The effects of expression of an activated rasG mutation on the differentiation of Dictyostelium.

Dictyostelium discoideum contains two ras genes, rasG and rasD, that are expressed during growth and differentiation, respectively. It was shown previously that Dictyostelium transformants expressing an activated rasD gene (a mutation producing a change in amino acid 12 from glycine to threonine) developed abnormally. When developed on filters these transformants formed multitipped aggregates, which did not go on to produce final fruiting bodies, but in a submerged culture assay on a plastic surface they either formed small aggregates or did not aggregate. In this study we transformed cells with the rasG gene, mutated to change amino acid 12 from glycine to threonine. The resulting transformants developed normally on filters, but aggregation under other conditions was impaired. In particular, in submerged culture on a plastic surface they either produced very small aggregates or did not aggregate, one of the phenotypes exhibited by the activated rasD transformants. Molecular analysis of the transformants revealed the presence of high copy numbers of the mutated rasG gene, but the level of expression of the mutant gene never exceeded the level of expression of the endogenous gene. These results indicate a powerful dominant effect of a relatively small amount of the activated RasG protein in Dictyostelium.     

How long should angiotensin-converting enzyme inhibitors be given to patients following myocardial infarction: implications of the HOPE trial

Long-term treatment with angiotensin-converting enzyme inhibitors reduces post-infarction morbidity and mortality in patients with left ventricular (LV) systolic dysfunction or symptomatic heart failure. Until recently, the effect of such treatment in patients with preserved LV function has not been known. The results from the Heart Outcome Prevention Evaluation trial have indicated that long-term treatment with ramipril leads to a significant reduction in cardiovascular events in patients with atherosclerotic disease, including those with prior myocardial infarction and preserved LV function. These results suggest that long-term angiotensin-converting enzyme inhibition should also be considered in post-infarction patients with normal cardiac function.     

Size Distribution of "Informational" RNA

Previous investigations suggested that the size of “informational” or “messenger” RNA was confined to sedimentation rates lying between 8 and 14S. These involved procedures permitting extended contact of the RNA with enzymatically active extracts. The present study re-examined the size distribution of T2-complementary RNA isolated by a method which minimized enzymatic degradation. A much greater diversity in size distribution (4S to 25S) was observed. Experiments are described indicating that 8 to 12S informational RNA does not readily attach to the 16S and 23S ribosomal components under the conditions used for sedimentation analysis.     

THE PREADAPTIVE OXIDATION OF GALACTOSE BY YEAST

A preadaptive purely aerobic utilization of galactose by yeast cells has been demonstrated. Hence, the adaptation by yeast to galactose is not to its utilization per se, but specifically to its metabolism by a glycolytic mechanism. An examination of this preadaptive oxidation of galactose reveals that it has many characteristics in common with the endogenous metabolism of yeast. Included among these are the similarities of the R.Q. values and the response of the Q _OO2 and Q_COCO2 ^O₂ to KCN and iodoacetic acid. Further, a competitive interaction appears to exist between the endogenous respiration and the preadaptive oxidation of the galactose. The latter can replace the endogenous respiration as a source of energy for the adaptation to the fermentation of the galactose. Carbon balance studies of the galactose oxidation revealed that polysaccharide could be formed as a result of this metabolism during the preadaptive period. Non-adaptable cells were also found to possess the capacity to oxidize galactose in the complete absence of any ability to metabolize it anaerobically. The significance of these findings for the biochemistry and physiology of the adaptation is discussed.     

THE RELATION OF ENZYMATIC ADAPTATION TO THE METABOLISM OF ENDOGENOUS AND EXOGENOUS SUBSTRATES

The source of energy for enzymatic adaptation has been investigated. Aerobically, it is found that the endogenous carbohydrate reserves may be used as such a source. In cells depleted of their reserves, the adaptive substrate itself can be oxidized even while it cannot be fermented, and so can serve as a source of energy for the adaptation to a fermentative mode of utilization. Anaerobically, adaptation may occur at the expense of stored energy-rich compounds, while the reserves and the adaptive substrate are now useless as fuel. Such compounds appear to be more plentiful in young than in old cells. The addition of any fermentable substrate, such as glucose, leads to rapid anaerobic adaptation. Experiments in which maltose-adapted cells are adapted anaerobically to galactose with the aid of a little added maltose, and conversely, show that fermentability is the criterion of usefulness for an exogenous substrate in aiding the adaptive process. None of the endogenous and exogenous energy sources which have been investigated will facilitate adaptation unless the adaptive substrate is present while they are being consumed. The significance of these findings and the adequacy of "activation" hypotheses to explain enzymatic adaptation has been discussed.