Cytochrome P-455 nm complex formation in the metabolism of phenylalkylamines. XII. Enantioselectivity and temperature dependence in microsomes and reconstituted cytochrome P-450 systems from rat liver.

Formation of metabolic intermediate (MI) complexes was studied with the enantiomers of amphetamine, 1-phenyl-2-pentanamine, N-hydroxyamphetamine, and 2-nitroso-1-phenylpropane (the C-nitroso analogue of amphetamine). Three different enzyme systems were used; liver microsomes from phenobarbital pretreated rats and two reconstituted systems containing the P450 2B1 and P450 2C11 forms of cytochrome P-450. Enantioselective complex formation in microsomes was shown for the amines and the nitroso compound, but not for the hydroxylamine. The highly purified P450 2B1 system formed the MI complex with all substrates tested, and the enantioselectivity observed with the microsomal system was reproduced. In the P450 2C11 system the nitroso compounds were completely inactive, whereas the enantiomers of N-hydroxyamphetamine still produced the complex at a high rate. Changes in temperature were shown to affect (R)-2-nitroso-1-phenylpropane more than its enantiomer. Both enantiomers showed biphasic Arrhenius plots for MI complex formation in microsomes (breaks around 22 degrees C), but the activation energies of the (R)-isomer were about five times higher than those of the (S)-isomer. A theory is presented which suggests different modes of interaction with the active site of P-450 to account for the different behaviour of the various substrates.     

Studies of the relationship between isoprene emission rate and CO2 or photon-flux density using a real-time isoprene analyser

Abstract. Studies of the isoprene emission rate in response to changes in photon-flux density and CO₂ partial pressure were conducted using a recently developed on-line isoprene analyser combined with a gas exchange system and chlorophyll fluorometer. Upon darkening, the isoprene emission rate from leaves of aspen ( Populus tremuloides Michaux.) began to decline immediately, demonstrating that the internal pool of isoprene, or its precursors, is small and that the instantaneous emission rate is tightly coupled to the rate of synthesis. A post-illumination burst of isoprene was observed within 5 min after darkening and lasted for 15–20 min in four isoprene-emitting species that were examined. In leaves of eucalyptus ( Eucalyptus globulus Labill.), the magnitude of the post-illumination burst was dependent on the photon-flux density that existed before darkening, but not on ambient CO₂ partial pressure. The dependence of the post-illumination burst on photon-flux density paralleled that for the steady-state rate of isoprene emission. A step-wise increase in intercellular CO₂ partial pressure from 24.5 to 60 Pa resulted in an immediate decrease in isoprene emission rate and non-photochemical fluorescence quenching, but an increase in CO₂ assimilation rate. Given the several recent studies that link isoprene emission to chloroplastic processes, the results of this study indicate that the linkage is not dependent on the rate of CO₂ flux through the reductive pentose phosphate pathway, but rather on more complex relationships involving metabolites not appreciably influenced by CO₂ partial pressure.     

The appearance of an enzyme activity catalysing the conversion of norsolorinic acid to averantin in Aspergillus parasiticus cultures

The activity of the enzyme responsible for the conversion of norsolorinic acid to averantin was studied in two strains of Aspergillus parasiticus. Cell-free extracts of the enzyme were purified from different aged mycelia and little activity was found prior to 24 hours after inoculation but this quickly reached a maximum at 48 hours and declined thereafter. Both strains of A. parasiticus, one in aflatoxin producing strain, the other a versicolorin A accumulating mutant, showed this trend. It was concluded that the enzyme responsible for this conversion was a secondary metabolic enzyme and was distinct from alcohol and mannitol dehydrogenases.     

Acclimatory responses in enzymatic activity and metabolite concentrations in the tropical scorpion Heterometrus fulvipes

1. 1.|Changes in tissue metabolite concentrations and enzyme activities in the pedipalpal (PM) and heart (HM) muscles of the tropical scorpion Heterometrus fulvipes show that the metabolism in PM and HM is fundamentally reorganized following low (18°C) and high (38°C) temperature acclimation.

2. 2.|Changes in metabolite concentrations show that metabolite biosynthesis showed increases after cold acclimation but decreases after warm acclimation.

3. 3.|Similarly, changes in enzyme activities show a preponderance of glycolysis and HMP shunt activity after cold acclimation, while after warm acclimation glycogenolysis, oxidative metabolism and gluconeogenesis predominated.

4. 4.|Higher metabolite concentrations and enzyme activities both before and after thermal acclimation in HM reflect its greater compensatory abilities.

根田鼠静止代谢率特征的研究

通过实地测定分布于青海高原的根田鼠的RMR,发现其RMR水平明显高于与其体重相似的其它田鼠,而平均最小热传导值并不比后者低,说明可能是以保持高的RMR水平来适应高原寒冷气候的。同时发现,根田鼠以化学体温调节为主,以适应高原气温较大的波动。     

Molecular epidemiology of plasmid patterns in Shigella dysenteriae type I obtained from an outbreak in West Bengal (India)

Abstract Multiple antibiotic-resistant Shigella dysenteriae type 1 isolates from a recent epidemic in West Bengal (India) showed identical plasmid patterns. All isolates were resistant to ampicillin (Am), chloramphenicol (Cm), tetracycline (Tc), streptomycin (Sm) and trimethoprim (Tp) and contained 6 plasmids, ranging from 2.5–120 kb. The Am resistance determinant was located on the 120 kb plasmid. This plasmid was unstable when the S. dysenteriae strains were grown above 37°C. The Bangladesh strains of S. dysenteriae type 1 showed identical plasmid patterns, except that many isolates were Am-sensitive and lacked the 120 kb plasmid. In strains from both Bangladesh and West Bengal, predominantly group-B plasmids conferred resistance to Cm and Tc. Comparisons of Eco R1 fragments generated from the total plasmid DNA content of each strain support the view that the plasmids present in the S. dysenteriae type 1 strains isolated from all recent epidemics in India and Bangladesh were identical.     

Effects of phorbol myristate acetate,phorbol dibutyrate,ethanol, dimethylsulfoxide,phenol, and seven metabolities of phenol on metabolic cooperation between chinese hamster v79 lung fibroblasts

The effect of phorbol myristate acetate, phorbol dibutyrate, ethanol, dimethylsulfoxide, phenol, and seven metabolites of phenol on metabolic cooperation were assessed as a function of mutant cell recovery from populations of cocultivated hypoxanthine-guanine phosphoribosyl transferase-deficient mutant (HGPRT–) and wild-type (HGPRT+) Chinese hamster V79 lung fibroblasts. Phorbol myristate acetate and phorbol diputyrate, two established tumor promoters, were potent inhibitors of metabolic cooperation. Ethanol and dimethylsulfoxide, solvents commonly used to prepare chemicals for testing, weakly inhibited metabolic cooperation. Phenol and phenylglucuronide had no effect on metabolic cooperation. Four oxidative metabolites (1,4-benzoquinone, catechol, hydroxyquinol and quinol) inhibited metabolic cooperation. Phenylsulfate weakly inhibited metabolic cooperation. Conversely, 2-methoxyphenol, a methylated derivative of catechol, appeared to enhance metabolic cooperation. These results generallyAbbreviations CAS Chemical Abstracts Service - DMSO dimethylsulfoxide - ETOH ethanol - HGPRT hypoxanthine-guanine phosphoribosyl transferase - HGPRT+ HGPRT-competent - HGPRT– HGPRT-te]deficient - MC metabolic cooperation - MC+ metabolic cooperation-competent - MC– metabolic cooperation-deficient - MEM minimum essential medium - PDBu phorbol dibutyrate - PMA phorbol myristate acetate - 6TG 6-thioguanine - 6TG^r 6-thioguanine-resistant - 6TG^s 6-thioguanine-sensitive - V79/MC assay Chinese hamster V79 lung fibroblast assay for metabolic cooperation     

Age-dependent metabolic changes in cultured human fibroblasts

Summary The effects of metabolic poisons on the ATP content of cultured human skin fibroblasts at selected in vitro and in vivo ages were studied. Potassium cyanide, iodacetemide, and Arsenate were used to inhibit ATP restoration by glycolysis and oxidative phosphorylation. Cells treated with these metabolic poisons showed an age-dependent change in their ATP content. The decrease in cellular ATP content after exposure to these drugs was taken as an estimate of ATP turnover. It was found that there was a decrease in the ATP turnover with increasing population doubling level (i.e. in vitro age), and cells cultured from a 68-yr-old donor had a lower ATP turnover than those cultured from a neonatal donor. This decreased ATP turnover correlates with a previous finding of a decreased ability of “older” cells to be stimulated to migrate in culture and suggests that there is a metabolic component to this age-related functional deficiency. This work was supported by National Institutes of Health grants 2, RO1 EY02523 and 1 RO1 1, AGO 1212 awarded to A.L. Muggleton-Harris.