Isoko Takazono

Quantitative analysis of hyaluronic acid by high-performance liquid chromatography of streptomyces hyalu-ronidase 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.

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 Benedicts 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.

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.

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.

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.

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.

Change of glycosaminoglycan in pus of patiens 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.