Inhibitor effect of products of metabolism on growth of Clostridium acetobutylicum

Summary The growth of Clostridium acetobutylicum was studied by three ways. 1. In batch fermentation, referred to as the control. 2. Fermentation in dialysis which permits elimination of all the products of metabolism: acids, solvents and gases. In order to test the toxic effect of acids, cultures were dialysed against 2 g l^-1 acetic acid or 2 g l^-1 butyric acid. 3. To test the toxic effect of gases only, batch fermentations were carried out under vacuum or with a continuous bubbling of nitrogen. The first method resulted in a productivity of 1.2 g l^-1 dry cell weight and a maximal specific growth rate of 0.2 h^-1; the second, 20 g l^-1 dry cell weight and a constant maximum specific growth rate (μ=0.39 h^-1) between 14 and 20 h. The toxic effect of acetic and butyric acids, starts at low concentrations and about 4 g l^-1 of both acids results in a decrease of 50% of maximal specific growth rate. The third series of experiments showed that gases produced by the bacteria have a high toxic effect, comparable to that of 5 g l^-1 of acid.     

Nebivolol enhances the effect of alendronate against methylprednisolone‐induced osteoporosis in rats

This study aimed to assess the possible modulatory effect of nebivolol against methylprednisolone‐induced osteoporosis in rats. Weekly administration of methylprednisolone (7 mg/kg), for six consecutive weeks caused significant increases in serum calcium, bone malondialdehyde, and hydroxyproline as well as serum alkaline phosphatase, but it significantly decreased serum phosphorous and osteocalcin, bone reduced glutathione, and nitric oxide (NO) as well as bone antioxidant enzymes activities compared with the control group. The results were confirmed by histopathological findings of femur bone. On the other hand, administration of alendronate (1 mg/kg) with nebivolol (1.5 mg/kg) orally and daily for seven consecutive days after methylprednisolone treatment caused marked mitigation in the above‐mentioned parameters compared with methylprednisolone group. In conclusion, nebivolol proved to enhance the effect of alendronate in modulating methylprednisolone osteoporotic effect, which might be attributed to its release of NO together with its profound reducing capability in the oxidative cascade of bone tissue.     

The effect of methylation on DNA replication in Salmonella enterica serovar typhimurium

The DNA adenine methylase of Salmonella typhimurium methylates adenine at GATC sequences. Strains deficient in this methylase are not well transformed by methylated plasmids, but unmethylated plasmids transform them at high frequencies. Hemimethylated daughter molecules accumulate after the transformation of dam(-) strains with fully methylated plasmids, suggesting that hemimethylation prevents DNA replication. It will also be shown that plasmids isolated from dam(-) bacteria are hemimethylated by restriction enzyme digestion. These results may explain why newly formed daughter molecules are not substrates for immediate reinitiation of DNA replication in dam(-) bacteria.     

Description of Lamellodiscus confusus n. sp, (Monogenea: Diplectanidae), parasite of Sarpa salpa (Teleostei: Sparidae)

Lamellodiscus confusus n. sp., a diplectanid gill parasite of Sarpa salpa Linnaeus, 1758 (Sparidae) from the coast of Algeria, is described. This monogenean has been previously reported in Sarpa salpa, by various authors, under the name L. ignoratus. L. confusus n. sp. is included (among other Lamellodiscus of Mediterranean sparids) in the "ignoratus" sub-group (Amine & Euzet, 2005). L. confusus n. sp. can be distinguished from L. ignoratus by the shape and size of the haptoral ventral bar and the male copulatory organ. For comparison, illustrations of the ventral bar and the male copulatory organ of all Mediterranean species of the "ignoratus" sub-group are provided. The question of the host specificity of L. ignoratus is discussed.     

Protein Disulfide Isomerase Is Required for Platelet-derived Growth Factor-induced Vascular Smooth Muscle Cell Migration, Nox1 NADPH Oxidase Expression, and RhoGTPase Activation

Vascular Smooth Muscle Cell (VSMC) migration into vessel neointima is a therapeutic target for atherosclerosis and postinjury restenosis. Nox1 NADPH oxidase-derived oxidants synergize with growth factors to support VSMC migration. We previously described the interaction between NADPH oxidases and the endoplasmic reticulum redox chaperone protein disulfide isomerase (PDI) in many cell types. However, physiological implications, as well as mechanisms of such association, are yet unclear. We show here that platelet-derived growth factor (PDGF) promoted subcellular redistribution of PDI concomitant to Nox1-dependent reactive oxygen species production and that siRNA-mediated PDI silencing inhibited such reactive oxygen species production, while nearly totally suppressing the increase in Nox1 expression, with no change in Nox4. Furthermore, PDI silencing inhibited PDGF-induced VSMC migration assessed by distinct methods, whereas PDI overexpression increased spontaneous basal VSMC migration. To address possible mechanisms of PDI effects, we searched for PDI interactome by systems biology analysis of physical protein-protein interaction networks, which indicated convergence with small GTPases and their regulator RhoGDI. PDI silencing decreased PDGF-induced Rac1 and RhoA activities, without changing their expression. PDI co-immunoprecipitated with RhoGDI at base line, whereas such association was decreased after PDGF. Also, PDI co-immunoprecipitated with Rac1 and RhoA in a PDGF-independent way and displayed detectable spots of perinuclear co-localization with Rac1 and RhoGDI. Moreover, PDI silencing promoted strong cytoskeletal changes: disorganization of stress fibers, decreased number of focal adhesions, and reduced number of RhoGDI-containing vesicular recycling adhesion structures. Overall, these data suggest that PDI is required to support Nox1/redox and GTPase-dependent VSMC migration.     

MVL-PLA2, a phospholipase A2 from Macrovipera lebetina transmediterranea venom,inhibits tumor cells adhesion and migration

Here, we report the purification and characterization of an acidic Asp49 phospholipase A2, named MVL-PLA2, with a molecular mass of 13,626.64 Da. The complete MVL-PLA2 cDNA was cloned from Macrovipera lebetina transmediterranea venom gland cDNA library. MVL-PLA2 possesses 122 amino acid residues, including 14 cysteines, and belongs to group II snake venom phospholipase A2 enzymes. MVL-PLA2 was not cytotoxic up to 2 μM and completely abolished cell adhesion and migration of various human tumor cells. Chemical modification with p-bromophenacyl bromide abolished the enzymatic activity of MVL-PLA2 without affecting its anti-tumor effect, suggesting the presence of ‘pharmacological sites’ distinct from the catalytic site in snake venom phospholipase A2. We demonstrated for the first time that the anti-tumor effect of MVL-PLA2 was mediated by α5β1 and αv-containing integrins. This finding may serve as starting point for structure–function relationship studies leading to design a new generation of specific anti-cancer drugs.     

Differences in pharmacokinetic and electroencephalographic responses to caffeine in sleep-sensitive and non-sensitive subjects

The present study investigated pharmacokinetic and electroencephalographic responses to caffeine (140 mg) in two groups of healthy volunteers reporting, or not, caffeine-related sleep disturbances. Significant differences in caffeine consumption and smoking habits were observed between the two groups. Plasma samples were taken from each subject before (T0) and after caffeine intake at 0.5, 1, 2, 4, 6 and 24 h. Three pharmacokinetic parameters: half-life (t1/2), maximum time (Tmax) and maximum plasma concentration (Cmax) were calculated from caffeine plasma concentration measurements determined by reversed phase HPLC analysis. Caffeine-sensitive subjects showed significantly greater half-life values when calculated on 24 h after the administration than tolerant subjects (p<0.05). Since the elimination kinetics were similar on the first 6 h after caffeine administration, the increased caffeine clearance observed overnight, when smoking was resumed in the control group, may indicate a short delay for the induction of hepatic cytochrome, reported here for the first time. Electrophysiological responses to caffeine, including vigilance and cortical activity, were assessed by ambulatory electroencephalographic (EEG) recorded during a period of 6 h before and after caffeine consumption. Following caffeine intake, the caffeine-intolerant subjects presented an increase in vigilance levels with faster peak alpha, beta frequency and lower delta and theta power when compared to tolerant subjects. Pharmacokinetic parameters and EEG data showed significant differences between sleep-sensitive and control subjects. These variations may be, in part, explained by cigarette smoking and the higher caffeine intake observed in the subjects of the control groups while caffeine sleep-sensitive subjects have a significantly lower caffeine intake, as already reported in previous studies on patients with sleep disturbances.