Inhibition of aldose reductases from rat and bovine lenses by hydantoin derivatives.
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Biomedical subjects
Publications and source records attributed to I Miwa.
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gamma-Cyclodextrin was found to be hydrolyzed by human salivary and pancreatic alpha-amylases (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1) at appreciable rates. The optimum pH for the enzyme reactions at 37 degrees C in the presence of 0.1 M NaCl was at around pH 5, which was remarkably different from the optimum pH (pH 6.9) of the enzymes for starch. The Km value (2.9 mg/ml) of pancreatic alpha-amylase for gamma-cyclodextrin was smaller than that (5.3 mg/ml) of salivary alpha-amylase at pH 5.3, while the V value of the former was 3.7-times larger than that of the latter. The hydrolyses of gamma-cyclodextrin by both enzymes took place via the multiple attack mechanism. The degrees of multiple attack by salivary and pancreatic alpha-amylases for gamma-cyclodextrin at pH 5.3 were 2.0 and 1.1, respectively. The distribution of maltodextrins produced by hydrolysis of gamma-cyclodextrin by salivary alpha-amylase was suggested to be independent of the substrate concentration, while that produced by pancreatic alpha-amylase was presumably dependent on the substrate concentration.
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The localization of mutarotase in rat kidney was investigated by fluorescein-labelled and peroxidase-labelled antibody techniques, and by method of isolation of the nuclei and cytoplasm in non-aqueous solvents. In these immunohistochemical studies, mutarotase was almost exclusively recognized in the nuclei of epithelial cells of renal tubules and glomeruli in rat. The specific activity of mutarotase was found to be 1.5 times higher in the nuclei (122 units/g dry wt) than that in the cytoplasm (80 units/g dry wt) isolated with non-aqueous solvents. These results suggest that mutarotase may be involved in the metabolism of D-glucose in nuclei.
Effects of alpha- or beta-D-glucose on the respiration of germinated spores (only germinated spores not including swollen spores and elongated spores) of Bacillus subtilis and B. megaterium were studied. In our conditions, net amount of oxygen consumed by 10(10) germinated spores of B. subtilis per min after addition of alpha- or beta-D-glucose was 1.6 microgram or 6.6 microgram (beta/alpha = 4.13), while that by B.megaterium was 4.5 microgram of 6.8 microgram (beta/alpha = 1.51), respectively. However, the net amounts of oxygen consumed by 10(10) vegetative cells per min after addition of alpha- or beta-D-glucose were identical, for B.subtillis in both cases 443.0 microgram and for B.megaterium in both cases 604.4 microgram.
On aerobic incubation of rat cerebral cortex slices with anomers of D-glucose and with 2-deoxy-D-glucose (2DG) for 5 min, the disappearance of beta-D-glucose from the incubation mixture was greater than that of alpha-D-glucose and both anomers had a greater rate of disappearance than that of 2DG. In addition, there were significantly greater consumption of oxygen and production of lactate with the beta-anomer than with the alpha-anomer. In similar experiments with 3H-labeled D-glucose anomers and [1-3H]-3-O-methyl-D-glucose (3MG), the accumulation of [1-3H]-beta-D-glucose (up to 5 min) by rat cerebral cortex slices was greater than that of [1-3H]-alpha-D-glucose. Although initially lower than that of the anomers, the accumulation of [1-3H]-3MG increased at a greater rate and, by 5 min of incubation, was greater than that of both glucose anomers. This preferential accumulation was seen to disappear when the slices were preincubated with 2DG (hexokinase inhibitor) or when the temperature of incubation was reduced to 20 degrees C. Under those conditions the data with the glucose anomers were similar to those obtained with 3MG. Our data then suggested that the greater accumulation of beta-D-glucose than of alpha-D-glucose by the slices was probably not due to differences in transport through brain cell membranes but rather to the preferential metabolism of the beta-D-glucose.
The effects of D-glucose anomers on the germination of dormant spores of Bacillus megaterium QM B1551 were studied, alpha-D-Glucose (1 mM) slightly initiated the germination of the dormant spores during 10 min incubation at 37 degrees C, while about 60% of the dormant spores became germinated with beta-D-glucose (1 mM) in the same conditions. From the above observations and the finding that only a trace amount of alpha- or beta-D-glucose may bind with the dormant spores, it is speculated that the beta-D-glucose-stereospecific receptor site for the germination exists on the surface of the dormant spores of the bacillus.
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Cytoplasm of Paramecium caudatum syngen 3 in the period of sexual immaturity was effective in inducing immaturity when injected into mating-reactive cells of other syngens of P. caudatum. When immature cytoplasm of P. caudatum was injected into mature cells of P. multimicronucleatum, immaturity was also induced in the latter cells. When cytoplasm of P. bursaria in the immaturity period was injected into mature cells of the same species, the latter became immature. But immature cytoplasm of P. caudatum was ineffective when injected into mature cells of P. bursaria. When immature cytoplasm of P. busaria was injected into mature cells of P. caudatum, no difference from the control was observed. These results suggest that a difference in specificity of the immaturity substances exists bewteen paramecia of the 'aurelia' group and those of the 'bursaria' group.
Cells of Paramecium primaurelia in the immaturity period of the life cycle contain substances which repress the expression of mating ability when introduced into mature cells. When the immature cytoplasm of P. primaurelia was injected into mating-reactive cells of P. tetraurelia, P. caudatum and P. multimicronucleatum, immaturity was also induced in them. But the immature cytoplasm of P. primaurelia was ineffective when injected into mature cells of P. bursaria. When the immature cytoplasm of P. bursaria was injected into mating-reactive cells of P. primaurelia and P. tetraurelia, no difference from the control was observed. When the immature cytoplasm of P. caudatum was injected into mating-reactive cells of P. primaurelia and P. tetraurelia, immaturity was induced in cells of the latter 2 species.
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4 forms of mutarotase from human kidney were demonstrated by DEAE-cellulose column chromatography. A major form of them was purified to homogeneity.
The anomeric compositions of D-glucose in the liver, kidney, heart, blood and plasma of rat were determined by our methods for the assay of D-glucose anomers and the percentages of the beta-anomer were found to be 61.8, 61.0, 62.4, 62.7 and 62.9, respectively.
The effects on insulin release of alpha-D-mannose and equilibrated D-mannose were studied in the isolated perfused rat pancreas. Although phosphomannose isomerase, in contrast to phosphoglucose isomerase, is stereospecific for beta-D-mannose-6-phosphate, the alpha anomer of D-mannose (2 mg/ml) was significantly more effective than equilibrated D-mannose in stimulating insulin release. The finding does not seem to accord with the view that the signal for hexose-induced insulin release arises from the glycolytic pathway, and suggests the existence of the stereospecific glucoreceptor(s) on the pancreatic B-cell membrane.
The D-glucose anomeric preference of hexokinases isolated from rat liver, brain, and skeletal muscle, and bovine retina was studied using the glucose-6-phosphate dehydrogenase-NADP system. The ratios of maximum phosphorylation rates of beta-D-glucose to those of alpha-D-glucose were 1.33, 1.46, and 1.54 for hexokinase types I, II, and III from rat liver, 1.45 and 1.63 for type I from rat brain and bovine retina, 1.53 for type II from rat skeletal muscle, and 0.55 (when determined at 5 mM) for type IV (glucokinase) from rat liver, respectively.
Uptake of D-glucose anomers by isolated rat retina was studied. After 3 min incubation at 37 degrees C in the presence of alpha or beta anomer (750 mug/ml), a significantly greater uptake (1.32 mg/g wet tissue) of beta-anomer was observed compared with that of alpha-D-glucose (1.11 mg/g wet tissue). This result and other data suggest that the carrier for D-glucose transport in the retina prefers the beta-anomer stereospecifically.