1989 Fiscal Year Final Research Report Summary
Role of Urinary Acid Glycosaminoglycans on Calcium Oxalate Crystal Growth
| 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 Ehime University, Associated Professor, 医学部, 助教授 (40108379)
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| Co-Investigator(Kenkyū-buntansha) |
KIN Masahiro Ehime University, Assistant, 医学部附属病院, 助手 (20214926)
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| Project Period (FY) |
1987 – 1989
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| Keywords | Urinary glycasaminoglycan / calcium oxalate crystal / inhibitory effect / gel column / anion-exchange chromatography |
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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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