Serum Glycosaminoglycans Research Articles

The isolation and characterization of glycosaminoglycans in normal human serum

The optimization of conditions for the isolation and characterization of human serum glycosaminoglycans (GAG) is described, together with studies of the accuracy and reproducibility of the method. The principle of the method is proteolytic digestion of serum using papain followed by precipitation of GAGs from the digested sample with cetyl pyridinium chloride (CPC). The uronic acid level and electrophoretic separations can be obtained from a 5 ml serum sample. The mean CPC-precipitable uronic acid level in pooled normal serum was 10.8 mg l(-1) serum. Using enzymatic and chemical analysis the major serum GAG was shown to be chondroitin sulphate (CS). Two distinct electrophoretic fractions were identified both consisting of CS but differing in their degree of sulphation. Dermatan sulphate, heparan sulphate and hyaluronic acid were not detected.

A method for the determination of glycosaminoglycans in serum

A method for the determination of glycosaminoglycans in serum

Quantitative and qualitative characterization of serum and urine glycosaminoglycans in hyperthyroidism

Quantitative and qualitative characterization of serum and urine glycosaminoglycans in hyperthyroidism

Elevated chondroitin 6-sulfotransferase activity in fetal bovine serum

Assay conditions for chondroitin sulfotransferase in serum were established using a rapid and simple paper disk method. Sulfotransferase activities towards endogenous serum glycosaminoglycans and exogenously added chondroitin were determined for fetal, newborn and adult bovine sera. The results indicate that 3'-phosphoadenosine 5'-phosphosulfate (PAPS); chondroitin 6-sulfotransferase activity in fetal calf serum is several times that in newborn or adult bovine sera. The enzyme transfers sulfate onto position 6 of internal nonsulfated galactosamine units of endogenous and exogenous chondroitin. Sulfation Chondroitin sulfate Sulfotransferase

Serum glycoconjugates in children with schizophrenia and conduct and adjustment disorders

The serum glycoproteins represented by the individual protein-bound carbohydrate components and glycosaminoglycans represented by the hexuronic acid contents were determined in the sera of black and Caucasian normal children and children with diagnoses of schizophrenia, conduct disorder, and adjustment disorder. There were no race-related or sexrelated differences in glycoproteins and glycosaminoglycans in the sera of normal children. Although the serum glycosaminoglycans were significantly elevated in children with a diagnosis of schizophrenia, the levels were in normal range in children with conduct and adjustment disorders. All of the protein-bound carbohydrates were elevated in schizophrenic children. However, only arabinose and galactosamine were significantly elevated in children with a diagnosis of conduct disorder, while only galactosamine was elevated in children with adjustment disorder. The presence of arabinose in serum glycoprotein was confirmed by chemical ionization-mass spectrometry. The possible causes of the differential elevation of the glycoconjugates in psychiatric disorders in relation to the effect of stress and environment are discussed.

Increased glycosaminoglycan excretion after chymopapain injection of intervertebral discs

The urinary excretion of glycosaminoglycans during the 24 hours after intradiscal injection of chymopapain was found to be greater than during the 24 hours before injection when comparison was made either on the basis of a complete 24-hour urine collection or by the use of a glycosaminoglycan/creatinine ratio. Each individual in a group of 14 patients showed this increase after injection. In contrast, no significant increase over the pre-injection levels was detected in serum glycosaminoglycans during the post-injection time periods for which blood samples were available.

Measurement of serum glycosaminoglycans by laser nephelometry

A simple, sensitive micromethod for the assay of serum hyaluronic acid and chondroitin sulphates is presented. The method is based on the binding of the quaternary ammonium salt, cetylpyridinium chloride (CPC) to serum polyanions, and quantitation of the complexes by laser nephelometry. Measurement of the CPC complexes in serum before and after digestion with specific enzymes enables quantitation of hyaluronic acid and chondroitin sulphates in less than 100 μl serum. Using this technique, hyaluronic acid is detectable in a small number of normal human sera at concentrations up to 4 mg/l, and chondroitin sulphates are consistently present at concentrations ranging from 2 to 25 mg/l.

A semi-quantitative micromethod for the determination of free glycosaminoglycans in serum. Results from studies on serum of healthy children of various age and patients affected by mucopolysaccharidosis

A method is presented for the determination of free glycosaminoglycan (GAG) concentration in as little as 30 μl serum. By filtration of the serum through DEAE-cellulose paper, the free GAG fraction is selectively adsorbed and concentrated on a circular area of 15.9 mm 2; these GAG spots are stained with Alcian Blue. The relationship between the amounts of adsorbed GAG and the optical density of the Alcian Blue spots is linear with a certain range; e.g. for chondroitin sulfate (mixed isomers), from 0.25 to 1.00 μg. With this method — which we will refer to as the “DEAE Alcian Blue” method — we estimated the free GAG concentration in sera of individuals of various ages including newborns.

Acidic glycosaminoglycans in urine, serum and myocardium of aged patients with myocardial infarction

The acidic glycosaminoglycans in urine, serum and myocardial tissue of aged patients with myocardial infarction have been analyzed by two-dimensional electrophoresis on cellulose acetate membranes in order to study the mechanism of repair of myocardium after infarction. Patients tested 4 years or more after infarction showed excretion of urinary glycosaminoglycans within the range normally encountered for the aged. However, in cases of acute myocardial infarction, the excretion of glycosaminoglycans was found to be increased 3 to 4 times in the early acute phase and then gradually to decrease to within the normal range during recovery. A significant increase in excretion of low sulfated chondroitin sulfate was observed in the acute phase. Low sulfated chondroitin sulfate was the major glycosaminoglycan in sera from the patients, regardless of stages. In normal myocardial tissue, hyaluronic acid was the major component and heparan sulfate, chondroitin sulfate and dermatan sulfate were the minor components. In myocardium from patients with acute infarction, a decrease in heparan sulfate was observed, while in that of old infarction, dermatan sulfate was the main component and in advanced aneurysm, increase in chondroitin sulfate was observed along with dermatan sulfate. These findings suggest that the repair process of infarcted myocardium is very similar to that occurring during healing of hepatic fibrosis and damaged arterial walls.

Micromethod for the determination of glycosaminoglycans in the serum. results from the serum of healthy and varicose subjects

Serum glycosaminoglycans (GAG) were isolated from 1 ml of serum by a modification of the method of Y. Emura and T. Mukuda (J. Jap. Biochem. Soc., 45 (1973) 30). The extraction was carried out as quantitatively as possible and the hexuronic acids in the GAG were measured with m-hydroxydiphenyl. This method for determination of the GAG is relatively easy, easily reproducible and requires no more than two days. Its application in clinical chemistry may prove to be useful in the detection of diseases involving connective tissue. A study was carried out with serum samples from 34 patients suffering from serious varicose conditions, compared with samples from 30 healthy subjects. The mean concentration of GAG in the serum of healthy persons was 0.602 ± 0.066 mg per 100 ml of serum. In the varicose patients, this concentration reached a level of 0.751 ± 0.108 ( p < 0.0005). There were not, however, any significant variations between patients suffering from serious varices without trophic changes and those with comparable varices accompanied by trophic changes, such as phlebitis, thrombosis and ulcers.