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Mode of action of oxathiin systemic fungicides. V. Effect on electron transport system of Ustilago maydis and Saccharomyces cerevisiae.

The systemic fungicide carboxin (5,6-dihydro-2-methyl-1,4-oxathiin-3-carboxanilide) at 100 mum inhibited succinate cytochrome c reductase in mitochondria from Ustilago maydis and Saccharomyces cerevisiae. It did not have any effect on reduced nicotinamide adenine dinucleotide (NADH) cytochrome c reductase. Succinate coenzyme Q reductase was also inhibited, but NADH coenzyme Q reductase was not. When dichlorophenolindophenol (DCIP) was used as the terminal acceptor of electrons from the oxidation of succinate, carboxin was very effective in inhibiting succinate-DCIP reductase. Carboxin was inhibitory to succinic dehydrogenase assayed with phenazine methosulfate plus DCIP when intact mitochondria were used as the enzyme source but not when solubilized enzyme was used. The main site of action of carboxin, therefore, appears to lie between succinate and coenzyme Q. The dioxide analogue of carboxin was also effective in inhibiting succinate-cytochrome c reductase, succinate-coenzyme Q reductase, or succinate-DCIP reductase, whereas the monoxide analogue was less effective in inhibiting these enzymes.

Anilides↗

Cyanide formation from oxidation of glycine of Pseudomonas species.

With whole cells of a hydrogen cyanide-producing bacterium strain C, of the genus Pseudomonas, it was found that the oxygen necessary for the oxidation of glycine to cyanide could be replaced by various artificial electron acceptors. The order of reactivity was: oxygen > phenazine methosulphate > methylene blue > 2,6-dichlorophenolindophenol > ferricyanide. Cyanide production was inhibited by pyrrolnitrin, a well-known inhibitor of many flavine enzymes. The molar ratio of added glycine to cyanide produced was found to be 1.09. With whole bacteria the apparent K(m) (glycine) for the cyanide production was found to be 5.0 x 10(-4) M.

Acridines↗

Purification and properties of pyridine nucleotide-independent L-lactate dehydrogenase from Polyporus circinatus.

Cell extracts of Polyporus circinatus grown on lactate catalyze the reduction of 2,6-dichlorophenolindophenol by l-lactate without the participation of nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate. The enzyme has been purified 78-fold and was homogenous by disc gel electrophoresis. The optimal pH was found to be 6.7. The Michaelis constant for l-lactate was 5.9 x 10(-4) M and the oxalate inhibition constant was 1.5 x 10(-4) M. The nature of the prosthetic group is discussed.

Alcohol Oxidoreductases↗

Properties of somatic cell hybrids between mouse cells and simian virus 40-transformed rat cells.

Hybrids between mouse cells and simian virus 40 (SV40)-transformed rat cells were made, and their properties and chromosome constitution were investigated over many generations. Their hybrid nature was confirmed by enzyme studies. During a period of 1 year a loss of 10 to 20% of the total number of chromosomes was observed. The SV40 tumor antigen was present and remained present in the hybrids. The parental and hybrid cells were studied for agglutination with concanavalin A, for growth in soft agar, and for serum requirement. These growth and surface characteristics of the transformed cells appeared in the hybrids.

Agglutination↗

The non-light-dependent reduction of 2,6-dichlorophenolindophenol by cells of the blue-green alga Anacystis nidulans.

Whole cells of the blue-green alga Anacystis nidulans reduced, in the dark, the oxidation-reduction dye, 2,6-dichlorophenolindophenol at rates severalfold higher than those of the other algae tested. Under anaerobiosis, the endogenous reductant was depleted after up to 80 nmol of dye were reduced per microliter of cells. Cells held in darkness for several hours exhibited lowered dark reduction rates relative to cells held in light. Treatment with lysozyme and ethylenediaminetetraacetic acid yielded cells that would photoreduce the dye, whereas untreated cells would not. Comparisons of photoreduction and dark reduction revealed that the dark reduction proceeded independently of the photoreduction. It was concluded that the dark reduction represents a pool of endogenous reductant of sufficiently low oxidation-reduction potenital to reduce completely 2,6-dichlorophenolindophenol. Additionally, untreated cells were shown to be permeable to the dye although they did not photoreduce it; thus lysozyme/ethylenediaminetetraacetic acid treatment was considered to make the oxidant accessible to the photosynthetic machinery.

Anaerobiosis↗

Optimal conditions for the estimation of ammonium by the Berthelot reaction.

A technique for optimising reagent concentrations on the AutoAnalyzer has been applied to the estimation of ammonia by the Berthelot reaction in the determination of urea and organic nitrogen. Comparison of the use of phenol and salicylate revealed that the optimum concentration of the latter is about four times that of the former. The optimum concentration of hypochlorite is five times greater with salicylate than with phenol, and for the catalyst, sodium nitroprusside, the factor is two. The precision obtained with the different methods is similar.

Ammonia↗

Assessment of Peroheme-40 for the presence of occult blood in the faeces.

Peroheme 40, a chemical test for occult blood in faeces, has been assessed by comparing it with results obtained using 51Cr labelled red cells. The study was carried out in two parts. Initially testing was carried out on 75 stool samples from 20 patients on a restricted diet, and in the second part, testing was performed on 40 samples from 10 patients on a normal ward diet. All patients were suspected of losing blood from the gastrointestinal tract. On the restricted diet there were 29 per cent false positive and 13 per cent false negative results, while on the normal diet, there were 86 per cent false positive and 8 per cent false negative results. Peroheme-40 has therefore been shown, on a restricted diet, to compare favourably with previously available chemical tests.

2,6-Dichloroindophenol↗