Influence of Temperature on Glucose Metabolism of Round Spermatids of Rats

Glucose utilization by spermatids was found to be 17.37±0.37 nmoles/hr/10⁶ cells at 34°C and 28.94±1.12nmoles/hr/10⁶ cells at 40°C. A good parallelism was observed between the increased rate of glucose utilization and lactate production at 40°C. There was no significant change in the levels of glycolytic intermediates in the cells, except for marked accumulations of fructose-1, 6-diphosphate, dihydroxyacetone phosphate and glyceraldehyde-3-phosphate in the presence of glucose (1 mM). Glucose oxidation in the citrate cycle by spermatids was higher at 40°C than at 34°C, but was never greater than 2% of the overall rate of glucose utilization. In addition, glucose did not prevent decrease of ATP at either 34 or 40°C. The effects of temperature on the activities of 11 glycolytic enzymes were examined. The activities of aldolase and phosphoglyceromutase were similar between 30 and 34°C, but increased markedly at 40°C. The higher temperature increased the V_max values, without affecting the Kms. The activities of other glycolytic enzymes were similar at the different temperatures. These findings indicate that the increased overall rate of glucose utilization in glycolysis at higher temperature is due to increased Vmax values of aldolase and phosphoglyceromutase.     

Studies on the metabolism of a sulfur-oxidizing bacterium VII. Oxidation of sulfite by a cell-free extract of Thiobacillus thiooxidans

Properties of the cell-free extract, prepared from a strainof Thiobacillus thiooxidans by sonic disruption followed byfractionation with centrifugatiori, were investigated with referenceto its sulfite-oxidizing activity. Without the addition of cofactors the particulate fraction(F-P)catalyzed oxidation of sulfite with oxygen or bacterial cytochromec-552 obtained from Pseudomonas stutzeri as electron acceptor.TMPD reduced by ascorbic acid was also oxidized by F-P. Thesoluble fraction(F-S) showed no activity in oxidizing sulfiteand TMPD, but stimulated TMPD oxidation by F-P. Oxygen uptake with either sulfite or TMPD as substrate was inhibitedby KCN, NaN₃, CO and c-phenanthroline. CO-Inhibition was reversedby light. Reduction of cytochrome c-552 by sulfite was insensitiveto these agents. Antimycin A markedly inhibited sulfite oxidation with eitheroxygen or cytochrome c-552 as electron acceptor, but was withouteffect on TMPD oxidation. DDC and SAO, both strong inhibitors of sulfur oxidation, didnot affect sulfite and TMPD oxidations. Cytochromes of the a, b and c types were contained in F-P. Thesecytochromes were rapidly reduced when F-P was incubated withsulfite. Cytochrome(s) of the c type was present in F-S, too. ¹VI.=References (3) ²Partly supported by a grant from the Ministry of Education ³Present address: Sanyo Women's College, Hatsukaichi, Hiroshima738, Japan ⁴Present address: Department of Biochemistry, Hiroshima UniversitySchool of Dentistry, Hiroshima 734, Japan (Received May 15, 1970; )     

PHYSIOLOGICAL RESPONSES OF LIGHT-GROWN CUCUMBER HYPOCOTYLS TO HELMINTHOSPOROL AND ITS DERIVATIVES

Helminthosporol, helminthosporic acid and dihydrohelminthosporicacid stimulated hypocotyl elongation of light-grown cucumberseedlings. The relative activities of the three compounds werein the order H₂-H-acid> H-acid> H-ol. The higher the dosage,the longer the promotion of the hypocotyl elongation lasted.The response of the hypocotyl to H-acid depended on the presenceof cotyledons. As the amount of the cotyledon attached to thehypocotyl was reduced, the response decreased. When IAA-oxidase was estimated as IAA metabolized per dry weightof the hypocotyl, there was an inverse relationship betweenIAAoxidase activity and H-ol-induced elongation of the hypocotyl.However, there was no difference between control and H-ol treatedmaterials when IAA metabolized per plant was compared. It isunlikely that the mechanism of H-ol action is closely relatedto IAA-oxidase activity of the plant. (Received April 5, 1967; )     

STUDIES ON THE METABOLISM OF A SULFUR-OXIDIZING BACTERIUM: I. OXIDATION OF SULFUR

1. Thiobacillus thiooxidans, isolated in our laboratory, was foundto oxidize sulfur, but not thiosulfate. Tetrathionate is alsooxidized slightly. Its ability to oxidize sulfur is inactivatedeven by such a mild treatment as keeping the cells in a frozenstate.
2. Inhibitory action of alcohols on the sulfur oxidationincreasesas the length of carbon chain of alcohols increases.Carboxylicacids do not inhibit the sulfur oxidation at pH abovetheirpK, while they strongly inhibit the reaction at pH belowthepK.
3. The sulfur oxidation is inhibited by cyanide, azide,diethyldithiocarbamateand carbon monoxide, and the inhibitionby carbon monoxide isnot reversed by light. These results suggestthe presence ofmetal enzymes in the sulfur oxidation system.The terminal enzymeof this reaction appears to be differentfrom the usual cytochromeoxidase.

(Received May 13, 1960; )     

Light Microscopic Observations of Infraciliature and Morphogenesis in Six Species of Rumen Ostracodinium Ciliates

Infraciliature and morphogenesis of six rumen ciliates, Ostracodinium mammosum, O. munham, O. dilobum, O. rugoloricatum, O. iwawoi, and O. tiete are described from pyridinated silver carbonate-impregnated specimens. These six Ostracodinium have a similar polybrachykinety arrangement, distinct from that of other ophryoscolecid ciliates and intermediate between those of O. gracile and 0. damaliscus. Buccal infraciliature is composed of three polybrachykineties, a kinety loop, and paralabial kineties. Small dorso-adoral polybrachykinety, slender vestibular polybrachykinety, and kinety loop are characteristic. Dorsal infraciliature consists of the dorsal polybrachykinety that extends laterally along the dorsal side of the body. Morphogenesis is different from that of O. gracile, because the right end of ventral primordium extends dorsally to grow into a right primordium without separation.     

Effects of Organic Matter on Solubilities and Crystal Form of Carbonates

The results of a study of the role of organic compounds in theformation of carlxmate crystals in marine biological systemsare reported. In an increasing concentration of certain organiccompounds which complex calcium ions, the proportion of aragonitedecreases and that of calcite increases. In increasing concentrationsof magnesium ions the proportion of aragonite increases andthat of calcite and vaterite decreases. When the influence oforganic compounds is greater or smaller than that of magnesiumions, only calcite or only aragonite is formed, respectively.Organic compounds forming a strong complex with calcium ionscause the formation of magnesium-rich calcite, and with an increasein temperature and the concentration of magnesium ions, themagnesium carbonate content of precipitated magnesian calciteincreases. When the influence of organic compounds is almostequivalent to that of magnesium ions, in increasing or decreasingtemperatures, the proportion of calcite decreases or increases,respectively, and the proportion of aragonite increases or decreases,respectively. The concentration of magnesium ions in the bodyfluids of marine calcareous organisms seems to differ littlefrom that of other organisms, and seems to be similar to thatof sea water. Only the presence of certain organic compoundsbrings about the formation of the carbonate crystals observedin marine biological systems. The very important role of organicmatter in the formation of crystals found in skeletal carbonatesis emphasized.     

Melanoma Antigen and Transforming Gene

We summarize our recent studies on the analysis of melanoma antigen and the melanoma gene recently cloned. The melanoma antigen analyzed by syngeneic monoclonal antibodies is composed of a complex of GM3 ganglioside and protein molecules. The epitope recognized by antimelanoma cytotoxic T lymphocytes seemed to be the combinatorial determinants of GM3 and proteins, whereas the epitope for the suppressor T cells was found to be solely GM3 (NeuAc). The genomic DNA controlling the melanoma antigen was recently isolated and was found to possess transforming activity. The structure of this transforming gene is discussed based on the sequence data of the corresponding cDNA clone.