Clinico-Biochemical and Molecular Studies of Purine Nucleoside Phosphorylase Deficiency
| Project/Area Number |
62570444
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pediatrics
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| Research Institution | Nihon University School of Medicine |
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Principal Investigator |
AKATSUKA A. Instructor, 医学部小児科, 助手 (60183133)
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| Co-Investigator(Kenkyū-buntansha) |
IWASE M. Instructor, 医学部小児科, 助手 (90223396)
SAKIYAMA T. Assistant Professor, 医学部小児科, 講師 (20130510)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1988: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1987: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | PNP deficiency / Northern blot / PCR / Western blot / invitro translation / 点突然変異 / PCR法 / point mutation / PNP合成能 / PNP活性 |
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| Research Abstract |
Purine nucleoside phosphorylase (PNP) deficiency is characterized by severe comgined immunodeficiency. Although cases of adenosine deaminase deficiency have been reported, this first case has been the only one ever reported in Japan. The patient was a 7-year-old girl who had developed recurrent respiratory infections since 3 years old and died from varicella infection. She had significantly low levels of serum uric acid and reduced T-lymphocytes function, which lead us to examine the PNP activity in erythrocytes. The activity was only 4.8% of the controls. In addition, PNP activity in the parents showed only about 50% of normal controls. The residual PNP in liver was immunologically cross reactive to the antibody which was determined by Ochtalony and Western blotting. The physico-chemical characteristics of PNP activities from the patient's liver showed no remarkable difference when compred to that of the controls except for the slightly different optimal pH. To determine the reason why the PNP activity in the patient was deficient, the molecular approach was necessary. For this purpose the mRNA was extracted from the patient's as well as control livers and Northern blotting method was used. The result showed that the patient had the same molecular RNA size and the intact volume of mRNA. Using the PCR method, the patient's genomic DNA was amplified and compared to controls' DNA by primer extension. These data strongly suggests that the patient has point mutation in PNP-DNA.
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Report
(3 results)
Research Products
(14 results)
