DEPARTMENT OF PATHOLOGY & MICROBIOLOGY SEED MONEY PROGRAMME FOR RESEARCH DEVELOPMENTPROPOSAL FORM* Part 1. ADMINISTRATIVE INFORMATION Please print or type in English Full name of the Principal Investigator(s) and Mentors Surname:MANSOORI Residency Training:FIRST YEAR First name(s) HUMA Year of Telephone Ext:1548 Fax or Email: (If different from AKUH) Title of the Project (120 characters maximum) ESTIMATION OF FREQUENCY OF RARE G6PD MUTATIONS IN PAKISTANI POPULATION THROUGH SEQUENCING OF EXONS 2-8 OF G6PD GENE Funds Requested (US $) 1000$ Proposed start of Project: As soon as possible after the approval of grant Expected duration of Project: ONE YEAR 1 . is a Resident at this department. I agree to abide by them. Responsible Administrative Authority (Departmental Head) Signature:___________________________ Department :Pathology and Microbiology Date: 24th May2011 Post: Resident 2 .Acceptance of general conditions by the Principal Investigator I have read the project for the submission for seed money funds which were provided with this proposal form and. the work will be accommodated and administered in the Department in accordance with the general conditions stated above. I shall be actively engaged in the project.HUMA MANSOORI. if support is granted. if my application is successful.. Dr/Mr/Mrs/Ms.. Signature of the Principal Investigator: _________________________ Date:24th May 2011 Signature of Mentor:_____________________________ Date :24th May 2011 _____________________________ Date ____________ _____________________________ Date ____________ Declaration of Department endorsement I confirm that I have read this application and that.. I also confirm that the Principal Investigator. statistical analysis.5 ml Case of 500 . 250ul (50 lanes)Cat # G2101.29 Rs 8000/= Rs 6000/= Rs 4000/= Rs 12000/= $94.Part II..17 Other Expenditures (Specify e.g.8 $235.58 $47 $141.11 $58.1 Budget request $1000 SUPPLIES Eppendorf Tubes 0.Promega Agarose Freezer boxes Primers(3set) DNA Sequencing(6 pairs of primers) 5 $ per sample $58 Rs 25000/= Rs 5000/= Rs 20000/ $294.1-10 ul cat # 161632 Gilson 100bp DNA Ladder. BUDGET: 2. data management. transport) Total Other Expenditures 3 .Eppendorf Cat # 951010057Eppendorf Taq Polymerase Enzyme D 10 Tips (10 packs /96 tips) 0.11 $70. 058 $999.08 4 . GRAND TOTAL Rs 80. 2.2 Budget justification Briefly relate each item in the budget to the activities outlined in the research proposal Eppendorf tubes.Taq polymerase enzymes.freezer boxes and primers are required for PCR while 6 pairs of primers for confirmation of PCR results.gml eppendorf. give the name of the organization(s) and summarize the amount and duration of the support. with dates Yes O No √O Is this or a substantially similar proposal currently being considered elsewhere? if "yes". 2.D10 tips.100bp DNA ladder.3 Other support for the proposed project (do not exceed the space provided for each item) No Is this research currently being supported by any other funding agency or by industry? if "yes".1. by what organization(s)? By what date is a decision expected? Yes O No √O 5 .agarose. 4 Other Research Activities of the Principal Investigator ( Please give titles of the projects) Nil 2. 2.5Other grants applied for previously (please give title) Nil 2.6Any grants awarded (please give duration and details of research award) Nil 6 . Mutational analysis of G6PD gene as identified through this research work can serve as a useful diagnostic tool to screen and diagnose G6PF deficiency in cases as described above. during active hemolysis and post transfusion. Discuss how the parent department will benefit from this research G6PD levels cannot be evaluated properly through routine G6PD assay in female carriers. PART III. PROJECT LINKS 3.2. Nil 3.1 Collaboration with other Research Projects and Institutions.The research work will be shared through abstracts during scientific meetings and publications which will increase the scientific database on behalf of department. 7 . This research is promising to reveal rare or novel G6PD genes which may act as a database for future work in G6PD deficiency for our country. Results of the study were as follows: G6PD Mediterranean (563C-T) was the most common genetic variant (78%) followed by G6PD Chatham (1003A-G) and G6PD Orissa (131C-G) showing respectively a frequency of 5 and 0. which do not shift the open reading frame.. Further to this credence it is necessary to identify rare or novel G6PD variants in diverse multi-ethnic population in Pakistan on behalf of department EXPERIMENTAL DESIGN: Descriptive study was conducted in 2006-2008 at AKU to identify G6PD deficient individuals through G6PD quantitative assay. The latter is significant for the survival of red cells as unchecked production of glutathione and hydrogen peroxide results in loss of red cell membrane integrity. rationale.during active hemolysis and post transfusion. SUMMARY (including brief description of objectives. Polymorphism in position 1311C/T was uniformly observed with all variants.The research work will be shared through abstracts during scientific meetings and publications which will increase the scientific database on behalf of department RELEVANCE: The information regarding rare and novel G6PD variants in our population remained fragmentary hitherto. RATIONALE:G6PD levels cannot be evaluated properly through routine G6PD assay in female carrriers.1. relevance and experimental design) Name of Principal Investigator::HUMA MANSOORI Title of the Project:Estimation the frequency of rare G6PD mutations in Pakistani population through sequencing of exons 2-8 of G6PD gene. We are proposing to work further on 43 uncharacterized samples by amplifying exons 28 through PCR and gene sequencing to identify the rare and novel G6PD genes in our population 8 . OBJECTIVE:To estimate the frequency of rare G6PD mutations in Pakistani population through sequencing of exons 2-8 of G6PD gene. This research is promising to reveal rare or novel G6PD genes which may act as a database for future work in G6PD deficiency for our country.. G6PD deficiency is the most common and is clinically significant. • SUMMARY:Background: Although several red cell enzymopathies have been discovered to date.Part IV: PROJECT DESCRIPTION 4. The sequence of entire G6PD gene has been determined and 140 mutations have been documented. Samples were subjected to RFLP-PCR to determine the common variants reported from South Asia. G6PD is the first and the most crucial enzyme of the hexose monophosphate pathway producing NADPH. Mutational analysis of G6PD gene as identified through this research work can serve as a useful diagnostic tool to screen and diagnose G6PD deficiency in cases as described above. Forty three or 17% of DNA samples remained uncharacterized. A number of Asian Countries have demonstrated a high prevalence of G6PD 563C-T. 276 subjects [237 males and 39 females] were enrolled in the research project funded through seed money grant [SM#050905]. Samples remaining uncharacterized were subjected to exon wise amplification of 9-12 exons through PCR with subsequent gene sequencing.7%. year) MBBS Dow University Of Health Sciences 2009 FCPS part 1 in Pathology in Sept 2010 3. 9 . Most recent posts held (type of post. title. Surname:MANSOORI First name(s): HUMA Date of birth:10th Jan 1986 Sex:Female Nationality:PAKISTANI 2. volume. dates) Nil 4. journal. page numbers. year] Nil Note: Use photocopies if additional Scientist involved. university or school. Please give full bibliographic reference [authors. institution/authority. CURRICULUM VITAE OF Principal Investigator (1 page maximum*) 1. Degree(s) (subjects. Recent publications: list only five most important and relevant publications or presentations over the last five years (papers in press or submitted for publication are also acceptable).