The Occurrence of Acid Mucopolysaccharides in Human Leukocytes and Urine 1

Abstract

Acid mucopolysaccharides have been found in urine (1) but have not been consistently demonstrated in the blood of normal subjects. Bassiouni (2) recently isolated a substance from leukocytes which he noted to be similar to chondroitin sulfate. In our laboratory leukocytes have been found to contain appreciable quantities of a substance resembling chondroitin sulfate. A direct relationship between this substance and the similar substance found in the urine has not been established. METHODSSubjects. Human subjects were used. These were hospital and laboratory personnel and patients who were unselected except for the omission of individuals with primary hematologic, metabolic, or rheumatic diseases.Preparation of leukocytes. Siliconed needles, syringes, and glassware were used throughout the preparation until the step in which the cells were deliberately disintegrated. Venous blood (40 to 70 ml.) was drawn and distributed immediately in 10-ml. amounts to tubes containing 1.0 ml. of 5 per cent sodium citrate in a balanced salt solution.2 The red blood cells were sedimented rapidly by mixing 2.5 ml. of 5 per cent dextran with the blood in each tube.3 The supernatant leukocyte-platelet suspension in plasma was centrifuged at 85 g for 7 minutes, and the sedimented leukocytes were washed gently one time in 11.0 ml. of physiologic saline. Following recentrifuging, they were then pooled in 20 ml. of saline, counted, made up to 25 ml. with saline in process of transferring to a Raytheon Sonic Oscillator, and oscillated at 9 Kc. per second for 8 minutes.Isolation of acid mucopolysaccharides from leukocytes.

The Distribution of Acid Mucopolysaccharides in Cohn Fractions of Human Plasma Proteins1

“…Acid mucopolysaccharides behaving similarly to chondroitin sulfate on paper electrophoresis and paper chromatography have been demonstrated previously in human leukocytes (1,2). A chondroitin sulfate-like substance has been recovered also from blood platelets of rats (3).…”

The Effect of Inflammation on the Hexuronate-Containing Polysaccharides of Human Plasma1

“…Samples of the material recovered were compared on paper chromatography with commercial chondroitin sulfate (General Biochemicals, Inc.) and with heparin (Liquaemin®g, Organon, Inc.), using methods previously described (8). Metachromasia developed with toluidine blue.…”

Total Nondialyzable Solids (Tnds) in Human Urine. I. The Amount and Composition of TNDS From Normal Subjects1

“…Similarly, hexuronic acid (26,27) and hexosamine (28,29) exist in urine in both dialyzable and nondialyzable4 forms; accordingly, the detection and estimation of these compounds in the TNDS will depend to some extent upon the conditions of dialysis. It is impossible to determine by conventional techniques the exact amount of hexuronic acid in the TNDS since it is relatively small, particularly susceptible to hydrolytic destruction, and is also progressively removed by dialysis.…”