The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) pathway regulates developmental cerebral-vascular stability via prenylation-dependent signalling pathway

Spontaneous intracranial hemorrhage is a debilitating form of stroke, often leading to death or permanent cognitive impairment. Many of the causative genes and the underlying mechanisms implicated in developmental cerebral-vascular malformations are unknown. Recent in vitro and in vivo studies in mice have shown inhibition of the 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) pathway to be effective in stabilizing cranial vessels. Using a combination of pharmacological and genetic approaches to specifically inhibit the HMGCR pathway in zebrafish (Danio rerio), we demonstrate a requirement for this metabolic pathway in developmental vascular stability. Here we report that inhibition of HMGCR function perturbs cerebral-vascular stability, resulting in progressive dilation of blood vessels, followed by vessel rupture, mimicking cerebral cavernous malformation (CCM)-like lesions in humans and murine models. The hemorrhages in the brain are rescued by prior exogenous supplementation with geranylgeranyl pyrophosphate (GGPP), a 20-carbon metabolite of the HMGCR pathway, required for the membrane localization and activation of Rho GTPases. Consistent with this observation, morpholino-induced depletion of the β-subunit of geranylgeranyltransferase I (GGTase I), an enzyme that facilitates the post-translational transfer of the GGPP moiety to the C-terminus of Rho family of GTPases, mimics the cerebral hemorrhaging induced by the pharmacological and genetic ablation of HMGCR. In embryos with cerebral hemorrhage, the endothelial-specific expression of cdc42, a Rho GTPase involved in the regulation of vascular permeability, was significantly reduced. Taken together, our data reveal a metabolic contribution to the stabilization of nascent cranial vessels, requiring protein geranylgeranylation acting downstream of the HMGCR pathway.     

In silico and in vitro comparative activity of novel experimental derivatives against Leishmania major and Leishmania infantum promastigotes

This in silico and in vitro comparative study was designed to evaluate the effectiveness of some biurets (K1 to K8) and glucantime against Leishmania major and Leishmania infantum promastigotes. Overall, eight experimental ligands and glucantime were docked using AutoDock 4.3 program into the active sites of Leishmania major and Leishmania infantum pteridine reductase 1, which were modeled using homology modeling programs. The colorimetric MTT assay was used to find L. major and L. infantum promastigotes viability at different concentrations of biuret derivatives in a concentration and time-dependent manner and the obtained results were expressed as 50% and 90% of inhibitory concentration (IC₅₀ and IC₉₀). In silico method showed that out of eight experimental ligands, four compounds were more active on pteridine reductase 1. K3 was the most active against L. major promastigotes with an IC₅₀ of 6.8 μM and an IC₉₀ of 40.2 μM, whereas for L. infantum promastigotes was K8 with IC₅₀ of 7.8 μM. The phenylethyl derivative (K7) showed less toxicity (IC₅₀s > 60 μM) in both Leishmania strains. Glucantime displayed less growth inhibition in concentration of about 20 μM. In silico and especially docking results in a recent study were in accordance with the in vitro activity of these compounds in presented study and compound K3, K2 and K8 showed reasonable levels of selectivity for the Leishmania pteridine reductase 1.     

Investigation on flight and walking activity of Oomyzus sokolowskii Kurdjumov (Hymenoptera: Eulophidae) at different generations under laboratory conditions

The flight and walking activities of second, third, fourth, fifth, sixth and seventh generations of Oomyzus sokolowskii reared on Plutella xylostella L. pupas were compared under laboratory conditions. The parasitoid wasps were obtained by collecting pupa masses of diamondback moth from cabbage fields. The experiments of flight and walking activities were carried out using black PVC tubes and the percentages of inactive, flying and walking wasps to light were calculated. Results showed a reduced flight and walking activities and increased percentage of inactive wasps with the increasing number of generations. In second- third- and fourth-generation average, 52.1 ± 1.27 and 29.6 ± 1.51 of population had flight and walking ability, respectively, and 17.1 ± 1.55 of them were not able to fly and walk. The highest percentage of inactive wasps was observed at seventh generation, whereas wasps showed significantly better flight and walking activities at second, third and fourth generations. Based on the results, flight and walking abilities of O. sokolowskii gradually decreased at continuous laboratory rearing.     

Growth-promoting bacteria and natural regulators mitigate salt toxicity and improve rapeseed plant performance

Salinity is a major environmental stress that limits plant production and portraits a critical challenge to food security in the world. In this research, the impacts of plant growth–promoting bacteria (Pseudomonas RS-198 and Azospirillum brasilense RS-SP7) and foliar application of plant hormones (salicylic acid 1 mM and jasmonic acid 0.5 mM) on alleviating the harmful effects of salt stress in rapeseed plants (Brassica napus cv. okapi) were examined under greenhouse condition. Salt stress diminished rapeseed biomass, leaf area, water content, nitrogen, phosphorus, potassium, calcium, magnesium, and chlorophyll content, while it increased sodium content, endogenous salicylic and jasmonic acids, osmolyte production, H₂O₂ and O₂^•− generations, TBARS content, and antioxidant enzyme activities. Plant growth, nutrient content, leaf expansion, osmolyte production, and antioxidant enzyme activities were increased, but oxidative and osmotic stress indicators were decreased by bacteria inoculation + salicylic acid under salt stress. Antioxidant enzyme activities were amplified by jasmonic acid treatments under salt stress, although rapeseed growth was not generally affected by jasmonic acid. Bacterial + hormonal treatments were superior to individual treatments in reducing detrimental effects of salt stress. The best treatment in rectifying rapeseed growth under salt stress was combination of Pseudomonas and salicylic acid. This combination attenuated destructive salinity properties and subsequently amended rapeseed growth via enhancing endogenous salicylic acid content and some essential nutrients such as potassium, phosphorus, and magnesium.

     

Dehydrogenation of 2-imidazolines with sodium periodate catalyzed by manganese(III) tetraphenylporphyrin

In the present work, dehydrogenation of 2-substituted imidazolines with sodium periodate in the presence of tetraphenylporphyrinatomanganese(III) chloride, [Mn(TPP)Cl], is reported. A wide variety of 2-imidazolines efficiently converted to their corresponding imidazoles by [Mn(TPP)Cl]/NaIO₄ catalytic system at room temperature in 1:2, CH₃CN/H₂O mixture. The effect of reaction parameters such as kind of solvent and catalyst amount was also investigated.     

B cell-specific expression of B7-2 is required for follicular Th cell function in response to vaccinia virus

Follicular Th (T(FH)) cells are specialized in provision of help to B cells that is essential for promoting protective Ab responses. CD28/B7 (B7-1 and B7-2) interactions are required for germinal center (GC) formation, but it is not clear if they simply support activation of naive CD4 T cells during initiation of responses by dendritic cells or if they directly control T(FH) cells and/or directly influence follicular B cell differentiation. Using a model of vaccinia virus infection, we show that B7-2 but not B7-1 deficiency profoundly impaired T(FH) cell development but did not affect CD4 T cell priming and Th1 differentiation. Consistent with this, B7-2 but not B7-1 was required for acquisition of GC B cell phenotype, plasma cell generation, and virus-specific neutralizing Ab responses. Mixed adoptive transfer experiments indicated that bidirectional interactions between CD28 expressed on activated T cells and B7-2 expressed on follicular B cells were essential for maintenance of the T(FH) phenotype and GC B cell development. Our data provide new insight into the source and nature of molecules required for T(FH) cells to direct GC B cell responses.     

Pannexin-1 is required for ATP release during apoptosis but not for inflammasome activation

Apoptotic cell death is important for embryonic development, immune cell homeostasis, and pathogen elimination. Innate immune cells also undergo a very rapid form of cell death termed pyroptosis after activating the protease caspase-1. The hemichannel pannexin-1 has been implicated in both processes. In this study, we describe the characterization of pannexin-1-deficient mice. LPS-primed bone marrow-derived macrophages lacking pannexin-1 activated caspase-1 and secreted its substrates IL-1β and IL-18 normally after stimulation with ATP, nigericin, alum, silica, flagellin, or cytoplasmic DNA, indicating that pannexin-1 is dispensable for assembly of caspase-1-activating inflammasome complexes. Instead, thymocytes lacking pannexin-1, but not the P2X7R purinergic receptor, were defective in their uptake of the nucleic acid dye YO-PRO-1 during early apoptosis. Cell death was not delayed but, unlike their wild-type counterparts, Panx1(-/-) thymocytes failed to recruit wild-type peritoneal macrophages in a Transwell migration assay. These data are consistent with pannexin-1 liberating ATP and other yet to be defined "find me" signals necessary for macrophage recruitment to apoptotic cells.     

The evaluation of cyclic uniaxial strain on myogenic differentiation of adipose-derived stem cells

It has been revealed that skeletal muscle cells have the potential to generate, sense and respond to biomechanical signals and that, mechanical force is one of the important factors influencing proliferation, differentiation, regeneration and homeostasis of skeletal muscle cells and myoblasts. The aim of this study was to illustrate the effect of cyclic uniaxial strain on myogenic differentiation of adipose-derived stem cells (ASCs). This study was designed to investigate this effect within 3 days in 4 groups: control (untreated), chemical, chemical-mechanical and mechanical based on exposure of ASCs to chemical growth factors for 3 days or to mechanical strain just on the 2nd day. Finally, cell orientation, muscle-related gene expression, myosin protein synthesis and the number of myosin-positive cells were examined to estimate the rate of differentiation. By studying the cells before and after exposure to uniaxial strain, it could be observed that by exerting the load, the cells were organized almost perpendicularly to strain direction. Real-time RT-PCR demonstrated that uniaxial strain had a significant effect on up-regulation of muscle-related genes in chemical–mechanical group (P < 0.001) as compared to mechanical or chemical groups. Immunocytochemistry confirmed the myosin-positive cells in treated groups and the numbers of these cells were enumerated by flow cytometry. These data suggest that uniaxial cyclic strain could affect ASCs and cause their myogenic differentiation and that the combination of chemical myogenic differentiation factors with mechanical signals promotes differentiation much more than differentiation by chemical myogenic differentiation factors or mechanical signals alone.     

In Vitro Interaction of Geldanamycin with Triazoles and Echinocandins Against Common and Emerging Candida Species

Mycopathologia - The aim of this study was to determine the in vitro interactions of geldanamycin (Hsp90-inhibitor) with triazoles and echinocandins against common and emerging Candida species....