| Project/Area Number |
10557149
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Keio University |
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Principal Investigator |
YOSHIMURA Yashimura Keio University School of Medicine, Dept of OB/GYN, Professor, 医学部, 教授 (10129736)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIMOTO Hitoshi Keio University School of Medicine, Dept of OB/GYN, Instructor, 医学部, 助手 (10212937)
MIYAZAKI Toyohiko Keio University School of Medicine, Dept of OB/GYN, Assistant Professor, 医学部, 講師 (20174162)
SUEOKA Kou Keio University School of Medicine, Dept of OB/GYN, Associate Professor, 医学部, 助教授 (90162833)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥13,000,000 (Direct Cost: ¥13,000,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1999: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1998: ¥9,100,000 (Direct Cost: ¥9,100,000)
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| Keywords | preimplantation genetic diagnosis / Duchenne muscular dystrophy / fetal angiography / Dystrophin / power Doppler |
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| Research Abstract |
In preimplantation genetic diagnosis (PGD) for X-linked genetic diseases, gender determination has been used to have clinically unaffected female baby. However, couples undergoing PGD by gender determination can not have boys by the selection procedure. Half of unaffected male embryos must be abandoned. The highly efficient diagnostic technologies is desired to develop for reaching the mutated genes instead of gender determination. We developed novel DNA extraction method "Spanning protocol" from single cell and disease-specific assay using multiplex nested PCR for Duchenne muscular dystrophy (DMD) and the combined genetic diagnosis of blastomere and polar body from the embryo. Three different DNA extraction procedures of (A), (B) and (C) were compared for efficiency ; (A) lysis in distilled water and freeze-thawing, then boiling, (B) a two-step lysis protocol involving an initial treatment in potassium hydroxide and dithiothreitol, and (C) Spanning protocol involving N-Lauroylsarcosin
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e solution. Amplification failure occurred in (A) : 47.5% , (B) : 16.25% and (C) : 5%, respectively. Spanning protocol was significantly more efficient as compared to all other methods. The efficiency of blastomere diagnosis was about 85%, whereas low false-negative rate was 5%. In final results, efficiency was increased to 99% while false-negative rate was decreased to O.1% by combined diagnosis of polar body with that of blastomere.
In another study, evaluated the clinical significance of "fetal angiography", the combined use of color and power Doppler ultrasound modalities to picture fetal vessels, from various angles. Transvaginal scanning was superior to transabdominal scanning in performing fetal angiography in pregnant patients with cephalic fetuses at the mid-trimester of gestation. Addition of fetal angiography to the conventional B-mode ultrasound was more effective in diagnosing most of fetal anomalies than the conventional B-mode alone. Another study was undertaken to determine whether introduction of B-flow, a newly-developed ultrasound modality, could lead to more understanding of the fetal placental circulation. We succeeded in three dimensional rendering of B-flow images of the fetal vasculature within the placenta for the first time. Although the quality of B-flow images of the placental vessels was satisfactory, there still remain several technical challenges to overcome for successful imaging of the fetal body vessels. However, future advances in this new technique will allow clinicians a better comprehension of the fetal vasculature. Less
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