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  • 7/30/2019 FINAL PATH-MICRO Seed Money Grant

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    DEPARTMENT OF PATHOLOGY & MICROBIOLOGY

    SEED MONEY PROGRAMME FOR RESEARCH DEVELOPMENT

    PROPOSAL FORM*

    Part 1. ADMINISTRATIVE INFORMATION Please print or type in English

    Full name of the Principal Investigator(s) and Mentors

    Surname:MANSOORI First name(s) HUMA Year ofResidency Training:FIRST YEAR

    Telephone Ext:1548 Fax orE-mail:(If different from AKUH)

    Title of the Project (120 characters maximum) ESTIMATION OF FREQUENCY OFRARE G6PD MUTATIONS IN PAKISTANI POPULATION THROUGHSEQUENCING 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

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    Acceptance of general conditions by the Principal Investigator

    I have read the project for the submission for seed money funds which were providedwith this proposal form and, if my application is successful, I agree to abide by them. Ishall be actively engaged in the project.

    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, if support is granted, the work willbe accommodated and administered in the Department in accordance with the generalconditions stated above. I also confirm that the Principal Investigator,Dr/Mr/Mrs/Ms...HUMA MANSOORI.., is a Resident at this department.

    Responsible Administrative Authority (Departmental Head)

    Signature:___________________________ Date: 24th May2011

    Department :Pathology and Microbiology Post: Resident

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    Part II. BUDGET:

    2.1 Budget request $1000

    SUPPLIESEppendorf Tubes 0.5 ml Case of 500 ,EppendorfCat # 951010057Eppendorf

    Taq Polymerase Enzyme

    D 10 Tips (10 packs /96 tips) 0.1-10 ul cat #161632 Gilson

    100bp DNA Ladder, 250ul (50 lanes)Cat #

    G2101,Promega

    Agarose

    Freezer boxes

    Primers(3set)

    DNA Sequencing(6 pairs of primers)

    Rs 8000/= $94.11

    Rs 6000/= $70.58

    Rs 4000/= $47

    Rs 12000/= $141.17

    Rs 25000/= $294.11

    Rs 5000/= $58.8

    Rs 20000/ $235.29

    5 $ per sample $58

    Other Expenditures(Specify e.g., data management, statistical analysis,

    transport)

    Total Other Expenditures

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    GRAND TOTAL Rs 80,058 $999.08

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    2.2 Budget justificationBriefly relate each item in the budget to the activities outlined in the research proposal

    Eppendorf tubes,Taq polymerase enzymes,D10 tips,100bp DNAladder,agarose,1.gml eppendorf,freezer boxes and primers are required for PCRwhile 6 pairs of primers for confirmation of PCR results.

    2.3 Other support for the proposed project (do not exceed the space provided for eachitem) No

    Is this research currently being supported by any other funding agency or by industry?

    if "yes", 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", by what organization(s)? By what date is a decision expected? Yes O No O

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    2.4 Other Research Activities of the Principal Investigator ( Please give titles of theprojects)

    Nil

    2.5Other grants applied for previously (please give title)

    Nil

    2.6Any grants awarded (please give duration and details of research award)Nil

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    PART III. PROJECT LINKS

    3.1 Collaboration with other Research Projects and Institutions;Nil

    3.2. Discuss how the parent department will benefit from this research

    G6PD levels cannot be evaluated properly through routine G6PD assay in female

    carriers, during active hemolysis and post transfusion. 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.

    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.The research work will be

    shared through abstracts during scientific meetings and publications which will increase

    the scientific database on behalf of department.

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    4.1. SUMMARY

    (including brief description of objectives, rationale, relevance and experimentaldesign)

    Name of Principal Investigator::HUMA MANSOORI

    Title of the Project:Estimation the frequency of rare G6PD mutations in Pakistanipopulation through sequencing of exons 2-8 of G6PD gene.

    SUMMARY:Background:Although several red cell enzymopathies have been discovered todate, G6PD deficiency is the most common and is clinically significant. G6PD is the first and themost crucial enzyme of the hexose monophosphate pathway producing NADPH. The latter is

    significant for the survival of red cells as unchecked production of glutathione and hydrogenperoxide results in loss of red cell membrane integrity.The sequence of entire G6PD gene has beendeterminedand 140 mutations have been documented.. A number of Asian Countries havedemonstrated a high prevalence of G6PD 563C-T. which do not shift the open reading frame.OBJECTIVE:To estimate the frequency of rare G6PD mutations in Pakistani population throughsequencing of exons 2-8 of G6PD gene.RATIONALE:G6PD levels cannot be evaluated properly through routine G6PD assay infemale carrriers,during active hemolysis and post transfusion. Mutational analysis of G6PDgene as identified through this research work can serve as a useful diagnostic tool to screen anddiagnose G6PD deficiency in cases as described above.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.The research work will be shared throughabstracts during scientific meetings and publications which will increase the scientific database onbehalf of department

    RELEVANCE: The information regarding rare and novelG6PD variants inour population remained fragmentary hitherto. Further to this credence itis necessary to identify rare or novel G6PD variants in diverse multi-ethnicpopulation in Pakistanon behalf of departmentEXPERIMENTAL DESIGN: Descriptive study was conducted in 2006-2008 at AKU toidentify G6PD deficient individuals through G6PD quantitative assay. 276 subjects [237 malesand 39 females] were enrolled in the research project funded through seed money grant[SM#050905]. Samples were subjected to RFLP-PCR to determine the common variantsreported from South Asia. Samples remaining uncharacterized were subjected to exon wiseamplification of 9-12 exons through PCR with subsequent gene sequencing. Results of thestudy 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) showingrespectively a frequency of 5 and 0.7%.. Polymorphism in position 1311C/T was uniformlyobserved with all variants. Forty three or 17% of DNA samples remained uncharacterized.We are proposing to work further on 43 uncharacterized samples by amplifying exons 2-8 through PCR and gene sequencing to identify the rare and novel G6PD genes in ourpopulation

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    Part IV: PROJECT DESCRIPTION

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    CURRICULUM VITAE OF Principal Investigator(1 page maximum*)

    1. Surname:MANSOORI Date of birth:10thJan 1986

    First name(s): HUMA Sex:Female Nationality:PAKISTANI

    2. Degree(s) (subjects, university or school, year)

    MBBS Dow University Of Health Sciences 2009

    FCPS part 1 in Pathology in Sept 2010

    3. Most recent posts held (type of post, institution/authority, dates)

    Nil

    4. Recent publications: list only five most important and relevant publications orpresentations over the last five years (papers in press or submitted for publication arealso acceptable). Please give full bibliographic reference [authors, title, journal, volume, page numbers, year]

    Nil

    Note: Use photocopies if additional Scientist involved.

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