Kinetic behaviour of succinate dehydrogenase of three fibre types in skeletal muscle. I. Effects of temperature and a competitive inhibitor

Summary The kinetic behaviour of succinate dehydrogenase [EC 1.3.99.1] in three fibre types of rat gastrocnemius was examined by a quantitative histochemical method without disruption of the cellular structure. 2-(2-Benzothiazolyl)-3-(4-phthalhydrazidyl)-5-styryl-tetrazolium chloride (BPST) and phenazine methosulphate were used as electron acceptors. On measurement of the absorbance value at 530 nm of BPST formazan, produced by the succinate dehydrogenase reaction in sections, it was found that the staining intensity of succinate dehydrogenase was linearly proportional to both the incubation time and the thickness of the slice therefore, the initial velocity of the staining could be calculated. By Michaelis-Menten (1913) treatment of the dependence of the initial velocity on the substrate concentration in the absence and the presence of a competitive inhibitor, malonate, the Km andVmax values for succinate and the Ki value for malonate were obtained. The Km and Ki values of the three fibre types were similar. The ratio of theVmax values of type A, B and C fibres was 1.02.03.3. The temperature dependence of the kinetic parameters was very similar in the three fibre types. These findings confirm that the differences in the staining intensity of the three fibre types reflect differences in the amounts, but not the properties, of succinate dehydrogenase.     

High concentration cultivation of Bifidobacterium longum in fermenter with cross-flow filtration

Summary High concentration cultivation of Bifidobacterium longum in a fermenter with cross-flow filtration using a ceramic filter is described. Continuous cross-flow filtration allowed complete recycling of the cells to the fermenter and also continuous separation of inhibitory metabolites. The final cell concentration attained in the cultivation was 54.4 g dry wt./l; this was seven times as high as that without cross-flow filtration. The time course of the cultivation with cross-flow filtration was predicted, based on the assumption that the specific growth rate can be expressed only as a function of concentrations of metabolites (acetate and lactate) in a culture broth.Nomenclature D dilution rate (h^-1) - m maintenance coefficient (h^-1) - OD ₅₇₀ optimal density at 570 nm - P _A acetate concentration (g/l) - P _A0 initial acetate concentration (g/l) - P _L lactate concentration (g/l) - P _L0 initial lactate concentration (g/l) - S lactose (substrate) concentration (g/l) - S ₀ initial lactose (substrate) concentration (g/l) - t cultivation time (h) - Y _x/s growth yield (g/g) - X dry cell concentration (g/l) - X ₀ initial dry cell concentration (g/l) - constant - constant     

The roles of glucagon and adrenal epinephrine in mediating hyperglycemia induced by third cerebroventricular injection of bombesin

The roles of glucagon and adrenal epinephrine in mediating bombesin-induced central hyperglycemia were further studied in anesthetized rats. Bombesin (10(-9) mol) injected into the third cerebral ventricle produced an increase in plasma concentrations of glucose, glucagon, and epinephrine. Prior bilateral adrenalectomy completely prevented the hyperglucagonemic and hyperglycemic responses to third cerebral ventricle injection of bombesin. These results support the view that bombesin-induced increases in plasma glucose and glucagon are fully dependent on adrenal epinephrine secretion. Furthermore, during constant intravenous infusion of somatostatin, the hyperglycemic response to third cerebral ventricle injection of bombesin was not significantly influenced despite complete inhibition of the increase in plasma glucagon. Therefore, it is suggested that bombesin-induced central hyperglycemia is mainly mediated by epinephrine itself rather than via epinephrine-stimulated glucagon secretion.     

Mercury pollution in Tokuyama Bay

The sediments and aquatic life of Tokuyama Bay, Japan, have been polluted by mercury effluent from chloro-alkali plants. In total, about 380 tons mercury were released from these plants and 6.64 tons of mercury were discharged into the bay in waste waters between 1952 and 1975, when mercury cells were employed. A number of surveys to study mercury pollution and the effectiveness of control measures in this area were conducted in the early 1970's by our laboroatory and other agencies. Analysis of human hair from Tokuyama Bay residents contained less mercury than those in Minamata and Agano districts, Japan, where serious mercury poisoning had occurred, but were contaminated with more mercury than those in other unpolluted areas. No occurrence of Minamata disease has been reported in the Tokuyama district.Reclamation of mercury contaminated sediments began in 1975; dredging of the bay continued until 1977. Since then, the levels of mercury contamination in sediments and aquatic life have gradually decreased. Today there are no problems with respect to mercury pollution.In this paper, we describe and discuss mercury pollution in Tokuyama Bay with regard to the following aspects of research and pollution control: the history of mercury pollution; mercury discharge and its accumulation in sediments; behaviour of mercury in sediments; mercury contamination of fish; mercury and the health of local residents; and remedial actions.     

Botryococcus braunii carbon/nitrogen metabolism as affected by ammonia addition

Carbon metabolism in photosynthesizing and respiring cells of Botryococcus braunii was radically changed by the presence of 1 mM NH₄Cl in the medium, when the so-called resting state previously had been subjected to a nitrogen-deficient medium. Ammonia addition to the algae photosynthesizing with ¹⁴C-labelled HCO ₃ ^- almost completely inhibited the synthesis of ¹⁴C-labelled botryococcenes and other hexane-extractable compounds, and also inhibited the formation of insoluble compounds; however, it resulted in a large increase in the synthesis of alanine, glutamine, other amino acids, and especially of 5-aminolevulinic acid. Total CO₂ fixation decreased about 60% and O₂ evolution decreased more than 50%.CO₂ fixation in the dark with ammonia present led to labelled products derived from phosphoenolpyruvate carboxylation, such as glutamine, glutamate, and malate. Respiratory uptake of O₂ increased by about 70%.The inhibition of terpenoid synthesis and increased synthesis of C₅ amino acids by Botryococcus upon ammonia addition indicates 1) a diversion of acetyl coenzyme A from synthetic pathways leading to terpenoids and 2) increased operation of pathways leading to the synthesis of amino acids, especially 5-aminolevulinic acid, a precursor to chlorophyll biosynthesis.This work was supported in part by the Office of Energy Research, Office of Basic Energy Sciences, Biological Energy Research Division of the U.S. Department of Energy under Contract No. DE-AC03-76SF00098, in part by a grant from SOHIO, and, in part, by a grant from the Japan-U.S. Cooperative Science Program (The Japan Society for the Promotion of Science, National Science Foundation, Division of International Programs)     

Isolation of an Arabidopsis thaliana Mutant in Which the Multiplication of both Cucumber Mosaic Virus and Turnip Crinkle Virus Is Affected

During the systemic infection of plants by viruses, host factors play an important role in supporting virus multiplication. To identify and characterize the host factors involved in this process, we isolated an Arabidopsis thaliana mutant named RB663, in which accumulation of the coat protein (CP) of cucumber mosaic virus (CMV) in upper uninoculated leaves was delayed. Genetic analyses suggested that the phenotype of delayed accumulation of CMV CP in RB663 plants was controlled by a monogenic, recessive mutation designated cum2-1, which is located on chromosome III and is distinct from the previously characterized cum1 mutation. Multiplication of CMV was delayed in inoculated leaves of RB663 plants, whereas the multiplication in RB663 protoplasts was similar to that in wild-type protoplasts. This suggests that the cum2-1 mutation affects the cell-to-cell movement of CMV rather than CMV replication within a single cell. In RB663 plants, the multiplication of turnip crinkle virus (TCV) was also delayed but that of tobacco mosaic virus was not affected. As observed with CMV, the multiplication of TCV was normal in protoplasts and delayed in inoculated leaves of RB663 plants compared to that in wild-type plants. Furthermore, the phenotype of delayed TCV multiplication cosegregated with the cum2-1 mutation as far as we examined. Therefore, the cum2-1 mutation is likely to affect the cell-to-cell movement of both CMV and TCV, implying a common aspect to the mechanisms of cell-to-cell movement in these two distinct viruses.     

Japanese Rice Wine can reduce psychophysical stress-induced depression-like behaviors and Fos expression in the trigeminal subnucleus caudalis evoked by masseter muscle injury in the rats

We determined if Japanese Rice Wine (Sake) had inhibitory effects on stress-induced enhancement of masseter muscle (MM) nociception in the rats. Male rats were subjected to the repeated forced swim stress (FS) or sham conditionings from Day ?3 to ?1. Daily administration of Sake or saline was conducted after each stress conditioning. At Day 0 the number of Fos positive cells, a marker for neural activity, was quantified at the trigeminal subnucleus caudalis (Vc) region by MM injury with formalin. FS increased MM-evoked Fos expression in the Vc region, which was inhibited by Sake compared to saline administration. Sake did not alter the number of Fos positive cells under sham conditions, indicating that inhibitory roles of Sake on neural activity in the Vc region were seen under FS conditions. These findings indicated that Sake had inhibitory roles on stress-induced MM nociception at the Vc region in our experimental conditions.