| Project/Area Number |
62570729
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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 |
Urology
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| Research Institution | Ehime University |
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Principal Investigator |
IWATA Hidenobu (1989) Ehime University, Associated Professor, 医学部, 助教授 (40108379)
松本、充司 アツシ (1988) 愛媛大学, 医学部, 助教授 (80110831)
西尾 俊治 (1987) 愛媛大学, 医学部, 助手 (20136328)
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| Co-Investigator(Kenkyū-buntansha) |
KIN Masahiro Ehime University, Assistant, 医学部附属病院, 助手 (20214926)
岩田 英信 愛媛大学, 医学部附属病院, 講師 (40108379)
松本 充司 愛媛大学, 医学部, 助教授 (80110831)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1989: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1988: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1987: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Urinary glycasaminoglycan / calcium oxalate crystal / inhibitory effect / gel column / anion-exchange chromatography / ヘパラン硫酸 / コンドロイチン硫酸 / エタノ-ル分画法 / 蓚酸カルシウム / 成長抑制 / GAG / 尿蛋白 / 酸性ムコ多糖 / 原発性副甲状腺機能亢進症 / 2次元電気泳動法 / ヒアルロン酸 / シュウ酸カルシウム / 結晶凝集 |
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| Research Abstract |
To evaluate the effect of urinary acid glycosaminoglycans(GAGs) in natural form on calcium oxalate crystal growth, we developed the method of isolation of urinary GAGs. The inhibitory activity on calcium oxalate crystal growth was examined using the 14C- oxalate-seeded crystal growth assay in a metastable solution.
First, normal control urine was precipitated by cetylpyridinium chrolide, which is a usual method for isolation of crude GAGs. Tamm-Horsfall mucoprotein was removed from the crude GAGs. The remaining sample was fractionated through a Sephacryl S-300 gel column and furthermore isolated using anion-exchange chromatography on DEAE-cellulose.
Second, cetylpyridinium chrolide was not used to collect the GAGs in natural form. After urine was cut off by a membrane of 30,000 M.W., the remaining urine was fractionated by anion-exchange chromatography on DEAE-cellulose and Sephacryl S-200 gel column.
Finally, ethanol method by Meyer was used after isolations of anion-exchange chromatography and gel column. Heparan sulfate(HS) was collected at 30% of ethanol concentration. Both HS and Chondroitin sulfate(Chs) were contained at 40%. Chs was collected at 50%.
We also examined the inhibitory effect of urinary macromolecular proteins on crystal growth. Our studies showed that urinary GAGs in natural form have high inhibitory effect on calcium oxalate crystals although the effect is smaller than that of urinary some kinds of proteins.
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