PubMed · 3473030
The source did not provide an abstract. Follow the original record for more information.
Explore connections, maps & timelines
1986. Amitrole.. https://pubmed.ncbi.nlm.nih.gov/3473030/
Cite the original work for its findings. Save a collection to share your selection of sources.
KEEP EXPLORING
The relative importance in human red blood cells of the plasma membrane as a site of redox activation of anthracyclines as compared to hemoglobin was evaluated by assaying the H2O2 produced upon exposure to daunomycin. The method of H2O2-dependent irreversible inhibition of catalase (EC 1.11.1.6) activity by 3-amino-1,2,4-triazole was applied to intact erythrocytes, as well as to isolated membranes with added purified catalase. The results obtained indicate a secondary role in daunomycin activation for the plasma membrane from a quantitative point of view, although membrane pathways can be more harmful than cytosolic pathways, especially towards extracellular targets, when the high efficiency of the cytosolic antioxidative defences and the external location of the membrane activation site are considered.
Quantification of intracellular and extracellular levels and production rates of reactive oxygen species is crucial to understanding their contribution to tissue pathophysiology. We measured basal rates of oxidant production and the activity of xanthine oxidase, proposed to be a key source of O2- and H2O2, in endothelial cells. Then we examined the influence of tumor necrosis factor-alpha and lipopolysaccharide on endothelial cell oxidant metabolism, in response to the proposal that these inflammatory mediators initiate vascular injury in part by stimulating endothelial xanthine oxidase-mediated production of O2- and H2O2. We determined a basal intracellular H2O2 concentration of 32.8 +/- 10.7 pM in cultured bovine aortic endothelial cells by kinetic analysis of aminotriazole-mediated inactivation of endogenous catalase. Catalase activity was 5.72 +/- 1.61 U/mg cell protein and glutathione peroxidase activity was much lower, 8.13 +/- 3.79 mU/mg protein. Only 0.48 +/- 0.18% of total glucose metabolism occurred via the pentose phosphate pathway. The rate of extracellular H2O2 release was 75 +/- 12 pmol.min-1.mg cell protein-1. Intracellular xanthine dehydrogenase/oxidase activity determined by pterin oxidation was 2.32 +/- 0.75 microU/mg with 47.1 +/- 11.7% in the oxidase form. Intracellular purine levels of 1.19 +/- 1.04 nmol hypoxanthine/mg protein, 0.13 +/- 0.17 nmol xanthine/mg protein, and undetectable uric acid were consistent with a low activity of xanthine dehydrogenase/oxidase. Exposure of endothelial cells to 1000 U/ml tumor necrosis factor (TNF) or 1 microgram/ml lipopolysaccharide (LPS) for 1-12 h did not alter basal endothelial cell oxidant production or xanthine dehydrogenase/oxidase activity. These results do not support a casual role for H2O2 in the direct endothelial toxicity of TNF and LPS.
Amitrole is a herbicide which has been found to induce thyroid and liver tumours in rodents, yet demonstrates limited genotoxic activity. The lack of mutagenicity of this compound in Salmonella typhimurium when employing a standard liver microsomal fraction, combined with evidence of activation of amitrole by peroxidases, warranted an investigation employing this other pathway of metabolic activation. Using prostaglandin H synthase as the activating system, the aromatic amine 2-aminofluorene provided a convenient positive control for optimisation of the metabolising system. Under such conditions, amitrole did not induce elevated numbers of revertant colonies in Salmonella typhimurium TA98, neither did it display evidence of interference with histidine biosynthesis as had been reported. Amitrole also remained nonmutagenic when preincubated at varying pHs. Thus, it has been shown that the alternative activation system, prostaglandin H synthase, does not produce metabolites which are mutagenic in the Ames test.