Urinary glycosaminoglycans in different phases of Balkan endemic nephropathy.

The relationship between glycosaminoglycans and beta 2-microglobulin, glycosaminoglycans and N-acetyl-beta-D-glucosaminidase as well as the relationship between the chondroitin sulfate/heparan sulfate ratio and SDS electrophoresis in the urine of subjects from the endemic area of Balkan nephropathy was studied in order to establish a method for early detection of this disease. The results show an unquestionable increase in urinary excretion of total glycosaminoglycans in subjects with or suspected of having Balkan endemic nephropathy while the chondroitin sulfate/heparan sulfate ratio was not statistically different between the groups studied. Thus, the chondroitin sulfate/heparan sulfate ratio cannot be used as a cheap and quick semiquantitative method for the diagnosis of early tubular damage in Balkan endemic nephropathy. However, the determination of total glycosaminoglycans in the urine of subjects from endemic areas proved to be valuable additional information helping with the diagnosis of Balkan endemic nephropathy.

Application of the monodimensional electrophoresis for the separation of glycosaminoglycans on Cellogel cellulose acetate strip.

A monodimensional electrophoretic method for the separation of glycosaminoglycans on Titan III Zip Zone cellulose acetate plate based on their different electrophoretic mobilities in barium acetate and different solubilities in ethanol was applied to the Chemetron electrophoretic equipment. Improved timing of individual steps of electrophoretic run, additional cooling and pressure must be introduced for optimal separation of glycosaminoglycans mixture (dermatan sulphate, heparan sulphate, hyaluronic acid, chondroitin-4-sulphate, chondroitin-6-sulphate and keratan sulphate) resulting in five well separated sharp bands. By all these changes in the original procedure of Hopwood and Harrison, the separation of chondroitin-4-sulphate and chondroitin-6-sulphate was not achieved. The modified procedure on Cellogel strip is suitable for the screening of mucopolysaccharidoses.

Effects of neuroleptic phenothiazines on the activities of aminotransferases and gamma-glutamyltransferase in serum and liver.

Serum alanine aminotransferase, aspartate aminotransferase and gamma-glutamyltransferase activities were monitored in psychiatric patients receiving normal doses of phenothiazine neuroleptics over a 30-day period. The first two enzymes showed slight initial increases and a subsequent return to normal, while the third showed a slight increase. In rats, dosage levels exceeding those used in human therapy produced much larger increases in the catalytic concentrations of all three enzymes in serum (1.4, 0.7 and 0.5 above the control value, respectively), and somewhat smaller increases in the liver homogenates of these animals.

Effect of the degree of hyperglycaemia on the catalytic activities of glycosidases in kidney and urine of diabetic rats.

The catalytic activities of N-acetyl-beta-D-glucosaminidase, beta-galactosidase and alpha-glucosidase in kidney and urine of diabetic rats were investigated in relation to the duration of diabetes, to the degree of constant hyperglycaemia and to the therapeutic control in the early stage of disease. The results suggest that the degree of constant hyperglycaemia and the duration of untreated diabetes are significant determining factors for the course of morphological changes. These changes are manifested as a decrease of the glycosidases in kidney (0.5 to 0.6 time the age-matched controls) and as moderate to severe enzymurias. Daily variation of blood glucose with inadequate insulin Lente therapy caused decreased N-acetyl-beta-D-glucosaminidase and beta-galactosidase activities in kidney as well as enzymuria. Since such changes can be correlated with histologically visible changes in the kidney, the measurement of these enzymes in urine is a simple way of monitoring the development of kidney damage in poorly controlled diabetes. When constant normoglycaemia was maintained for three weeks with insulin Ultralente in diabetic rats with a confirmed decrease of kidney glycosidases, the persisting morphological alteration of the organ was reflected by a urinary output of N-acetyl-beta-D-glucosaminidase.

Cytological and biochemical methods for the characterization of in vitro cultured cells.

The aim of our presentation was to show how we characterize cells cultured in monolayer system. Cytological and biochemical methods were used. Ovarian Krukenberg tumour fibroblasts were investigated and findings were correlated with normal human diploids (HDZ1) and with fibroblasts obtained from Blighted ovum. Cytomorphologically Malignancy associated changes in the tumour fibroblasts were found. Cytochemically acid phosphatase and alpha-naphtyl-esterase were positive (+++). PAS reaction was doubled in 18th passage. Cytogenetically normal human diploids were found. Biochemically enzymatic assay showed phosphopentose shunt is decreased in tumour fibroblasts and alpha-glucosidase and beta-galactosidase activities were significantly lower in these cells. A form of N-acetyl-beta-glucosaminidase fell during the investigation from normal 75% to lower percent (42% of the total activity). Much more parameters were obtained by different methods and Krukenberg tumour fibroblasts may be better understood. In vitro investigation makes a contribution to biomedical knowledge in cancer research.

A liver urocanase deficiency.

Two sisters with a rare inborn error of histidine metabolism resulting from urocanase deficiency are being presented. The more common form of familial histidinemia due to histidase deficiency is excluded. The urocanase deficiency is proven by demonstrating increased excretion of metabolites of the product of the urocanase enzyme action. Further, the strongest evidence for the urocanase defect rests on the demonstration of urocanase deficiency and normal histidase activity in liver.