Excretion patterns of glycosaminoglycans and glycoproteins in normal human urine

Manley, G.; Severn, M.; Hawksworth, Jason

doi:10.1136/jcp.21.3.339

Cited by 64 publications

(34 citation statements)

References 20 publications

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“…The methods currently available for determination of GAG components in biological fluids tend to be lengthy and complicated, and are subject to various limitations. The most common techniques are the measurement of overall GAG content by photometrical quantification of uronic acids [14, 151, ion-exchange chromatography [16] and gel electrophoresis, followed by quantification by densitometry [17]. In the present study, a specific HPLC method was applied to determine the concentration and biochemical composition of GAG and different GAG components in the urine of G O patients in comparison with controls, and compared with the classical techniques.…”

Section: Introductionmentioning

confidence: 99%

HPLC Glycosaminoglycan Analysis in Patients with Graves' Disease

et al. 1997

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1. Orbital accumulation of hydrophilic, interstitial glycosaminoglycans (GAGs) and subsequent expansion of retrobulbar tissue lead to the clinical manifestation of exophthalmos in patients with Graves' eye disease. 2. A highly specific method to determine the concentration and biochemical composition of different GAGs was developed in order to obtain a sensitive test system for the activity of the disease. By means of this method, GAG excretion in 24 h urine collections of 56 patients and 21 controls was analysed by precipitation with cetylpyridinium chloride and potassium acetate in ethanol, followed by sequential enzymic hydrolysis with chondroitin AC lyase, chondroitin ABC lyase and hyaluronate lyase, with HPLC analysis of the resulting alpha, beta-unsaturated disaccharides by anion-exchange chromatography. 3. Concentrations of GAG, chondroitin sulphate A (CA), dermatan sulphate (DS) and hyaluronic acid (HA) were determined in patients and controls, with high recovery rates [72.2 +/- 5.3%, mean +/- SEM; detection limit, 4.2 micrograms/l (0.01 mumol/l)], revealing marked differences in urinary concentrations of total GAG and HA, as well as an elevation of CA in patients compared with controls. 4. Method sensitivity was 0.86 for patients with active Graves' eye disease, and 0.87 for patients with untreated ophthalmopathy, whereas specificity was 1.0 for patients with inactive disease. Patients with increased GAG concentration responded well to steroids and/or orbital irradiation (before therapy: GAG, 111.49 +/- 40.32; CA, 59.58 +/- 21.34; DS, 25.05 +/- 8.12; HA, 26.88 +/- 11.63 mg/24 h; during therapy: GAG, 54.22 +/- 10.94; CA, 20.52 +/- 4.58; DS, 17.65 +/- 3.46; HA, 16.05 +/- 3.69 mg/24 h), whereas GAG excretion increased markedly 2-3 months after stopping prednisone therapy in patients with still active eye disease (GAG, 109.9 +/- 10.51; CA, 63.8 +/- 7.34; DS, 24.1 +/- 5.07; HA, 22.0 +/- 6.28 mg/24 h). 5. This sensitive method determines the nature of renally excreted GAGs, reflecting the aberrant synthesis pattern of fibroblasts in patients with Graves' disease.

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Section: Introductionmentioning

confidence: 99%

HPLC Glycosaminoglycan Analysis in Patients with Graves' Disease

et al. 1997

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“…GAG excretion in urine from normal subjects has been extensively studied and urinary GAGS have been well characterized (1)(2)(3). The presence ofGAGS in human plasma has been demonstrated in several laboratories (4)(5)(6); however, these GAGs have not been fully identified and characterized mainly due to their low blood concentration and to a deficiency of adequate methodology.…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of glycosaminoglycans in human plasma.

Staprãns¹,

Felts²

1985

J. Clin. Invest.

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We have described methodology for the isolation and quantitation of glycosaminoglycans present in human plasma. Plasma glycosaminoglycans can be quantitatively adsorbed on a DEAESephacel ion exchanger and eluted with a salt gradient as two groups: a low-charge fraction and a high-charge fraction. The low-charge fraction consists of chondroitin sulfate with a low sulfate content and the high-charge fraction consists of heparan sulfate, chondroitin sulfate, and keratan sulfate (type I). We have determined the plasma concentration of each of these glycosaminoglycans in six normal human subjects.We have established that none of the glycosaminoglycans in plasma are covalently linked to plasma proteins. All are isolated as complexes with plasma proteins in noncovalent linkages. The glycosaminoglycans in the low-charge fraction are bound with high affinity to a single plasma glycoprotein by a lectin-type bond that can be disrupted by a simple glycoside. The highcharge fraction contains three major proteins and several minor proteins associated with the glycosaminoglycans by both lectintype and ionic bonding. The plasma proteins associated with glycosaminoglycans represent <0.5% of the total plasma proteins.Little is known about the physiologic role of the plasma glycosaminoglycans as components of metabolic processes. Because glycosaminoglycans have been implicated in lipid metabolism and atherosclerosis, we tested all of these compounds, isolated in free form, on the in vitro hydrolysis of triglycerides by lipoprotein lipase. Plasma heparan sulfate stimulated the rate of this reaction severalfold. All other plasma glycosaminoglycans were inactive. Thus, plasma heparan sulfate may play an important role in plasma lipoprotein metabolism.

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“…It was determined by electrophoresis that chondroitin sulfate was the major mucopoiysaccharide in the urine from normal individuals. Minor components of normal urine (heparan sulfate, keratan sulfate, dermatan sulfate, and products of partial desulfation of chondroitin sulfate) described previously [4,7] were not detected by this electrophoretic system. A minor Alcian blue-positive fraction (A' in Fig.…”

Section: Resultsmentioning

confidence: 63%

Mucopolysaccharides in Urine during Normal Human Development

1973

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ExtractTo determine the pattern of urinary mucopolysaccharides during normal human development, the urine samples from 92 normal individuals (aged from birth to 25 years) were analyzed for degradation ratio (large molecular weight mucopolysaccharides/fragments) and for the electrophoretic pattern of the different mucopolysaccharides present. The degradation ratio averaged 0.42 ± 0.21 (range 0.13-0.75) and was independent of age and sex. It was determined by electrophoresis that the major mucopolysaccharide was chondroitin sulfate. In the eight patients with a genetic mucopolysaccharidosis the degradation ratio was above 1.0 and electrophoresis revealed the presence of large quantities of dermatan sulfate and heparan sulfate. The two tests, therefore, permit distinction of urine from a normal individual from that of a patient with a genetic mucopolysaccharidosis. SpeculationOn the basis of studies done on cultured fibroblasts, it is apparent that the term genetic mucopolysaccharidosis encompasses a much broader base of cellular pathology than that previously considered. Normal values for urinary mucopolysaccharides throughout development in conjunction with cell culture studies are needed by clinicians trying to determine whether a patient with some but not all the stigmata suggestive oi a genetic mucopolysaccharidosis has a disorder of mucopolysaccharide metabolism.

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Excretion patterns of glycosaminoglycans and glycoproteins in normal human urine

Cited by 64 publications

References 20 publications

HPLC Glycosaminoglycan Analysis in Patients with Graves' Disease

HPLC Glycosaminoglycan Analysis in Patients with Graves' Disease

Isolation and characterization of glycosaminoglycans in human plasma.

Mucopolysaccharides in Urine during Normal Human Development

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