Tissue content of glycosaminoglycans in the diffuse idiopathic interstitial fibrosis patient.

Quantitative changes of glycosaminoglycans in lung tissues of patients with diffuse idiopathic interstitial fibrosis and patients with bronchial asthma were studied. The total amount of glycosaminoglycans in lung tissue of patients with diffuse idiopathic interstitial fibrosis increased in comparison to the lung tissue of those patients with bronchial asthma. The increase in tissue content of glycosaminoglycans was accompanied by an increase in dermatan sulfate level. The increases in total amount of glycosaminoglycans in relative proportion of dermatan sulfate were closely related to the clinical findings, including vital capacity of the lung, PaO2, serum lactate dehydrogenase, and to the pathologic pictures of the lung.

Extracellular matrix in hepatic granulomas of mice infected with Schistosoma mansoni. Qualitative and quantitative analysis.

To investigate the role of extracellular matrix molecules in the granulomatous inflammation, cercariae of Schistosoma mansoni were injected subcutaneously into BALB/c mice. Well-organized granulomas consisting mainly of stimulated macrophages and epithelioid cells developed in the liver at 11 weeks after infection, thereafter showing a tendency to heal. Fibronectin and heparan sulfate proteoglycan deposition appeared around parasite eggs, then increased distinctly at 11 weeks after infection, and subsequently diminished. Quantities of glycosaminoglycans and hydroxyproline in the egg lesion increased significantly at 11 weeks after infection. Thereafter, the amounts of glycosaminoglycans decreased, whereas hydroxyproline content did not. The data suggest that fibronectin and other macromolecules interact to form granuloma extracellular matrix, and that these extracellular events participate in the development of granulomatous inflammation and subsequent fibrosis induced by schistosome eggs.

Quantitative analysis of hyaluronic acid by high-performance liquid chromatography of streptomyces hyaluronidase digests.

The separation and quantitative analysis of streptomyces hyaluronidase ( HAase ) degradation products of hyaluronic acid (HA) by high-performance liquid chromatography are described. The substituted 4,5-unsaturated tetrasaccharide and hexasaccharide which result from the digestion of HA are quickly separated on a silica gel (Zorbax SIL) column. The assay of HA is linear between 5 and 250 micrograms of HA. This procedure is suitable for some weakly acidic glycosaminoglycan contaminants having similar properties to those of HA.

Isolation and characterization of peptidoglycans in urine from patients with mucopolysaccharidoses.

Urinary dermatan sulfate (DS) and heparan sulfate (HS) were purified from mucopolysaccharidosis patients. DS shows average mol. wt 8600-12,000 (approx. one half of tissue DS), iduronic acid content 99.1-99.6% (81.8% in tissue DS), core peptide mostly di- or tri-peptide (--Ser--Gly--or--Ser--Gly--Glu--). Molecular weight of HS ranged from 2500 to 20,000, averaging about 5000. Highly sulfated HS was found in the low molecular weight fraction, and no bound core peptide. By contrast, HS in the high molecular weight fraction bound one sulfate per repeating unit, which include core peptide.

Change of glycosaminoglycan in pus of patients with purulent pleurisy.

The glycosaminoglycan content in pus from patients with purulent pleurisy was studied. The uronic acid content rose in the first 4 hospital days, continued at a high level during hospital days 5-8, and then fell to a low level after 9 hospital days. Four glycosaminoglycans were isolated from the preparation; they were identified as hyaluronic acid, chondroitin 4-sulfate, chondroitin 6-sulfate, and dermatan sulfate. Hyaluronic acid was the main component and its relative proportion increased with increasing hospital days. The relative proportions of chondroitin 4-sulfate and chondroitin 6-sulfate were low during the first 4 day and during Days 10-21, whereas they were high during Days 5-9. The proportion of dermatan sulfate was high during the early hospital days, and thereafter decreased with increasing hospital days.

Glycopeptide in sputum from allergic asthma patients.

The glycopeptide and glycosaminoglycan content of sputa from allergic asthma, bronchiectasis, and common cold patients was assayed. The glycopeptide content was higher in sputum from allergic asthma patients than that in bronchiectasis and common cold patients, while no significant difference in the glycosaminoglycan content was detected among these materials. Fractionation of the glycopeptide by DEAE-cellulose column chromatography yielded four glycopeptide fractions at concentrations of 0.05 to 0.3 M NaCl from the allergic asthma samples, whereas it yielded three fractions at concentrations of 0.05 to 0.2 M NaCl from the bronchiectasis and common cold samples. They were characterized by increases in sialic acid and sulfate as the molarity of NaCl increased. Hexose was the main component and hexosamine was the next in each fraction from all materials. The increase in sputum glycopeptide in the allergic asthma samples was due to a large increase in sialic acid- and sulfate-rich glycopeptide.

Analyses of glycosaminoglycans in human lung cancer.

Studies were conducted on the total amount of glycosaminoglycans and glycosaminoglycan composition in adenocarcinoma tissue of human lung. The glycosaminoglycans were prepared by exhaustive proteinase digestion of adenocarcinoma tissue from human lungs and of lung tissue without pulmonary diseases taken at autopsy as a control. The glycosaminoglycan classes were characterized by biochemical, enzymatic, and electrophoretic methods. The presence of heparin, which has until now not been found in lung cancer tissue, was demonstrated on both carcinoma and control tissues. The levels of whole glycosaminoglycans were markedly increased in cancer tissue compared to the controls. The classes of glycosaminoglycans which increased in lung carcinoma tissue were predominantly chondroitin-4-sulfate and chondroitin-6-sulfate. Both hyaluronic acid and heparin were slightly increased in cancer tissue.

Composition of glycosaminoglycans in human pancreatic cancer.

Five glycosaminoglycans were isolated from tryptic digestion of both cancerous and normal tissues of the human pancreas and were assayed by determining the carbohydrate content of materials. Separation of these five polymers was achieved by Dowex 1-X2 column chromatography and fractionation with Benedict's solution. They were identified as hyaluronic acid, heparan sulfate, dermatan sulfate, chondroitin-4-sulfate, and chondroitin-6-sulfate, respectively. The total amount of glycosaminoglycans in cancer tissue increased in comparison to the controls. The increase in tissue content of glycosaminoglycans was accompanied by increases in chondroitin-4-sulfate and chondroitin-6-sulfate levels.

Glycopeptides in pus of acute pleurisy patients.

Glycopeptides were isolated from tryptic digests of pus from acute pleurisy patients. The hexose, hexosamine, and sialic acid contents rose over the first 2-4 days after admission to the hospital, continued at high levels for 5-8 hospital days, and fell to low levels after 9 hospital days. The course of duration after admission to the hospital was divided into three stages: 1-4 days after admission to the hospital, 5-8 hospital days, and 9-21 hospital days. Materials corresponding to these three stages were then collected for fractionation by DEAE-cellulose column chromatography. Fractionation of glycopeptides by DEAE-cellulose column chromatography yielded three glycopeptide fractions at 0.05 to 0.2 M NaCl. Column chromatography of crude material on DEAE-cellulose showed an increase in the 0.2 M NaCl fraction from the concentrate over a period of 4-8 hospital days due to a large increase in sialic acid-rich glycopeptide fraction.

Analysis of glycosaminoglycans in human prostate by high-performance liquid chromatography.

OBJECTIVE: To investigate changes in glycosaminoglycan (GAG) profiles in human prostatic tissue. MATERIALS AND METHODS: Seventeen human prostatic samples were examined; five were normal, six hyperplastic and six were cancerous. Prostatic proteoglycans were extracted with 4 mol/L guanidine-HCl containing protease inhibitors. After digestion of the freeze-dried proteoglycan extract with papain, the prostatic GAGs were purified. Total GAGs were measured by a modified dimethylmethylene blue (DMB) method. High-performance liquid chromatography (HPLC) was used to quantify the extracted GAGs. RESULTS: Six types of GAGs, i.e. chondroitin 4-sulphate (Ch-4S), chondroitin 6-sulphate (Ch-6S), dermatan sulphate (DS), chondroitin, heparan sulphate and hyaluronic acid, were analysed qualitatively and quantitatively. The total amount of GAGs was increased in hyperplastic and cancerous prostates, with the predominant components being DS and Ch-6S in these specimens. The Ch-S:DS ratio in cancerous prostate was significantly higher than that in normal and BPH tissue (P < 0.05). Moreover, chondroitin was increased in hyperplastic prostatic tissue (P < 0.01). CONCLUSIONS: These results suggest that an increase in chondroitin levels may be associated with hyperplastic change and an increase in the Ch-S:DS ratio may be related to the development of malignancy.