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Pho300007 Risk Modeling and Screening for Brcai ?4 101111111111 PHO300007 RISK MODELING AND SCREENING FOR BRCAI MUTATIONS AMONG FILIPINO BREAST CANCER PATIENTS by ALEJANDRO Q. NAT09 JR. A Master's Thesis Submitted to the National Institute of Molecular Biology and Biotechnology College of Science University of the Philippines Diliman, Quezon City As Partial Fulfillment of the Requirements for the Degree of MASTER OF SCIENCE IN MOLECULAR BIOLOGY AND BIOTECHNOLOGY March 2003 In memory of my gelovedmother Mrs. josefina Q -Vato who passedaway while waitingfor the accomplishment of this thesis... Thankyouvery inuchfor aff the tremendous rove andsupport during the beautifil'30yearstfiatyou were udth me... Wom, you are he greatest! I fi)ve you very much! .And.. in memory of 4 collaborating 6reast cancerpatients who passedaway during te course of this study ... I e.Vress my deepest condolence to your (overtones... Tou have my heartfeligratitude! 'This tesis is dedicatedtoa(the 37 cofla6oratingpatients who aftruisticaffyjbinedthisstudyfor te sake offuture generations... iii This is to certify that this master's thesis entitled "Risk Modeling and Screening for BRCAI Mutations among Filipino Breast Cancer Patients" and submitted by Alejandro Q. Nato, Jr. to fulfill part of the requirements for the degree of Master of Science in Molecular Biology and Biotechnology was successfully defended and approved on 28 March 2003. VIRGINIA D. M Ph.D. Thesis Ad RIO SUSA B. TANAEL JR., M.Sc., M.D. Thesis Co-.A. r Thesis Reader The National Institute of Molecular Biology and Biotechnology endorses acceptance of this master's thesis as partial fulfillment of the requirements for the degree of Master of Science in Molecular Biology and Biotechnology. ,_all CYNTHI YDA, Ph.D. Director National Institute of Molecular Biology and Biotechnology This master's thesis is hereby officially accepted as partial fulfillment of the requirements for the degree of Master of Science in Molecular Biology and Biotechnology. RHODORA Ph.D. Dean, Ilege of Science iv PLEASE POST PLEASE POST THE NATIONAL INSTITUTE OF MOLECULAR BIOLOGY AND BIOTECHNOLOGY College of Science University of the Philippines Diliman, Quezon City ANNOUNCEMENT OF THE MASTERAL EXAMINATION Of ALEJANDRO Q. NATO, JR. In defense of his Masteral Thesis "Screening by PTT, SSCP, DHPLC and Bayesian Modeling Techniques for,6RCAI Mutations Among Early-Onset and Familial Filipino Breast Cancer Cases" For the degree of Master of Science in Molecular Biology and Biotechnology 9:00 A.M., Friday, 28 March 2003 Room 102, Albert Hall THESIS ADVISER THESIS CO-ADVISER VIRGINIA D. MONJE, Ph.D. APOLINARIO D. NAZAREA, Ph.D. Professor Professor National Institute of Molecular Biology Science and Society Program and Biotechnology College of Science College of Science University of the Philippines University of the Philippines Diliman, Quezon City Diliman, Quezon City THESIS READER SUSANO B. TANAEL, JR., M.Sc., M.D. Associate Professor Biology Department College of Science De La Salle University Taft Avenue, Manila THESIS EXAMINER THESIS EXAMINER AMEUIRFINA D. SANTOS, Ph.D. DR. CYNTHIA P. SALOMA Professor Associate Professor National Institute of Molecular Biology National Institute of Molecular Biology and Biotechnology and Biotechnology College of Science College of Science University of the Philippines University of the Philippines Diliman, Quezon City Diliman, Quezon City ENDORSED BY: AUTHORIZED BY: VIRGINIA D. RHODO ZA Ph.D. Director M* 1 Dean National Institute of Molecular Biology and College c ence Biotechnology University of the Philippines College o Science Diliman, Quezon City University of the Philippines Diliman, Quezon City V BIOGRAPHICAL DATA Adwbiwh Alejandro Q. Nato, Jr. Science Research Specialist 1 Health Physics Research Section, Atomic Research Division Philippine Nuclear Research Institute, Commonwealth Avenue, Difirrian I 01 Quezon City, Philippines aqnatopnri dost.gov.ph / aqnjpworld.nel.ph laqnjryalioo.coin Tel 632-9296011 loc. 235 Fax 632-9201646 Summary of Qualifications Molecular Biology Molecular Genetics Radiation Biochemistry Performing (a) radioactive protein truncation test (using '5S) utilizing SDS-PAGE visualized by autoradiography and (b) single-sirand confiannation polymorphism analysis utilizing PAGE visualized by silver staining, in a pioneering endeavor to detect BRCAJ truncating mutations and/or putative polymorphisins among eaTly-onset and familial Filipino breast cancer patients Utilizing BRCAPRO, to determine apriori and aposterioriBRCA112 carr ier probabilities of breast cancer patients Performing DNA Analysis aer extracting genornic DNA from whole blood of familial and early-onset breast cancer patients Knowledgeable in the role of RRCA I gene mutation and the pathogenesis of breast cancer Utilized silver staining procedure for visualizing AFLP-PCR profile of radiation-inducedvariants and compared it with profile obtained sing `P Performing ALP-PCR Mutational Screening of "Co y radiation-induced variants of onamentals and foliage plants (e.g., Hurraya exotica and Dracaena sanderiona)after extracting plant genorruc DNA using ethanol precipitation or plant DNAzol as part of te IAEA TC Project PHI/5/027 Participated in reporting accomplishments for te IAEA TC Project on Mutation Breeding Studied te effect of different doses of Radiation on te proteins of Vigna ungulculaia Utilized 2-inelhyl DOPA as sbstrate to detect tyrosinase hrough production of dopachl'Orne found to have a peak at 484 nm Determined radioresistance of P. acruginosautilizing different doses of y-rays Drafted a synthesis of te recent trends in Hepatitis C Virus HCV) studies Performed a initial sudy on te tilization of AtB and AtC insect mosquito) cell lines in ile propagation of dengue virus and sbsequent detection usirig RT- PCR nd agarose gel clectrophoresis Determined CTT Tplex (CSFIPO, THOI, TPOX) STR loci fingerprint in a Filipino (Bicol) population utilizing PCR and PAGE vsualized hrough silver stainingof DNA bands Performed TXRF to detect trace elements present in isolated Gs-protein Performed gel electrophoresis (SDS-PAGE) as an initial sep in he isolation of Gs-protein of ile Oriental fruit fly (Bociroceraphilippinunsts) and sbsequent Iyophilization of isolated Gs-protein for electrophoretic-rerunning purposes Performed chromophore experiment and tyrosinase assay (using oxidized L-tyrosine as the substrate) for the Pupal homogenate of B. philippmenw to determine other possible effects of irradiation a 100 Gy Performed Bradford assay to determine aount of Gs-protein per pupa of B. philippinensis Performed human peripheral blood ymphocyte isolation and sbsequent trichloroacetic acid DNA extraction to determine the HGPRT mutation indices of Valerizuela and Las Pirlas residents followed by performing statistical calculations to analyze te database obtained Health Pysics Environmental Participated as Pilippine Representative in te IAEA/RCA Poject Formulation Meeting on ]improving Regional Capacity for Assessment, Planning ad Responding to Aquatic Environmental Emergencies, 21-25 July 2003 Astralia Produced contour, wirefirarrie, and post aps of inarine radioactivity for te whole Asia-Pacific Region sing Surficr 7 for Windows s pail ofthe compilation of te Asia-Pacific Marine Radioactivity Database (ASPAMARD I and 2) Performed environmental and radiological surveillance in (a) former US Bases: Clark ad Sbic, (b) anufacturing corporation in the NCR, ad (c) oillerareas of interest in tile Pilippines (eg_ Palawan, Tanay, and Subic) sing the Berkeley Nucleonics Surveillance and Measurement Systems (BNC SAM935) coupled With QUantuin MCA Gold Soflware for quantitative radionuclide analysis and sared knowledge with colleagues Performed sediment sampling sediment core grab sampling) in Sbic Bay, murtian Bay Palawan), and Sulu Sea Performed initial study on radionuclide igration in Kara Sea sing tree-dimensional modeling (FLESCOT) as part of OJT on Dose Assessment in Battelle Pacific Northwest National Laboratory, Richland, WA, USA Environmental Srveillance within PNRI. at the perimeter of PNRI, and 25 km radius of PNR1 sing file igh pressure ionization camber and carborne y spectrometer (Exploranium 650) to easure abient y-radiation dose rate Participated as member of the Secretarial in the IAEA Project Formulation Meetings on ) Marine Coastal Environment and Its Pollution, 23-27 February 1998 Pilippines and 2) Reference Asian Man, Phase 11,June 1996 Pilippines Knowledgeable in the preparation of mixed radionuclide standards to be sed for HPGe detector Advisorship Supervised and trained (year-, month-, or week-long) on-tlie-job-training and tesis students both local and international- graduate, undergrad. or igh school students) regarding vst range of laboratory echniques and topics Computer Work Gathered PDF files offinportant scientific primary journal articles by downloading from te internet and interacting with the corresponding aLlihors Gathering ofhuge aount ofscientific data(i.e., binary files) and literature fro te Interne trough UNIX ad Windms-based systerns Made vrious templates aving appropriate formulae to aid computation of data for both environmental ad somatic mutation projects and lso for tile different administrative ad euipment request forms of te Health Pysics Research Section (HPRS) Gathered scientific research data through CD-ROM and University libraries (UPEPSF) Computed statistical antities concerning, srident needs at the national scale (UP OCG) Vi Administrative Fine-tuned HPR procurement and onthly stock requisition from 1997-present by
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