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Biomedical subjects

A Hara

Publications and source records attributed to A Hara.

At least 595 records · Page 33Linked to original sources
PubMed1981

Microsomal reductase for aromatic aldehydes and ketones in guinea pig liver. Purification, characterization, and functional relationship to hexose-6-phosphate dehydrogenase.

An NADPH-specific aromatic aldehyde-ketone reductase located in guinea pig liver microsomes can be effectively solubilized with nonionic detergents, but not with bile salts and hydrolytic enzymes. Destruction of microsomal membranes by nonionic detergents or acetone treatment leads to significant activation of the reductase, indicating that the enzyme is partly latent in intact microsomes. After solubilization with Triton X-100, the reductase has been highly purified. The purified enzyme catalyzes the NADPH-linked reduction of xenobiotic aromatic aldehydes and ketones as well as 3-ketosteroids, notably 5 alpha- and 5 beta-dihydrotestosterones. The reductase activities for xenobiotic carbonyl compounds and for 3-ketosteroids are each inhibited by addition of the other type of substrate and show the same pH optimum, cofactor requirement, and heat stability, indicating the same enzyme is responsible for the reduction of the two types of substrates. Hexose-6-phosphate dehydrogenase, purified from guinea pig liver microsomes, acts as a more effective NADPH generator for the reductase than yeast and guinea pig liver cytosolic glucose-6-phosphate dehydrogenase. Evidence has been obtained that hexose-6-phosphate dehydrogenase undergoes a functional interaction with the reductase, facilitating the provision of NADPH to the reductase activity both in the reconstituted system and in microsomes.

Alcohol Oxidoreductases↗
PubMed1981

Continuous recording of microvascular dimensions with a rotating mirror microscope.

A system which permits continuous recording of the dimensions of microscopic blood vessels is described. The system uses a combination of a rotating mirror and a pinhole aperture with a microscope to scan microvessel image signals at a rate of one per second. Image signals from the gate circuit are converted to digital signals; further conversion to analogue signals is obtained by integrating the digital signals. The analogue voltages are proportional to the vessel dimensions. A linear relationship was observed over a range of 20-200 micrometer (standard deviation +/- 3.0%). The contrast between vessels and surrounding tissue is amplified by using a B-12 optical filter. By using this system, the mesenteric arteriole was observed after reaction with adrenaline and noradrenaline in rats. The contractile response of the arteriole to adrenaline was greater than with noradrenaline. A maximum response of 20% decrease of the vessel outer diameter is induced by 5.5 x 10(-7) mmol adrenaline and 5.9 x 10(-6) mmol noradrenaline.

Animals↗
PubMed1980

A possible functional relationship between microsomal aromatic aldehyde-ketone reductase and hexose-6-phosphate dehydrogenase.

Aromatic ketone reductase activity of microsomes showed a unique cofactor requirement: Addition of NADP and glucose-6-phosphate was as effective as that of an artificial NADPH generating system, whereas NADPH alone served as a cofactor less efficiently. Microsomal aromatic ketone reductase, purified partially from guinea pig liver microsomes after solubilization with Triton X-100, reduced 5 beta-dihydrotestosterone, aromatic aldehydes, and ketones with NADPH as a cofactor. However, addition of hexose-6-phosphate dehydrogenase, purified from the same source, as an NADPH generator produced about 2 times higher activity than that of yeast glucose-6-phosphate dehydrogenase or NADPH alone.

Alcohol Oxidoreductases↗
PubMed1980

Renal function, plasma renin, and spontaneous diuresis in an advanced stage of hypertension in rats.

Relationship of renal function and plasma renin level to a spontaneous diuresis in an advanced stage of hypertension was examined in the stroke-prone spontaneously hypertensive rats (SHRSP). For a comparison, it was also investigated in 2 kidney 1 clip (1CR) and 2 kidney 2 clip (2CR) models of renovascular hypertensive rats. The spontaneous diuresis was usually associated with an elevation of plasma renin level and an impairment of renal function in SHRSP and 1CR. In SHRSP, renal plasma flow showed an inverse correlation to the plasma renin level. The diuresis, however, is not prerequisite to the increase in plasma renin or to the renal impairment, since the latter changes were not always accompanied by diuresis. No correlations were found between these factors in 2CR. Remittent diuresis was observed in many 2CRs and 2 SHRSPs with normal renal function, indicating that renal damage underlies the continuous progression of the diuresis observed in most cases of SHRSP and 1CR. Both sodium and potassium balances tended to be negative in 1CR and 2CR in the diuretic phase, whereas potassium alone showed this tendency in SHRSP. The negative balances, if any, were not due to an excessive loss of the electrolytes but attributed mainly to a decrease in intake. The critical level of blood pressure for the spontaneous diuresis was highest in SHRSP and lowest in 1CR. It is probable that an interaction and cooperation between high blood pressure, impaired renal function and high plasma renin participate in the mechanism of the spontaneous diuresis during the development of malignant phase of hypertension of the rat.

Aldosterone↗
PubMed1980

[Effect of anti-human alpha-fetoprotein serum on human endodermal sinus tumor transplanted in nude mice (author's transl)].

The effects of horse anti-human alpha-fetoprotein (AFP) immunoglobulin (IgG) on growth, serum AFP concentration and histopathology of human endodermal sinus tumor serially xeno-transplanted in nude mice were examined. Anti-AFP IgG given in amounts sufficient to neutralize the serum AFP secreted by the tumor moderately reduced tumor size and significantly decreased the serum AFP levels of treated nude mice. On histopathologic examination, spindle shaped tumor cells of reticular pattern often fell into the state of necrosis. But little change was observed on the cells consisting of both solid patterns and Schiller-Duval bodies. With a growth rate of re-transplanted tissues respectively, which were obtained from both antibody administrated and control tumors, no disparity was found between two groups. Indeed this result is preliminary report, but inhibitory effects of anti-AFP IgG against tumor growth and AFP producing ability of endodermal sinus tumor xeno-transplanted in nude mouse is hopeful to apply these data on clinical cancer immunochemotherapy.

Animals↗
PubMed1980

Reductases for aromatic aldehydes and ketones from rabbit liver. Purification and characterization.

Four aldehyde reductases, F1, F2, F3, and F4, were isolated from rabbit liver cytosol to homogeneity by various chromatographic techniques. F2 is an aldehyde reductase with a molecular weight of 32,000, which resembled aldehyde reductases of human liver and pig kidney in properties. It was inhibited in a noncompetitive way by alpha-ketoglutarate and oxaloacetate with Ki values of 4 x 10(-5) M. The other three enzymes were NADPH-dependent aromatic aldehyde-ketone reductases. F1 and F3 were monomeric enzymes with molecular weights of 38,000 and 29,000, respectively. F4 showed a molecular weight of 78,000 on gel filtration, but sodium dodecyl sulfate gel electrophoresis revealed two different subunits with molecular weights of 26,000 and 24,000. The molar ratio of NADPH : F1, F2, or F3 binding was 1 : 1, whereas that of NADPH:F4 binding was 3 : 1. The number of thiol groups in order molecule of F1, F2, F3, and F4, was 4, 4, 4, and 5, respectively. The enzyme activity of F3 was inhibited by addition of an equal mole amount of PCMB. Only F4 was inhibited by metal chelating agents. F1 also catalyzed the interconversion of 3(17)-keto and 3(17) beta-hydroxysteroids, whereas F3 catalyzed oxidoreduction of some 3 alpha-hydroxysteroids of the 5 alpha-series. These results suggest that F1 and F3 are 3(17) beta-hydroxysteroid dehydrogenase and 3 alpha-hydroxysteroid dehydrogenase, respectively. Endogenous substrates for F4 could not be identified in this work. F1 showed the lowest Km value for reduction of aldehydes and ketones among the four reductases. It has been suggested that F1 is the primary enzyme responsible for the reduction of endogeneous aldehydes and xenobiotic aldehydes and ketones in vivo.

Aldehyde Oxidoreductases↗
PubMed1979

Destruction and resynthesis of mouse beta-glucosidases.

1. Injection of a single dose of conduritol B epoxide into mice produced almost complete destruction of glucocerebrosidase (D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45) in liver, spleen, brain, and kidney within 5 h. Restoration of activity became noticeable within 1 day (2 days in the case of brain) and was about 80% of normal within 16 days. 2. The same injection produced less destruction of aryl beta-glucosidase (EC 3.2.1.21), measured at pH 5.4 with methylumbelliferyl glucoside in the absence of taurocholate. Brain showed the least amount of destruction, about 50%, but measurements of activity at lower pH values revealed complete loss of activity. This suggests that brain contains two different aryl glucosidases with differing sensitivity to the inhibitor. Liver, on the other hand, did not show differential destruction when assayed at different pH values. Resynthesis of the enzyme activities was almost complete by 16 days. 3. Injection of phenylhydrazine produced hemolysis and spleen enlargement, with concomitant increases in specific activities of glucocerebrosidase and aryl glucosidase in liver and spleen (but not in kidney). When this experiment was done in mice previously treated with conduritol B expoxide, the reappearance of cerebrosidase was found to be accelerated. This is interpreted to mean that the increased load of glucolipids from the erythrocytes had induced an enhanced synthesis of the glucohydrolase. A similar explanation may apply to aryl glucosidase and glucopeptides in the cells.

Animals↗
PubMed1979

Enzymic effects of beta-glucosidase destruction in mice. Changes in glucuronidase levels.

1. The injection into mice of a single dose of conduritol B epoxide, a covalent inhibitor of glucosidases, quickly produced changes in tissue levels of beta-D-glucuronidase (EC 3.2.1.31). The specific activity of the enzyme decreased in liver, spleen and kidney while brain showed little change. The inhibitor did not act on glucuronidase in vitro, so the effect of the inhibitor is complex, possibly a result of the loss of glucosidase activity. Since glucuronidase contains glucose, we suggest that the transport of the enzyme between subcellular regions and tissues involves loss of part of the glucose moieties. 2. Levels of glucocerebrosidase (D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45) dropped very rapidly after epoxide injection, reaching a minimum at 1 h in liver. There was a noticeable restoration of activity within the next 1--2 h. Aryl beta-glucosidase (EC 3.2.1.21) decrease somewhat less than cerebrosidase, reaching a minimum within 2 h. It too showed some recovery of activity within 3 h. 3. Acid phosphatase rose slightly in liver but not in brain. alpha-L-Fucosidase and angiotensin-converting enzyme were not affected by the epoxide injection. The latter two enzymes are known to contain glucose. 4. Injection of a hemolyzing agent, phenylhydrazine, produced an increased level of glucuronidase in liver and spleen within 6 days, but not in kidney. This enhancement was a little less in mice previously injected with the glucosidase inhibitor. 5. Mice injected with the epoxide once a day eight times showed a distinct rise in brain glucuronidase level, as well as a rise in brain weight. However, the other organs showed only the same decrease in glucuronidase specific activity noted with the single injection protocol. It is suggested that the difference is due to the blood-brain barrier, which could slow the loss of brain glucuronidase from the extracellular fluid.

Acid Phosphatase↗
PubMed1979

Purification and properties of reductases for aromatic aldehydes and ketones from guinea pig liver.

NADPH-dependent enzymatic reduction of aromatic aldehydes and ketones observed in the cytosol of guinea pig liver was mediated by at least three distinct reductases (AR 1, AR 2, and AR 3), which were separated by DEAE-cellulose chromatography. By several procedures AR 2 and AR 3 were purified to homogeneity, but AR 1 could be purified only 30-fold because of the small amount. These enzymes were found to have similar molecular weights of 34,000 to 36,000 and similar Stokes radii of about 2.5 nm. AR 3 was identical to aldehyde reductase [EC 1.1.1.2] in substrate specificity for aromatic aldehydes and D-glucuronate and specific inhibition by barbiturates. AR 1 and AR 2 acted on aromatic ketones and cyclohexanone as well as aromatic aldehydes at optimal pHs of 5.4 and 6.0, respectively, and were immunochemically distinguished from AR 3. AR 1 was the most sensitive to sulfhydryl reagents, and AR 2 was more stable at 50 degrees C than the other enzymes. Similar heterogeneity was observed in the kidney enzymes, but other tissues had little aldehyde reductase activity and contained only AR 3. In addition, lung contained a high molecular weight aromatic ketone reductase different from the above reductases.

Aldehyde Oxidoreductases↗