Sterol requirements of T-strain mycoplasmas

T-strain mycoplasmas are very sensitive to digitonin, amphotericin B, and progesterone. This sensitivity and the relatively high content of cholesterol found in the cells indicated a possible requirement of T-strain mycoplasmas for sterols. This suspected requirement was demonstrated directly in a lipid-poor medium and can be met by cholesterol, as well as by beta-sitosterol and to a lesser degree by 7-dehydrocholesterol, cholestanol, stigmasterol, and ergosterol but not by cholesterol laurate or cholestan-3-one. Coprostanol, epicoprostanol, and epicholestanol inhibited cell growth. This inhibition could be partially reversed by increasing the cholesterol concentration in the growth medium. Because of their sterol requirement and their unique requirement for urea, T-strain mycoplasmas might be classified as the third genus in the order Mycoplasmatales.     

LARGE-SCALE ISOLATION AND FRACTIONATION OF ORGANS OF DROSOPHILA MELANOGASTER LARVAE

Methods for the mass isolation of diverse organs from small animals are described. They involve novel devices: a mechanical dissecting system, a centrifugal agitator for the separation of fibrillar from globular particles, and a settling chamber for the fractionation at unit gravity of particles with sedimentation velocities above the useful range for centrifugation. The application of these methods to the isolation of polytene and nonpolytene nuclei from Drosophila melanogaster larvae is described.     

Regulation of the pentose phosphate cycle

1. A search was made for mechanisms which may exert a ;fine' control of the glucose 6-phosphate dehydrogenase reaction in rat liver, the rate-limiting step of the oxidative pentose phosphate cycle. 2. The glucose 6-phosphate dehydrogenase reaction is expected to go virtually to completion because the primary product (6-phosphogluconate lactone) is rapidly hydrolysed and the equilibrium of the joint dehydrogenase and lactonase reactions is in favour of virtually complete formation of phosphogluconate. However, the reaction does not go to completion, because glucose 6-phosphate dehydrogenase is inhibited by NADPH (Neglein & Haas, 1935). 3. Measurements of the inhibition (which is competitive with NADP(+)) show that at physiological concentrations of free NADP(+) and free NADPH the enzyme is almost completely inhibited. This indicates that the regulation of the enzyme activity is a matter of de-inhibition. 4. Among over 100 cell constituents tested only GSSG and AMP counteracted the inhibition by NADPH; only GSSG was highly effective at concentrations that may be taken to occur physiologically. 5. The effect of GSSG was not due to the GSSG reductase activity of liver extracts, because under the test conditions the activity of this enzyme was very weak, and complete inhibition of the reductase by Zn(2+) did not abolish the GSSG effect. 6. Preincubation of the enzyme preparation with GSSG in the presence of Mg(2+) and NADP(+) before the addition of glucose 6-phosphate and NADPH much increased the GSSG effect. 7. Dialysis of liver extracts and purification of glucose 6-phosphate dehydrogenase abolished the GSSG effect, indicating the participation of a cofactor in the action of GSSG. 8. The cofactor removed by dialysis or purification is very unstable. The cofactor could be separated from glucose 6-phosphate dehydrogenase by ultrafiltration of liver homogenates. Some properties of the cofactor are described. 9. The hypothesis that GSSG exerts a fine control of the pentose phosphate cycle by counteracting the NADPH inhibition of glucose 6-phosphate dehydrogenase is discussed.     

Iris Seed Dormancy

Iris embryos were treated in several ways to study the cause of delayed germination. The results of this work indicate that a chemical inhibitor is present in the endosperm and mechanical inhibition of embryo growth also occurs. The mechanical inhibition reduces excised Iris embryo growth to the same extent as does the chemical inhibitor.