The study on excretion of hyaluronan (HA) in urine samples

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The study on excretion of hyaluronan (HA) in urine samples

 

Author: Wong, Wa-sang Watson
Title: The study on excretion of hyaluronan (HA) in urine samples
Year: 1999
Subject: Urine -- Analysis
Urine -- Examination
Kidneys -- Calculi -- Diagnosis
Hyaluronic acid
Glycosaminoglycans
Hong Kong Polytechnic University -- Dissertations
Department: Multi-disciplinary Studies
Dept. of Nursing and Health Sciences
Pages: ix, 101 leaves, [4] leaves of plates : ill. ; 30 cm
Language: English
InnoPac Record: http://library.polyu.edu.hk/record=b1477328
URI: http://theses.lib.polyu.edu.hk/handle/200/1368
Abstract: Urinary polyanionic macromolecules were recovered from urine samples of 10 normal controls and 10 stone formers by cetylpyridinium chloride (CPC) precipitation. CPC-polyanion complexes were analyzed individually. Separation of the urinary polyanions was monitored based on like mobilities to tissue derived reference GAGs on cellulose acetate electrophoresis and identified by susceptibilites to specific GAG degrading enzymes and chemical treatment. Chondroitin sulphate was identified as the major GAG subclass in the urinary polyanionic macromolecules recovered. The urinary GAG concentration in samples was determined by uronic acid assay. The average urinary GAG concentration of samples from normal controls was 2264 ug hexauronate/L (S.D.=+-2348 ug hexauronate/L), that was significantly higher than 657 ug hexauronate/L (S.D.=+- 395ug hexauronate/L) in that of stone formers (p=0.0185). Hyaluronan (HA) in urine samples can be determined as their unsaturated (Δ) disaccharide derivatives following digestion with hyaluronidase SD (from Streptococcus dysgalactiae). After the digestion products were separated, those disaccharide derivatives permit the determination of GAG distribution patterns by a weak anion exchange HPLC analysis. Two isocratic elutions form the same column (Econosphere NH2) separated urinary Δdi-nonSHA from Δdi-nonSCS and Δdi-monoSCS with high response sensitivity (0.71 V.s.ug-1, 0.25-20 ug) and good reproducibility (SD=+-0.12 min). Unsaturated Δ-HA tetra- and hexasaccharides were found precipitated along with the digestion products in ethanol which were identified during HPLC analysis. The response sensitivity due to these HA oligosaccharides were however much lower (0.12 V.s.ug-1, 0.6-4.6 ug). Urinary HA in each of the urinary extracts was determined as an aggregate of integrated responses generated from all unsaturated oligosaccharides of HA with them. Results of the urinary disaccharide analysis by HPLC indicated that HA was detected in all urinary extracts. There was no significant difference in HA concentration between urine samples of stone formers and normal controls. However, when taking the GAG concentration together for comparison, a statistical significant higher (p=0.043) percentage of HA within GAGs (HA / total GAG %) in urine extracts of on stone former group (11.56%) was observed when compared to that of normal control group (6.21%). Thus excretion of a lower GAG concentration accompanied by a high percentage of HA present appears to be a characteristic of stone formers which favor the calcium oxalate stone formation, since urinary HA in stone formers was found to be crystal-active. In that sense, urinary HA might be a very useful marker to identify individuals at risk of renal stone disease.

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