Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:1.6.99.1 (NADPH-diaphorase)
3,903 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Cell-free extracts of a streptomycin-bleached strain of Euglena gracilis var. bacillaris have been examined for enzyme systems primarily responsible for the oxidation of reduced pyridine nucelotides. NADH lipoyl dehydrogenase, NADH and NADPH oxidase, NADH and NADPH diaphorase, and NADH and NADPH cytochrome c reductase have been demonstrated. The NADPH-linked enzymes had lower activity rates and were less sensitive to N-ethyl maleimide and p-hydroxymercuribenzoate than their NADH-linked counterparts. NADH cytochrome c reductase was the most sensitive to antimycin A. Michaelis-Menten constants (Km) determined were as follows: NADH diaphorase, 350 muM; NADPH oxidase 150 muM ; NADH lipoyl dehydrogenase, 0.35 muM. Enzyme activities after storage at -5 C indicate that the diaphorases are less labile than the other tested enzymes, and the differential activities of the NADH and NADPH linked enzymes suggest that functionally they may have different roles.
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PMID:Reduced pyridine nucleotide oxidases of Eugena gracilis var. bacillaris. 40 56

A study of the ultrastructural localization of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), glucose-6-phosphate dehydrogenase (G-6-PD), beta-hydroxybutyrate dehydrogenase (beta-HBD), NADH diaphorase (NADH-D) and NADPH diaphorase (NADPH-D) in the guinea-pig testis is reported. The procedures employed included short immersion or perfusion fixation with aldehydes followed by incubation of small blocks in a tetrazolium salt or a ferricyanide medium. The effects of incubation conditions were investigated, and a reaction medium for the ultracytochemical demonstration of 11 beta-HSD is described. Using suitable controls, evidence for the specificity of the cytochemical reactions is presented. It was found that all the enzymes studied were present in both the Leydig and Sertoli cells of the guinea-pig testis and that the intracellular distribution pattern for each enzyme was independent of the cell type. Using tetrazolium salt techniques, both 3 beta-HSD and 11 beta-HSD activities were localized on or in membranes of smooth endoplasmic reticulum and within the mitochondria. With the ferricyanide techniques, G-6-PD activity was found to be associated mainly with the smooth endoplasmic reticulum membranes, while beta-HBD activity was limited to mitochondria. With both the tetrazolium salt and ferricyanide techniques, the reaction products for NADH-D and NADPH-D activities showed localizations which were similar to those observed for the steroid dehydrogenases.
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PMID:The ultrastructural localization of the enzymes related to steroid hormone metabolism in the guinea-pig testis. 42 99

(1) Various buffer systems for the starch gel electrophoresis of human diaphorase isozymes have been explored. Electrophoresis in a Tris/Borate system at pH 8.6 which includes 70 micron NADH in the gel and cathodal electrode buffers, provides good resolution of the six DIA3 phenotypes previously resolved by isoelectric focusing. (2) The variant genes DIA13, DIA23 and DIA33 occur with frequencies of about 0.76, 0.23 and 0.01 respectively in the English population. (3) The isozymes determined by the least common gene, DIA33, are markedly different from the isozymes determined by DIA13 and DIA23 in their relatively low heat stability, high affinity for Blue Sepharose and slow anodal electrophoretic mobility in buffer systems containing borate. The DIA3 1 and DIA3 2 isozymes are similar to one another in these characteristics.
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PMID:A comparison of the biochemical properties of the human diaphorase (DIA3) isozymes determined by the common alleles DIA13, DIA23 and DIA33. 43 72

The usual techniques for determination to total 3 alpha-hydroxy bile acids in serum involving liquid-solid extraction of the bile acids with the adsorbent XAD-2 and fluorimetric measurement of NADH generated from the reaction with a NAD-linked 3 alpha-hydroxysteroid dehydrogenase are evaluated and improved. The influence of different types of enzyme preparations on the results is examined. The results with the improved technique are compared to the results obtained with another method, avoiding extraction of the bile acids before the enzymatic reaction which is followed by fluorimetric measurement of resorufin, produced by transfer of the hydrogen of the generated NADH by diaphorase to resazurin. No significant difference between the results with the two types of methods was found. The concentration of total 3 alpha-hydroxy bile acids in serum of 46 fasting 'healthy' individuals aged 17 to 82 years is estimated. 30 were females, of whom 10 were taking estrogen-containing oral contraceptives, and 16 were males. Mean +/- standard deviation in all the females was 3.0 +/- 1.1 micromol/l, and in the males 4.0 +/- 1.9 micromol/l. There was no significant difference between any of the groups.
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PMID:Determination of total 3 alpha-hydroxy bile acids in serum. 43 89

The distribution and activities of several oxidative enzymes in the urinary apparatus of five freshwater fish species (river lamprey, lobe finned eel, Prussian carp, rainbow trout and three-spined stickleback) have been studied. Species were selected from three main taxonomic groups: Cyclostomata, Polypterini, Teleostei. Distinctly positive enzyme reactions were only found in the tubular elements of the kidney and the collecting duct-archinephric duct system, with the exception of the generally weak staining intensities of lactate dehydrogenase. The distal tubule normally showed strong to very strong reactions for most of the enzymes investigated. In the epithelial cells of the collecting tubule-collecting duct system, stronger reactions were observed for most of the mitochondrial-bound enzymes, especially succinate dehydrogenase and NADH-diaphorase. For these enzymes, the cells of the archinephric duct reacted strongly positive in Lampetra, Carassius and Gasterosteus. The enzyme patterns of various types of urinary tubules and ducts are compared with results of several morphological studies. In addition, the histochemical findings are discussed in relation to kidney function in different vertebrate groups.
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PMID:Oxidative enzymes in the urinary apparatus of several freshwater fishes. 43 99

The ovary of the domestic pigeon, Columba livia, has been assayed histochemically for the localization of delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSDH), 17 beta-hydroxysteroid dehydrogenase (17 beta-HSDA), 11 beta-hydroxysteroid dehydrogenase (11 beta-HSDH), glucose-6-phosphate dehydrogenase (G6P-DH) and NADH-diaphorase activities during different periods of the reproductive cycle. delta 5-3 beta-HSDH, 17 beta-HSDH, 11 beta-HSDH, G6P-DH and NADH-diaphorase activity was found in the theca interna of growing, atretic and postovulatory follicles, the granulosa of ovulatory, atretic and postovulatory follicles, and interstitial gland cells during the pre-incubation and the laying periods. During the incubation and squab feeding periods only delta 5-3 beta-HSDH, G6P-DH and NADH-diaphorase activities were observed in the above mentioned cells. The steroidogenic potential of atretic follicles depends upon the type of atresia a follicle undergoes.
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PMID:Steroid synthesizing cellular sites in the ovary of the domestic pigeon Columba livia (Gmelin): a histochemical study. 45 38

Techniques for the ultrastructural demonstration of dehydrogenases in cerebral cortex are described. The best fixation for good fine structural preservation and retention of LDH and NADH-diphorase was obtained by perfusion with a misture of formaldehyde and glutaraldehyde and for SDH by perfusion with formaldehyde. Comparison of incubation conditions showed that consistent results were obtained using enzyme markers NBT and DS-NBT for LDH and NADH-diaphorase: DS-NBT was more satisfactory than NBT and BSPT for SDH. Penetration of incubation media was improved by Triton X-100: DMSO and ultrasonic treatment were less effective. The techniques enabled the first electron cytochemical demonstration of dehydrogenases in different elements of prefixed cerebral cortex. Ultrastructural sites of enzyme activities were localized within cristae and inter-membrane spaces of mitochondria in nerve cell cytoplasm and its processes, oligodendrocytes and astrocytes. Authenticity of the ultrastructural sites was confirmed by four different control experiments.
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PMID:Ultrastructural demonstration of dehydrogenases in rat cerebral cortex. 47 91

The organic phosphate allosteric effectors of hemoglobin, inositol hexaphosphate, 2,3-diphosphoglycerate, and ATP, interact with NADH-methemoglobin reductase (NADH-diaphorase). Significant inhibitory effects on the enzyme were found when dichlorophenolindophenol, or ferricyanide were used as electron acceptors in place of methemoglobin. In contrast, apparent stimulation of enzyme activity was observed when adult human methemoglobin was used as the electroganic phosphate on the rate of reaction due to its interaction with the substrate methemoglobin to produce the favored T type of quaternary conformation. The inhibitory effect of inositol hexaphosphate on the enzyme is associated with a perturbation in the reactivity of essential sulfhydryl group(s) on the enzyme. It is suggested that the interaction of the organic phosphate with the enzyme as well as with the substrate is significant in determining the overall rate of methemoglobin reduction.
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PMID:Inhibition of NADH-methemoglobin reductase by organic phosphates. 49 34

The effect of 4'-demethyl-epipodophyllotoxin-beta-D-thenylidene glucoside (VM-26) , a semi-synthetic derivative of podophyllotoxin, on the cell cycle was studied with chick embryo fibroblasts cultivated in vitro. DNA, RNA and protein content, as well as NADH-diaphorase activity were determined by quantitative microdensitometry and cytofluorometry. The incorporation of [3H]thymidine and [3H]leucine into DNA and proteins were analysed by autoradiography. These metabolic data correlated with morphological observation showed that VM-26 blocks the cell cycle at different moments of its kinetics depending on both the dose and the time exposure. NADH-diaphorase activity is the first to be affected, then biochemical changes (involving the metabolism of RNA and proteins) and morphological alterations (especially of mitochondria) follow. This suggests that VM-26 may act primarily upon the mechanism of respiration of the cell.
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PMID:A quantitative microdensitometric and autoradiographic study of the effect of 4'-demethyl-epipodophyllotoxin-beta-D-thenylidene glucoside (VM-26) on the cell cycle of cultured fibroblasts. 53 43

Histochemical and biochemical studies on the folate metabolism (folic acid an its principal enzyme-dihydrofolate-reductase) in bovine cortex - gyrus marginallis in the process of ageing were performed, in parallel with NADH2-cytocrom-C-reductase (diaphorase). Folic acid and folate enzyme, weak positive in neurons in young age, increased in old age in nerve cells and especially in their processes and in capillaries. The diaphorase strongly increased in all cells, glia and vessels, in old age.
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PMID:Histochemical study on the dihydrofolatereductase and folic acid in mammalian brain cortex. 54 85


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