| Project/Area Number |
01571182
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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 |
Physical pharmacy
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| Research Institution | Higashi Nippon Gakuen University |
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Principal Investigator |
IKEGAWA Shigeo Higashi Nippon Gakuen University, Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (90111301)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIMURA Teruki Higashi Nippon Gakuen University, Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (60220737)
MAHARA Reijiro Higashi Nippon Gakuen University, Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (30192347)
TOHMA Masahiko Higashi Nippon Gakuen University, Pharmaceutical Sciences, Professor, 薬学部, 教授 (30001043)
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| Project Period (FY) |
1989 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1991: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1990: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1989: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Congenital biliary atresia / Bile acids / Radioimmunoassay / High-performance liquid chromatography / Conjugated 1beta-hydroxycholic acid / Mass screening / 化学発光 / 3α-hydroxysteroid dehydrogenase / 1β-hydroxycholic acid / 固定化酵素 |
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| Research Abstract |
To develop a diagnostic technique for congenital hepatobiliary disorders, qualitative and quantitative analysis of the conjugated 1beta- and 6alpha-hydroxy bile acids including common bile acids has been performed by high-performance liquid chromatography with chemiluminescence detection. Analysis of urine samples from newborns, normal women and women with pregnancy showed a large defderence in bile acid composition and conjugation mode, suggesting the distinctiveness of the bile acid metabolism during fetal and neonetal periods.
Anti-tauro 1beta-hydroxycholic acid antisera have been prepared by immunizing rabbits with bovine serum albumin conjugates, in which the glycine and beta-alanine derivatives of 1 -hydroxycholic acid were coupled at the C-24 position. The antisera had high affinity and specificity for conjugated 1beta-hydroxycholic acid ; cross-reactivities for glyco 1beta-hydroxycholic acid were 100% and those for glycine and taurine conjugates ranged from 43.1 to 0.23%. Concentrations of conjugated 1beta-hydroxycholic acid were determined by radioimmunoassay with the more specific antiserum to be significantly large amounts in newborns at 0 - 20 days after birth, ranging from 0.29 to 18.51 mug/ml, when compared to normal women (<0.12 mug/ml) and women with late pregnancy (<1.13 mug/ml).
To facilitate mass screening for congenenital biliary atresia, radioimmunoassay of conjugated 1 -hydroxycholic acid in dried blood spotted on filter paper was developed. The assay method was sufficiently sensitive and specific to determine the bile acid in one disc (3mm i. d., 4mul of whole blood). Preliminary tests suggested that the efficiency of this screening is acceptable, in which the levels of the patients (2.35 ng/disc) were significantly higher values than those of normal newborns (0.97 ng/disc).
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