1996 Fiscal Year Final Research Report Summary
A diagnostic study on cardiomyopathy by using DNA analysis in the autopsy cases of sudden death and its application to a medicolegal judgment
| Project/Area Number |
07670498
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Legal medicine
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| Research Institution | Yokohama City University |
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Principal Investigator |
FUJIWARA Satoshi Yokohama City University, School of Medicine, Professor, 医学部, 教授 (20173487)
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| Co-Investigator(Kenkyū-buntansha) |
INAMURA Keiji Yokohama City University, School of Medicine, Assistant, 医学部, 助手 (60117736)
AIHARA Hitsunori Yokohama City University, School of Medicine, Assistant, 医学部, 助手 (80046086)
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| Project Period (FY) |
1995 – 1996
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| Keywords | Molecular Biology / Cardiomyopahy / beta-Myosin Heavy Chain Gene / Sudden Death / Forensic Pathology / DNA / PCR-SSCP / Point Mutation |
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| Research Abstract |
Methods
In each case which was autopsied within 24 hours after sudden cardiac death, a 10 ml blood sample was freezed at -80ﾟC and two pieces of the left cardiac ventricle and two ones of the right ventricle were fixed in 10% formaline solution.
A blod sample was defrosted and DNA was extracted from isolated nuclei of peripheral white cells by procedures of sodium iodide method. A PCR amplification of exons 13 in human beta-Myosin Heavy Chain gene was done by using a set of primers. PCR-amplified fragments were broken by Dde I restriction enzyme and resolved on a 10% polyacrylamide gel run at room temperature. The gel was stained with ethidium bromide and gel bands were identified under ultraviolet light.
Each piece of the cardiac ventricle was stained with hematoxylin-eosin (H-E) and azan stains and examined histopathologically.
Results
In the first case (59-year-old male), the heart weighed 920 g. Next, in the 56-year-old male, the heart weighed 590 g. In the 63-year-old male, the one did 700 g. In the 48-year-old male, the one did 530 g. In the 27-year-old male, the one did 450 g. In the 65-year-old male, the one did 920 g. In the 25-year-old male, the one did 680 g. In the 48-year-old male, the one did 400 g.
In a DNA analysis, 161 basepair fragment was identified and 129 basepair fragment was not in each case. In histopathological study on the cardiac ventricles, we found myocardial disarray.
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