Superoxide constricts rat pulmonary arteries via Rho-kinase-mediated Ca2+ sensitization

Reactive oxygen species play a key role in vascular disease, pulmonary hypertension, and hypoxic pulmonary vasoconstriction. We investigated contractile responses, intracellular Ca²⁺ ([Ca²⁺]_i), Rho-kinase translocation, and phosphorylation of the regulatory subunit of myosin phosphatase (MYPT-1) and of myosin light chain (MLC₂₀) in response to LY83583, a generator of superoxide anion, in small intrapulmonary arteries (IPA) of rat. LY83583 caused concentration-dependent constrictions in IPA and greatly enhanced submaximal PGF_2α-mediated preconstriction. In small femoral or mesenteric arteries of rat, LY83583 alone was without effect, but it relaxed a PGF₂α-mediated preconstriction. Constrictions in IPA were inhibited by superoxide dismutase and tempol, but not catalase, and were endothelium and guanylate cyclase independent. Constrictions were also inhibited by the Rho-kinase inhibitor Y27632 and the Src-family kinase inhibitor SU6656. LY83583 did not raise [Ca²⁺]_i, but caused a Y27632-sensitive constriction in α-toxin-permeabilized IPA. LY83583 triggered translocation of Rho-kinase from the nucleus to the cytosol in pulmonary artery smooth muscle cells and enhanced phosphorylation of MYPT-1 at Thr-855 and of MLC₂₀ at Ser-19 in IPA. This enhancement was inhibited by superoxide dismutase and abolished by Y27632. Hydrogen peroxide did not activate Rho-kinase. We conclude that in rat small pulmonary artery, superoxide triggers Rho-kinase-mediated Ca²⁺ sensitization and vasoconstriction independent of hydrogen peroxide.     

Structural and energetic profiling of SARS-CoV-2 receptor binding domain antibody recognition and the impact of circulating variants

The SARS-CoV-2 pandemic highlights the need for a detailed molecular understanding of protective antibody responses. This is underscored by the emergence and spread of SARS-CoV-2 variants, including Alpha (B.1.1.7) and Delta (B.1.617.2), some of which appear to be less effectively targeted by current monoclonal antibodies and vaccines. Here we report a high resolution and comprehensive map of antibody recognition of the SARS-CoV-2 spike receptor binding domain (RBD), which is the target of most neutralizing antibodies, using computational structural analysis. With a dataset of nonredundant experimentally determined antibody-RBD structures, we classified antibodies by RBD residue binding determinants using unsupervised clustering. We also identified the energetic and conservation features of epitope residues and assessed the capacity of viral variant mutations to disrupt antibody recognition, revealing sets of antibodies predicted to effectively target recently described viral variants. This detailed structure-based reference of antibody RBD recognition signatures can inform therapeutic and vaccine design strategies.     

Synapse Density and Dendritic Complexity Are Reduced in the Prefrontal Cortex following Seven Days of Forced Abstinence from Cocaine Self-Administration

Chronic cocaine exposure in both human addicts and in rodent models of addiction reduces prefrontal cortical activity, which subsequently dysregulates reward processing and higher order executive function. The net effect of this impaired gating of behavior is enhanced vulnerability to relapse. Previously we have shown that cocaine-induced increases in brain-derived neurotrophic factor (BDNF) expression in the medial prefrontal cortex (PFC) is a neuroadaptive mechanism that blunts the reinforcing efficacy of cocaine. As BDNF is known to affect neuronal survival and synaptic plasticity, we tested the hypothesis that abstinence from cocaine self-administration would lead to alterations in neuronal morphology and synaptic density in the PFC. Using a novel technique, array tomography and Golgi staining, morphological changes in the rat PFC were analyzed following 14 days of cocaine self-administration and 7 days of forced abstinence. Our results indicate that overall dendritic branching and total synaptic density are significantly reduced in the rat PFC. In contrast, the density of thin dendritic spines are significantly increased on layer V pyramidal neurons of the PFC. These findings indicate that dynamic structural changes occur during cocaine abstinence that may contribute to the observed hypo-activity of the PFC in cocaine-addicted individuals.     

Identification of MAMDC1 as a Candidate Susceptibility Gene for Systemic Lupus Erythematosus (SLE)

Background

Systemic lupus erythematosus (SLE) is a complex autoimmune disorder with multiple susceptibility genes. We have previously reported suggestive linkage to the chromosomal region 14q21-q23 in Finnish SLE families.

Principal Findings

Genetic fine mapping of this region in the same family material, together with a large collection of parent affected trios from UK and two independent case-control cohorts from Finland and Sweden, indicated that a novel uncharacterized gene, MAMDC1 (MAM domain containing glycosylphosphatidylinositol anchor 2, also known as MDGA2, MIM 611128), represents a putative susceptibility gene for SLE. In a combined analysis of the whole dataset, significant evidence of association was detected for the MAMDC1 intronic single nucleotide polymorphisms (SNP) rs961616 (P –value = 0.001, Odds Ratio (OR) = 1.292, 95% CI 1.103–1.513) and rs2297926 (P –value = 0.003, OR = 1.349, 95% CI 1.109–1.640). By Northern blot, real-time PCR (qRT-PCR) and immunohistochemical (IHC) analyses, we show that MAMDC1 is expressed in several tissues and cell types, and that the corresponding mRNA is up-regulated by the pro-inflammatory cytokines tumour necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) in THP-1 monocytes. Based on its homology to known proteins with similar structure, MAMDC1 appears to be a novel member of the adhesion molecules of the immunoglobulin superfamily (IgCAM), which is involved in cell adhesion, migration, and recruitment to inflammatory sites. Remarkably, some IgCAMs have been shown to interact with ITGAM, the product of another SLE susceptibility gene recently discovered in two independent genome wide association (GWA) scans.

Significance

Further studies focused on MAMDC1 and other molecules involved in these pathways might thus provide new insight into the pathogenesis of SLE.     

Analysis of Micro-RNA-144 Expression Profile in Patients with Multiple Sclerosis in Comparison with Healthy Individuals.

Background:Etiology of multiple sclerosis is non-clarified. It seems that environmental factors impact epigenetic in this disease. Micro-RNAs (MIR) as epigenetic factors are one of the most important factors in non-genetically neurodegenerative diseases. It has been found MIR-144 plays a main role in the regulation of many processes in the central nervous system. Here, we aimed to investigation of MIR-144 expression alteration in Multiple sclerosis (MS) patients.Methods:In this study 32 healthy and 32 MS patient''s blood sample were analyzed by quantitative Real-Time PCR method and obtained data analyzed by REST 2009 software.Results:Analysis of Real-Time PCR data revealed that miR-144 Increase significantly in MS patients compared to healthy controls.Conclusion:The increase of MIR-144 expression in MS patients is obvious. MIR-144 can be used as a biomarker of MS and help to early diagnosis and treatment of this disease.Key Words: MicroRNA (miRNA), MiRNA-144, Multiple Sclerosis (MS)     

Enhancing compost quality by using whey-grown biosurfactant-producing bacteria as inocula

Wild screening of bacterial strains from compost materials was performed. Several biosurfactant-producer strains were isolated and then cultured in whey as a low-cost medium for biosurfactant production. Two strains, identified as Bacillus sp. and Streptomyces sp., were the best biosurfactant producers and were selected for determination of compost quality enhancing. The effect of cell biomass, cell-free supernatant, and a consortium of these two strains on compost quality were determined and specific parameters of compost were analyzed. The results showed that using these bacteria (or supernatants) in compost processing have slight stimulatory effect on bacterial population (8.08 log₁₀ CFU/g), surface tension reduction (to 42.6 mN/m at 24 h), and heavy metal bioremediation (>50 % in most treatments), speeding up the decomposition rate of organic matter (42.3 % OM at the end of experiment), accelerating the stabilization process by reduction of NH ₄ ⁺ to NO ₃ ^? ratio (reduced from 0.2 to 0.026), decreasing the biotoxicity (tested by seed germination and root length of germinated seed), and also reduction of pathogens (reduced from 2100 to 120 MPN/g in fecal coliform).     

Effects of Cymbopogon citratus and Ferula assa-foetida extracts on glutamate-induced neurotoxicity

Many of CNS diseases can lead to a great quantity of release of glutamate and the extreme glutamate induces neuronal cell damage and death. Here, we wanted to investigate the effects of Cymbopogon citratus essential oil and Ferula assa-foetida extracts treatment on glutamate-induced cell damage in a primary culture of rat cerebellar granule neurons. Cerebellums were collected from 7-d rat brains and cerebellar granule neurons were obtained after 8-d culture. CGN cells were treated with C. citratus essential oil and F. assa-foetida extracts at concentration of 100 μg/ml before, after, and during exposure to 30 μM glutamate. The cellular viability was evaluated by 3-(4, 5-dimethytthiazol-2-yl)-2, 5-diphenyltetrazoliumbromide (MTT) staining. The flow cytometry assay was used to examine cell cycle and apoptosis. MTT assay showed a glutamate-induced reduction in cellular viability while treatment with C. citratus essential oil and F. assa-foetida extracts before, during, and after exposure to glutamate was increased. Flow cytometric analysis indicated that F. assa-foetida extracts treatment significantly (p?<?0.001) attenuated glutamate-induced apoptotic/necrotic cell death and the necrotic rate was decreased by C. citratus essential oil treatment compared to glutamate group, significantly (p?<?0.001). The results show that C. citratus essential oil and F. assa-foetida extracts display neuroprotective effects in glutamate-induced neurotoxicity. These extracts exert antiapoptotic activity in cerebellar granule neurons due to cell cycle arrest in G0G1 phase, which explain the beneficial effects of C. citratus essential oil and F. assa-foetida extracts as therapies for neurologic disorders.