Determination of urinary hexosamines for diagnosis of bladder pain syndrome.

INTRODUCTION AND HYPOTHESIS: Bladder pain syndrome (BPS) is a chronic disease characterized by urgency, bladder pain, and frequency, and urinary glycosaminoglycans are thought to reflect bladder epithelial deficiency in BPS. Sensitive and specific evaluation of total urinary glycosaminoglycans may be useful for the clinical diagnosis of BPS and its treatment. METHODS: A procedure for the simultaneous determination of glucosamine and galactosamine produced from urinary glycosaminoglycans has been performed in BPS patients and healthy subjects. RESULTS: The total content of urinary hexosamines in BPS patients significantly increased by ~130% with the increase in glucosamine greater than galactosamine. CONCLUSIONS: A significant increase in total hexosamines content and in particular in glucosamine belonging to urinary heparan sulfate was determined in BPS patients compared with controls. We propose HS and in particular its low-molecular mass fragments and glucosamine assay as useful markers for a biochemical diagnosis of BPS and for monitoring this syndrome.

High-throughput determination of urinary hexosamines for diagnosis of mucopolysaccharidoses by capillary electrophoresis and high-performance liquid chromatography.

Mucopolysaccharidoses (MPS) diagnosis is often delayed and irreversible organ damage can occur, making possible therapies less effective. This highlights the importance of early and accurate diagnosis. A high-throughput procedure for the simultaneous determination of glucosamine and galactosamine produced from urinary galactosaminoglycans and glucosaminoglycans by capillary electrophoresis (CE) and HPLC has been performed and validated in subjects affected by various MPS including their mild and severe forms, Hurler and Hurler-Scheie, Hunter, Sanfilippo, Morquio, and Maroteaux-Lamy. Contrary to other analytical approaches, the present single analytical procedure, which is able to measure total abnormal amounts of urinary GAGs, high molecular mass, and related fragments, as well as specific hexosamines belonging to a group of GAGs, would be useful for possible application in their early diagnosis. After a rapid urine pretreatment, free hexosamines are generated by acidic hydrolysis, derivatized with 2-aminobenzoic acid and separated by CE/UV in ∼10min and reverse-phase (RP)-HPLC in fluorescence in ∼21min. The total content of hexosamines was found to be indicative of abnormal urinary excretion of GAGs in patients compared to the controls, and the galactosamine/glucosamine ratio was observed to be related to specific MPS syndromes in regard to both their mild and severe forms. As a consequence, important correlations between analytical response and clinical diagnosis and the severity of the disorders were observed. Furthermore, we can assume that the severity of the syndrome may be ascribed to the quantity of total GAGs, as high-molecular-mass polymers and fragments, accumulated in cells and directly excreted in the urine. Finally, due to the high-throughput nature of this approach and to the equipment commonly available in laboratories, this method is suitable for newborn screening in preventive public health programs for early detection of MPS disorders, diagnosis, and their treatment.

Increased urinary excretion of acidic mucopolysaccharides in exophthalmos.

The excretion of mucopolysaccharides normally found in urine (chondroitin, chondroitin sulfates A and C, keratosulfate, and heparitin sulfate) is increased approximately twofold in patients with progresive exophthalmos. A threefold elevation of total serum mucopolysaccharides is also found. These increases are unrelated to thyroid function.

2-Acetamidoglucal, a new metabolite isolated from the urine of a patient with sialuria.

A new metabolite, namely 2-acetamidoglucal, has been found in the urine of a patient with sialuria in addition to the metabolites N-acetylneuraminic acid, N-acetylmannosamine, N-acetylglucosamine and 2-deoxy-2,3-dehydro-N-acetylneuraminic acid reported earlier. the structure has been identified by mass spectrometry and 360 MHz proton nuclear magnetic resonance spectroscopy and verified by synthesis. All accumulated compounds fit into the metabolic pathway for the biosynthesis of CMP-N-acetylneuraminic acid. Sialuria is discussed in terms of a failure of regulation of UDP-N-acetylglucosamine 2-epimerase.

Urinary excretion of glycosaminoglycans in disseminated neoplasm.

Urinary glycosaminoglycan excretion was studied in 24 cases of disseminated neoplasm, 12 of which had unequivocal evidence of skeletal involvement. Urinary hydroxyproline, cetylpyridinium chloride (CPC)-precipitable uronic acid, and CPC-precipitable hexosamine were expressed as a ratio to urinary creatinine. Glycosaminoglycans contained in urine concentrated x 1000 by vacuum-dialysis were separated by electrophoresis on cellulose acetate and stained with alcian blue. Of the 12 cases with clear evidence of skeletal involvement, eight (66%) showed elevation of serum alkaline phosphatase, five (42%) showed elevation of urinary hydroxyproline, and three (25%) showed elevation of urinary uronic acid. It is concluded that urinary uronic acid is not a sensitive index of skeletal involvement in disseminated neoplasm. The most striking feature of the study was the identification of a well-defined fraction indist inguishable from hyaluronic acid in seven (58%) of the cases with evidence of skeletal involvement. Hyaluronic acid is not normally identifiable in adult human urine. The hyaluronic acid excretors showed more consistent biochemical evidence of bone disease (elevation of serum alkaline phosphatase and urinary hydroxyproline) than the non-excretors. The possibility that the urinary hyaluronic acid is derived from degradation of skeletal hyaluronic acid is discussed. An alternative explanation is that the hyaluronic acid is derived from neoplastic cells as part of a reversion of glycosaminoglycan synthesis to a more ;fetal' state, a glycosaminoglycan counterpart of the production of oncofetal antigens by neoplastic cells.

Significance of variation in turnover of glycated albumin on indices of diabetic control.

We measured the half-time of disappearance of 125I-labelled glycated albumins in a rat model of diabetes with continuous infusion of physiological saline and insulin. Our results indicate that (i) in non-diabetic rats, continuous infusion of saline per se did not affect the concentrations of glucose or of fructosamine, and the half-time of disappearance of albumin was unaffected by degree of glycation; (ii) hyperglycaemia (mean plasma glucose concentration of 18-27 mmol/l) caused a small but significant increase in half-time of labelled glycated albumin disappearance from a mean of 42 h to a mean of 47 h; (iii) this effect of hyperglycaemia outweighed any effect of increase in albumin excretion detected in poorly controlled diabetic rats without infusion. We conclude that the effect of hyperglycaemia in slowing turnover of glycated albumin is likely to be insignificant in relation to its effect in promoting glycation, and may be species-dependent. However, in nondiabetics, variation of turnover of glycated albumin may well be significant in explaining the wide interindividual variation in concentration of glycated protein.

Glycosaminoglycan fractions in normal human urine.

A study has been made of glycosaminoglycans in normal urine to determine which are non-dialysable, which are ultrafilterable, and which are precipitable with cetypyridinium chloride or alcohol. The main fractions present in human urine are: (1) High molecular weight material which is non-dialysable and precipitated by alcohol and by cetylpyridinium chloride. (2) High molecular weight material which is non-dialysable and precipitated by alcohol but not by cetylpyridinium chloride. (3) Low molecular weight material which is ultrafilterable and precipitated by alcohol and by cetylpyridinium chloride. (4) Low molecular weight material which is ultrafilterable and precipitated by alcohol but not by cetyipyridinium chloride. The heavy losses of glycosaminoglycans on dialysis have led us to conclude that this method should not be used to study the excretion of these polymers and that direct precipitation with cetylpyridinium chloride, which can be used to isolate both non-dialysable and ultrafilterable macromolecular fractions relevant to the diagnosis of the mucopolysaccharidoses, is the method of choice.

Effect of bioflavonoids on the urinary excretion of hydroxyproline, hydroxylysyl glycosides and hexosamine in adjuvant arthritis.

The effects of (+)--Catechin (AC) and 0--(beta hydroxyethyl) rutosides (HR) on the urinary collagen metabolites were studied up to 49 days in rats with adjuvant-induced arthritis. The elevated levels of urinary total, non-dialysable and dialysable hydroxyproline, hydroxylysyl glycosides and total hexosamine in the arthritic animals were found to be slightly decreased in the acute phase and significantly decreased in the chronic phase of the disease due to the administration of bioflavonoids. Of the two bioflavonoids tests, CA was found to afford more protective action than HR.

[Effect of age on the urinary excretion of mucopolysaccharides]. / O vliianii vozrasta na ékskretsiiu mukopolisakharidov s mochoi

Content of mucopolysaccharides was studied by a test for hexuronic acids and hexosamines in day urine of men and women at the age of 20-80, and older. Mucopolysaccharides were precipitated with ethanol. In urine content of mucopolysaccharides was slightly increased up to the age tof 40-54 and later on it was decreased. Content of acid mucopolysaccharides was lower in urine of women aged 20-29 than in urine of men of the same age group.

[Assessment of criteria for forming follow-up groups of patients at high risk for ulcerogenesis in ambulatory care]. / Otsenka kriteriev formirovaniia grupp nabliudeniia sredi lits s faktorami vysokogo riska ul'tserogeneza v usloviiakh polikliniki.

A method of clinico-laboratory examinations to reveal persons at high risk for the development of duodenal ulcer, suitable for wide use during prophylactic medical screenings was devised. The rise of the levels of serum pepsinogen-I, pepsin and hexosamines in the urine, being of prognostic importance as applicable to ulcerogenesis, was the most significant indicator in screening risk group patients. During 3 to 5 years of the screened group follow-up and carrying out health measures, peptic ulcer was ascertained in 8.4% of the patients with chronic gastroduodenitis. Of these, 66.6% had initially suffered from pylorobulbitis. It is shown that there is a real opportunity of preventing ulcer formation in patients with chronic primary gastroduodenitis under outpatient conditions.