A marked inhibition of the incorporation of S(35)-sulfate by normal calf costal cartilage was produced by potassium ascorbate in the presence of catalytic amounts of cupric ions. The effect of the various components of the ascorbic acid oxidizing system (potassium ascorbate, cupric ions, cuprous ions, hydrogen peroxide, dehydroascorbic acid) was investigated. The results of experiments in which hydrogen peroxide, catalase, or sodium azide were used singly or in combination suggest that the inhibition produced by the ascorbic acid oxidizing system is due, to a considerable extent, to the production of hydrogen peroxide. Dehydroascorbic acid was also found to inhibit the incorporation of S(35)-sulfate by cartilage slices. However, the gradual fall in pH which resulted from the addition of dehydroascorbic acid could account to a large extent for the inhibitory effect observed because the incorporation of S(35)-sulfate by cartilage slices decreases sharply as the pH is lowered. The incorporation of S(35)-sulfate by cartilage slices is inhibited also by increasing the concentration of phosphate.

Animals↗

PubMed1958

The effect of x-radiation on the glutathione metabolism of intact erythrocytes in vitro.

The x-irradiation of intact washed erythrocytes results in an inhibition of the glyoxalase activity of the cells chiefly as a result of a decrease in the reduced glutathione level. The percentage inhibition is markedly increased by an increase in the dilution of the cells in physiological saline suggesting that the effect of radiation is indirect, via the production in the aqueous medium of free radicals, H(2)O(2), etc. This is supported by the decrease in the inhibition produced by lowering the oxygen tension or by the addition of catalase. The inhibition of glyoxalase activity is also decreased by the addition of methylglyoxal, plasma, adenosine, inosine, glucose, and a number of other sugars to the erythrocyte suspension prior to radiation. Furthermore, some reactivation of the glyoxalase system results from the addition of plasma, glucose, adenosine, and inosine following radiation. These results are discussed in relation to the role of SH compounds, particularly glutathione, in the toxicity of ionizing radiations.

Erythrocytes↗

PubMed1958

The relationship of hydrogen peroxide to the inhibition of the glyoxalase activity of intact erythrocytes by x-radiation.

The x-irradiation of a dilute suspension of erythrocytes results in a decrease in the glyoxalase activity of the cells as a result of a fall in the reduced glutathione level. The present paper deals with the possible role of H(2)O(2) in this reaction. The addition of intact erythrocytes to physiological saline previously irradiated with 150,000 r or 225,000 r results in a fall in the glyoxalase activity of the cells. The inhibition is prevented by the preincubation of the irradiated saline with catalase and is reversed by the addition of plasma, glucose, adenosine, and inosine to the cell suspension. An inhibition of the glyoxalase activity is also produced by the addition of H(2)O(2) to the suspension of erythrocytes. The inhibitory effect of H(2)O(2) can be prevented and largely reversed by plasma, glucose, adenosine, and inosine. Methylglyoxal is also protective under these conditions. Hydrogen peroxide formed continuously and in low concentrations by enzyme systems appears to be more effective than added H(2)O(2) in inhibiting the glyoxalase system. The inhibition by H(2)O(2)-producing enzyme systems is minimized by the addition of catalase, plasma, glucose, methylglyoxal, and to a lesser extent, by adenosine and inosine, and is accentuated by the addition of sodium azide. The results are discussed in relation to the role of H(2)O(2) and catalase in the toxicity of ionizing radiations.

Catalase↗

PubMed1957

Glutathione metabolism. II. The oxidation and reduction of glutathione in intact erythrocytes.

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Erythrocytes↗