Effect of oxygen on ubiquinone-10 production by Paracoccus denitrificans

Summary Ubiquinone-10 (Q₁₀) production was measured in batch cultures of Paracoccus denitrificans grown for 8 h at increasing oxygen concentrations (0–21 % O₂ in the sparging gas). Whereas the cellular level of Q₁₀ decreased monotonically from 1.2 to 0.5 mol/g d.w., the total yield of Q₁₀ was maximal at 2.5 % O₂ and amounted to 350 nmol (0.3 mg) per L of culture.     

The Chemiluminescence Assay of Lipid Peroxidation Products in Human Blood Plasma Lipoproteins

A chemiluminescence (CL) flash kinetics on the addition of Fe²⁺ ions into oxidized low density lipoprotein (LDL) suspension has been studied. LDL oxidation was carried out at 37°C without and in the presence of 5 or 50 μM of Cu.²⁺ It has been found that under certain experimental conditions (the addition of excess iron ions, more than 1 mM) the amplitude of CL flash depended almost linearly (1) on the concentration of oxidized LDL and (2) on the extent of LDL oxidation measured as diene conjugates (DC) and 2-thiobarbituric acid-reactive substance (TBARS) accumulation. The corresponding correlation coefficients were: for TBARS - 0.94 and for DC - 0.97, in the case of LDL autooxidation; 0.72 and 0.98, in the case of copper-induced LDL oxidation. A sensitivity of the CL method was shown to be significantly enhanced (by more than two orders) in the presence of CL sensitizer - 2, 3,5, 6-lH,4H-tetrahydro-9-(2' -benzoimidazolyl)-quinolizin-(9, 9a, 1 -gh)coumarin.     

Differential induction of regulatory genes during mesoderm formation in Xenopus laevis embryos

Mesoderm development in Xenopus laevis depends on inductive cell interactions mediated by diffusible molecules. The mesoderm inducer activin is capable of redirecting the development of animal explants both morphologically and biochemically. We have studied the induction of four regulatory genes, Mix. 1, goosecoid (gsc), Xlim-1 and Xbra in such explants by activin, and the influence of other factors on this induction. Activin induction of gsc is strongly enhanced by dorsalization of the embryo by LiCl, while expression of the other genes is only slightly enhanced. The protein synthesis inhibitor cycloheximide (CHX) inhibits the activin-dependent induction of Xbra partially, while induction of Mix. 1 and Xlim- 1 is essentially unaffected. In contrast, gsc shows strong superinduction in the presence of activin and CHX, and can be induced in animal explants by CHX alone. Induction and superinduction by CHX have previously been observed for immediate early genes in a variety of systems, notably for the activation of c-fos expression by serum stimulation, but have not been reported in early amphibian embryos. © 1993Wiley-Liss, Inc.     

Stereoselectivity in β-cyclodextrin complexation of 1,4-dihydropyridine derivatives

Structure–interaction relationships, stereoselectivity, and solubility enhancement in inclusion compexation of β-cyclodextrins (CDs) with some racemic and enantiomerically pure 1,4-dihydropyridine derivatives (DHPs) were investigated. 1:1 and 1:2 (mole ratio) complexes were prepared and characterized by X-ray powder diffraction, differential scanning calorimetry (DSC), MS-FAB spectrometry, ¹H-NMR spectroscopy, water and phase solubility. The solubility studies have revealed different complexation equilibria for optically pure DHP enantiomers, and corresponding racemic mixtures in water solutions. By means of ¹H-NMR chemical shift measurements, the inclusion of aromatic fragments of racemic and enantiomerically pure DHP molecules within the cavities of different CDs was elucidated. Considerable stereoselectivity in complexation interactions was observed. The results indicate the potential use of cyclodextrins as chiral selectors for enantiomeric resolution of 1,4-DHP calcium antagonists. © 1993 Wiley-Liss, Inc.     

Characterization of the serotoninergic system in the C57BL/6 mouse skin.

We showed expression of the tryptophan hydroxylase gene and of tryptophan hydroxylase protein immunoreactivity in mouse skin and skin cells. Extracts from skin and melanocyte samples acetylated serotonin to N-acetylserotonin and tryptamine to N-acetyltryptamine. A different enzyme from arylalkylamine N-acetyltransferase mediated this reaction, as this gene was defective in the C57BL6 mouse, coding predominantly for a protein without enzymatic activity. Serotonin (but not tryptamine) acetylation varied according to hair cycle phase and anatomic location. Serotonin was also metabolized to 5-hydroxytryptophol and 5-hydroxyindole acetic acid, probably through stepwise transformation catalyzed by monoamine oxidase, aldehyde dehydrogenase and aldehyde reductase. Activity of the melatonin-forming enzyme hydroxyindole-O-methyltransferase was notably below detectable levels in all samples of mouse corporal skin, although it was detectable at low levels in the ears and in Cloudman melanoma (derived from the DBA/2 J mouse strain). In conclusion, mouse skin has the molecular and biochemical apparatus necessary to produce and metabolize serotonin and N-acetylserotonin, and its activity is determined by topography, physiological status of the skin, cell type and mouse strain.     

Osmotic water permeability of cell membranes from Mesembryanthemum crystallinum leaves: effects of age and salinity

The osmotic water permeability ( P _os) of cell membranes isolated from leaves of 40-, 50- and 60-day-old Mesembryanthemum crystallinum plants was estimated by measuring light-scattering kinetics using stopped-flow spectrophotometry. The measurements were performed on the plasma membrane (PM), purified tonoplast (TP), and TP-enriched vesicles. The PM and TP-enriched vesicles were obtained by partitioning the microsomal fraction in an aqueous polymer two-phase system, whereas the purified TP vesicles were prepared by microsomal vesicle flotation on a sucrose cushion. The P _os of isolated membranes declined with plant age. The kinetic experiments showed that there was no difference between the P _os of the PM and TP isolated from plants of all ages. A 24-h exposure of plants to 400 m M NaCl caused a decline in the P _os as well. These findings suggest that, during M. crystallinum transition to CAM, which was induced by plant ageing or salinity, plant osmoregulatory responses included changes in the P _os of the leaf-cell membranes. These variations in the P _os are discussed in the context of adaptive mechanisms responsible for the maintenance of the water balance in the common ice plant.     

INTRAPOPULATION DIFFERENTIATION IN ANNUAL PLANTS. III. THE CONTRASTING EFFECTS OF INTRA- AND INTERSPECIFIC COMPETITION

The roles of intraspecific and interspecific competition in producing differentiation within populations of Veronica peregrina were studied in two populations under controlled, greenhouse conditions. In nature, each population spans an environmental gradient across the center and sides of a temporary, vernal pool in California. Individuals at the center are subjected to intense intraspecific competition produced by high densities (to 30 seedlings/cm²) generated by quasi-simultaneous germination (90% of seeds germinate in one week). Individuals at the periphery are subjected to interspecific competition with grasses, which shade out the Veronica 4–6 weeks after the onset of winter growth. I predicted that 1) when grown under immediate intraspecific competition in the greenhouse, offspring of plants from the central subpopulation (C) would perform better (i.e., grow larger and produce more seeds) than those from the periphery (P) and that 2) when grown under delayed interspecific competition provided by Agrostis tenuis and Lollium multiflorum, offspring of plants from the periphery would perform better than those from the center. Both predictions were confirmed. The center-periphery differences were pronounced and statistically significant in an undisturbed population (V-2), while in a population disturbed by yearly plowing (V-3), the differences tended to be consistent with those in V-2 but seldom significant. Distribution of variability tended to be positively skewed and/or leptokurtic in subpopulations grown under “foreign” competition (i.e., intraspecific for P plants and interspecific for C plants) but was normally distributed following exposure to “familiar” competition. Timing of competition affected many results. There were four additional significant differences between the central and peripheral subpopulations. 1) Germination rate: the faster rate in central plants can be advantageous under immediate intraspecific competition. The slower rate in peripheral plants can be advantageous under conditions of erratic and unpredictable soil moisture. 2) Response to nutrient competition: central plants were more sensitive to N-deficiency and peripheral plants were more sensitive to P-deficiency. 3) Allocation of biomass: central plants allocated a greater proportion of biomass to seeds, while peripheral plants allocated a greater proportion of biomass to leaves under all growing conditions. 4) Root elongation: at the seedling stage, central plants have longer roots, while at the adult stage, peripheral plants have longer roots (but not more root mass). Most components of this complex pattern of differentiation are interpretable in an adaptive context. Other results defy simple explanations and underline the importance of phenotypic plasticity, which was pronounced in the competition experiments.     

Similarity Between Naturally Occurring Modified Desialylated, Electronegative and Aortic Low Density Lipoprotein

The sialic acid content of electronegative low density lipoprotein (LDL) and LDL isolated from human aortic intima was measured. Sialic acid level in electronegative LDL of healthy subjects was 1.7-fold lower than in native LDL. Sialic acid content in electronegative LDL of coronary atherosclerosis patients was 3-fold lower than in native LDL. Lipoproteins isolated from grossly normal human aortic intima and from fatty streaks contained 20-56% less sialic acid as compared to blood plasma LDL. A negative correlation was established between the ability of electronegative and aortic LDL to stimulate lipid accumulation in cells cultured from uninvolved human aortic intima and lipoprotein sialic acid content. The results obtained indicate that electronegative and aortic LDLs have a low sialic acid content, i.e., are desialylated lipoproteins. Considered together with the fact that all known atherogenic LDLs have similar characteristics, our findings suggest that modified LDLs are the same lipoprotein particles subjected to multiple modification.