Glutathione peroxidase induction protects Saccharomyces cerevisiae sod1deltasod2delta double mutants against oxidative damage

Saccharomyces cerevisiae mutants deficient in superoxide dismutase genes (sod1delta, sod2delta and the double mutant) were subjected to H2O2 stress in the stationary phase. The highest sensitivity was observed in the sod2delta mutant, while the sod1deltasod2delta double mutant was not sensitive. Sod mutants had lower catalase activity (44 percent) than wild-type cells, independent of H2O2 stress. Untreated cells of sod1deltasod2delta double mutants showed increased glutathione peroxidase activity (126 percent), while sod1delta had lower activity (77 percent) than the wild type. Glutathione levels in sod1delta were increased (200-260 percent) after exposure to various H2O2 concentrations. In addition, the highest malondialdehyde levels could be observed without H2O2 treatment in sod1delta (167 percent) and sod2delta (225 percent) mutants. In contrast, the level of malondialdehyde in the sod1deltasod2delta double mutant was indistinguishable from that of the wild type. These results suggest that resistance to H2O2 by sod1deltasod2delta cells depends on the induction of glutathione peroxidase and is independent of catalase, and that glutathione is a primary antioxidant in the defense against H2O2 in stationary phase sod1delta mutants.

Effect of IL-3 and E. coli LPS on the proliferation of mouse bone marrow cells in vitro

Bone marrow cells from adult BALB/c mice were cultured at 37-C, with 5% CO2 in air, in RPMI 1640 medium complemented with fetal serum. The addition of IL-3 (5% of WEHI-3-conditioned medium) or E. coli lipopolysaccharides (LPS, 50 µg/ml) to the cultures stimulated cell proliferation (1.29 and 1.22-fold, respectively, relative to control culture), whereas the simultaneous addition of the two factors reduced the number of cells recovered by 38% relative those from control cultures (which were around 2.83 x 10***5 cells for each 10***6 plated cells). The frequency of blasts and cells with surface Ig presented the same pattern of variation (o.07 and 0.02%, respectively, in control cultures). The inhibitory effect of IL-3+LPS on cell proliferation was evident from the first day of culture, but more apparent on day 3. Macrophage-colony stimulating factor (M-CSF, L929-conditioned medium) and LPS each given alone stimulated proliferation but reduced it when given together. In contrast, fetal liver cells were not affected by the simultaneous addition of IL-3 and LPS or by M-CSF and LPS. The mechanism of action of the cumulative effect of these two factors in unknown. Since crude cell-conditioned medium was used as the source of IL-3, it is possible that another factor present in this medium interacts with LPS to cause the inhibitory effect on cell proliferation