The overall objectives of she research are: (a) the detailed biochemical characterization of the glycoconjugates of the mammalian bladder epithelium; (b) to determine the role of the bladder mucosal glycoconjugates in the pathogenesis of urinary bladder disorders of women such as interstitial cystitis, urinary tract infections and urinary incontinence. Because of the limitations of the availability of human bladders, parallel basic studies on both human and rabbit bladders will be conducted to elucidate the biochemical nature of mammalian bladder glycoconjugates. The focus will be on mucins, to test the hypothesis that they, rather than the other ladder glycoconjugates (proteoglycans/glycosaminoglycans, N-linked glycoproteins and glycolipids), play a crucial role in the protection of bladder against invasion by pathogenic microorganisms and injury from 'toxic' substances in the urine. Previously, we have discovered and partially characterized a mucin glycoprotein (named epitectin) which is a component of human urothelium and which can be readily isolated from human urine. The specific experiments proposed are: 1) to determine the glycoconjugate composition as well as to purify and biochemically characterize the mucin(s) of rabbit bladder mucosal layer; 2) to investigate the relationship between rabbit urine and bladder glycoconjugates. The information gathered will enable us to determine whether all human bladder glycoconjugates are likely to be represented in human urine (a readily available source for research); 3) quantitative analysis of mucins and glycosaminoglycans of urine from patients with various bladder disorders and normal adult controls. Epitectin levels will be estimated by ELISA, other mucins and glycosaminoglycans will be quantitated by gel and cellulose acetate electrophoresis; 4) to biochemically characterize the mucin(s) and glycosaminoglycans purified from urine of interstitial cystitis female patients and bladder disease-free women; 5) to examine normal and pathologic human bladder glycoconjugates by immunocytochemical and metabolic labeling techniques using biopsy specimens; 6) to measure the levels of glycosaminoglycan and mucin catabolic enzymes in the urine of patients with bladder disorders and of normal adult controls and determine whether the decreased concentration of urinary glycosaminoglycans in IC patients noted by other investigators could be the result of increased degradation.
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