A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases.
01 Jul 1957-American Journal of Clinical Pathology (AMERICAN JOURNAL OF CLINICAL PATHOLOGY)-Vol. 28, Iss: 1, pp 56-63
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TL;DR: The newer salicylic acid congeners inhibited the normal alanine aminotransferase but did not significantly alter the normal aspartate aminotsferase, which may be related to their anti-inflammatory property.
Abstract: Sm,-The exact mechanism of anti-inflammatory activity of drugs is not well understood. The anti-inflammatory agents like salicylates, butazolidine, resorcylic acid, hydrocortisone, glycyrrhetic acid and imipramine have been shown to inhibit aminotransferases (Moses & Smith, 1961 ; Huggins, Bryant & Smith, 1961 ; Huggins, Smith & Moses, 1961 ; Tangri, Seth, Parmar & Bhargava, 1965 ; Tangri, Saxena, Seth & Bhargava, 1966), uncouple oxidative phosphorylation (Whitehouse & Haslam, 1962) and stimulate ATP phosphohydrolase activity (Whitehouse & Haslam, 1962; Tangri & others, 1965, 1966); all of these actions may be related to their anti-inflammatory property. Some newer salicylic acid congeners like 2,4-diacetoxybenzoic acid, m-cresotinic acid and 5-ethyl-2hydroxybenzoic acid were reported to have potent anti-inflammatory activity (Tangri & Bhargava, 1964). The effect of these agents on serum aminotransferases and tissue ATP phosphohydrolase has been examined to correlate their biochemical effects with their anti-inflammatory activity. Enzyme assays were made both in normal and arthritic albino rats with or without drug treatment. Serum for the estimation of aminotransferase activity was obtained from the blood of decapitated rats. The livers were removed immediately and pooled for the estimation of ATP phosphohydrolase (EC. No. 3.6.1.4.) activity. Serum L-aspartate : 2-oxoglutarate aminotransferase (EC. No. 2.6.1 .l, aspartate aminotransferase) and serum L-alanine : 2-oxoglutarate aminotransferase (EC. No. 2.6.1.2, alanine aminotransferase) were assayed by the method of Reitman & Frankel (1957). One unit of enzyme activity was the change in the extinction of O.OOl/min/ml of serum, which was measured in a Bausch & Lomb Spectronic '20' colorimeter at 505 mp. ATP phosphohydrolase activity was estimated in 10% (w/v) pooled liver homogenate prepared in 0.25 M sucrose. The reaction mixture consisted of 0.05 M Tris pH 8.0, 1 mM ATP and 0.1 ml of 10% tissue homogenate in a final volume of 2 ml. Release of Pi (inorganic phosphorus) from ATP was measured according to Fiske & Subbarow (1925). The release of 1 PM of Pi/lOO mg of tissue in 15 min at 37\" was considered as one unit of enzyme activity. The arthritis in the ankle joints of albino rats (100-1 10 g) was produced by injecting 0.1 ml of 2% (v/v) formaldehyde subcutaneously under the plantar aponeurosis according to Brownlee (1950). The animals were treated with daily intraperitoneal injections of 2,4-diacetoxybenzoic acid, m-cresotinic acid, 5-ethyl-2-hydroxybenzoic acid (2 mg/100 g bodyweight) and hydrocortisone (0.5/100 g bodyweight) for 10 consecutive days. The results are in Tables 1 and 2. The newer salicylic acid congeners inhibited the normal alanine aminotransferase but did not significantly alter the normal aspartate aminotransferase. A greater sensitivity of alanine aminotransferase to anti-inflammatory agents (viz. salicylates, glycyrrhetic acid, hydrocortisone and imipramine) compared to aspartate aminotransferase has been reported (Huggins & others, 1961a, b; Tangri & others, 1965, 1966). Furthermore, the inflammatory reaction increased the levels of both serum aminotransferases but particularly aspartate aminotransferase. This increase in the enzymic activities due to inflammation was prevented by the salicylic acid congeners to the same degree as hydrocortisone. Other anti-inflammatory agents, glycyrrhetic acid, methyl glycyrrhetic acid and imipramine, also prevented the rise in the serum aminotransferases caused by inflammation (Tangri & others 1965, 1966). Since the anti-inflammatory agents significantly reduced the enhanced serum aspartate aminotransferase level induced by the inflammatory stimulus and since they failed to alter the normal aspartate aminotransferase activity, it may be
61 citations
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TL;DR: Pretreatment of hydroalcoholic extract of Black grapes to lead exposed rats significantly ameliorated lead-induced oxidative stress in tissues and produced improvement in hematological parameters over lead-exposed rats, indicating the beneficial role of Black grape to counteract the lead- induced oxidative stress.
61 citations
TL;DR: Potassium considerably improved nitrogen metabolism under normal water supply conditions and also resulted in amelioration of the negative impact of water and osmotic stresses indicating that potassium supplementation can be used as a potential tool for enhancing the nitrogen use efficiency in wheat for exploiting its genetic potential.
Abstract: Present communication reports laboratory and pot experiments conducted to study the influence of water and osmotic stress on nitrogen uptake and metabolism in two wheat (Triticum aestivum L) cultivars with and without potassium supplementation. Polyethylene glycol 6000-induced osmotic stress/restricted irrigation caused a considerable decline in the activity of nitrate reductase, glutamate synthase, alanine and aspartate aminotransferases, and glutamate dehydrogenase. Potassium considerably improved nitrogen metabolism under normal water supply conditions and also resulted in amelioration of the negative impact of water and osmotic stresses indicating that potassium supplementation can be used as a potential tool for enhancing the nitrogen use efficiency in wheat for exploiting its genetic potential.
61 citations
TL;DR: The purified L-methioninase from solid cultures of A. flavipes displayed unique biochemical and catalytic properties over the currently applied Pseudomonad enzyme.
Abstract: L-Methioninase was purified to electrophoretic homogeneity from cultures of Aspergillus flavipes using anion-exchange and gel filtration chromatography by 12.1 fold compared to the crude enzyme preparation. The purified enzyme had a molecular mass of 47 kDa under denaturing conditions and an isoelectric point of 5.8 with no structural glycosyl residues. The enzyme had optimum activity at pH 7.8 and pH stability from 6.8–8.0 at 35°C. The enzyme appeared to be catalytically stable below 40°C. The enzyme activity was strongly inhibited by DL-propargylglycine, hydroxylamine, PMSF, 2-mercaptoethanol, Hg+, Cu2+, and Fe2+, with slight inhibition by Triton X-100. A flavipes L-methioninase has a higher catalytic affinity towards L-methionine (Km, 6.5 mM and Kcat, 14.1 S−1) followed by a relative demethiolating activity to L-homo-cysteine (Km, 12 mM and Kcat, 9.3 S−1). The enzyme has two absorption maxima at 280 and 420 nm, typical of other PLP-enzymes. Apo-L-methioninase has the ability to reconstitute its structural catalytic state completely upon addition of 0.15 mM PLP. L-Methioninase has neither an appreciable effect on liver function, platelet aggregation, nor hemolysis of human blood. The purified L-methioninase from solid cultures of A. flavipes displayed unique biochemical and catalytic properties over the currently applied Pseudomonad enzyme.
61 citations
TL;DR: In conclusion, HaE exhibits good hepatoprotective, curative, and antioxidant potential against DMBA-induced hepatorenal dysfunction in rats that might be due to decreased free radical generation.
Abstract: Oxidative stress is a common mechanism contributing to the initiation and progression of hepatic damage. Hence there is a great demand for the development of agents with potent antioxidant effect. The aim of the present study is to evaluate the efficacy of Holothuria atra extract (HaE) as an antioxidant against 7,12-dimethylbenz[a]anthracene- (DMBA-) induced hepatorenal dysfunction. Experimental animals were divided into two main groups: protective and curative. Each group was then divided into five subgroups pre- or posttreated either with distilled water (DMBA subgroups) or with HaE (200 mg/kg body weight) for seven and fourteen days. Single oral administration of DMBA (15 mg/kg body weight) to Wistar rats resulted in a significant increase in the serum liver enzymes and kidney function's parameters. DMBA increased level of liver malondialdehyde (MDA), decreased levels of reduced glutathione (GSH), glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) in the liver tissue, and induced liver histopathological alterations. Pre- or posttreatment with HaE orally for 14 days significantly reversed the hepatorenal alterations induced following DMBA administration. In conclusion, HaE exhibits good hepatoprotective, curative, and antioxidant potential against DMBA-induced hepatorenal dysfunction in rats that might be due to decreased free radical generation.
61 citations