Presentation number: FG 39 Abstract number: ABS-1653-ISABS-2007
ADDITIONAL ANALYSIS OF CROATIAN POPULATION STRUCTURE AND COMMENTS ON IDENTIFICATION OF REMAINS IN MASS CASUALTY DATABASES: POWERPLEX 16 DATABASES FOR SOUTHERN CROATIANS
Schanfield M1, Marjanović D2,3, Škaro V3, Anđelinović S4, Primorac D5,6
1George Washington University, Department of Forensic Sciences, Washington, DC, USA; 2Institute for Genetic Engineering and Biotechnology Sarajevo, Sarajevo, Bosnia and Herzegovina; 3Ruđer Bošković Institute, Department of Molecular Medicine, Forensic Genetics Group, Zagreb, Croatia; 4 Clinical Hospital Split, Department of Forensic Genetics, Split, Croatia; 5Medical School at University of Split, Split, Croatia; 6Medical School, Josip Juraj Strossmayer University of Osijek, Osijek, Croatia. [email protected]
The validation of a Powerplex 16 (PP16) database for southern Croatians provided the opportunity to address questions raised by Birus et al (1) regarding the occurrence of linkage disequilibrium and false matches. The PP16 database had no significant deviations from HWE and were not significantly different from the previous study of the 13 shared CODIS loci (2) indicating that the two data sets could be pooled. A test for two locus linkage disequilibrium did not yield any unexpected significant correlations precluding any higher order disequilibrium. The power of exclusion (RPNE) (3) in a parentage test assumes that a single comparison is being performed. When using parents or children to identify remains the exclusionary power using either allele is very low compared to using separate alleles, e.g. using the pooled Croatian database with the two most common alleles at each locus the combined RPNE for 15 loci when using two alleles is only 0.011, compared to an RPNE of 3.24E-11 when using separate alleles, or a random match likelihood using the same alleles of 9.3E-14. Thus, the likelihood of false matches when making large numbers of comparisons is very high, making it necessary to either compare multiple relatives to profiles, when a hit is made, or correct the expected RPNE for the number of comparisons made to get an estimate of the true RPNE. 1. Birus et al. CMJ 44:322-326, 2003. 2. Schanfield et al. JFS 47 :669-670, 2002. 3. Primorac et al. CMJ 41 :32-46, 2000.
Keywords: Population, Structure, PP16, Croatia, Power of exclusion
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 118 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Forensic Genetics DNA Identification of Victims of Mass Disaster – the Latest Approach Presentation number: FG 40 Abstract number: ABS-666-ISABS-2007
SUCCESS RATES OF NUCLEAR STR TYPING FROM DIFFERENT SKELETAL ELEMENTS
Selmanović A1, Miloš A1, Smajlović L1, Huel R1, Katzmarzyk C1, Rizvić A1, Parsons T1
1International Commission on Missing Persons, Sarajevo, Bosnia and Herzegovina. [email protected]
The International Commission on Missing Persons (ICMP) performs high throughput short tandem repeat (STR) typing to assist with the identification of skeletal remains exhumed from mass graves originating from conflicts that occurred in the former Yugoslavia during the 1990’s. Using a very large data set, we have evaluated DNA typing success rates of different skeletal elements. Samples representing different categories with respect to time since death have permitted an assessment of relative rate of degradation of DNA over time from different skeletal elements. These data are presented and discussed with the aim of increasing understanding of the factors that affect DNA preservation in bone, and how this information may be useful in guiding sampling strategies in DNA identification casework involving degraded bone.
Keywords: DNA Typing, STR, missing persons, skeletal elements, success rates
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 119 POSTER PRESENTATIONS – Molecular Anthropology
POSTER PRESENTATIONS Molecular Anthropology
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 120 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Y-Chromosome Polymorphisms
Presentation number: MA 9 Abstract number: ABS-8648-ISABS-2007
INTRA-ETHNIC VARIATION OF THE Y CHROMOSOME IN EUROPEAN COUNTRIES: A COMPARATIVE STUDY
Balanovsky O1, Pshenichnov A1, Rootsi S2, Churnosov M3, Atramentova L4, Tegako L5, Yankovsky N6
1Research Centre for Medical Genetics, Moscow, Russia; 2Estonian Biocenter, Tartu, Estonia; 3Belgorod State University, Belgorod, Russia; 4Kharkov State University, Kharkov, Ukraine; 5Institute of ethnography and folklore, Minsk, Belarus; 6Vavilov Institute for General Genetics, Moscow, Russia [email protected]
To reveal degree of the regional Y chromosomal variation in Europe we compared our data on Russians (14 regional populations), Ukrainians (4 populations) and Belorussians (2 populations, and 2 populations from Behar et al., 2003) with the published regional data on other European countries. For reliable calculations neighbour populations were pooled to reach sample sizes above 70. To measure the intra-ethnic variation we calculated (i) the average genetic distance between regional populations of every group and (ii) Gst (Fst) variation. Gst value was considered as preferable measure, as it was found to be less sensitive to level of phylogenetic resolution in the data. Croatians, Finns, Russians and Italians were proved to be the most diverse (genetically subdivided) groups; Swedes and Germans demonstrated moderate variation; Greeks, Turks, Poles, Belorussians and Ukrainians were more genetically homogenous, showing lower geographic variation of the paternal lineages inside their countries. However, even lower variation of the Y chromosome is significantly higher as compared with analogous values calculated from mitochondrial DNA and autosomal data. This finding stresses that forensic studies may demand not only country-specific, but province- specific databases (at least for listed above highly genetically subdivided countries), since haplogroup profiles differ significantly from one province to another, inside the same country. Despite the high intra-ethnic variation (Gst=0.03 on average), the inter-ethnic differences were five times higher (Gst=0.15), revealing dominance of inter-ethnic variation in structuring the paternal gene pool in Europe.
Keywords: Y chromosome, intra-ethnic variation, regional variation, genetic distances, subdivided population
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 121 POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 10 Abstract number: ABS-9631-ISABS-2007
Y CHROMOSOME ABERRATIONS AND POLYMORPHISMS IN PATIENTS WITH REPRODUCTIVE FAILURE
Bellovits O1, Rusz A2, Fodor F3, Csonka E4, Hadlaczky G4, Bujdosó G1
1Semmelweis University, Department of Forensic Medicine, Budapest, Hungary, 2Semmelweis University, Department of Urology, Budapest, Hungary, 3LabOrigo Molecular Diagnostics, Budapest, Hungary, 4Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary [email protected]
Data from the Hungarian Central Statistical Office (KSH) shows that infertility affects about 150,000 couples in Hungary, which means that one in every seven couples has problems in conceiving, and this number increases continuously. Nowadays the significance of genetic examination is to diagnose the cause of infertility and to help making conception by assisted reproductive techniques for many couples with severe male factor infertility. The aim of our examination was to investigate Y chromosome aberrations and polymorphisms in infertile male with compared to fertile control men. Sixty men applied for assisted reproduction technique and undertook the genetic examination during 2003 and 2006. The genetical examination included classical cytogenetic methods (GTG, QFQ, CBG and NOR banding techniques), FISH analyses and molecular characterisation of Y chromosome. The infertile male group was compared with 568 fertile male (control group) who took part in paternity cases in our laboratory. Our results confirmed that beside the Klinefelter’s syndrome (8,3%) the extra Y chromosome (3,3%), and Y chromosome microdeletion (3,3%) were the most frequent disorders. Examination of the incidence of fluorescence polymorphisms of Y chromosome did not show significant difference between the fertile and infertile men. Incidence of extreme size (large and small) polymorphisms of Y chromosome was higher in the infertile male group than the expected value; hence the Y chromosome with extreme size can be in connection with infertility. In addition to the routine andrological examination of infertile men, the necessity of genetic investigations can be proved by the numerous chromosome disorders.
Keywords: infertility, azoospermia, Y chromosome disorders, cytogenetics, molecular genetics
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 122 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 11 Abstract number: ABS-2988-ISABS-2007
ALLELE FREQUENCIES OF 17 STR Y CHROMOSOME LOCI IN THE POPULATION OF REBUBLIC OF MACEDONIA
Jakovski Z1, Nikolova K 1, Jankova R 1, Janeska B 1, Marjanovic D 2, Aleksej D1
1Institute of forensic medicine and criminology, Medical faculty, Univesrsitu St. Ciril and Metodious, Skopje; Republic of Macedonia; 2ISABS, Zagreb, Croatia [email protected]
Allele frequencies of 17 STR loci (DYS19, DYS189I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385I/II, DYS438, DYS439, DYS437, DYS448, DYS456, DYS458, DYS635 and Y GATA H4) were determined in a sample of 100 unrelated individuals from the Republic of Macedonia. AmpFISTRYFiler Kit (Applied Biosystems) was used for PCR amplification. Allele frequencies of these loci have been compared with data obtained from geographically close European populations (Serbia, Kosovo, Bulgaria, Bosnia and Herzegovina and coastal part of Croatia). The significance level after Bonferroni’s correction was considered as p<0.01 except for DYS437 and DYS438 loci since for these comparisons only two populations was used because of lack of data. Exact tests of populations differentiation show statistically significant differences of allele frequencies of DSY385 locus between Macedonian and Serbia, Kosovo, Bosnia and Herzegovina populations (population from coastal part of Croatia was not considered in calculation). Almost the same result (excluding Kosovo population and including population from coastal part of Croatia) was noticed for DYS389- II locus. Macedonian and Kosovo populations show significant differences of allele frequencies for DYS392 loci as well as for DYS19. The significant difference result for latest mentioned locus was also recorded between Macedonian and population from coastal part of Croatia and for DYS390, DYS391 as well as DYS393 loci. Macedonian and Bosnian and Herzegovnian populations have significant difference in allele frequencies for DYS438 locus.
Keywords: Y chromosome STR, Macedonian Population, Frequencies, Statistic, Comaparative
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 123 POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 12 Abstract number: ABS-8046-ISABS-2007
MOBILITY OF MALES IN AN ADMIXED MATRILINEATE USING Y- CHROMOSOMAL STRS AND SNPS
Langstieh BT1, Reddy BM2
1Department of Anthropology, North-Eastern Hill University, Meghalaya, India; 2Molecular Anthropology Group, Biological Anthropology Unit, Indian Statistical Institute, Habsiguda, Hyderabad [email protected]
To trace the ethnic origin of an assumed admix group ‘the Lyngngam’, an Austro- Asiatic population residing in the sub-Himalayan belt of North-East India. DNA was extracted from 423 male individuals belonging to the broader Austro-Asiatic (Mon- Khmuic) linguistic group, of which 71 Tibeto-Burman speakers were also included in the above sampled group. The analytical approach was to reconstruct population history, admixture estimate and also possibly trace the time since most common recent ancestor (TMRC). Ethnic identity of the studied population had been questioned from the socio-cultural perspective, from language as well as traditional genetic markers with no conclusive evidence. The contention was as to whether they were Austro-Asiatic or Tibeto-Burman in ethnic origin. The reason that they might have been an admixed group could not be ruled out. Following the matrilineal line of descent and having matrilocal residence results from Y chromosomal polymorphism showed convincing remarks that genetically mobility of males from the neighbouring groups in particular from the Tibeto- Burman Garo was dominant. The findings of the present study revealed that more mobility of males is operating in matrilineal societies whereas; female mobility has been proved time and again to be a characteristic trait of the patrilineal societies of the world. Comparative analyses do conform that the only Austro-Asiatic (Mon- Khmuic) population of India constitute the bulk of migratory bands from genetic affinities to S.E. Asia. Amidst the multitude of Tibeto-Burman tribes in the region, the Austro-Asiatic trait not only of language but genes seemed to have survived the onslaught of admixture.
Keywords: Austro-Asiatic (Mon-Khmuic), Tibeto-Burman, Lyngngam, Y-STRs, Y SNPs
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 124 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 13 Abstract number: ABS-7193-ISABS-2007
Y CHROMOSOME HAPLOTYPES IN ROMA AND CAUCASIAN POPULATIONS FROM SOUTHERN-WEST HUNGARY
Nagy G1, Könczöl F1, Márk L2, Bajnóczky I1
1Institute of Forensic Medicine, Medical School, University of Pécs, Pécs, Hungary; 2 Depertment of Biochemistry and Medical Chemistry, Medical School, University of Pécs, Pécs, Hungary [email protected]
To enlarge our understanding of genetic variation in Hungarian population, a population genetic study was carried out on unrelated 50 Hungarian Caucasian and 50 Hungarian Roma male from South-West Hungary area (Baranya, Somogy and Tolna)-. Sixteen Y chromosome STR polymorphisms (DYS456, DYS389I, DYS390 DYS389II, DYS458, DYS19, DYS385 a/b, DYS393, DYS391, DYS439, DYS635, DYS392, Y GATA H4, DYS437, DYS438, DYS448.) were analyzed in the samples. Genomic DNA was extracted according to standard techniques (Chelex-100 method) and amplified utilizing different amplification approaches (Y filer® produced by Applied Biosystem). PCR products were separated by capillary gel electrophoresis on an ABI PRISM 310 Genetic Analyzer and typed by comparison against standard allelic ladders. Row data were analyzed by GeneMapper ID software. A general STR allelic frequency pattern in the Roma and Caucasian population from Hungary corresponds to other European and Indo- Asian populations. In Roma population 42 haplotypes, in Caucasian population 48 haplotypes were observed in single copy. 24 Caucasian Hungarian haplotype and 21 Roma haplotype were not previously observed in the Y STR Haplotype reference Database among the set of 447 populations. We analyze our samples with minHt (9 loci) and minHt+ SWGDAM core set (11 loci).
Keywords: Y chromosome, haplotypes, Hungary, Y Filer, Yhrd
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 125 POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 14 Abstract number: ABS-4319-ISABS-2007
THE APPLICATION OF Y-STRS AND MITOCHONDRIAL DNA TO FAMILIAL SEARCHING
1 1 1 1 Noor J , Underwood S , Evans C , Bates F
1The Forensic Science Service, Trident Court, Birmingham, UK [email protected]
When a full SGM/SGM+ profile is obtained from a crime scene, but no matches against the National DNA Database (NDNAD) are obtained, it is possible to identify possible relatives of an offender, whose profiles are already on the NDNAD. Familial searching is carried out using an algorithm that compares the number of matching alleles between the crime sample and the sample on the NDNAD. DNA profiles of relatives are more likely to be similar than unrelated individuals. Typically a search would result in 2000 to 3000 individuals being identified as potential relatives (nominals). However, other information can be used to restrict the number of nominals, such as ethnicity and location. It is possible to reduce the number of nominals even further by Y- chromosome STR’s and mitochondrial DNA (mtDNA) analysis. Nominal Y- STR’s are analysed using Applied Biosystems AmpFℓSTR Yfiler™ system. The mtDNA analysis involves sequencing of hypervariable regions HVI and HVII. To date, the FSS has successfully processed fifteen cases using Y- STR familial screening, one case using mtDNA sequencing and one case using both Y-STR and mtDNA sequencing.
Keywords: Y-STR, mitochondrial DNA, familial searching, relatives, SGM+
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 126 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 15 Abstract number: ABS-3398-ISABS-2007
Y-CHROMOSOMAL SHORT TANDEMS REPEATS (Y-STRs) AND THEIR APPLICATIONS FOR POPULATION STUDIES AND FORENSIC CASEWORK IN LATVIA
Pliss L1, Puzuka A2, Rozane S3, Sabule A3, Baumanis V1, Krumina A2
1Latvian Biomedical Research and Study Centre; University of Latvia, Riga, Latvia; 2Riga Stradins University, Riga, Latvia; 3Centre of Legal Medicine, Riga, Latvia [email protected]
A series of polymorphic Y-specific microsatellites show high levels of heterogeneity within and between populations and thus seem to be very useful for population genetic, evolutionary and forensic studies. The aims of our research were: to characterize Y-STRs variability in the Latvians; to validate the application of a 17-locus Y-STR system for forensic. A sample of 140 paternally unrelated Latvians were analyzed at 12 Y-STR loci using PowerPlex Y System (Promega, USA). Haplotype diversity was calculated using the Arlequin 2.0 package. AmpFlSTR Yfiler PCR amplification kit (Applied Biosystems, USA) was validated according to the SWGDAM (the Scientific Working Group on DNA Analysis Methods) guidelines for forensic. Y-STR analyses of the haplogroup (hg) I revealed 12 different haplotypes (0.957±0.016). In contrast to hg I, the haplotype diversity of hgs R1a and N3a was 0.992±0.003, and 93 distinct haplotypes were detected. We have found two predominant alleles at DYS392 and DYS 393 loci that make up 94% within hg R1a samples. However, for hg N3a the most frequent alleles were 10 at DYS438 (95.5%) and 14 at DYS437 (95.5%). Based on 17- locus Y-STR system our results showed that full profiles are attainable with low levels of male DNA and no detectable cross-reactive products were obtained on female DNA, bacteria, and commonly encountered animal species. The analysis of Y-STRs revealed genetic variability within predominant phylogenetic branches in the Latvian population. Application of a 17-locus Y-STR system in forensic is very useful, especially in cases where typing autosomal STRs has met with limited success.
Keywords: Y chromosome, Short tandem repeats (STR), Haplotypes, Forensic, Latvia
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 127 POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 16 Abstract number: ABS-3786-ISABS-2007
VARIATION OF PATERNAL, MATERNAL AND AUTOSOMAL GENETIC MARKERS ON INTRA-ETHNIC (UKRAINIANS) AND INTER-ETHIC (EUROPE) LEVEL SUPPORTS Y CHROMOSOMAL MARKER BIAS
Pshenichnov A1, Balanovska E1, Balanovsky O1, Atramentova L2, Churnosov M3, Soloviova D1, Zaporozhchenko V1
1Research Centre for Medical Genetics, RAMS Moscow, Russia; 2 Kharkov National University, Kharkov, Ukraine; 3 Belgorod State University, Belgorod, Russia [email protected]
It was judged that interpopulational variation of Y is higher then for mtDNA and authosomes (e.g. Kittles 1999, Jorde 2000). But following study of Y chromosome Alu insertion sequences (Wilder 2004) yielded equal estimates of variation. It was proposed then that Y polymorphisms utilized in earlier estimates overrepresented intercontinental differences which led to higher diversity estimates. If Y SNPs really overrepresented intercontinental variation, they wouldn’t satisfactorily distinguish closely related populations. To check this, we genotyped Y SNPs, mtDNA haplogroups and five autosomal loci in Ukrainian populations: Western, Central, Eastern (N=511). For further comparisons we gathered Y, mtDNA and autosomal data for other 17 European populations. Nei genetic distances between 20 populations were calculated for Y, mtDNA and autosomes. We found that correlation of genetic and geographic distances depends on size of area for which correlation is estimated. For mtDNA and autosomes correlation is same when estimated for populations closer then 2,000km and closer than 4,000km: r=0.2-0.3. For Y, correlation grows with area (2,000-3,000-4,000km) and finally reaches 0.7. This reflects growing of Y variation compared to mtDNA and autosomes for distant populations and supports the idea that Y-SNPs overrepresent interregional variation. But even at the ethnic level, Y inter-population variation is higher then for mtDNA and autosomes. Even being biased to inter-regional variation, Y chromosome catches ethnic traits more accurately then mtDNA and autosomes, at least in Europe. “Сorrecting” the bias of Y supposedly wouldn’t reduce its distinguishing capacity.
Keywords: Y chromosome, mtDNA, autosomal DNA markers, Ukrainians, genetic diversity
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 128 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 17 Abstract number: ABS-3945-ISABS-2007
ORIGIN AND PHYLOGEOGRAPHY OF Y-HAPLOGROUP N
Rootsi S1
1Department of Evolutionary Biology, University of Tartu and Estonian Biocentre, Tartu, Estonia [email protected]
One of the most widespread and frequent branches of the Y phylogeny in Eurasia is haplogroup NO. It entails a low number of NO* lineages and two frequent sister clades, N and O. While the phylogeography of clade O has drawn considerable scrutiny, knowledge about hg N was relatively impoverished. To study the origin, phylogeographic patterning and demographic significance of haplogroup N, 5389 samples from 58 populations in different geographical regions were genotyped (M9 derived samples with ancestral allele of 92R7 were typed for markers M214, M231, M128, P43, Tat, M175) and analyzed together with data from literature. Results presented in this abstract are based on study by Rootsi et al. (2007). Analysis of hg N suggests that its high frequency in East Europe is due to its expansion westward on a counter-clock northern route from Inner Asia/ Southern Siberia, approximately 12-14 thousand years ago. The widespread presence of haplogroup N in Siberia and its absence in Native Americans implies its spread happened after the founder event for the Americas. The most frequent sub-clade N3, arose probably in the region of present-day China, and subsequently experienced serial bottlenecks in Siberia and secondary expansions in Eastern Europe. Another branch, N2, forms two distinctive sub-clusters of STR haplotypes, Asian (N2-A) and European (N2-E), the latter now mostly distributed in Finno- Ugric and related populations. These phylogeographic patterns provide evidence consistent with male-mediated counter-clockwise Late Pleistocene - Holocene migratory trajectories towards Northwestern Europe from an ancestral East Asian source of Palaeolithic heritage.
Keywords: Y chromosome, haplogroup origin, phylogeography, northern Eurasia, expansion
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 129 POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 18 Abstract number: ABS-575-ISABS-2007
PHYLOGEOGRAPHY OF Y-CHROMOSOMAL LINEAGES IN NORTH EURASIA
Stepanov V1, Kharkov V1, Medvedeva O1, Puzyrev V1
1Institute for Medical Genetics, Tomsk, Russian Federation. [email protected]
Aim of the study was to reconstruct the evolution of paternal lineages in populations of Siberia, Central Asia and Eastern Europe. Forty population samples from 22 ethnic groups were studied. Totally 1600 Y chromosomes were genotyped for 40 biallelic markers according to Y chromosome consortium (YCC) classification. The microsatellite haplotypes within HG were constructed using 7 STR loci. Thirty one haplogroups were observed, but frequencies of only 7 of them (N3a, R1a1, Q*, C3xC3c, N2, C3c, O3) were higher than 3 percent. In sum these 7 haplogroups comprise 86% of Y-chromosomal gene pool in North Eurasia. The proportion of inter- population differences in the total genetic variability of region's population according to the analysis of molecular variance is 19%. Analysis of genetic relationships between populations reveals three main clusters of populations in space of two first PCs reflecting the differential presence of ancient West-Eurasian Caucasoid, Proto-Uralic and Paleoasiatic components. Based on analysis of microsatellite haplotypes within main Y- chromosomal haplogroups, molecular diversity within monophyletic lineages were calculated and phylogenetic trees for most common haplogroups were reconstructed. Western-Eurasian lineages (R1a1, R1b) are characterized by the maximal diversity in Eastern European populations. Eastern-Eurasian lineages have the high level of diversity in populations of Eastern Siberia and North-East Asia. The age of genetic diversity generation and time of population differentiation (Td) shows that most lineages which are common in North Eurasian populations dated back to Upper Paleolithic period before the last glacial maximum.
Keywords: Y chromosome, genetic diversity, North Eurasia, phylogeography, population genetics
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 130 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Y-Chromosome Polymorphisms Presentation number: MA 19 Abstract number: ABS-6278-ISABS-2007
HAPLOTYPE ANALYSIS OF 17 Y-STR LOCI IN SOUTHERN CROATIAN POPULATION
Sutlović D1, Ljubković J1, Definis-Gojanović M1, Anđelinović Š1
1Department of Pathology and Forensic Medicine, Split University Hospital and School of Medicine, Split, Croatia [email protected]
To comparate analytical methods to evaluate nuclear DNA concentrations of forensic and ancient bone samples. Three different methods were used: agarose gel electrophoresis, spectrophotometry and Quantitative Real-time PCR. QRT- PCR offers several advantages with respect to other compared methods. Twenty four DNA extracts from bone samples: eight DNA extracts from ancient bone samples (3 DNA samples retrieved from 10-50 years old bones and 5 DNA samples obtained from about 1300 years old bones recovered from an old church burrial sight), eight DNA were extracted from mass graves bones and eight were extracted from fresh bones. The quantification assay was performed in total volume of L of DNA extract, Quantifiler human primer mix andμL containing 2 μ25 Quantifiler PCR reaction mix with thermal cycling conditions according to the manufacture’s protocols In the following trials total DNA was evaluated by gel electrophoresis (1.5% agarose) and ethidium bromide staining by UV illuminations (312 nm) of the gel . Absorbencies at 260 and 280 nm were measured by spectrophotometer. Highly sensitive methods for quantification of human DNA retrieved from forensic and ancient DNA samples are important because they ensure the consistency of low copy number DNA typing. RT- PCR offers several advantages with respect to other compared methods. In samples which contain high amounts of DNA all of the tested methods gave positive results with good agreement. All samples that possibly contain small amount of DNA or inhibitors therefore need to be quantitated by QRT-PCR as method of choice.
Keywords: Y-chromosomal STR, population data, forensic science, Haplotype, Southern Croatia
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 131 POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Mitochondrial DNA Polymorphisms
Presentation number: MA 20 Abstract number: ABS-1820-ISABS-2007
DECIPHERING THE PAST: A MOLECULAR GENETIC STUDY ON OLD THRACIAN POPULATIONS FROM ROMANIA, DATING FROM THE BRONZE AND IRON AGE
Cardos G1,2, Miritoiu N3, Soficaru A3, Kroll A1 and Rodewald A1
1Institute of Human Biology, University of Hamburg, Hamburg, Germany; 2Department of Genetics, University of Bucharest, Bucharest, Romania; 3Institute of Anthropology, Romanian Academy, Bucharest, Romania. [email protected]
We carried out this paleogenetic study on skeletal samples of 50 old individuals from Thracian populations dating from the Bronze and Iron Age, found in Romania. In order to show the degree of their genetic kinship with nowadays Romanian population and other old and modern human populations, we analyzed mtDNA (HVR I and HVR II regions) and nuclear DNA (vWA31A Microsatellite) polymorphisms. The genetic sex of the old individuals was identified by amplifying a sequence of the Amelogenin gene. We extracted ancient DNA from skeletal remains of 50 old individuals by three methods, as follows: a phenol-chloroform DNA extraction method, a guanidine-tiocianate and silica–based method and an Invisorb Forensic Kit-based DNA extraction method. After amplifying by PCR, the mtDNA HVR I and II regions were sequenced by the Sanger method. The nuclear DNA sequences were separated and identified on PAA gel, Ag-stained. The mtDNA sequences of the old Thracian individuals were compared with similar DNA sequences of individual samples from the present-day Romanian population and other old and modern European populations. The data about vWA31A DNA marker in the Thracian population were also compared with similar genetic data of other human populations. Our results reflected an evident genetic similarity between the old Thracian populations and other modern populations from South- East Europe. Moreover, our genetic analysis suggested that the old Thracian populations who lived in Romania during the Bronze and Iron Age might have contributed to the foundation of the modern Romanian genetic pool.
Keywords: aDNA, mtDNA, Thracian population, vWA31A, Amelogenin
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 132 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 21 Abstract number: ABS-896-ISABS-2007
RELIABILITY IN PHYLOGEOGRAPHIC SIGNALS: A CASE STUDY OF AN mtDNA SUBHAPLOGROUP
Dulik MC1, Gökçümen O1, Schurr TG1
1Department of Anthropology, University of Pennsylvania, Philadelphia, PA, USA. [email protected]
The lack of evidence for recombination in mitochondrial DNA (mtDNA) provides support for the assumption that a direct correspondence between polymorphisms found in the coding and control regions of the mtDNA exist. This has lead to a near total reliance on motifs in the hypervariable segments (HVS) 1 and 2 of the mitochondrial control region for anthropological studies. The aim in this analysis was to assess the reliability of phylogeographic patterns of a subhaplogroup based solely on HVS1 sequences versus complete sequence analysis. For this analysis, D5 sequences in published literature were combined into a database, and new complete sequences were generated and added to this database. Networks were created using HVS1 and complete sequences to compare the topographies of the networks, and maximum likelihood trees and likelihood ratio tests were used to evaluate the presence of a molecular clock model. Discrepancies between coding and control region mutations were evident, having considerable effect on phylogeographic signals and the dating of coalescent events. In the future, caution should be used in relying only on HVS1 motifs for examining population histories.
Keywords: mtDNA, phylogeography, complete sequences, molecular clock, likelihood ratio tests
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 133 POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 22 Abstract number: ABS-1750-ISABS-2007
CULTURAL AND GENETIC DIVERSITY IN CENTRAL ANATOLIA: A LOCAL PERSPECTIVE
Gokcumen O1, Gultekin T2, Alakoc Y3, Tug A3, Gulec E2, Schurr TG1
1Department of Anthropology, University of Pennsylvania, Philadelphia, United States; 2Department of Anthropology, Ankara University, Ankara, Turkey; 3Department of Forensic Medicine, Ankara University Medical Faculty, Ankara, Turkey. [email protected]
Anatolia has been an important crossroads for numerous populations since the Neolithic. Among these, the Hattis, Urartians, Lydians, Phyrigians and Ottomans emerged in Anatolia proper. In addition, although non-Anatolian in origin, the Hittites, Greeks, Romans and Byzantines influenced and were influenced by local Anatolian cultures. These dynamics, as well as more recent events, such as the Turco-Ottoman War of late 19th century, the reconfiguration of the populations of the Ottoman Empire and the Greek- Turkish population exchange of 1920s, have made Anatolia a culturally and genetically complex region. Despite this complex history, Anatolia has been often been viewed as a uniform cultural landscape. Working from this perspective, previous studies of genetic variation in Anatolia analyzed samples from Turkish populations obtained from mostly urban hospitals or universities. Such studies not only overlooked the regional variation within Anatolia, but also treated contemporary Turkish populations as the direct representatives of Medieval and Neolithic Anatolian populations. To address these problems, we collected ~125 samples and extensive ethnographic data from a location in Central Anatolia southeast of Ankara. The samples were analyzed for mtDNA and NRY diversity, and the resulting data compared with those from previous genetic analyses of Turkish populations. We observed that several ethnic and cultural groups having different population histories co-existed in this location. This pattern likely represents the typical picture of Anatolian variation. We are using our genetic data to help us clarify these distinct population histories in greater detail.
Keywords: Turkey, Kinship, Haplotypes, DNA, Lineages
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 134 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 23 Abstract number: ABS-3634-ISABS-2007
THE MITOCHONDRIAL DNA POLYMORPHISM IN UKRAINIAN POPULATION
Grechanina EY1, Gusar VA1, Villems R2
1Institute of Clinical Genetics, Kharkiv, Ukraine; 2Biocenter, Tartu, Estonia [email protected]
Study of mtDNA polymorphism for estimation genetic diversity of Ukrainian population. There are 239 samples of Ukrainians from different regions of Ukraine. There are sequention of hypervariable segment HVS I in combination with RFLP- analysis of coding sites of mtDNA and phylogeographical analysis. This research was in Estonian Biocenter. Length of sequencing fragment was 377 bp 102 positions from 377 were polymorphic. From these 91 nucleotide substitutions are transitions, with prevalence pyrimidine under purine (69:22). Transversions were in 11 sites. Were determined 157 haplotypes. The most common haplotype (10.0%) corresponds to CRS. Indexes of genetic diversity for Ukrainian population are H=0.986 and Di=5.19. Screening of polymorphic sites established following haplogroups of mtDNA, which have all-European spreading: H-33.5%, V-5.4%, HV-3.7%, J- 11.7%, T-6.7%, U-20.9% (U2, U3, U4, U5, U7, U8), K-2.9%, I-2.1%, W- 2.1%, X-2.5%. Subhaplogroup U3 (2.5%) may testify the presence Iranian component. Subhaplogroup U4 was detected with frequency 3.7%. Subhaplogroup U5 has maximal frequency in Scandinavian people also was detected in Ukrainian population with high frequency 10.8%. Haplogroup V, as marker of Finnish-Hungarian people, has high frequency in Ukrainian population. There were found Asian lines (A, B, C, D, Z) with frequency 2.0%. Data indicate on complicated ethnical formation of modern Ukrainian population, where assimilation processes and inter-ethnic interactions played considerable role. They will be important additions in context about polymorphism of European populations mtDNA.
Keywords: mitochondrial DNA, Ukrainian population, polymorphism, genetic diversity, phylogeographical analysis
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 135 POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 24 Abstract number: ABS-6191-ISABS-2007
GENETIC STRUCTURE OF THE ISLAND OF RAB REVEALED BY MITOCHONDRIAL DNA ANALYSIS
Havaš D1, Jeran N1
1Institute for Anthropological Research, Zagreb, Croatia [email protected]
The aim of this study was to determine genetic structure of the Croatian Adriatic Island of Rab population through analysis of mitochondrial DNA polymorphisms. Mitochondrial DNA variation was analyzed in a sample of 163 randomly chosen adults from 5 different towns and villages of the Island of Rab (Rab, Lopar, Barbat, Banjol, Supetarska Draga). The hypervariable segment I (HVS-I) of the control region of mtDNA was sequenced between nps 16024 and 16383 and together with RFLP sites diagnostic for main Eurasian haplogroups was analyzed in order to confirm the exact haplogroup affiliation of mtDNA HVS-I lineages. The analysis revealed the highest predominance of haplogroup H (36.8%), as in majority of European populations. Haplogroup U (32.5%) is the second most prevalent haplogroup in the investigated population. Subhaplogroup U5 (3.0%) has a decreased frequency comparing to Croatian mainland and other Island population. The most predominant subhaplogroup is U4 (24.5%), with much higher frequency than in other Croatian or European populations. Furthermore, the phylogenetic analysis showed very small number of haplotypes in most of the haplogroups in investigated population. All this findings, uniformity of haplotypes which can be seen in one or even more villages and very elevated frequency of subhaplogroup U4 confirm the effects of evolutionary forces, especially genetic drift caused by founder effect, which mediated a specific distribution of mtDNA haplogroups and shaped genetic structure in the isolated population of the Island of Rab.
Keywords: isolated population, mitochondrial DNA, Island of Rab, genetic drift, founder effect
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 136 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 25 Abstract number: ABS-205-ISABS-2007
MITOCHONDRIAL DNA HAPLOGROUP DIVERSITY OF THE ISLAND OF VIS
Jeran N1, Havaš D 1
1Institute for Anthropological Research, Zagreb, Croatia [email protected]
The aim of this study was to determine population genetic structure of the mitochondrial haplogroups in an isolated island population as well as the overall level of genetic diversity. MtDNA variation was explored in a sample of 130 individuals from the Island of Vis (eastern Adriatic) by sequencing HVS-I region between nps 16024 and 16383 and screening of selected RFLP sites diagnostic for main Eurasian mtDNA haplogroups. Similar to other European populations, the most frequent mtDNA haplogroups were H (30.0%) and U (26.15%). However, as in other previously investigated Croatian islands, the Island of Vis shows departures of certain (sub) haplogroups from the expected frequencies implying to genetic drift: the increased frequency of haplogroup HV (9.23%), subhaplogroup U5 (22.3%) and preV (3.85%); absence of haplogroup I and decreased frequency of haplogroup J (4.62%). Furthermore, the frequencies of haplogroups V and T are increased comparing to general Croatian sample. The phylogenetic analysis of haplogroups showed relatively high number of haplotypes in some haplogroups (H and U) that confirm their heterogeneity and their early age ancestry. On the other side, homogenity of haplotypes in some haplogroups (HV) confirms the effect of genetic drift in a small and isolated population. As in other Croatian islands the genetic drift caused by founder effect has been the main evolutionary force included in shaping the observed genetic diversity of mtDNA on the Island of Vis.
Keywords: isolated population, mitochondrial DNA, Island of Vis, genetic drift, founder effect
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 137 POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 26 Abstract number: ABS-1683-ISABS-2007
HIGH RESOLUTION SURVEY OF ANDAMAN ISLANDERS mtDNA VARIATION SUGGESTS COMPLEX PATTERN FOR THE PEOPLING OF THE ARCHIPELAGO AND HINT FOR A BACK-MIGRATION EVENT FROM SOUTHEAST ASIA INTO SOUTH ASIA
Metspalu M1, Endicott P2, Barik SS3, Sanchez JJ4, Chandrasekar A3, Rao VR3
1Institute of Molecular and Cell Biology, Tartu University and Estonian Biocentre, Tartu, Estonia, 2 Department of Zoology, Oxford, United Kingdom, 3 Anthropological Survey of India, Kolkata, India, 4 National Institute of Toxicology and Forensic Science, Tenerife, Spain [email protected]
Previous genetic research using mtDNA has shown isolation of the region- specific genetic pools in Eurasia and Oceania soon after the initial Out of Africa migration during the late Pleistocene. Comparable, very early isolation has been proposed for the physically isolated Andaman Islands populations of Island South-East Asia. Using whole-genome sequencing, combined with multiplexed SNP typing, we investigate here the deep structure of mtDNA haplogroups M31 and M32 that are ubiquitous to the archipelago. We identify a, thus far unnoticed, rare polymorphism shared between these two lineages, what suggests, that these lineages actually form a single clade – M31’32 - which evolved in South-East Asia or the Andaman Islands after the rapid dispersal of modern humans from Africa to Australia at least 45 thousand years ago. The enhanced resolution of M31a indicates a back migration from South-East Asia into an area that now contains most of the Austro- Asiatic speakers of India, presenting a far more complex picture of inter-regional interaction for mtDNA than is usually assumed.
Keywords: mtDNA, Andaman Islands, M31, M32, phylogeography
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 138 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 27 Abstract number: ABS-1357-ISABS-2007
MITOCHONDRIAL CONTROL REGION POLYMORPHISMS REMAIN CONSTANT DURING AGEING
Hewakapuge SK1, van Oorschot R2, Baindur-Hudson S1
1School of Molecular Science, Victoria University, Werribee VIC Australia, 2Victoria Police Forensic Science Services Department, Macleod VIC Australia. [email protected]
Ageing is a predictable, universal and detrimental process in humans. The ‘mitochondrial theory of ageing’ implies that the accumulation of mitochondrial DNA mutations, and the subsequent cytoplasmic segregation of these mutations during life, is an important contributor to the ageing process. Mitochondria have a separate autonomously replicating DNA genome and are maternally inherited. Previous research has identified accumulation of mutations in the control region of the mitochondrial genome with age. In this study we analysed two most variable sequences in control region (hypervariable region 1 and 2) within twelve separate families, eleven of three unbroken maternal lineages and one with four maternal lineages (ages 5 to 96). Since mitochondria are maternally inherited, any sequence difference between the youngest member of the family compared with the others was considered to be age related. No age related mitochondrial mutation/s observed in the twelve families analysed. Therefore this methodology of direct sequencing of mitochondrial hypervariable regions does not appear to be a useful tool to assist in the prediction of the age of the person from whom a biological sample was collected at a crime scene.
Keywords: Ageing, mitochondrial DNA, prediction, Control region, Maternal three generations
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 139 POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 28 Abstract number: ABS-3927-ISABS-2007
TWO UNIVERSAL PRIMERS ON THE 12S AND 16S RIBOSOMAL RNA LOCI FOR SPECIES IDENTIFICATION
Osawa M1, Kitano T2, Hasegawa I1, Sato F1, Umetsu K2
1Department of Forensic Medicine, Tokai University School of Medicine, Isehara, Kanagawa; 2Department of Experimental and Forensic Pathology, Yamagata University Faculty of Medicine, Yamagata, Japan [email protected]
To identify animal species from a variety of remains such as the bone and hair shafts, molecular procedures to the mitochondria genome have been developed, in which most have utilized the cytochrome b locus to design universal primers. In the present study, extensive multiple alignments to the whole genome of 30 mammals revealed highly conserved regions in the 12S and 16S ribosomal RNA genes. Two sets of oligonucleotide primers directed to the conserved tags of the loci were introduced to amplify equivalent segments of less than 250 bp in length from degraded material. PCR employing the primers yielded single products using DNA extracted from tissues of nine non-mammalian vertebrates of avis, reptilia, amphibia, and pisces, in addition to common mammals. Further, both segments contained sufficient nucleotide substitutions to determine animal species. This PCR-direct sequencing system upon the 12S and 16S ribosomal RNA loci will be attributable to the species identification of a wide variety of biological remains in forensic laboratories.
Keywords: mitochondria, species identification, sequence, forensic remains, ribosomal RNA
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 140 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 29 Abstract number: ABS-2248-ISABS-2007
A GLIMPSE AT THE FRENCH mtDNA GENETIC POOL
Pennarun E1,2,, Richard C1, Metspalu E2, Rudan P3, Villems R2, Chaventré A1, Moisan JP1
1Laboratoire d´Etude du Polymorphisme de l´ADN, Faculté de Médecine, Nantes, France; 2Department of Evolutionary Biology, Institute of Molecular and Cell Biology, University of Tartu and Estonian Biocentre, Tartu, Estonia [email protected]
In the coverage of the genetic pool of Europe, some major cavities were left, hence to fill one of them, namely the French mtDNA pool we collected 868 samples from twelve different locations of France. Those samples were sequenced for the hypervariable segment I (HVS-I) and then typed for SNPs in the coding region, either by RFLP or 5' nuclease allelic discrimination, in order to assign them to the right haplogroup. Then the mtDNA gene pools of French Basques and Bretons were compared in terms of frequency and composition with relevant neighbouring populations. The French Basques’ mtDNA pool shares some common cardinal features with that of the Spanish Basques, represented in the high prevalence of haplogroup H. However, the French Basques do show a number of distinct features, most notably expressed in the much higher frequency of haplogroups linked with the Neolithic diffusion in Europe. In Brittany, Finistère shows closer affinities with Britain and Scandinavia than the two other departments of Brittany. The mtDNA haplogroup composition of the French does not differ significantly from the surrounding European genetic landscape. In a finer grain, microgeographical differentiation can be revealed as shown for the French Basque country and for Brittany.
Keywords: France, microgeographical differentiation, phylogeography, Basque, Breton
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 141 POSTER PRESENTATIONS – Molecular Anthropology Mitochondrial DNA Polymorphisms Presentation number: MA 30 Abstract number: ABS-5763-ISABS-2007
BERINGIAN STANDSTILL AND SPREAD OF NATIVE AMERICAN FOUNDERS
Tamm E1, Kivisild T1, Reidla M1, Metspalu M1, Smith DG2, Mulligan CJ3, Bravi CM4, Rickards O5, Martinez-Labarga C5, Khusnutdinova EK6, Fedorova SA1,7, Golubenko MA1,8, Stepanov VA8, Gubina MA1,9, Zhadanov SI1,9, Ossipova LP9, Damba L1,9, Voevoda MI9, Dipierri JE10, Villems R1, Malhi RS11
1Department of Evolution Biology, University of Tartu and Estonian Biocentre, Tartu, Estonia; 2Department of Anthropology, University of California, Davis, CA, USA; 3Department of Anthropology, University of Florida, Gainesville, FL, USA; 4IMBICE, La Plata, Argentina; 5Department of Biology, University of Rome “Tor Vergata” Italy; 6Institute of Biochemistry and Genetics, Ufa Research Center, Russian Academy of Sciences, Ufa, Russia; 7Department of Molecular Genetics, Yakut Research Center, Russian Academy of Medical Sciences, Yakutia, Russia; 8Institute of Medical Genetics, Tomsk Research Center, Russian Academy of Medical Sciences, Tomsk, Russia; 9Institute of Genetics and Cytology, Siberian Branch of Russian Academy of Sciences, Novosibirsk, Russia; 10INBIAL-UNJu, Jujuy, Argentina; 11Department of Anthropology and Institute for Genomic Biology, University of Illinois Urbana-Champaign, IL, USA. [email protected]
The genetic studies on Native Americans have convincingly established four major pan American (A2, B2, C1 and D1) and three minor North American founding mtDNA haplotypes (X2a, D2 and D3). The paucity of established founding mtDNAs suggests that a small number of migrants initially peopled the Americas. However, previous investigations are based on low-resolution sequence data, preventing accurately estimate the possible scenarios of initial peopling of the Americas. We analyzed 110 Native American and 513 Asian complete mtDNA genomes (including 20 new mtDNAs from the Americas and 7 from Asia generated in current study). Further, we used these sequence data to direct the high-resolution genotyping of 20 American and 26 Asian populations of distant origin and language affiliation and analyzed the combined dataset along with publicly available data. We determined the hidden diversity within the Native American genepool by identifying several new founding lineages (C1b, C1c, C1d, C4 and D4h3). The New World founder lineages are separated from their Asian sister-clades by several mutations and show similar coalescence time estimates, they lack nested structure inside haplogroups, but have largely autochthonous variation of sub-lineages. These results suggest that the ancestors of Native Americans had to pause in Beringia long enough for the mutations to accumulate the subsequent simultaneous movement southward was likely swift followed by long-term isolation. Besides, finding Native American A2 haplotypes in southern and western Siberia and distribution pattern of D2 sub-clades in Siberia and North American arctic allows identifying more recent gene flow around and across Beringia.
Keywords: Native Americans, Beringia, complete sequences, founder lineages, mtDNA
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 142 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Population Genetics Studies on Other Molecular and Phenotypical Markers
Presentation number: MA 31 Abstract number: ABS-1146-ISABS-2007
GENETIC POLYMORPHISMS OF 15 STR LOCI IN THREE EAST SLOVAKIAN POPULATIONS
Bernasovská J1, Bernasovský I 1, Boroňová I1, Petrejčíková E1, Soták M1
1Institute of Biology, Faculty of Humanities and Natural Sciences, University of Prešov, Prešov, Slovakia [email protected]
Romanies are a peculiar ethnic group whose members are scattered throughout the world. The exact number of Romanies is not known, but it is estimated to be about 10 million in the world. 350,000 of them living in Slovakia. This study provides additional population genetic data of the three different East Slovakian populations. We studied 645 unrelated and healthy individuals from East Slovakia: Slovak population (355) and two Romany populations: Romanies from region Spiš (127) and Olachian Romanies (163).Allele frequencies have been calculated in 15 STR loci. The comparison of the allele frequencies between the Romany populations revealed statistically significant differences in 11 out of the 15 loci (except for FGA, THO1, D5S818 and D18S51). Significant differences were observed between the Romanies from region Spiš and Slovak population (except for THO1 and D7S820) and between Olachian Romanies and Slovak population (except for D13S317). The selected kit PowerPlex® 16 system (Promega) includes amelogenin for sex determination; two penta- nucleotide repeats: Penta- D and Penta-E; and 13 tetra-nucleotide repeats: D3S1358, THO1, D21S11, D18S51, VWA, D8S1179, TPOX, FGA, D5S818, D13S317, D7S820, D16S539, CSF1PO. Due to the thirteen of these STR loci are present in the combined DNA index system (CODIS) PowerPlex® 16 is sufficient for human identity testing and ethno-genetic studies. The P-values of the exact test for Hardy-Weinberg equilibrium probabilities, observed and expected heterozygosity, matching probability, power of discrimination and exclusion, poly-morphic information content, typical paternity index, genetic diversity and the other population-genetic indices were calculated.
Keywords: Genetic polymorphism, Short tandem repeats, Romanies, East Slovakia, Population data
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 143 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 32 Abstract number: ABS-2433-ISABS-2007
NOVEL POLYMORPHISM IN INTRON 3 FOR TP53 GENE IN IRANIAN POPULATION
Biramijamal F1, Nazari E1, Soltani MS 1, Iravanloo G2
1National Institute for Genetic Engineering and Biotechnology NIGEB, Tehran, Iran; 2Cancer Institute, Tehran, Iran [email protected]
The p53 protein is considered a key combination in countering stress messages such as DNA damage. The p53 protein induces apoptosis and suppresses cancer cells containing multiple genetic alterations. This protein is a transcription factor and it can induce cell growth arrest or apoptosis to protect genome and cell against genotoxic damage. Also, The p53 tumor suppressor gene is involved in the etiology of malignant disease. It is recorded that most cancers carry p53 gene mutations. Several studies were described the polymorphism in intron 3 of the p53 gene and susceptibility to several types of cancer and diseases. To investigate the P53 polymorphisms among Iranian populations, we collected samples from healthy population. The P53 genotypes were determined by polymorphism chain reaction and direct DNA sequencing in healthy individuals. Among the healthy subjects, we found one novel polymorphism in intron 3 at nucleotides -23 (C → A). It indicates that a significant proportion of P53 polymorphisms currently unrecognized. So a better understanding of polymorphic metabolizing and protein expression of the gene can contribute in the future. This finding can help in molecular anthropology to detect Iranian population.
Keywords: p53 gene, novel polymorphism, population, Iran, molecular anthropology
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 144 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 33 Abstract number: ABS-6670-ISABS-2007
DISTRIBUTION OF THREE POLYMORPHIC MARKERS AND HAPLOTYPES WITHIN THE HUMAN TNF GENE CLUSTER AMONG CROATIANS
Car H1, Štingl K1, Žunec R1, Čečuk-Jeličić E1, Kerhin-Brkljačić V1, Grubić Z1
1Tissue Typing Centre, University Hospital Centre Zagreb, Zagreb, Croatia [email protected]
The TNF gene cluster is located within highly polymorphic human leukocyte antigens (HLA) region on chromosome 6. TNF locus contains many polymorphisms including microsatellites (TNFa, TNFb, TNFc, TNFd, TNFe and TNFf) and single nucleotide polymorphisms. Many of these polymorphisms are in linkage disequilibrium with HLA class I and II alleles. Polymorphism of TNFa, TNFb and TNFd microsatellites, linkage disequilibrium (LD) within TNF region and their relationship with alleles at HLA-A, -B and -DRB1 loci were investigated in this study. Sample of 160 unrelated Croatians, previously typed for HLA-A, -B and DRB1 loci, was tested. Analysis was performed using electrophoresis of PCR amplified products on polyacrylamide gel in ALFexpress sequencer. The most frequent alleles at each TNF microsatellite were: TNFa10 (22.4%), TNFb4 (44.4%) and TNFd4 (56.6%). LD between TNFa, TNFb and TNFd loci was investigated analysing two- and three-locus haplotypic associations. Strongest two-locus haplotypic associations were TNFa2-TNFb1 (LD=3.67; P<0.00001), TNFa2- TNFd2 (LD=3.22; P<0.00001), and TNFb1-TNFd5 (LD=2.72; P=0.0008). Among three-locus haplotypic associations highest LD was observed for TNFa10-TNFb4- TNFd4 (LD=3.36; P=0.0010), while the most significant P value was calculated for TNFa2-TNFb3-TNFd2 (LD=2.82; P<0.00001). Analysis of HLA extended haplotypes demonstrated presence of 3 highly conserved haplotypes: HLA-A*01, - B*08, -TNFa2, -TNFb3, -TNFd2, - DRB1*03; HLA-A*03, -B*07, -TNFa11, -TNFb4, TNFd4, -DRB1*15 and HLA- A*02, -B*18, -TNFa10, -TNFb4, -TNFd4, -DRB1*11. In conclusion, this study revealed that differences and similarities regarding HLA region apparently exist between our and other populations. These results should be considered in future studies about association between TNF microsatellite alleles as well as HLA/TNF haplotypes and diseases.
Keywords: TNF, microsatellites, haplotypic associations, linkage disequilibrium, Croatians
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 145 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 34 Abstract number: ABS-3681-ISABS-2007
ABSENCE OF ASSOCIATION BETWEEN GENETIC POLYMORPISMS IN CRFR1 AND PTSD
Ćurić G1, Pavlinić D1, Wagner J1, Đurković M2, Braš M3, Lauc G1
1DNA Laboratory, School of Medicine, J.J.Strossmayer University, Osijek, Croatia; 2Department of Psychiatry, School of Medicine, J.J.Strossmayer University, Osijek, Croatia; 3Clinic for Psychological Medicine, School of Medicine, University of Zagreb [email protected]
One of the main mechanisms in the activation of the hypothalamo– pituitary– adrenocortical (HPA) axis is the release of corticotropin releasing hormone hormone (CRH) from the hypothalamus and it’s binding to the corticotropin releasing hormone receptor (CRHR1) in the pituitary gland. Recently it was reported that two haplotype tagging SNPs (htSNP) in CRHR1 are associated with patterns of human alcohol drinking and could potentially contribute to the development of alcohol dependence. Aiming to see whether same genetic variations are associated with potential predisposition for the development of post- traumatic stress disorder (PTSD) we have investigated association of these two haplotypes of CRHR1 and PTSD. DNA was isolated from blood of 200 patients with diagnosed PTSD and 200 matching control individuals. TaqMan Pre- designed SNP genotyping assays were used to genotype two htSNPs (rs242939 corresponding to T to C exchange at position 44371356 and rs1876830 corresponding to C to T exchange at position 44386772 of Chromosome 17). Contrary to the situation observed in alcoholism, we were not able to find any association of CRFR1 genotype with PTSD since frequencies of all alleles were nearly the same in both studied populations. Interestingly, the observed frequency of the major allele at rs242939 corresponded to the frequency reported to correlate with low alcohol consumption, while the observed frequency of the major allele at rs1876830 was between the reported frequencies for low and high alcohol consumers.
Keywords: CRFR1 gene, PTSD, genetic polymorphism, SNPs, HPA axis
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 146 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 35 Abstract number: ABS-7953-ISABS-2007
THE UTILITY OF SNPs IN INFERENCES OF BIOGEOGRAPHICAL ANCESTRY
Daniel R1, Sanchez JJ2, Nassif NT1, Walsh SJ3
1Centre for Forensic Science, University of Technology Sydney, NSW, Australia; 2National Institute of Toxicology and Forensic Science, Tenerife, Spain; 3Biological Criminalistics Forensic and Technical, Australian Federal Police, ACT, Australia [email protected]
Single nucleotide polymorphisms (SNPs) have been widely investigated as candidate markers of DNA-based assessment of biogeographical ancestry. The ability to infer an individual’s biogeographical ancestry can be instrumental in narrowing a pool of potential suspects in the initial stages of a criminal investigation. Currently available commercial systems for the SNP-based inference of biogeographical ancestry and physical traits are useful to forensic investigators who are unable to match crime scene samples with database profiles. However, such systems are not directly relevant to the Australian population. This research aims to investigate autosomal SNPs which can be used to infer biogeographical origin in a multiplex assay. Based on an extensive literature survey, sixteen SNPs, including membrane-associated transporter protein (MATP), were specifically selected for their potential to distinguish major sub-populations in Australia. Allele and genotype frequency data were used to assess the usefulness of this multiplex PCR assay as a predictor of biogeographical ancestry for investigative purposes. Results were cross-compared to genealogical information self-declared by participants over three generations. Preliminary analysis of over 200 individuals using Linear Discriminate Function and Maximum Likelihood Estimates indicates a high degree of accuracy for the inference of Asian, African and Caucasian sub- populations. MATP was also investigated for its association to pigmentation phenotypes. The development of rapid and robust tests suitable for inference of biogeographical ancestry in the Australian population can provide a valuable intelligence tool for forensic investigators.
Keywords: SNPs, biogeographical ancestry, SNaPshot, multiplex PCR assay, Phenotype
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 147 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 36 Abstract number: ABS-3094-ISABS-2007
THE STUDY OF HUMAN SKELETAL REMAINS FROM THE CROATIAN EARLY MEDIEVEL GRAVEYARD NAKLICE-GREBČINE
Grkovic S1, Penovic A1, Ljubkovic J1, Definis Gojanovic M1, Delonga V2, Anđelinović Š1
1Department of Pathology, Forensic Medicine and Citology, University Hospital Split, Split, Croatia; 2Museum of Croatian Archaeological Monuments, Split, Croatia. [email protected]
Anthropological analyses of the Croatian early medieval graveyard Naklice- Grebčine includes 12 skeleton remains that were found within five graves. This graveyard was positioned on the prehistorical cult graves (tumulus) where used to be a vineyard. These graves, dated from the time of Croatian immigration to Dalmatia in the 9th century AC, were made from the stone blocks, rectangular- shaped and oriented east-west. Old jewellery and parts of ceramics characteristic for this region of Croatia were found in the graves along with skeletons. Due to vineyard’s cultivation throughout the history most skeletal remains were destroyed. Long bones without proximal and distal parts, and the scull bones were mostly preserved. The age analyses of the skeleton remains revealed that the most of them were younger than 45 years. Anthropometrical analyses couldn’t precisely determine the gender of each skeleton therefore DNA analyses were performed. The DNA results for grave number 16 confirmed that two adults were buried in this grave, a female and a male. Among the poorly preserved maxilla and mandibula remains caries was the most common dental disease as well as visible tooth abrasion. Cribra orbitalia, which points to anaemia, is evident on one preserved child scull. The change of temporal bone of the scull indicated that the child suffered scurvy. Long bones (femur, tibia and humerus) showed visible signs of periostitis and osteoarthritis. Schmorl's defects were visible on some vertebrates as a sign of degenerative changes.
Keywords: skeletal remains, early medieval, Dalmatia, anthropometrical analysis, DNA
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 148 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 37 Abstract number: ABS-4438-ISABS-2007
ITPA AND TPMT GENE POLYMORPHISMS IN CROATIAN POPULATION
Jokić M1, Poparić I1, Jurišić G1 , Kapitanović S1
1Ruđer Bošković Institute, Zagreb, Croatia; [email protected]
AZA, a thiopurine prodrug is widely used to treat IBD and rheumatic diseases. There is evidence that polymorphisms in the genes encoding thiopurine methyltransferase (TPMT) and inosine triphosphate pyrophosphatase (ITPase) predict adverse drug reactions to AZA therapy. Low TPMT activity plays a significant role in the occurence of myelosupression. Three identified TPMT alleles (TPMT*2, TPMT*3A and TPMT*3C) account for 80-95% of intermediate or low enzyme activity. Inosine triphosphate pyrophosphatase (ITPase) deficiency is characterized by an abnormal accumulation of ITP in erythrocytes. In ITPase- deficient patients treated with thiopurines, toxic 6-thio-ITP is predicted to accumulate. It's been reported that ITPA 94C>A missense mutation is associated with side effects to AZA therapy. The aim of our study was to determine allelic frequency for SNPs in TPMT and ITPA gene in the Croatian population. DNAs obtained from 350 unrelated individuals were genotyped for the TPMT*2, TPMT*3A, TPMT*3B and TPMT*3C SNPs using allele-specific PCR or PCR-RFLP method. The frequency of heterozygous TPMT genotype in Croatian population was 5.7%. The frequency of the three allelic variants of the TPMT gene was: 0.6% for TPMT*2, 4.2% for TPMT*3A, and 0.9% for TPMT*3C. A total of 127 unrelated individuals were genotyped for the ITPA 94C>A mutation using PCR-RFLP. The frequency of mutated heterozygous ITPA genotype in Croatian population was 9,5%. We can conclude that our results might be helpful for identifying patients at high risk of inadequate responses to thiopurine therapy due to association of polymorphisms in TPMT and ITPA genes with adverse thiopurine therapy.
Keywords: ITPA, TPMT, Croatian, population, AZA
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 149 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 38 Abstract number: ABS-2046-ISABS-2007
SOME FACTORS OF THE BIRTHRATE AT BASHKIRS
Kinjabaeva G1
1Institute of History, Language and Literature of the Ufa Scientific Center of the Russian Academy of Sciences [email protected]
The investigation of birth rate at the Bashkirs is lead on the analysis of censuses materials - VII (1816), VIII (1834), X (1859). Method of investigation is a statistic- mathematic one. The number of men was averaged 51, 7% from the general number of the population. It is known, that this prevalence was distinctive for all nomadic peoples. The most of Bashkirs’ women was married, and by the age about 45 years old each woman have had about 7-8 children. It was high level of the birth-rate across the ethnic groups of Russia. The greatest death rate at the Bashkirs population fell on the age from a birth till 1 year - 24, 5%, and across Russia this parameter was 33, 9%. There was a negative dynamics of the children\'s population share (till 15 years) by this period censuses: in the 1816th it was 40,7%; in the 1834th - 44,6%,in the 1859th –41,7% % The principal cause of the given demographic phenomenon consisted in the social, economic and political conditions of the Bashkir ethnos existence in the XIXth century. Nevertheless, high genetic potential of survival rate allowed to support high share of the young population in the general demographic structure and promoted a survival of ethnos during the critical periods of the history.
Keywords: Birth-rate, Bashkirs, survival, genetic potential, ethnic groups
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 150 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 39 Abstract number: ABS-6726-ISABS-2007
THE G1733A POLYMORPHISM OF THE ANDROGEN RECEPTOR GENE IS ASSOCIATED WITH RECURRENT SPONTANEOUS ABORTIONS
Karvela M 1, Papadopoulou S 1, Velissariou V 2, Lamnissou K 1
1 Dept of Biology, University of Athens, Athens, Greece, 2 Dept of Molecular & Cytogenetics, Mitera Maternity and Surgical Center, Athens, Greece [email protected]
The risk of miscarriage is enhanced by a variety of genetic and environmental factors. In the last years, interest has focused on association studies between different genetic polymorphisms and recurrent spontaneous abortions (RSA), in order to understand the contribution of single genes towards the development of recurrent miscarriage. The aim of our study was to determine whether the G1733A polymorphism of the androgen receptor (AR) gene, is associated with an increased risk for RSA. A total of 52 women who had at least two unexplained spontaneous abortions before 20 weeks of gestation, with the same partner, were included in the study group. The control group consisted of 144 women with at least two live births and without history of abortions. All patients and controls were investigated for the G1733A polymorphism. To genotype the cohorts we used the PCR-RFLPs method. The observed frequencies of GG, AG, AA genotypes were 0.41, 0.44, 0.14, respectively, for the patient group and 0.75, 0.16, 0.09, respectively, for the control group. Allele frequencies were 0.63 for the G allele (wild type) and 0.37 for the A allele (mutant), in the patient group, and 0.83 and 0.17, respectively, in the control group. Statistical analysis of the results indicates significant difference between the two groups. Thus, the results of this study lead to the conclusion that the A allele of the G1733A polymorphism of the AR gene is associated with an increased risk for RSA.
Keywords: G1733A polymorphism, androgen receptor gene, RSA, association studies, risk factor
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 151 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 40 Abstract number: ABS-1627-ISABS-2007
ARTIFICIAL INTELIGENCE APPLIED IN ANTHROPOLOGICAL DATABASE SEARCH
Laslo E1, Noje D1 Tomulescu IM2, Pusta CT3
1University of Oradea, Faculty of Sciences, Department of Mathematics and Informatics, Oradea, Romania 2University of Oradea, Faculty of Sciences, Department of Biology, Oradea, Romania 3University of Oradea, Faculty of Medicine and Pharmacy, Department of Anatomy, Oradea, Romania [email protected]
In this paper we propose a bridge between the genetic algorithms and the database structures to create a universal database searching model, virtually applicable for every arbitrary data. In the first stage, the method we are propose, we applied for each records of the database the method of the polynomial approximation with genetic algorithms (PAGA) witch results in the creation of an algebraic polynom for each of the records. When this stage is over we upgrade the original database with the table witch contain the coefficients of the polynoms. In analogy with the Neuronal Network concepts this stage is the learning stage. In the second stage we process the input data with the PAGA method witch means we also create an algebraic polynom that represent the input data. The search for the input data in the database means that we need to find all the associated polynoms (created in the stage one) witch approximate best the polynom representing the input data (created in the second stage). If the difference doesn’t satisfy the minimal required, we consider this data inexistent in the original database and we could save this data set like a new record. This method could be applied in the general sense for every database and in particular we built the method to interpret the 2D coordinates of the points which represent the particularities of the fingerprints.
Keywords: artificial intelligence, fingerprints, genetic algorithms, the database structures, anthropology
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 152 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 41 Abstract number: ABS-4758-ISABS-2007
GENETIC STUDY OF CHILDREN FROM BURIAL GROUND POHANSKO BY BŘECLAV
Mertlová K1, Drozdová E1
1Institute of Anthropology, Faculty of Science, Masaryk University, Brno, Czech Republic [email protected]
The South Outer Precinct is the second largest burial ground that was discovered on archaeological area Pohansko, which is localized near Břeclav in the Czech Republic. Bone material was excavated in the period of 1975 – 1979 during a rescue archaeological research. The burial ground is dated within the end of 8th century and the beginning of 10th century. The bone material from The South Outer Precinct is preserved in very poor quality, the bones are fragmentary and in most cases they are undeterminable. Altogether 189 skeletons were discovered, 27 were recognised as masculine, 40 as feminine and 88 as children. The sex could not be identified in 34 skeletons. The aim of this study is to determine the sex of children and non-identified skeletons using methods of Polymerase Chain Reaction (PCR). The sex is determined by amplifying a segment of the X-Y homologous gene amelogenin and by amplifying a short 93 bp fragment of the SRY gene. The study is at the beginning, but we have already got some positive results which will be very important for demography of the burial ground.
Keywords: Amelogenin gene, SRY gene, Sex determination, PCR, Pohansko
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 153 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 42 Abstract number: ABS-6988-ISABS-2007
ETHNIC ANTHROPOLOGY OF CAUCASUS PEOPLES AND PROBLEM OF CAUCASOID ORIGIN
Nazarova A1, Alkhutov S 1, Aslanishvili V2
1 Institute of Ecology and Evolution Problems, Russian Academy of Sciences, Moscow, Russia, 2 Institute of History, Georgian Academy of Sciences, Tbilisi, Georgia. [email protected]
There was studied the electrophoretical polymorphism of blood proteins of Talyshes populations of Pirasora situated on South-East of Azerbaidjan. We calculated gene frequencies of Talyshes population and determined the genetic distances of Talyshes to Iranians of North, Central and South Iran, to Afghanians, and to three populations of Azerbaidjanians. The Talyshes are most close to Iranians of Shiraz, and Azerbaidjanian populations are most far from Talyshes. Our anthropological investigations shoved that the different human races traits reresentatives in Caucasus populations, Caucasoid and Mongoloid, were lived in Aragvi basin from eneolithic. Those data are supported the opinion of academician V.P.Alexeev (1974) about mixed in race aspect of Caucasus population .We are calculated the matrix of genetic distances of populations of Caucasus: Georgians, Ossetians, Azerbaidjanians and others to populations of neighbour regions and to Basks by 28 alleles of 12 loci of proteins, enzymes and blood groups, and constructed the dendrogram of those populations. The position of Caucasus populations on both dendrogrammes: only Caucasus and neighbour populations, and 55 populations are corresponded on principle to data of native anthropologists. The clasterisation of another Caucasoid populations also is corresponded wholly to antropological and historical data, and supported of early putting forward our hypothesis (Nazarova, 1999) about inhabited in Asia while paleolythic.
Keywords: Anthropology, Genetics, Caucasus, Populations, Caucasoids
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 154 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 43 Abstract number: ABS-7853-ISABS-2007
THE ANTHROPOLOGIC, GENETIC AND ARCHEOLOGIC EVIDENCES OF PALAEOASIATIC ORIGIN OF NORTH MONGOLOIDS, AMERINDS AND CAUCASOIDS
Nazarova A1
1Inst of Ecology and Evolution Problems, Russian Academy of Sciences, Moscow, Russia [email protected]
We were distinguished the Caucasoids frequencies of genes of blood proteins and enzymes in populations of Altaians. The matrix of genetic distances of 28 alleles of 12 loci of proteins, enzymes and blood groups of 11 populations of Europe, Asia and America, and than the matrix of genetic distances of 55 human populations of Europe, Asia, America Africa and Oceania were calculated. On data of these matrixes we constructed the evolutional dendrograms. From this dendrograms we suggested that Caucasoids were differentiated with North Mongoloids and Amerinds from Ancient Asiatic population while Middle Palaeolithic in region of Altay and in neighbour regions. The investigations of mitochondrial DNA polymorphism are supported our hypothesis about paleoasiatic origin of North Mongoloids, Caucasoids and Amerinds. The haplogroups of mitochondrial DNA of different human populations of Eurasia and America were marked the way of ancient tribes in their Palaeolithic migrations on map constructed by us. The data of Russian anthropologists also supported Palaeoasiatic origin of Caucasoids, for example the distribution of frequencies of supraorbital canals in different human populations. Russian scientists decoded the petroglifs near Baikal Lake as ancient inscription. This inscription marked the holy places of the goddess Ama-terasu who belonged to the pantheon of ancient inhabitants of Nothern Asia (Siberia) who were ancestors both of the Shumers and the Khetts (ancient Caucasoids) as well as the Japanese (Mongoloids).
Keywords: Caucasoids, North Mongoloids, Amerinds, Origins, Paleoasiatic
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 155 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 44 Abstract number: ABS-816-ISABS-2007
GENETICS OF QUANTITATIVE DERMATOGLYPHICS AND ITS BILATERAL ASYMMETRY
Sengupta M1
1Biological Anthropology Unit, Indian Statistical Institute [email protected]
Dermatoglyphics is an important quantitative traits used by the biological anthropologists. As it has manifold applications in diverse field, a thorough understanding of its mode of inheritance is essential. The purpose of the present study is, therefore, to add some information regarding the relative contribution of genetic and environmental effects on dermatoglyphics and its bilateral asymmetry. Data comprised 200 families (824 individuals) living parents with at least two children consisting of from an endogamous caste- Vaidya. Finger and palm prints were collected by the ink method of Cummins and Midlo (1976) and analyzed after Holt (1968). Bilateral Fluctuating (FA) and Directional Asymmetry (DA) were calculated by Jantz and Webb (1980). Statistical analyses include intraclass and interclass Correlation, Heritability Estimation, Principal Component Analysis and Segregation Analysis using Pedigree Analysis Package (PAP). Major findings included that: i) There is strong genetic basis of dermatoglyphics (heritability 32- 77%) ii) But there is weak involvement of family factor and a considerable amount of environmental effect of both types asymmetry iii) Five principal factors were extracted from the asymmetric traits explaining 74.207% of overall cumulative variance iv) By segregation analysis, Sporadic, Environmental, No Major Gene (MG) effect and no polygenic component were strongly rejected v) Most parsimonious Mendelian model clearly indicated the contribution of MG with either dominant or additive effect vi) Any effect of MG could not be detected for any kind of asymmetry vii) The result supports the idea postulated by several authors that asymmetry is a measure of developmental instability in man.
Keywords: Genetics, Dermatoglyphics, Bilateral Asymmetry, Vaidya, India
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 156 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 45 Abstract number: ABS-762-ISABS-2007
ARTIFICIAL INTELIGENCE APPLIED IN ANTHROPOLOGICAL DATABASE SEARCH
Laslo E1, Noje D1 Tomulescu IM2, Pusta CT3
1University of Oradea, Faculty of Sciences, Department of Mathematics and Informatics, Oradea, Romania 2University of Oradea, Faculty of Sciences, Department of Biology, Oradea, Romania; 3University of Oradea, Faculty of Medicine and Pharmacy, Department of Anatomy, Oradea, Romania [email protected]
In this paper we propose a bridge between the genetic algorithms and the database structures to create a universal database searching model, virtually applicable for every arbitrary data. In the first stage, the method we are propose, we applied for each records of the database the method of the polynomial approximation with genetic algorithms (PAGA) witch results in the creation of an algebraic polynom for each of the records. When this stage is over we upgrade the original database with the table witch contain the coefficients of the polynoms. In analogy with the Neuronal Network concepts this stage is the learning stage. In the second stage we process the input data with the PAGA method witch means we also create an algebraic polynom that represent the input data. The search for the input data in the database means that we need to find all the associated polynoms (created in the stage one) witch approximate best the polynom representing the input data (created in the second stage). If the difference doesn’t satisfy the minimal required, we consider this data inexistent in the original database and we could save this data set like a new record. This method could be applied in the general sense for every database and in particular we built the method to interpret the 2D coordinates of the points which represent the particularities of the fingerprints.
Keywords: genetic algorithms, database structures, artificial intelligence, 2D coordinates, fingerprints
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 157 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 46 Abstract number: ABS-7352-ISABS-2007
CRANIAL AND FACIAL DYSMORPHISM AS PHYSICAL MARKERS IN A POPULATION WITH MENTAL DEFICIENCY
Tomulescu IM1
1University of Oradea, Faculty of Sciences, Department of Biology, Oradea, Bihor County, Romania [email protected] [email protected]
The number and types of traits in mentally affected person varies in large limits. In legal medicine is very important to establish the physical and psychiatric diagnosis. Our study is about the incidence of cranial and facial dysmorphism in a population with mental deficiency. We decided to make this research because of different etiology of mental deficiency and we wanted to establish if the cranial and facial dysmorphism is an important element in description of different types of mental deficiency. Cranial and facial dysmorphism is very often observed in the delinquent individuals, too. This research was realised on 393 children with diverse types of mental deficiency. The children were interned in Neuropsychiatry Infantile Section of Neurology and Psychiatry Clinical Hospital of Oradea, during the 1999-2001 periods. The children were clinical examinated. Also, we used the methods for chromosomes analysis. We observed that the incidence of cranial and facial dysmorphism increase in accordance with the level of the mental deficiency. In the group with severe mental deficiency, we obtained an incidence over the 40% of this abnormality. The explanation for these results may be the presence of some genes and chromosomes syndromes which associate cranial and facial dysmorphism, too. The cranial and facial dysmorphism is a common feature in description of mental deficiency disease. The incidence of this abnormality increase in accordance with the worsening of the mental deficiency.
Keywords: cranial dysmorphism, facial dysmorphism, mental deficiency, physical marker, malformative diseases
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 158 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 47 Abstract number: ABS-555-ISABS-2007
SPECIFIC FEATURES OF DERMATOGLYPHICS AND DIGIT RATIO IN A DELINQUENT POPULATION
Tomulescu IM1
1University of Oradea, Faculty of Sciences, Department of Biology Bihor County, Romania [email protected]
The scientific study of papillary ridges of the hands and foot is credited as beginning with the work of Joannes Evangelista Purkinje, in 1823. in 1892, Sir Francis Galton published his classic treaties on fingerprints. Dermal palmer and plantar ridges are highly useful in biological studies. Noel Jaquin began to speculate about the psychological connections of fingerprints and individual subjects in print in 1933 as he wondered whether the worl pattern, then commonly found on the prints of certain types of criminals, indicated some defect of moral perception that he would attribute to some psychological deficiency. In this research we proposed to study the specific features of dermatoglyphics in a delinquent population. We investigated the hands (fingers and palms) of 500 delinquents. We studied the dermatoglyphics characteristics. Also, we realised statistics about the average of ridges, types of prints on each finger, values of atd angle, digit ratio. We calculated the variation coefficient. The results are important. The types of prints varied in different fingers, the number of ridges and atd angle values are different than in control group. We may suggest that in a delinquent population the dermatoglyphics are modified, perhaps because of aberrant behaviour sometimes determined by genetic factors. We may say that there is a connection between the behaviour and dermatoglyphics.
Keywords: dermatoglyphics, digit ratio, TRC, delinquent population, abberant behavior
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 159 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 48 Abstract number: ABS-8421-ISABS-2007
THE IMPORTANCE OF CHROMOSOME BANDING IN FORENSIC MEDICINE
Tomulescu IM1, Pusta CT2
1University of Oradea, Faculty of Sciences, Department of Biology, Oradea, Romania 2University of Oradea, Faculty of Medicine and Pharmacy, Department of Anatomy, Oradea, Romania [email protected], [email protected], [email protected]
Patau syndrome, also known as trisomy 13, is a chromosomal aberration. The effects of this abnormality are often letal. Many unborned children die in the different period of prenatal development. Patau syndrome due to a translocation can be inherited. In establishing the cause of unborned children we need a specific genetic test called chromosome banding. Also, we can establish the provenience of abnormal chromosome. In this material we purposed to demonstrate the importance of chromosome banding in forensic medicine examination. Our case is a one year old girl who was borned with all the physical features of Patau syndrome and died at the age of 16 months. We made the clinical and paraclinical examinations and we partially established the cause of death. We also made the caryotype and then the chromosomes banding to the child and her parents. The child had abnormalities such as: developmental delay, profound mental retardation, lack of eyes, cleft lip, cleft palate, polydactyly, syndactyly, deafness, hypotonia, neurological disability, bound malformations, heart malformation and others. After many repetitions, the caryotype was still normal. So, we assumed that could be a Patau syndrome occurred due to a chromosome translocation which determined such a dramatically malformations. After chromosome banding was made we had the certainty of the Patau syndrome. We also obtained that father was the carrier of the translocation. The chromosome banding was very important to establish the cause of many malformations that determined the death.
Keywords: chromosome banding, cause of death, Patau syndrome, congenital abnormalities, translocation
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 160 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 49 Abstract number: ABS-7395-ISABS-2007
THE IMPORTANCE OF THE FAMILY HEREDITARY ANTECEDENTS IN APPEARANCE OF MENTAL DEFICIENCY
Tomulescu IM1
1University of Oradea, Faculty of Sciences, Department of Biology Oradea, Romania [email protected]
Mental deficiency is a very severe and common disease. There are many causes of mental deficiency: the congenital abnormalities, metabolic diseases, cranial traumatisms, X fragil syndrome, monogenic or polygenic syndromes and chromosomes syndromes, the critical age of parents (especially of mother’s), some harmful factors in appearance of mental deficiency. In this study we show the importance of hereditary antecedents in appearance of mental deficiency. We investigated 596 children interned in Neuropsychiatry Infantile Section of Neurology and Psychiatry Clinical Hospital of Oradea, during the 1999-2001 period. In 596 children that were examined, 393 presented different types of mental deficiency. We realised family investigations and genealogical tree. More than 65% of children with mental deficiency have one or more affected relation in family. The relations may be affected by congenital abnormalities and/ or mental diseases. The incidences of affected relations are important in groups with mild and moderate mental deficiency. In the group with severe mental deficiency, the incidence of affected relation is lower. This result may be an argument for the hypothesis that genetic factors are very important in the inheritance of mental deficiency. The increased incidence in groups with mild and moderate mental deficiency may be explained by harmful factors, critical age of parents and the presence of X fragile syndrome. It seems that, elementary, severe mental deficiency appears because of genes and chromosomes disorders, and secondary because of dominant or recessive inheritance.
Keywords: mental deficiency, family hereditary antecedents, genetic factors, harmful factors, congenital abnormalities
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 161 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 50 Abstract number: ABS-690-ISABS-2007
THE IMPORTANCE OF THE PHYSICAL AND SOME MORPHOLOGICAL TRAITS ANALYSIS IN FORENSIC MEDICINE
Tomulescu IM1, Pusta CT2, Laslo E3
1University of Oradea, Faculty of Sciences, Department of Biology, Oradea, Romania 2University of Oradea, Faculty of Medicine and Pharmacy, Department of Anatomy, Oradea, Romania 3University of Oradea, Faculty of Sciences, Department of Mathematics and Informatics, Oradea, Romania [email protected]
Many physical and morphological traits are determined by the effects of many genes and by the environment. In this research we purpose to show the high genetic homogenity of some phenotypical traits in a female population of Seghistea locality, in Bihor county, Romania, comparatively with a genetic heterogenous control group from Oradea locality. These data are important to identify an unknown female dead body. We studied 300 women of Seghistea locality and 300 of Oradea locality. We investigated the type of hair, eyes pigmentation, aspect of lips, hair and the profile of nose, cranial diameters, body and facial height, weight. To establish the certain aspect of lips, hair and the profile of nose, the colour of eyes and hair, we used the visual exploration. The aspect of hair is established by using the scheme realised by Rudolf Martin in 1914. He also realised a scheme to establish the lip aspect. We utilised a vernier callipers for diameters measurements. The results are high significant. All of variation coefficients have very small values that demonstrate a small variation of these traits. We already made a database with data from different localities. If we introduce the data of one unknown female dead body in the PC programme made by us, we can identify with a certain probability if she has the specific trait for certain area. Analysing some physical traits is important in legal medicine to identify the geographic area from unknown dead body proceeds.
Keywords: physical traits, female dead body, identification, Seghistea, geographic area
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 162 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 51 Abstract number: ABS-2724-ISABS-2007
DNA BASED SEX DETERMINATION IN CHILDREN SKELETAL REMAINS FROM ENEOLITHIC BURIAL SITE HOŠTICE ZA HANOU
Vaňharová M.1, Dozdová E.1
1Institute of Anthropology, Masaryk University, Brno, Czech Republic [email protected]
Sex determination belongs to essential anthropologic characteristics. Nevertheless, the reliability of morphologic and morphometric methods for sexing skeletal remains is limited by a number of diverse factors: pathological disposition, skeletal condition, environmental influences, diet, occupational stress and particularly by biological age, because in immature individuals there are not expressed effective sex indicating signs. The definite answer can provide the analysis of genes located on sex chromosomes. The aim of this study was to apply the modern genetic analysis to the children skeletal remains from burial site Hoštice za Hanou. This eneolithic burial site was discovered within rescue research in 2002. It is the biggest burial site of the people of the bell beaker period in the Czech Republic and probably in the Middle Europe. We apply two sex markers, the commonly used amelogenin gene and the specific sequence localised on Y chromosome – the SRY gene. The method was based on amplification the sequence from the first intron of the amelogenin gene, which includes X-chromosome specific 6 bp deletion, so we can differentiate the male and female genotype. As the second marker we amplified Y-chromosome specific 93 bp fragment from SRY gene. This marker appears more reliable, because of shorter, easily amplifiable nucleotide sequence. Using this molecular approach we examined 57 samples. The state of skeletal preservation was mainly poor and fragmentary. This study demonstrates the applicability of the molecular methods for sex determination in children skeletal remains from eneolithic archaeological findings.
Keywords: eneolith, sex, children, amelogenin, SRY
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 163 POSTER PRESENTATIONS – Molecular Anthropology Population Genetics Studies on Other Molecular and Phenotypical Markers Presentation number: MA 52 Abstract number: ABS-0852-ISABS-2007
EFFECTS OF DETERGENTS ON THE REDISTRIBUTION OF GANGLIOSIDES AND GPI-ANCHORED PROTEINS IN BRAIN TISSUE SECTIONS
Viljetić B1, Vajn K1, Heffer-Lauc M1, Lauc G1
1School of Medicine, J.J.Strossmayer University, Osijek, Croatia [email protected]
Gangliosides and glycosylphosphatidylinositol (GPI)-anchored proteins contain lipid tails that tether them to the outer side of the cell membrane. This mode of association with the cell membrane enables them to take part in the organization of lipid rafts, but it also permits gangliosides and GPI-anchored proteins to be actively released from one cell and inserted into the membrane of an another cell. Recently we have reported that under conditions of lipid raft isolation Triton X-100 causes significant redistribution of both gangliosides and GPI-anchored proteins. Aiming to find a less disruptive detergent we have evaluated effects of CHAPS, Saponin, deoxycholic acid, Trappsol, Tween 20, Triton X-100, Brij 96V, Brij 98, and SDS on the distribution of GM1 and GD1a gangliosides and Thy-1 (GPI- anchored protein) in brain tissue sections at room temperature and at 4ºC. At room temperature all detergents (in 1% concentration and incubation for 2 h) extracted significant amounts of both gangliosides and Thy-1 from the membranes. At 4ºC the extraction was weaker, but Triton X- 100 CHAPS and deoxycholic acid caused significant redistribution of GD1a and Thy-1 from grey matter into the white matter. Both redistribution and extraction were significantly augmented if sections were incubated with detergents in the presence of primary antibodies. Other detergents like Trappsol and Tween 20 apparently cause less redistribution but they are also less efficient in revelation of cryptic epitopes. Neither one of the nine tested detergents is the ideal choice. However, Brij 96V appears to be able to sufficiently reveal myelin epitopes, while causing the least amount of artifacts.
Keywords: GPI-anchored proteins, gangliosides, detergents, lipid rafts, imunohistochemistry
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 164 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Usage of the Latest Methods in Molecular Anthropology Usage of the Latest Methods in Molecular Anthropology
Presentation number: MA 53 Abstract number: ABS-9192-ISABS-2007
DNA TYPING FROM SKELETON REMAINS, CASE STUDY IN NAKHONNAYOK PROVICE, THAILAND
Imtang S1, Machaopa J1, Riproumsup K 1
1Forensic DNA Service Center, Central Institute of Forensic Science, Bangkok, Thailand [email protected]
As a result of the tsunami in South East Asia in December 2004, a very large number of corpses have to be identified. DNA analysis was used successfully in mass casualty incidents to identify the victims. DNA experts from Central Institute of Forensic Science (CIFS) were worked together with Thailand Tsunami Victim Identification (TTVI) team. After this situation we recognize to develop knowledge our scientist and techniques about extraction DNA and statistic for identification. This is the first identification of skeleton remains. There are present individual body efforts to identify skeleton remains and relatives of missing persons in Nakhonnayok Province, Thailand. The police officer found unknown skeleton remains on October 21, 2005. The autopsy information of this remains was an old woman and wear traditional Thai-skirt. We extracted DNA from femur by organic extraction and Microcon YM-100. The 15 short tandem repeat (STR) loci typing were obtained and compared with missing person relative. This case was identified by prior probability 99.99% estimated. In conclusion, DNA-based were helpful identification skeleton remain for missing person if more information about AM-PM Data for Missing Database.
Keywords: short tandem repeat (STR), skeleton remains, DNA typing, Forensic Anthropology, Bone Extraction
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 165 POSTER PRESENTATIONS – Molecular Anthropology Usage of the Latest Methods in Molecular Anthropology Presentation number: MA 54 Abstract number: ABS-286-ISABS-2007
HIGH-THROUGHPUT SCREENING METHOD FOR DISTINCTION AND PALEOPATHOLOGICAL ANALYSIS OF FORENSIC AND ARCHAEOLOGICAL HUMAN FINDINGS BY FT-IR
Mark L1, Lorand T1, Nagy G2, Bajnoczky I2, Marcsik A3
1Institute of Biochemistry and Medical Chemistry, Faculty of Medicine, University of Pecs, Pecs, Hungary; 2Institute of Forensic Medicine, Faculty of Medicine, University of Pecs, Pecs, Hungary; 3Department of Anthropology, Faculty of Sciences, University of Szeged, Szeged, Hungary [email protected]
Human bone is a composite material. It contains by weight approximately 20-30% organic fractions and 70% mineralized carbonate-hydroxyapatite as its inorganic part. The structure and the composition of carbonated apatite is close to the mineral dahllite, the general formula Ca5(PO4)3-x(CO3)x(OH) x+1. The second most abundant component of bone is collagen, predominantly type I, which provides the tissue with elasticity and directs the organisation of matrix. In fibrils, molecules of the collagen are parallel and overlapping each other multiples of 67 nm, furthermore each molecule being 300 nm long. In bone tissue, inorganic crystals grow and lie in the 40 nm gaps between collagen fibers. In this study we report the chemical analyses of various non-pathological, tuberculosis and syphilis infected bone samples from different burial environments by Fourier transform infrared spectroscopy (FT-IR), in the framework of a general study of diagenesis. Dating human skeletal remains is one of the most important and yet unreliable aspects of forensic anthropology. Within this paper a new method has been suggested, using the crystallinity index and carbonate-phosphate index as a means of distinction between recent and archaeological anthropological bone samples. Pathologic bone samples were analyzed with the same method to see if changes in cristallinity interfere with the process of dating. Reference G. Nagy et al., Analysis of pathological and non-pathological human skeletal remains by FT- IR spectroscopy, Forensic Sci. Int. (2007), doi:10.1016/j.forsciint.2007.05.008
Keywords: FT-IR, relative dating, forensic anthropology, paleoanthropology, paleopathology
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 166 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Usage of the Latest Methods in Molecular Anthropology Presentation number: MA 55 Abstract number: ABS-2985-ISABS-2007
HORMONE MASS FINGERPRINTING: MOLECULAR SEX DETERMINATION OF ANCIENT HUMAN SKELETAL REMAINS BY MALDI TOF MASS SPECTROMETRY
Mark L1, Marcsik A2
1Institute of Biochemistry and Medical Chemistry, University of Pécs, Pécs, Hungary, 2Department of Anthropology, University of Szeged, Szeged, Hungary [email protected]
In this study, a rapid, high-throughput, sensitive matrix assisted laser desorption ionization time of flight (MALDI TOF) mass spectrometric technique has been developed for analysis of sexual steroid hormones as gender specific biomarkers in human skeletal findings. The hormone mass fingerprinting (HMF) method is extremely suitable for sex determination of fragmented forensic and paleoanthropological remains. We successfully extracted and analyzed sex specific hormones as estrone, estradiol, estriol and testosterone from 7000-year- old archaeological bone samples and used the method for gender determination of these human bones.
Keywords: gender determination, paleoanthropology, fornsic anthropology, sexual steroid, mass spectrometry
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 167 POSTER PRESENTATIONS – Molecular Anthropology Usage of the Latest Methods in Molecular Anthropology Presentation number: MA 56 Abstract number: ABS-08-ISABS-2007
COMPARATIVE STUDY ON TOOTH SAMPLES COLLECTED FROM DIFFERENT ARCHAEOLOGICAL SITES AND THE IMPLICATIONS OF FOSSILIZATION IN PRESERVATION OF DNA OVERTIME
Stanciu F1,3, Galan E1, Stoian G M1, Andreescu G1, Soficaru D A2, Miriţoiu N2
1Forensic Science Institute, General Inspectorate of Romanian Police, Bucharest, Romania; 2Laboratory of Paleoanthropology, “Francisc I. Rainer” Anthropological Research Centre, Bucharest, Romania; 3Department of Genetics and Biotechnologies, Faculty of Biology, University of Bucharest, Bucharest, Romania [email protected]
Several human tooth samples from Neolithic Period, Bronze Age, Iron Period and VIII-X Century Period, collected from different archaeological sites (belonging to the same geographic area - South-East of Romania) had been analyzed. Tooth DNA extracts had been obtained using a GuSCN DNA isolation protocol. Based on spectophotometric readings on DNA extracts, it was possible to determine a DNA degradation index using tooth samples from our days as etalon for DNA degradation occurred in the steps of isolation protocol. The results were correlated to elemental composition of tooth as well as their calcinations residues, in order to relieve the effect of fossilization process on DNA preservation. For qualitative and quantitative analysis, XRF and SEM/EDS techniques have been used. We consider this type of knowledge being of great interest because investigations about particularities of molecular processes involved in bone and tooth DNA conservation (or degradation) are just a few, insufficiently for problems rise by this matter.
Keywords: DNA degradation index, teeth, fossilization, paleogenetics, taphonomy
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 168 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Molecular Anthropology Usage of the Latest Methods in Molecular Anthropology Presentation number: MA 57 Abstract number: ABS-1742-ISABS-2007
A DETAILED STUDY OF mtDNA AND NRY HAPLOGROUPS IN AFRICAN AMERICAN PROSTATE CANCER CASES AND CONTROLS: BUILDING A MORE COMPREHENSIVE PICTURE IN EPIDEMIOLOGY STUDIES
Stefflova K1, Pai AA1, Zhadanov SI1, Zeigler-Johnson CM2, Rebbeck TR2, Schurr TG1
1Department of Anthropology, 2Department of Biostatistics and Epidemiology, University of Pennsylvania, Philadelphia, USA [email protected]
Ancestry is an important factor in epidemiology case-control studies. The knowledge of genetic background can help prevent associating markers that are unlinked to disease loci but instead reflect the population history and can therefore contribute to better matching of the cases and controls. We have focused on a case-control group of self-reported African- Americans that are known to be an admixed population suffering from a high incidence of prostate cancer. We have assessed maternally and paternally inherited genetic loci of 200 African American controls and cases by detailed sequence analysis and SNP typing of mtDNA and NRY SNP typing and compared the haplogroup distribution and the admixed ancestry both with 200 European American and 150 African (Senegalese) controls and cases. A sex biased admixture with higher contribution of non- African populations to the NRY vs. mtDNA haplogroups was observed. In addition, the detailed typing of both mtDNA and NRY haplogroups broadens the picture of African American ancestry that was previously drawn by mtDNA only (Salas et al. 2005). Finally, the recent burst of associations of mtDNA and NRY haplogroups with cancer is challenged.
Keywords: African Americans, mtDNA, NRY, admixture, prostate cancer
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 169 POSTER PRESENTATIONS – Molecular Anthropology Usage of the Latest Methods in Molecular Anthropology Presentation number: MA 58 Abstract number: ABS-852-ISABS-2007
THE IMPORTANCE OF DIGIT RATIO STUDIES IN FORENSIC AND POPULATIONAL GENETICS
Tomulescu IM1, Pusta CT2, Laslo E3, Noje D3
1University of Oradea, Faculty of Sciences, Department of Biology, Oradea, Romania 2University of Oradea, Faculty of Medicine and Pharmacy, Department of Anatomy, Oradea, Romania 3University of Oradea, Faculty of Sciences, Department of Mathematics and Informatics, Oradea, Romania [email protected]
The length of fingers and the ratio between them are studied in populational and forensic genetics too. Some authors suggested that the ratio of the length between the ring and index fingers is determined during early fetal development, and that it is influenced by sex hormones. If this is true, the fingers may provide a permanent, and easily visible, historic marker of important hormonal events that occurred during a critical time of fetal development. The effect size for digit ratio between the sexes varies as a function of geography and race. We measured the length of index, median and ring digit in a human population from Diosig locality, Romania. Were measured right and left hands of 200 men and, respectively, of 200 women. We also realised the digit ratio: 2D:4D, 2D:3D and 3D:4D. We compared the two groups with control groups from Oradea locality, where the population is very heterogenous genetic. The obtained results are significant. The differences between men and women from the studied localities are highly significant. We interpreted statistically the results, and the obtained results were significant different. We made a database with digital measurements of individuals from different areas of Bihor county and neighbouring counties. We also made a PC soft which allows us to identify a certain unknown individual by introducing the data of him in our database. The explanation for these results may be the genetic content of the population. The study because we may use this measurements in individual identification and in populational genetics.
Keywords: digit ratio, individual recognition, forensic genetics, populational genetics, fetal development
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 170 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Advances in Genomic Methods
POSTER PRESENTATIONS Advances in Genomic Methods
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 171 POSTER PRESENTATIONS – Advances in Genomic Methods Genes that predict cancer outcome Genes that predict cancer outcome
Presentation number: AM 1 Abstract number: ABS-8827-ISABS-2007
MOLECULAR GENETIC DIAGNOSTICS IN SPLIT-SEVEN YEARS OF EXPERIENCE
Drmić Hofman I1, Tomić I2, Kuret S2, Anđelinović Š1
1Split University Hospital & School of Medicine, Split, Croatia; 2Split University Hospital, Split, Croatia [email protected]
Thanks to the completion of Human Genome Project, genetic testing is becoming more important and is clearly increasing in all European countries, as well as in Croatia. The end of 2000 has established Laboratory for clinical and molecular genetics, at Split University Hospital. During that period were performed over 1,700 analyses, mostly tests in tumor tissue, molecular tests for leukemia and lymphoma, tests for risk factors, and recently tests for infectious diseases. It is evident the constant increase of the number of requests per test. Special attention is paid on current available accreditation standards used within Europe. Laboratory has just started the process of getting accreditation ISO 15 189 and participating in External Quality Assessment (EQA) schemes. It is very important for country with less developed system for genetic diagnosis to implement genetic tests for common diseases, referring clinicians, reduce the test costs, improve the quality of testing and apply ethical guidelines at all times.
Keywords: molecular diagnostics, genetic testing, standardization, quality, Croatia
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 172 September 3-7, 2007, Split, Croatia POSTER PRESENTATIONS – Advances in Genomic Methods Genes that predict cancer outcome Presentation number: AM 2 Abstract number: ABS-196-ISABS-2007
THE ASSOCIATION OF BRCA1 AND BRCA2 SNP WITH CANCER LOCALIZATION
Smirnova TY1, Pospekhova NI1, Loginova AN1, Lubchenko LN2, Ginter EK1, Karpukhin AV1
1Research Centre for Medical Genetics, Moscow, Russian Federation; 2Cancer Research Centre, Moscow, Russian Federation. [email protected]
The BRCA1 and BRCA2 are the breast and ovarian cancer susceptibility genes. But the molecular basis for cancer localization in either the breast or in the ovary is unclear. We have suggested that cancer localization may be modified by different variants of BRCA1/2 genes. We studied 4 samples of patients for SNPs in the BRCA1 and BRCA2 genes: patients with sporadic breast and ovarian cancer, patients with the same types of cancer with BRCA1 mutations and a control sample. The BRCA1 E1038G and BRCA2 N372H, H1256P coding variants, and a SNP located in the 5'- untranslated region were investigated. We received different haplotype profiles for the samples investigated. In a sample of sporadic breast cancer patients, an association of the 1O38EG variant with cancer risk was found (OR=2,11; P=0,016). There was no difference in four SNPs frequencies between BRCA1 mutation carriers with breast cancer and control sample. In sporadic ovarian cancer 203AA homozygous variant was associated with cancer risk (OR=6,59; P=0,0016). An association of 1O38GG (OR=0,12; P=0,019) and 203GA (OR=0,29; P=0,0033) were revealed for cases of ovarian cancer with BRCA1 mutations. The data suggest a possibility distinguish four groups of patients with breast or ovarian cancer by profiles of BRCA1/2 SNPs association with cancer risk. The risk of sporadic breast and ovarian cancer is associated with different SNPs. The BRCA1 mutations caused breast cancer independently of the SNPs studied. We conclude that BRCA1/2 polymorphic variants may act as modifiers of ovarian cancer risk under BRCA1 mutations.
Keywords: breast cancer, ovarian cancer, SNP, cancer localization, mutations
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 173 About Invited Speakers
ABOUT INVITED SPEAKERS
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 174 September 3-7, 2007, Split, Croatia About Invited Speakers
Plenary lecture
Svante Pääbo ● Max-Planck-Institute of Evolutionary Anthropology Leipzig, Germany "The Neandertal Genome Project"
Svante Pääbo is a biologist specializing in evolutionary genetics. He earned his PhD from Uppsala University in 1986. Since 1997, he has been director of the Department of Genetics at the Max Planck Institute for Evolutionary Anthropology in Leipzig, Germany. In 1992, he received the Gottfried Wilhelm Leibniz Prize of the Deutsche Forschungsgemeinschaft, which is the highest honour awarded in German research. Pääbo's department in August 2002 published findings about the "language gene", FOXP2, which is lacking or damaged in some individuals with language disabilities. Pääbo is known as one of the founders of paleogenetics, a discipline that uses the methods of genetics to study early humans and other ancient populations. In 2006, he announced a plan to reconstruct the entire genome of Neanderthals.
Forensic Genetics
Theodore Anderson ● Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD, USA
I possess a Bachelor's degree in Chemistry and a Master's degree in Forensic Sciences. I have been working in the field of forensic molecular biology for 13 years. I have applied both nuclear and mitochondrial DNA analysis technologies to human identification efforts. I have served as a (Supervisory) DNA Analyst, Training & Education Coordinator, University Consulting Faculty Member and Special Projects Manager and currently serve as the DNA Technical Leader for Laboratory Automation and Biometrics at the Armed Forces DNA Identification Laboratory in Rockville, Maryland, USA. I also served briefly as the Deputy DNA Program Director at the International Commission for Missing Persons in Sarajevo, Bosnia & Herzegovina. I am a Diplomat of the American Board of Criminalistics, a Fellow of the American Academy of Forensic Sciences, a Member of the Mid- Atlantic Association of Forensic Scientists and a regular participant in the European Academy of Forensic Sciences.
Frederick Bieber ● Harvard Medical School and Birgham and Women's Hospital, Boston, MA, USA
Dr. Frederick R. Bieber serves as Medical Geneticist at Brigham and Women\'s Hospital and as Associate Professor of Pathology at Harvard Medical School in Boston, MA. He has expertise and long-standing interest in human genetics and
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 175 About Invited Speakers forensic medicine and has participated in the publication of over 100 articles, chapters, and books in these fields. Dr. Bieber serves on advisory boards of the Federal Bureau of Investigation, the Royal Canadian Mounted Police, and the United States Department of Defense. He was appointed by Massachusetts Governor Mitt Romney to serve as Co-Chair of the Governors Council on Capital Punishment, which studied forensic and legal problems in the justice system. Their acclaimed report made landmark recommendations designed to prevent incorrect convictions of defendants charged with committing capital offenses.
Zoran Budimlija ● NYC Office of Chief Medical Examiner, New York, NY, USA
Dr Zoran M. Budimlija, MD, PhD, is forensic pathologist and DNA expert. Employee of NYC Office of Chief Medical Examiner, Department of Forensic Biology as leader of Pathology Research group. Fields of interest: DNA extraction from highly compromised tissues, genomic instability of tissues used as reference samples, DNA/RNA recovery from formalin/non-formalin fixed tissues, gene expression in forensics. Teaching appointments: New York University School of Medicine, Yeshiva University, PACE University. Lives and works in New York, New York, U.S.A.
Bruce Budowle ● FBI Laboratory, Quantico, VA, USA
Bruce Budowle is the Senior Scientist for the Laboratory Division of the FBI. Some of the methods he developed are: 1) analytical assays for typing a myriad of protein genetic marker systems, 2) designing electrophoretic instrumentation, 3) developing molecular biology analytical systems to include RFLP typing of VNTR loci and PCR-based SNP assays, VNTR and STR assays, and direct sequencing methods for mitochondrial DNA, and 4) designing image analysis systems. His current human DNA forensic research is on mass spectrometry and mitochondrial DNA analysis, use and interpretation of Y STRs, and selection of SNPs for identity testing. Some of Dr. Budowle’s more recent efforts are in counter terrorism, primarily in identification of victims from mass disasters and in efforts involving microbial forensics.
Katja Drobnič ● Forensic Science Centre, Ministry of Interior, Ljubljana, Slovenia
Holds a PhD in genetic recombination and gene expression of gene involved in steroid metabolism. In forensic genetic last 10 years. I'm also professor for forensic science and forensic genetic. Scientific work: - proper recovery and collection of different biological evidence at the crime scene (victim or suspect) - improvement of methods used for isolation of DNA from different biological samples (bones, hairs and evidence from sexual assault) - population studies - development of new sex determination marker and validation studies
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 176 September 3-7, 2007, Split, Croatia About Invited Speakers
Mitchell Holland ● Forensic Science, Pennsylvania State University, University Park, PA, USA
Holds a B.S. in Chemistry from Hobart Collage, Ph.D. in Biochemistry from the University of Maryland and a Postdoctoral Fellowship at Johns Hopkins University School of Medicine where he studied Human Genetics. Is a Fellow in the American Academy of Forensic Sciences and is on the Editorial Board of the Journal of Forensic Sciences. He has held positions on governmental and company advisory boards. Prior to being asked to help develop the Forensic Science program at Penn State, was the Senior Vice President, Operations and Laboratory Director of The Bode Technology Group from 2000-2005. From 1991- 2000, held various positions at the Armed Forces DNA Identification Laboratory, including Scientific Laboratory Director from 1993- 2000. Has more experience in the last 16 years with human remains identification cases than almost any other forensic scientist. Is currently an Associate Professor of Biochemistry & Molecular Biology, and is the Associate Director of the Forensic Science Program at Penn State University.
Edwin Huffine ● Bode Technology Group, Lorton, VA, USA
As vice president for international development and humanitarian services at Bode Technology Group, Ed Huffine has overall responsibility for providing identification and forensic services for regions experiencing conflict or natural disaster as well as assisting nations with forensic system development. His forensic career began in 1990 when he was hired by the Federal Aviation Administration and tasked with developing a DNA laboratory to assist in the identification of fatal air crash victims. From 1994 – 1999, he worked for the Armed Forces DNA Identification Laboratory where he became the Chief of the section responsible for the identification of missing American service members from the Vietnam, Korean, and World War II losses. Mr. Huffine served as the Director of the Forensic Sciences Program for the International Commission on Missing Persons (ICMP), headquartered in Sarajevo from 1999 - 2004.
Rebecca Just ● Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD, USA
Rebecca Just received her BA in Biology and Sociology from Hamilton College in 2000, and her MFS in Forensic Science from George Washington University in 2002. Since 2002, Ms. Just has been working in the Research section at the Armed Forces DNA Identification Laboratory (AFDIL). Her research projects have included entire mitochondrial DNA genome sequencing of common mtDNA types to identify forensically informative SNPs, design and optimization of multiplex mtDNA SNP assays, and, most recently, work on non-CODIS mini-STR multiplexes for use with highly degraded forensic samples.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 177 About Invited Speakers
Henry Lee ● University of New Haven, West Haven and Connecticut Forensic Science Laboratory, Meriden, CT, USA
Dr. Henry C. Lee is one of the world’s foremost forensic scientists. Dr. Lee’s work has made him a landmark in modern-day forensic sciences. He has been a prominent player in many of the most challenging cases of the last 40 years. Dr. Lee’s testimony figured prominently in the O. J. Simpson trial, and in convictions of the “Woodchipper” murderer. Dr. Lee has assisted in the investigations of other famous crimes, such as the murder of Jon Benet Ramsey in Boulder, Colorado, the 1993 suicide of White House Counsel Vincent Foster, and the reinvestigation of the Kennedy assassination. In 1975, Dr. Lee joined the University of New Haven, where he created the school’s Forensic Sciences program. He has taught at more than a dozen universities, law schools, and medical schools. Dr. Lee has authored hundreds of articles in professional journals and has co-authored more than 25 textbooks, covering the areas of; DNA, Fingerprints, Trace Evidence, Crime Scene Investigation and Crime Scene Reconstruction.
José A. Lorente ● Department of Legal Medicine, University of Granada, Granada, Spain
Director of the Laboratory of Genetic Identification at the University of Granada and President of the Latin American Working Group on DNA Analysis (GITAD). Our work has been focused on missing persons for many years, and recently we have been working to identify missing kids and to prevent human trafficking. I have published over 100 papers and attended over 200 meetings all over the world. Our research now is focused on Y-chromosome and autosomal SNPs and developing techniques to improve DNA analysis results from old and degraded samples.
Damir Marjanović ● Institute for Genetic Engineering and Biotechnology Sarajevo, Sarajevo, B&H and Ruđer Bošković Institute, Zagreb, Croatia
Positions: a) Head of the Laboratory for forensic genetics, Institute for genetic engineering and biotechnology, University of Sarajevo b) Assistant professor Forensic bio-anthropology, Faculty of Criminal Sciences , University of Sarajevo, Bosnia and Herzegovina Research field: a) forensic genetics c) molecular anthropology Personal data: 33 years old Bibliography: around 60 publications (scientific papers, reviews, proceedings, abstracts etc)
Marilyn Menotti-Raymond ● National Cancer Institute, Frederick, MD, USA
Dr. Menotti-Raymond\'s research focuses on development and application of genomic resources in the domestic cat to contribute to understanding of human hereditary disease analogues and genes of general biological interest. The development of comprehensive genetic maps has been a major focus. In the last two years, Dr. Menotti-Raymond\'s group have mapped and characterized
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 178 September 3-7, 2007, Split, Croatia About Invited Speakers mutations causative of feline spinal muscular atrophy (LIX1) and retinal atrophy (CEP290). They are additionally interested in genes responsible for coat color and pattern formation in the cat, and have recently characterized variants at the tyrosinase locus (siamese, burmese), tyrosinase related protein locus (brown and cinnamon coat color), and melanophilin (dilute). The development of an STR genotyping system for genetic individualization of cat specimens was initiated following their first introduction of an animal DNA fingerprint into court.
Rebecca Mikulasovich ● NYC Office of Chief Medical Examiner, New York, NY, USA
I have been a Criminalist in the Department of Forensic Biology since June 2001, and have recently been promoted to a supervisory position within the lab. During my tenure, I have worked on traditional sexual assault and homicide casework before moving to the Low Copy Number DNA lab of which I was one of the original casework analysts. I have played an active role in structuring this new lab and establishing the flow of laboratory work. In addition to my role as a rotation supervisor and casework analyst, I am also a member of the automation team supervising such projects as the implementation of liquid handling systems such as the Biomek NX and CAS 1200, laser microdissection for the collection of single cells for DNA analysis, and the optimization of the QIAgen BioRobot M48 for multiple forensic sample types.
Tahnee M. Nelson ● San Francisco Police Department, San Francisco, CA, USA
Tahnee Nelson graduated from Western Washington University in Bellingham, Washington with a Bachelor of Science in Cellular Biology and later received a Master of Science in Forensic Science from The George Washington University in Washington, DC. As part of her graduate work, Tahnee completed a thesis research project in collaboration with the Armed Forces DNA Identification Laboratory which entailed developing a multiplex assay able to type fourteen mitochondrial DNA haplogroups in a single reaction. The haplogroups were chosen based upon their ability to discriminate the individual into one of the major ancestral lineages. The intended utility of the assay is haplogroup typing of highly degraded human remains for either forensic casework or as a low cost high throughput alternative for sample processing of anthropological specimens. Tahnee is currently employed by the San Francisco Police Department Criminalistics Laboratory where she conducts forensic examinations of biological evidence.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 179 About Invited Speakers
Thomas Parsons ● International Commission on Missing Persons, Sarajevo, B&H
Dr. Thomas Parsons joined ICMP as Director of Forensic Sciences in 2006. He has served on the Scientific Advisory Board of the ICMP since 2000, and in 2005 was appointed Chair of the ICMP Steering Committee on Forensic Sciences. Before joining ICMP, Dr. Parsons was Chief Scientist at the U.S. Armed Forces DNA Identification Laboratory (AFDIL), where he had worked since 1994, and where one of his primary roles was to direct the AFDIL Research Section. He is also an adjunct faculty member in the Department of Genetics and the Department of Forensic Sciences at the George Washington University. His primary specialization is in forensic DNA applications and basic molecular genetics. Since September 2001, Dr. Parsons has served on a (U.S.) National Institute of Justice expert advisory panel for the World Trade Center DNA identification efforts, and is currently a member of the expert panel advising
Lutz Roewer ● Institute of Legal Medicine, Charité University of Medicine, Berlin, Germany
I am biochemist by profession and work as the head of the Department of Forensic Genetics in Berlin, Germany. We were the first who have cloned, sequenced and applied autosomal and Y-chromosomal microsatellites for forensic identification and anthropological research in the late 1980s. We established Y- STR haplotyping using a core set of markers in forensic genetics and molecular anthropology in the 1990s and work since than by means of international cooperation on a large reference database comprising Y chromosomal haplotypes (YHRD). Population genetics, especially research on genetic substructures and population history, as wells as its proper use in forensic genetics is my field of research.
Antti Sajantila ● Department of Forensic Medicine, University of Helsinki, Helsinki, Finland
Professor and Vice-director of Department of Forensic Medicine at University of Helsinki. The head of the unit of Forensic Pathology and the director of the Laboratory of Forensic Biology. Scientific interests include forensic DNA analysis and population genetics. In addition, classical forensic pathology and anthropology, particularly DVI work.
Brion C. Smith ● Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD, USA
Brion Smith is a 1978 graduate of the biology program at the Virginia Military Institute and received his doctorate of dental surgery degree from the Medical College of Virginia, Virginia Commonwealth University in 1982. He retired from the
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 180 September 3-7, 2007, Split, Croatia About Invited Speakers
US Army in 2006 after 24 years of service during which he completed a residency in oral and maxillofacial pathology and a fellowship in ear, nose and throat pathology. He is board certified in forensic odontology and oral and maxillofacial pathology. His current work focuses on laboratory management and obtaining reportable results from complex and challenging dental cases.
Molecular Anthropology
Ranjan Deka ● University of Cincinnati College of Medicine, Cincinnati, OH, USA
I am a Professor at the Center for Genome Information at the University of Cincinnati Medical Center. My primary areas of research are molecular population genetics and genetics of complex diseases. My laboratory is involved in finding genetic variants associated with metabolic syndrome, stroke, and prostate cancer. In the area of population genetics, we are interested in evaluating the linkage disequilibrium patterns in isolated populations and their usefulness in mapping complex traits.
Irena Martinović Klarić ● Institute for Anthropological Research, Zagreb, Croatia
My research interest has mostly been focused on studies of uniparental polymorphisms and autosomal microsatellites in Eastern Adriatic isolates as well as Croatian and other populations across Southeastern Europe. Most recently I am engaged in the research group at the Institute for Anthropological Research that is interested in studying the Roma population in Balkans as a model of transnational, isolated founder population in order to investigate its genetic and population structure as well as genetic (single-gene) disease heritage.
Giuseppe Passarino ● Department of Cell Biology, University of Calabria, Rende, Italy
Giuseppe Passarino is Professor of genetics at the University of Calabria in Italy. He has previously worked at the University Pavia (Italy), at the University of Leiden in the Netherlands, and at the University of Stanford in California. He studied for many years the structure and the history of human populations by means of mitochondrial DNA and Y chromosome markers. In this frame he participated to the definition of human Y chromosome phylogeny and to the discovery of important aspects of the early stages of the history of Homo sapiens. In the recent years he has studied the genetic basis of human aging and longevity in the frame of fast-evolving western societies. In particular he is interested to the different hereditability of the longevity trait in different populations. He is currently participating to different European Projects for the study of healthy aging.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 181 About Invited Speakers
Mario Pirastu ● Institute of Population Genetics, National Research Council, C.N.R., Alghero and Shardna Life Sciences Cagliari, Italy
Graduated in Medicine at the University of Cagliari, during a significant experience at Dpt. of Genetics, University of San Francisco, he has characterized molecular basis of Thalassemias and has identified several new mutations and their prevalence in Sardinia and in the Mediterranean area. He performed his research activity at the Institute of Talassemie ed Anemie Mediterranee, CNR Cagliari, pursuing studies on monogenic common diseases in Sardinia (i.e., Wilson disease). >From 1995 he has been Director at the Institute of Population Genetics, CNR Alghero, where he started a research project on complex disorders in genetically isolated populations of Ogliastra, regarded as “an Island in an Island” in Sardinia. In 1999 he created the Genetic Park of Ogliastra, a non-profit Consortium with public and private partners to support research activities carried out in the field. In 2000, together with other partners, he started up and was named Scientific Director of the first Italian genomic Company, Shardna.
Pavao Rudan ● Institute for Anthropological Research, Zagreb, Croatia
Professor of Anthropology, Director of the Institute for Anthropological Research, Zagreb, Croatia. MD and PhD - School of Medicine, University of Zagreb. Ordinary Member of the Croatian Academy of Sciences and Art; President of the Commission on Medical Anthropology and Epidemiology (IUAES); Secretary General of the International Association of Human Biologists; Member of the Executive Committee of the IUBS (UNESCO). Awards: Gorjanović-Kramberger Medal for Anthropology; Anthropology Award for Distinguished Service - College of William and Mary, Williamsburg, USA; Ruđer Bošković Award for Scientific Achievement of the Republic of Croatia; Aleš Hrdlička Memorial Medal for Anthropology, Czechoslovakia; First Mario D. Zamora Memorial Lecture and Mario D. Zamora Award, College of William and Mary, Williamsburg, USA. Main research topics: Anthropological research of population structure and genetic history of the Adriatic island isolates through holistic analytic approach. Analyses of biological and socio-cultural traits of current populations. Strategy of genetic epidemiology.
Peter Underhill ● Stanford University Medical Center, Stanford, CA, USA
I have worked in the Department of Genetics at Stanford University School of Medicine since 1990. My research involves the molecular analysis of human DNA sequence variation in human populations. I have been doing pioneering research on human Y chromosome diversification since 1992 that has lead to the development of a robust gene tree that elegantly defines numerous Y- chromosome varieties with distinctive geographic localization. The main focus of my research involves deciphering population affinity and substructure in contemporary populations using Y-chromosome compound SNP and STR lineages. I have coauthored numerous peer-reviewed publications on the subject.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 182 September 3-7, 2007, Split, Croatia About Invited Speakers
Richard Villems ● Institute of Molecular and Cell Biology, University of Tartu and Estonian Biocentre, Tartu, Estonia
My background is molecular biology, but about a dozen of years ago, I became so much interested in what is now sometimes identified as archaeogenetics, that I changed my research direction. Since then, I work, with my colleagues in Tartu, Estonia, and in a close collaboration with many all around the world, in this wide, largely interdisciplinary field.
Advances in Genomic Methods
Jean-Pierre Kocher ● Mayo Clinic College of Medicine, Rochester, MN, USA
Dr. Jean-Pierre Kocher joined Mayo Clinic in October 2005 to develop and direct the activities of the newly created Bioinformatics Core. Prior to his assignment at Mayo Clinic, Dr. Kocher served as CEO of Molecular Networks, a company focused on the development of software applications and services for the pharmaceutical industry. He also served as Director of Computational Chemistry at Camitro Corp., where his responsibilities included the development of computational models to predict ADME properties. Prior to joining Camitro, Dr. Kocher was a postdoctoral fellow in the Laboratory of Molecular Biology at the National Institutes of Health, National Cancer Institute. Dr. Kocher received his Ph.D. in Computational Chemistry from the University Libre de Bruxelles, Belgium, and his M.S. in Informatics and B.S. in Chemistry from the University Louis Pasteur, Strasbourg, France.
David Smith ● Mayo Clinic College of Medicine, Rochester, MN, USA
My research laboratory focuses on several projects. The first is the use of state-of- the art genomic technologies to better understand the genetic events that occur during cancer development. We are identifying and characterizing a whole new group of important regulatory transcripts which do not code for protein. These appear to also be targets of alteration during cancer development. We are also using gene expression profiling and complementary genomic technologies to comprehensively examine different head and neck cancers for diagnostic and prognostic purposes. I am also in charge of the committee at Mayo that oversees the Mayo Core Research Laboratories and to evaluate new technologies that could be employed to further facilitate research and its translation into clinical practice.
George Vasmatzis ● Mayo Clinic College of Medicine, Rochester, MN, USA
Dr. Vasmatzis is a Senior Associate Consultant in the Experimental Pathology division, in the Department of Laboratory Medicine and Pathology and a member of the Mayo Clinic Cancer Center. He is an Assistant Professor in the Department
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 183 About Invited Speakers of Laboratory Medicine at the Mayo Medical School. He has a Ph.D. in Biomedical Engineering and has acquired experience in diverse disciplines, including Bioinformatics, Molecular Biology, and Computational Biology. Dr. Vasmatzis’ research program consists of bioinformatics specialists, molecular biologists, epidemiologists, and pathologists. He combines computational and experimental techniques to facilitate discovery of genes that can be used as diagnostic and prognostic markers for cancer.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 184 September 3-7, 2007, Split, Croatia Sponsor's Information
SPONSOR'S INFORMATION
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 185 Sponsor's Information
Applied Biosystems
Organization Information Organization: Applied Biosystems Address: Frankfurter Strasse 129B City: Darmstadt State/Province: ZIP or Postal Code: 64293 Country: Germany Phone: +49 6151 9670 0 Fax: +49 6151 9670 5599 E-Mail: [email protected] URL: http://europe.appliedbiosystems.com Applied Biosystems is the leader in Human identification testing. We supply validated products, expertise and application support for forensic Keywords: laboratories. Since 1989, we have been the leader in DNA analysis technologies. Today we offer the most robust, reliable and validated forensic solutions for Human Identification testing. 310 Genetic Analyzer 3130 Genetic Analyzer 3130xl Genetic Analyzer 96-Well GeneAmp® PCR System 9700 7500 Real-Time PCR System AmpFLSTR® MiniFiler™ PCR Amplification Kit AmpFLSTR® Yfiler™ PCR Amplification Kit AmpFLSTR® Identifiler® PCR Amplification Kit Product(s): Quantifiler™ Human DNA Quantification Kit Quantifiler™ Y DNA Quantification Kit AmpFLSTR® Profiler Plus® PCR Amplification Kit AmpFLSTR® COfiler® PCR Amplification Kit AmpFLSTR® SEfiler™ PCR Amplification Kit AmpFLSTR® SGM Plus® PCR Amplification Kit GeneMapper® ID Software v3.2 VALID™ Software v1.0
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 186 September 3-7, 2007, Split, Croatia Sponsor's Information
Biosistemi
Organization Information Organization: Biosistemi Address: Pijavišče 32 City: Zagreb State/Province: ZIP or Postal Code: 10090 Country: Croatia Phone: +385 1 3460 838 Fax: +385 1 3460 840 E-Mail: [email protected] URL: www.biosistemi.hr Biosistemi and its daughter company Vivogen is representative of Applied Biosystems for Croatia, Bosnia Keywords: and Herzegovina, Serbia, Macedonia, Montenegro, Albania and Romania Instruments and reagents for: PCR, RT PCR, genetic analysis (forensics, etc.) Product(s): Mass spectroscopy and protein and peptides analysis
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 187 Sponsor's Information
BodeTechnology
Organization Information Organization: BodeTechnology Address: 10430 Furnace Rd, Suite 107 City: Lorton State/Province: Virginia ZIP or Postal Code: 22079 Country: United States Phone: 703-646-9740 Fax: 703-646-9741 E-Mail: [email protected] URL: www.Bodetech.com DNA Identification Laboratory and Products Keywords: mtDNA, STR and Y-STR DNA analysis Crime Scene Collector, Buccal DNA Collector and Product(s): BodEpen Digital Pen
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 188 September 3-7, 2007, Split, Croatia Sponsor's Information
Eppendorf AG
Organization Information Organization: Eppendorf AG Address: Barkhausenweg 1 City: Hamburg State/Province: Hamburg ZIP or Postal Code: 22339 Country: Germany Phone: 0049 (0) 40 538 01 0 Fax: 0049 (0) 40 538 01 556 E-Mail: [email protected] URL: www.eppendorf.com Eppendorf is a biotechnology company that develops, manufactures and distributes systems comprising instruments, consumables and reagents for use in laboratories worldwide. The company is focused on two business sectors: • Bio Tools, which includes tools such as pipettes, dispensers and centrifuges, and consumables such as test tubes and pipette tips. • Molecular Technologies, which includes instruments and systems for cell manipulation, automatic devices for high-throughput screening (HTS), complete systems for DNA multiplication, nucleic acid purification kits and biochips. A series of “Complementary Products" manufactured Keywords: both by Eppendorf and other companies rounds off the product range. On the basis of this product program and the core competencies of Eppendorf the department of Customized Solutions offers individual problem solutions to its partners. Such projects are accompanied from defining the specifications to serial production and logistic organisation. Eppendorf was founded in 1945 with headquarters in Hamburg and today has more than 1,500 employees. The group owns subsidiaries in strategically important countries and is represented in all other markets by distribution partners.
New! Eppendorf Plate Deepwell 96 und 384 Product(s): New! ep Dualfilter T.I.P.S - STOP AEROSOLS! News and Highlights New! BioPhotometer plus – a valuable addition to which will be your laboratory! represent on MixMate: efficient and versatile mixing on a microliter Biotechnica 2007 scale Multipette Xstream incl. illuminated display 5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 189 Sponsor's Information
Gorea Plus d.o.o.
Organization Information Organization: Gorea Plus d. o. o. Address: Svetonedeljska cesta 97 City: Kerestinec State/Province: N/A ZIP or Postal Code: 10431 Country: Croatia Phone: +385 1 3369 610 Fax: +385 1 3369 611 [email protected]; E-Mail: [email protected] URL: www.gorea-plus.hr Trading, export-import company, distributor of leading Keywords: worldwide manufactures and suppliers for all sorts of lab consumables Molecular biology and microbiology products, Product(s): Laboratory consumables, equipment and other labware
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 190 September 3-7, 2007, Split, Croatia Sponsor's Information
INEL - medicinska tehnika d.o.o.
Organization Information Organization: INEL - medicinska tehnika d.o.o. Address: Avenija Dubrovnik 10a City: Zagreb State/Province: ZIP or Postal Code: 10000 Country: Croatia Phone: +385 1 6520-546, 6520-556 Fax: +385 1 6520-966 E-Mail: [email protected] URL: www.inel-mt.hr distributor, Eppendorf, Qiagen, Tecan, Sarstedt, Keywords: Fermentas, biotest, forensic All products for biology, molecular biology, forensic and Product(s): proteomics laboratories.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 191 Sponsor's Information
Invitrogen
Organization Information Organization: Invitrogen Address: Inchinnan Business Park, 3 Fountain Drive City: Inchinnan State/Province: Paisley ZIP or Postal Code: PA4 9RF Country: UK Phone: +44 141 814 6100 Fax: E-Mail: [email protected] URL: www.invitrogen.com Invitrogen provides products and services that support academic, government, research institutions, Keywords: pharmaceutical and biotech companies worldwide in their efforts to improve the human condition. Nucleic Acid Purification & Quantification Product(s): Cloning & Protein Expression
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 192 September 3-7, 2007, Split, Croatia Sponsor's Information
MDS Analytical Technologies (formerly Molecular Devices)
Organization Information MDS Analytical Technologies (formerly Molecular Organization: Devices) Address: 1311 Orleans Drive City: Sunnyvale State/Province: CA ZIP or Postal Code: 94089 Country: USA Phone: (408) 747-1700 Fax: (408) 747-3601 E-Mail: [email protected] URL: www.moleculardevices.com Instrumentation and reagents for drug discovery and life Keywords: sciences research FLIPR Tetra® for aequorin, fluorescence & luminescence assays SpectraMax® microplate readers ImageXpress Micro/Ultra imaging systems Arcturus LCM instruments Product(s): Cell-Key Label Free technology Automated and Conventional Electrophysiology AquaMax Washers and Dispensers MetaMorph data analysis software FlexStation benchtop reader with fluidics
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 193 Sponsor's Information
PLIVA d.d.
Organization Information Organization: PLIVA d.d. Address: Ulica grada Vukovara 49 City: Zagreb State/Province: Zagreb ZIP or Postal Code: 10 000 Country: Croatia Phone: +385 1 612 09 99 Fax: +385 1 611 10 11 E-Mail: [email protected] URL: http://www.pliva.com The leading CEE pharmaceutical company and European generic arm of Barr Pharmaceuticals, Inc., which develops, Keywords: manufactures and markets generics, biopharmaceuticals and APIs; strong R&D. Finished dosage form branded generic pharmaceutical products and active pharmaceutical ingredients (APIs), with products including solid dose forms, injectables, Product(s): creams/ointments, Over-The-Counter (OTC) products, cytostatics, APIs and biopharmaceuticals.
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 194 September 3-7, 2007, Split, Croatia Sponsor's Information
SoftGenetics LLC
Organization Information Organization: SoftGenetics LLC Address: 200 Innovation Blvd Suite 235 City: State College State/Province: PA ZIP or Postal Code: 16801 Country: USA Phone: 888-791-1270 Fax: 814-237-9343 E-Mail: [email protected] URL: www.softgenetics.com DNA Fragment, STR, Sequence Analysis Software Keywords: SoftGenetics Human Identification GeneMarker Analytical software for: GeneMarker HID – forensic STR analysis GeneMarker – fragment analysis Mutation Surveyor – DNA Variant Analysis from Product(s): Sequence traces Mutation Explorer – clinical DNA Variant analysis JelMarker – gel image and conversion software
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 195 Author index
AUTHOR INDEX
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 196 September 3-7, 2007, Split, Croatia Author index
—A— Brajković J 74 Abdigoudarzi M 85 Braš M 146 Ahmetaš L 116 Bravermanová M 63 Akhmetova G 107 Bravi CM 142 Alakoc Y 134 Budimlija ZM 33 Aleksej D 123 Budowle B 34, 35 Alkhutov S 154 Bujdosó G 122 Alvarez JC 42 —C— Anđelinović S 118 Calandro L 81 Anđelinović Š104, 115, 131, 148, Calloway C 78 172 Car H 145 Anderson T 31 Caragine T 33, 45 Andreescu G 168 Cardos G 108, 132 Aslanishvili V 154 Čečuk-Jeličić E 145 Atramentova L 121, 128 Chandrasekar A 138 —B— Chaventré A 141 Baindur-Hudson S 139 Chiaroni J 75 Bajnoczky I 166 Churnosov M 121, 128 Bajnóczky I 94, 125 Ciciała A 93 Bajrovic K 43 Coble M 31, 40 Bakal N 43, 92 Crews J 39 Bakarić P 115 Crimmen O 113 Balanovska E 128 Csonka E 122 Balanovsky O 121, 128 Čukić D 109 Ballantyne KN 73, 86 Cukrov S 52 Balode I 68 Ćurić G 110, 117, 146 Barać Lauc L 52 —D— Barbisin M 81 Damba L 142 Barik SS 138 Daniel R 147 Barritt S 40 David V 44 Barritt-Ross S 31, 50 Davoren J 39, 43 Bartosik I 71 Definis Gojanovic M 148 Bašić L 87 Definis-Gojanović M 104, 131 Bates F 126 Deka R 51 Batrana A 108 Delonga V 148 Batrana L 108 Desnoyers A 31 Baum H 45 Dinparast-djadid N 85 Baumanis V 127 Dipierri JE 142 Bego T 116 Dmochowska G 93 Bellovits O 122 Dobosz T 72, 93 Bermisheva M 67 Dogan Alakoc Y 101 Bernasovská J 143 Dogra TD 97 Bernasovský I 143 Domazet-Lošo T 74 Bieber FR 32 Dozdová E 163 Biramijamal F 144 Drmić Hofman I 172 Biruš I 117 Drobnic K 43 Boroňová I 143 Drobnič K 36, 37 5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 197 Author index
Drozdová E 153 Hasegawa I 90, 140 Ducharme-Smith A 59 Havaš D 136, 137 Dulik MC 133 Haveric S 43 Đurković M 146 Hedman M 49 Durmic-Pasic A 43 Heffer-Lauc M 164 Džijan S 106, 110, 117 Hellmann A 64 —E— Hercog R 110 Edson S 31 Hewakapuge SK 139 Eminović I 87 Hodžić B 116 Endicott P 138 Holland M 38 Entrala C 42 Horinek A 91 Erlich HA 78, 79 Huel R 87, 119 Evans C 126 Huffine E 39, 43 —F— Hulce D 38 Fang R 81 Hung M 100 Fedorova SA 142 —I— Feldman M 69 Imtang S 165 Fernandez-Rosado F 42 Iravanloo G 144 Fodor F 122 Irwin J 31, 40 Fogt F 33 Ishida T 76 Frassati C 75 —J— Frolík J 63 Jakovski Z 123 Furtado M 81 Janeska B 123 —G— Janićijević B 52 Galan E 168 Jankova R 123 Garvin AM 88 Jeran N 136, 137 Gerke N 80 Jokić M 149 Giambra A 111 Jonkisz A 93 Ginter EK 173 Jukić-Pavan D 106 Goetz R 102 Jurišić G 149 Gokcumen O 101, 134 Just R 40 Gökçümen O 133 Just RS 46 Golubenko MA 142 —K— Grechanina EY 135 Kaczmarczyk G 71 Grkovic S 148 Kalamujic B 43 Grubić Z 145 Kapitanović S 149 Gubina MA 142 Karpukhin AV 173 Gugić D 104 Karvela M 151 Gulec E 101, 134 Kashima T 76 Gultekin T 101, 134 Kastelic V 36 Gunn P 102 Katzmarzyk C 119 Gusar VA 135 Kazakova SA 95 —H— Kazic A 113 Hadlaczky G 122 Kažoka D 68 Hadziselimovic R 43 Kerhin-Brkljačić V 145 Hajkova J 63, 89, 99, 111 Kharkov V 130 Halling M 59 Khayrullin R 107 Hammond JBW 113 Khusainova R 67 5th ISABS Conference in Forensic Genetics and Molecular Anthropology 198 September 3-7, 2007, Split, Croatia Author index
Khusnutdinova E 67 Marjanovic D 43, 92, 123 Khusnutdinova EK 142 Marjanović D 96, 116, 118 Kimura R 90 Mark L 66, 166, 167 Kinjabaeva G 150 Márk L 94 Kitano T 140 Martinez-Espin E 42 Kivisild T 142 Martinez-Gonzalez LJ 42 Knight A 69 Martinez-Labarga C 142 Kocher JP 58 Martinović Klarić I 52 Könczöl F 94, 125 Maynard K 31 Korabecna M 91 Medvedeva O 130 Kovacevic L 43, 92 Menotti-Raymond M 44 Kovačević L 116 Merrett NR 113 Kračun SK 117 Mertlová K 153 Kricskovics A 94 Metspalu E 141 Kroll A 132 Metspalu M 138, 142 Krumina A 127 Mikulasovich R 33, 45 Krzyżańska A 72, 93 Miloš A 87, 119 Kuret S 172 Miritoiu N 108, 132 Kutuev I 67 Miriţoiu N 168 —L— Mitchell RJ 73, 86 Lamnissou K 151 Mitchenko I 107 Langstieh BT 124 Moisan JP 141 Laslo E 152, 157, 162, 170 Morzfeld J 64 Lauc G 106, 110, 117, 146, 164 Mountain J 69 Lebioda A 93 Mulligan CJ 142 Lee D 50 —N— Lee H 41 Nagle E 68 Liu CSJ 38 Nagy G 94, 125, 166 Ljubkovic J 148 Nassif NT 147 Ljubković J 131 Nazari E 144 Lobov A 67 Nazarova A 154, 155 Loginova AN 173 Nelson TM 46 Lorand T 166 Nikolova K 123 Loreille O 31, 46, 50 Nikulin MV 95 Lorente JA 42 Noje D 152, 157, 170 Lu C 33 Noor J 126 Lubchenko LN 173 —O— Luna A 42 O’Brien SJ 44 —M— O’Shea C 81 MA Cywinski P 71 Oliver R 31 Machaopa J 165 Opolska-Bogusz B 71 Mack SJ 79 Osawa M 90, 140 Madacki-Todorović K 87 Ossipova LP 142 Makarevic E 92 Ozguven HD 105 Makino K 76 —P— Malhi RS 142 Pääbo S 30 Malikova R 114 Pai AA 169 Marcsik A 166, 167 Palo J 49 5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 199 Author index
Papadopoulou S 151 Rudan P 55, 141 Parsons T 47, 87, 119 Rusz A 122 Passarino G 53 Ryakhovsky M 107 Pavlinić D 110, 146 —S— Pennarun E 141 Sabule A 127 Penovic A 148 Sajantila A 49 Perez D 59 Šamija I 96 Peričić Salihović M 52 Sanak M 71 Petrejčíková E 143 Sanchez JJ 138, 147 Petričević S 115 Sanchez-Mazas A 75 Picard C 75 Saremi M 98 Piniewska D 71 Saremi MA 98 Pirastu M 54 Sarzyńska M 93 Pliss L 127 Saskova L 63, 89, 99, 111 Podini D 46, 100 Sato F 90, 140 Pojskic N 43 Schanfield M 46, 118 Pokupčić K 52 Schanfield MS 100 Polyakov AV 95 Schleenbecker U 64 Poparić I 149 Schurr TG 101, 133, 134, 169 Popiolek D 33 Sears A 102 Pospekhova NI 173 Selmanović A 119 Pospisek M 63, 99 Sengupta M 156 Potorac R 103 Shchagina OA 95 Primorac D 43, 96, 115, 118 Shewale J 81 Prinz M 33, 45 Simons, JL 112 Pritchett J 59 Skaro V 43 Projic P 43 Škaro V 96, 118 Projić P 96 Smajlović L 87, 119 Pshenichnov A 121, 128 Smirnova TY 173 Pusta CT 152, 157, 160, 162, 170 Smith B 50 Puzuka A 127 Smith D 59, 60 Puzyrev V 130 Smith DG 142 —R— Soemantri A 76 Raina A 97 Soficaru A 132 Ramic J 43 Soficaru D A 168 Rao VR 138 Soloviova D 128 Rebbeck TR 169 Soltani MS 144 Reddy BM 124 Soták M 143 Reidla M 142 Štambuk S 115 Richard C 141 Stanciu F 103, 168 Rickards O 142 Stefflova K 169 Riproumsup K 165 Stepanov V 130 Rizvić A 119 Stepanov VA 142 Rodewald A 108, 132 Štingl K 145 Roewer L 48 Stoian G M 168 Rootsi S 121, 129 Stoian V 103 Roux C 102 Stuart S 78 Rozane S 127 Sturk K 40
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 200 September 3-7, 2007, Split, Croatia Author index
Sutlović D 104, 115, 131 Vasmatzis G 61 —T— Vasserman NN 95 Tamm E 142 Velissariou V 151 Taneja P 82 Viljetić B 164 Tautz D 74 Villanueva E 42 Tavallaei M 98 Villems R 57, 67, 135, 141, 142 Tegako L 121 Vintiner, SK 112 Teibe U 68 Voevoda MI 142 Tomić I 172 —W— Tomulescu IM152, 157, 158, 159, Wagner J 106, 146 160, 161, 162, 170 Walsh SJ 102, 147 Tug A 101, 105, 134 Wan N 38 —U— Wehrhahn D 89 Udina I 69 West AB 33 Umetsu K 140 —Y— Underhill PA 56 Yadav B 97 Underwood S 126 Yankovsky N 121 —V— Yunusbayev B 67 Vajn K 164 Yusupov R 67 van Daal A 65 —Z— van Oers P 81 Zaporozhchenko V 128 van Oorschot R 139 Zeigler-Johnson CM 169 van Oorschot RAH 73, 86 Zhadanov SI 142, 169 Vanek D 63, 89, 99, 111 Zhivotovsky L 69 Vaňharová M 163 Žunec R 145
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 201 Keyword index
KEYWORD INDEX
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 202 September 3-7, 2007, Split, Croatia Keyword index
17025 99 autosomal DNA 42 2D coordinates 157 autosomal DNA markers 128 7500 Real Time PCR System 81 Autosomal SNPSTR systems 69 abberant behavior 159 Autosomal STR Analysis 31 acari 85 average expected heterozygosity 115 adaptation 74 AZA 149 admixture 169 azoospermia 122 aDNA 108, 132 backlog 88 AFDIL 50 Bashkirs 150 African Americans 169 Basque 141 age 107 Bayash 52 Ageing 139 Beringia 142 Aging 53 Bilateral Asymmetry 156 Alleged baby died cases 105 biogeographical ancestry 147 Allele frequencies 94 Bioinformatics 61 Alleles 101 Bioterrorism 34 amelogenin 163 Birth-rate 150 Amelogenin 108, 132 bisulphite 90 amelogenin gene 36 Bode 39 Amelogenin gene 153 bone 107 Americas 69 Bone Extraction 165 Amerinds 155 bone samples 104 AmpFℓSTR® system 81 bonepowdering 116 Amplification Bias 86 bones 72 analysis 111 breast cancer 173 Analysis 38 Breton 141 analytical methods 104 buried body 37 ancestry 46 calliper 107 Ancient bone sample 94 Cancer 61 ancient DNA 63, 72 cancer localization 173 Ancient DNA 30 cancer markers 59 Andaman Islands 138 case work 64 androgen receptor gene 151 cat breeds 44 anthropology 152 Caucasoids 154, 155 Anthropology 41, 55, 79, 154 Caucasus 154 anthropometrical analysis 148 cause of death 160 antropology 109 Centralized Repository System 58 archaeology 37 children 163 archeogenetics 63 chromosome banding 160 archive materials 71 cladistics 56 artificial intelligence 152, 157 coding region 78 Asia 69 CODIS 35, 95 association studies 53, 151 collecting sample 92 Attribution 34 Comaparative 123 Austro-Asiatic (Mon-Khmuic) 124 comparison 109 Austronesian 76 complete sequences 133, 142 automation 88, 89, 111 Complex Trait 51
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 203 Keyword index congenital abnormalities 160, 161 DNA IQ 89 conserved 59 DNA polymorphism 108 Control region 139 DNA quantitation 80 cranial dysmorphism 158 DNA typing 93, 96, 110, 117, 165 craniofacial indices 68 DNA Typing 119 craniofacial parameters 68 Domestic cat 44 CRFR1 gene 146 dysgenesis 114 Crime scene 82 early medieval 148 Crime Scene 41 East Slovakia 143 crime scene officer 37 Endogamy 101 Criminal Cases, Validation 45 eneolith 163 Croatia 52, 118, 172 Epidemiology 34 Croatian 149 Epithelial Cells 112 Croatian population 96 Eppendorf epMotion 89 Croatians 145 ethnic groups 150 cytogenetics 122 European haplogroups 100 Cytokeratins 112 evidence collection 37 Dalmatia 148 evolution 66 Dane's stain 112 exhumation 37 Data Analysis 58 expansion 129 Data Management 58 facial dysmorphism 158 database structures 157 familial searching 126 deamination 90 Familias 91 degraded DNA 63, 72, 93 family hereditary antecedents 161 Delhi 97 family searching 32 delinquent population 159 fatherhood 109 Demineralization 31 fatherless paternity cases 91 Dental 50 female dead body 162 dermatoglyphics 159 fetal cell free DNA 106 Dermatoglyphics 156 fetal development 170 detergents 164 fetal gender 106 Development Forensic Genetic 98 Fingerprint 94 different methods 92 fingerprints 152, 157 Differentiation 69 forensic 102 digit ratio 159, 170 Forensic 50, 97, 127 Disequilibrium 51 forensic anthropology 47, 49, 166 DNA39, 42, 49, 50, 71, 97, 102, 109, Forensic Anthropology 165 134, 148 forensic archeology 47 DNA analysis 92, 105, 109, 116 forensic biology 33 DNA Analysis 82 Forensic botany 115 DNA contamination 99 forensic DNA 32 DNA damage 30 forensic evidence 64 DNA database 32 forensic genetics 49, 170 DNA Database 35 Forensic Genetics 82 DNA degradation index 168 forensic pathology 49 DNA extraction 72, 89, 92, 116 forensic remains 140 DNA identification 43 forensic science 104, 131 DNA Identification 47 Forensic Sciences 105
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 204 September 3-7, 2007, Split, Croatia Keyword index forensic typing system 44 High quality & quantity 98 forensic, DNA 111 High Sensitivity DNA Testing 45 Forensics 38, 39 high throughput technologies 60 formalin-fixed fish 113 high-resolution analysis 52 fornsic anthropology 167 historical case 103 fossilization 168 Historical Migrations 55 founder effect 136, 137 HLA 79 founder lineages 142 HLA polymorphism 75 France 141 Homicide 41 Frequencies 123 hormonal 114 frequencies distribution 95 HPA axis 146 FT-IR 166 human 102 G1733A polymorphism 151 human demography 53 gangliosides 164 human DNA 104 GC content 90 human genetic diversity 75 gender determination 167 human identification 110, 117 genealogical tree 103 human migrations history 75 genealogy 108 Human origins 30 GeneMarker 38 Human populations 69 genetic algorithms 152, 157 Hungary 125 Genetic Analysis 41 HVI/HVII 78 genetic database 44 hyalomma anatolicum anatolicum 85 genetic distances 121 identification 71, 102, 162 genetic diversity 56, 128, 130, 135 identity testing 64 genetic drift 136, 137 immobilized SSO probes 78 Genetic epidemiology 55 Immunohistochemistry 112 genetic factors 161 imunohistochemistry 164 genetic match 110 inbreeding 117 genetic polymorphism 146 India 97, 156 Genetic polymorphism 143 individual recognition 170 genetic potential 150 infertility 122 genetic testing 172 Infrastructure Development 58 Genetics 34, 79, 154, 156 inhibitors 63 Genocide 39 Interpretation 35 Genome, sequencing 60 intra-ethnic variation 121 genomics 74 Investigation 41 geographic area 162 Iran 144 gonads 114 Island of Rab 136 GPI-anchored proteins 164 Island of Vis 137 guanidinium-based protocol 113 isolated population 136, 137 haplogroup origin 129 ITPA 149 haplogroups 46 ixodidae 85 Haplotype 48, 131 Kinship 134 haplotypes 125 kinship analysis 32 Haplotypes 127, 134 KIR polymorphism 75 haplotypic associations 145 laboratory environment 99 HapMap 51 Laser capture microdissection 82 harmful factors 161 Latvia 127
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 205 Keyword index likelihood ratio 40 molecular diagnostics 172 likelihood ratio tests 133 molecular genetics 122 Lineages 101, 134 most probable genetic profile 103 linear array 78 Mother-baby mix ups 105 linear measurements 68 mtDNA46, 78, 100, 128, 132, 133, Linkage 51 138, 142, 169 linkage disequilibrium 145 mtDNA genomes 57 lipid rafts 164 Multiple Displacement Amplification Locked Nucleic Acids 73 86 longevity 53 multiplex 40 low copy number 80 multiplex PCR assay 147 Low Copy Number 31 Multiplex TaqMan® assay 81 Low Copy Number DNA Testing 45 Multiplexing 73 low volume 80 museum collection 113 Lyngngam 124 mutation 36, 95 M31 138 mutations 173 M32 138 Native Americans 142 Macedonian Population 123 Neandertal 30 macroevolution 74 nested cladistics 57 male-female ratio 53 Newest Technic 98 malformative diseases 158 Next Generation 60 mass graves 43 NoC1 NoC2 94 Mass graves 47 non-coding transcripts 59 mass spectrometry 167 non-CODIS 40 maternal plasma 106 non-human DNA 64 Maternal three generations 139 noninvasive prenatal testing 106 mental deficiency 158, 161 nonsyndromic clefts 68 metazoa 74 North Eurasia 130 Microbial Forensics 34 North Mongoloids 155 Microevolution 55 northern Eurasia 129 microgeographical differentiation 141 novel polymorphism 144 microsatellite based genotyping 115 NRY 169 microsatellites 145 Oceania 76 mini-haplotypes 117 off-ladder allele 87 minisequencing 46 old bone samples 116 miniSTR 40, 93 Olea europaea L. 115 Mini-STR 63 optimization 80 miniSTRs 102 Origins 155 missing kids 42 out-of-Africa 57 missing persons 42, 119 ovarian cancer 173 mitochondria 140 p53 codon 72 polymorphism 76 mitochondrial DNA42, 126, 135, 136, p53 gene 144 137, 139 paleoanthropology 66, 166, 167 Mixtures 81 Paleoasiatic 155 Moldavia 108 paleogenetics 168 molecular anthropology 75, 144 paleopathology 66, 166 molecular clock 133 paleoproteomics 66 Molecular Crowding 86 Papuan 76
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 206 September 3-7, 2007, Split, Croatia Keyword index partial DNA profile 32 prostate cancer 169 Patau syndrome 160 Protein specificity 112 paternity test 96 PTSD 146 paternity testing 95 purification 88 pathology 33 QA/QC 33 PCR 100, 153 quality 172 PCR amplification 90 quality control 110 PCR multiplex 93 quantification 104 PCR protocol 116 Quantities 81 personal documents 103 RAPD approach 113 personalized medicine 60 RAPD-PCR 85 phenotype 65 Rapid DNA Extraction 98 Phenotype 147 rare allelic variant 96 phenotypic markers 68 real-time PCR 80 phylogeny 57 regional variation 121 Phylogeny 79 relative dating 166 phylogeographical analysis 135 relatives 126 phylogeography56, 57, 67, 129, 130, ribosomal RNA 140 133, 138, 141 risk factor 151 phylostratigraphy 74 Roma 52 physical marker 158 Romanies 143 physical traits 162 RSA 151 pigmentation 65 RT-qPCR 89, 99 Pohansko 153 saliva specimens 92 point mutation 87 Samoa 51 polymorphism 65, 135 sampling kits 111 population 67, 144, 149 Second World War 43 Population 118 Seghistea 162 population data 131 sequence 140 Population data 143 sequences 113 Population database 48 sex 163 population genetics 130 sex determination 36 Population history 30 Sex determination 153 population structure 56 Sexual assault 88 Population structure 55 sexual steroid 167 Population Structure 101 SGM+ 126 population substructure 117 short tandem repeat (STR) 165 populational genetics 170 short tandem repeats 64 Populations 79, 154 Short tandem repeats 143 Power of exclusion 118 Short tandem repeats (STR) 127 PP16 118 Shortest time 98 prediction 139 SIDS 90 Primer design 73 Single base extension 100 probability calculation 91 Single Nucleotide Polymorphism 51 Prognosis 61 skeletal elements 119 Project Support 58 skeletal remains 43, 148 proliferation 61 Skeletal Remains 31 prostate 61 skeleton remains 165
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia 207 Keyword index skin 71 Trace evidence 82 Slovenia 43 translocation 160 SNaPshot 147 TRC 159 SNP 46, 65, 100, 173 triallelic 87 SNPs 146, 147 tuberculosis 66 Software 38 tubular 107 Southeast Asia 76 Turkey 101, 134 Southern Croatia 131 Ukrainian population 135 Southern Urals 67 Ukrainians 128 species identification 140 Vaidya 156 Sperm DNA 88 validation 36 SRY 163 Validity 35 sry gene 36 variant allele 87 SRY gene 153 variation 65 SSR 115 vernier 107 standardization 172 Violence 39 Statistic 123 vWA31A 132 Statistics 35 war victims 49, 110 Stolen baby cases 105 whole genome 59 STR38, 40, 44, 48, 71, 73, 87, 93, Whole Genome Amplification 86 95, 99, 119 Y chromosome48, 52, 56, 121, 125, STR analysis 91 127, 128, 129, 130 STR databases 103 Y chromosome disorders 122 STR loci 96 Y chromosome STR 123 STR markers 97 Y Filer 125 STRs 47 Y SNPs 124 Structure 118 Y-chromosomal STR 91, 131 subdivided population 121 Y-chromosome 106, 114 substitutional 114 Y-Chromosome STR Analysis 31 Substructure 48 Y-chromosome, diversity 67 success rates 119 Yhrd 125 survival 150 Y-STR 126 swabs 111 Y-STRs 124 taphonomy 168 techniques 33 teeth 72, 168 Teeth 50 Telogen Hair 94 Testimony 45 the database structures 152 Thracian population 132 Tibeto-Burman 124 ticks 85 tiling arrays 59 tissue contamination 33 TNF 145 TPMT 149 Trace DNA 73, 86
5th ISABS Conference in Forensic Genetics and Molecular Anthropology 208 September 3-7, 2007, Split, Croatia
NOTES
5th ISABS Conference in Forensic Genetics and Molecular Anthropology September 3-7, 2007, Split, Croatia