Overexpressed Na(V)1.7 channels confer hyperexcitability to in vitro trigeminal sensory neurons of Ca(V)2.1 mutant hemiplegic migraine mice

Trigeminal sensory neurons of transgenic knock-in (KI) mice expressing the R192Q missense mutation in the alpha 1A subunit of neuronal voltage-gated Ca(V)2.1 Ca2+ channels, which leads to familial hemiplegic migraine type 1 (FHM1) in patients, exhibit a hyperexcitability phenotype. Here, we show that the expression of Na(V)1.7 channels, linked to pain states, is upregulated in KI primary cultures of trigeminal ganglia (TG), as shown by increased expression of its alpha 1 subunit. In the majority of TG neurons, Na(V)1.7 channels are co-expressed with ATP-gated P2X3 receptors (P2X3R), which are important nociceptive sensors. Reversing the trigeminal phenotype with selective Ca(V)2.1 channel inhibitor omega-agatoxin IVA inhibited Na(V)1.7 overexpression. Functionally, KI neurons revealed a TTX-sensitive inward current of larger amplitude that was partially inhibited by selective Na(V)1.7 blocker Tp1a. Under current-clamp condition, Tp1a raised the spike threshold of both wild-type (WT) and KI neurons with decreased firing rate in KI cells. Na(V)1.7 activator OD1 accelerated firing in WT and KI neurons, a phenomenon blocked by Tp1a. Enhanced expression and function of Na(V)1.7 channels in KI TG neurons resulted in higher excitability and facilitated nociceptive signaling. Co-expression of Na(V)1.7 channels and P2X3Rs in TGs may explain how hypersensitivity to local stimuli can be relevant to migraine.Paroxysmal Cerebral Disorder

Development of polymeric nanocomposite (Xyloglucan-co-Methacrylic acid/Hydroxyapatite/SiO 2 ) scaffold for bone tissue engineering applications—In-vitro antibacterial, cytotoxicity and cell culture evaluation

Advancement and innovation in bone regeneration, specifically polymeric composite scaffolds, are of high significance for the treatment of bone defects. Xyloglucan (XG) is a polysaccharide biopolymer having a wide variety of regenerative tissue therapeutic applications due to its biocompatibility, in-vitro degradation and cytocompatibility. Current research is focused on the fabrication of polymeric bioactive scaffolds by freeze drying method for nanocomposite materials. The nanocomposite materials have been synthesized from free radical polymerization using n-SiO2 and n-HAp XG and Methacrylic acid (MAAc). Functional group analysis, crystallinity and surface morphology were investigated by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD) and scanning electron microscopy (SEM) techniques, respectively. These bioactive polymeric scaffolds presented interconnected and well-organized porous morphology, controlled precisely by substantial ratios of n-SiO2. The swelling analysis was also performed in different media at varying temperatures (27, 37 and 47 °C) and the mechanical behavior of the dried scaffolds is also investigated. Antibacterial activities of these scaffolds were conducted against pathogenic gram-positive and gram-negative bacteria. Besides, the biological behavior of these scaffolds was evaluated by the Neutral Red dye assay against the MC3T3-E1 cell line. The scaffolds showed interesting properties for bone tissue engineering, including porosity with substantial mechanical strength, biodegradability, biocompatibility and cytocompatibility behavior. The reported polymeric bioactive scaffolds can be aspirant biomaterials for bone tissue engineering to regenerate defecated bone

Synthesis and Characterization of Silver-Coated Polymeric Scaffolds for Bone Tissue Engineering: Antibacterial and In Vitro Evaluation of Cytotoxicity and Biocompatibility

In bone tissue engineering, multifunctional composite materials are very challenging. Bone tissue engineering is an innovative technique to develop biocompatible scaffolds with suitable orthopedic applications with enhanced antibacterial and mechanical properties. This research introduces a polymeric nanocomposite scaffold based on arabinoxylan-co-acrylic acid, nano-hydroxyapatite (nHAp), nano-aluminum oxide (nAl₂O₃), and graphene oxide (GO) by free-radical polymerization for the development of porous scaffolds using the freeze-drying technique. These polymeric nanocomposite scaffolds were coated with silver (Ag) nanoparticles to improve antibacterial activities. Together, nHAp, nAl₂O₃, and GO enhance the multifunctional properties of materials, which regulate their physicochemical and biomechanical properties. Results revealed that the Ag-coated polymeric nanocomposite scaffolds had excellent antibacterial properties and better microstructural properties. Regulated morphological properties and maximal antibacterial inhibition zones were found in the porous scaffolds with the increasing amount of GO. Moreover, the nanosystem and the polymeric matrix have improved the compressive strength (18.89 MPa) and Young’s modulus (198.61 MPa) of scaffolds upon increasing the amount of GO. The biological activities of the scaffolds were investigated against the mouse preosteoblast cell lines (MC3T3-E1) and increasing the quantities of GO helps cell adherence and proliferation. Therefore, our findings showed that these silver-coated polymeric nanocomposite scaffolds have the potential for engineering bone tissue

A P53-TLR3 Axis Ameliorates Pulmonary Hypertension by Inducing BMPR2 Via IRF3

Pulmonary arterial hypertension (PAH) features pathogenic and abnormal endothelial cells (ECs), and one potential origin is clonal selection. We studied the role of p53 and toll-like receptor 3 (TLR3) in clonal expansion and pulmonary hypertension (PH) via regulation of bone morphogenetic protein (BMPR2) signaling. ECs of PAH patients had reduced p53 expression. EC-specific p53 knockout exaggerated PH, and clonal expansion reduced p53 and TLR3 expression in rat lung CD117+ ECs. Reduced p53 degradation (Nutlin 3a) abolished clonal EC expansion, induced TLR3 and BMPR2, and ameliorated PH. Polyinosinic/polycytidylic acid [Poly(I:C)] increased BMPR2 signaling in ECs via enhanced binding of interferon regulatory factor-3 (IRF3) to the BMPR2 promoter and reduced PH in p53−/− mice but not in mice with impaired TLR3 downstream signaling. Our data show that a p53/TLR3/IRF3 axis regulates BMPR2 expression and signaling in ECs. This link can be exploited for therapy of PH

Short Telomeres Compromise β-Cell Signaling and Survival

The genetic factors that underlie the increasing incidence of diabetes with age are poorly understood. We examined whether telomere length, which is inherited and known to shorten with age, plays a role in the age-dependent increased incidence of diabetes. We show that in mice with short telomeres, insulin secretion is impaired and leads to glucose intolerance despite the presence of an intact β-cell mass. In ex vivo studies, short telomeres induced cell-autonomous defects in β-cells including reduced mitochondrial membrane hyperpolarization and Ca2+ influx which limited insulin release. To examine the mechanism, we looked for evidence of apoptosis but found no baseline increase in β-cells with short telomeres. However, there was evidence of all the hallmarks of senescence including slower proliferation of β-cells and accumulation of p16INK4a. Specifically, we identified gene expression changes in pathways which are essential for Ca2+-mediated exocytosis. We also show that telomere length is additive to the damaging effect of endoplasmic reticulum stress which occurs in the late stages of type 2 diabetes. This additive effect manifests as more severe hyperglycemia in Akita mice with short telomeres which had a profound loss of β-cell mass and increased β-cell apoptosis. Our data indicate that short telomeres can affect β-cell metabolism even in the presence of intact β-cell number, thus identifying a novel mechanism of telomere-mediated disease. They implicate telomere length as a determinant of β-cell function and diabetes pathogenesis

Computational Study of the Human Dystrophin Repeats: Interaction Properties and Molecular Dynamics

Dystrophin is a large protein involved in the rare genetic disease Duchenne muscular dystrophy (DMD). It functions as a mechanical linker between the cytoskeleton and the sarcolemma, and is able to resist shear stresses during muscle activity. In all, 75% of the dystrophin molecule consists of a large central rod domain made up of 24 repeat units that share high structural homology with spectrin-like repeats. However, in the absence of any high-resolution structure of these repeats, the molecular basis of dystrophin central domain's functions has not yet been deciphered. In this context, we have performed a computational study of the whole dystrophin central rod domain based on the rational homology modeling of successive and overlapping tandem repeats and the analysis of their surface properties. Each tandem repeat has very specific surface properties that make it unique. However, the repeats share enough electrostatic-surface similarities to be grouped into four separate clusters. Molecular dynamics simulations of four representative tandem repeats reveal specific flexibility or bending properties depending on the repeat sequence. We thus suggest that the dystrophin central rod domain is constituted of seven biologically relevant sub-domains. Our results provide evidence for the role of the dystrophin central rod domain as a scaffold platform with a wide range of surface features and biophysical properties allowing it to interact with its various known partners such as proteins and membrane lipids. This new integrative view is strongly supported by the previous experimental works that investigated the isolated domains and the observed heterogeneity of the severity of dystrophin related pathologies, especially Becker muscular dystrophy

(Per)chlorate reduction by an acetogenic bacterium, Sporomusa sp., isolated from an underground gas storage

A mesophilic bacterium, strain An4, was isolated from an underground gas storage reservoir with methanol as substrate and perchlorate as electron acceptor. Cells were Gram-negative, spore-forming, straight to curved rods, 0.5–0.8 μm in diameter, and 2–8 μm in length, growing as single cells or in pairs. The cells grew optimally at 37°C, and the pH optimum was around 7. Strain An4 converted various alcohols, organic acids, fructose, acetoin, and H2/CO2 to acetate, usually as the only product. Succinate was decarboxylated to propionate. The isolate was able to respire with (per)chlorate, nitrate, and CO2. The G+C content of the DNA was 42.6 mol%. Based on the 16S rRNA gene sequence analysis, strain An4 was most closely related to Sporomusa ovata (98% similarity). The bacterium reduced perchlorate and chlorate completely to chloride. Key enzymes, perchlorate reductase and chlorite dismutase, were detected in cell-free extracts

A benzene-degrading nitrate-reducing microbial consortium displays aerobic and anaerobic benzene degradation pathways

All sequence data from this study were deposited at the European Bioinformatics Institute under the accession numbers ERS1670018 to ERS1670023. Further, all assigned genes, taxonomy, function, sequences of contigs, genes and proteins can be found in Table S3.In this study, we report transcription of genes involved in aerobic and anaerobic benzene degradation pathways in a benzene-degrading denitrifying continuous culture. Transcripts associated with the family Peptococcaceae dominated all samples (2136% relative abundance) indicating their key role in the community. We found a highly transcribed gene cluster encoding a presumed anaerobic benzene carboxylase (AbcA and AbcD) and a benzoate-coenzyme A ligase (BzlA). Predicted gene products showed >96% amino acid identity and similar gene order to the corresponding benzene degradation gene cluster described previously, providing further evidence for anaerobic benzene activation via carboxylation. For subsequent benzoyl-CoA dearomatization, bam-like genes analogous to the ones found in other strict anaerobes were transcribed, whereas gene transcripts involved in downstream benzoyl-CoA degradation were mostly analogous to the ones described in facultative anaerobes. The concurrent transcription of genes encoding enzymes involved in oxygenase-mediated aerobic benzene degradation suggested oxygen presence in the culture, possibly formed via a recently identified nitric oxide dismutase (Nod). Although we were unable to detect transcription of Nod-encoding genes, addition of nitrite and formate to the continuous culture showed indication for oxygen production. Such an oxygen production would enable aerobic microbes to thrive in oxygen-depleted and nitrate-containing subsurface environments contaminated with hydrocarbons.This study was supported by a grant of BE-Basic-FES funds from the Dutch Ministry of Economic Affairs. The research of A.J.M. Stams is supported by an ERC grant (project 323009) and the gravitation grant “Microbes for Health and Environment” (project 024.002.002) of the Netherlands Ministry of Education, Culture and Science. F. Hugenholtz was supported by the same gravitation grant (project 024.002.002). B. Hornung is supported by Wageningen University and the Wageningen Institute for Environment and Climate Research (WIMEK) through the IP/OP program Systems Biology (project KB-17-003.02-023).info:eu-repo/semantics/publishedVersio