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Mechanosensitive ion channels (MSCs) are key molecules in the mechano-electrical transduction of arterial baroreceptors. Among them, acid-sensing ion channel 2 (ASIC2) and transient receptor potential vanilloid subfamily member 1 (TRPV1) have been studied extensively and documented to play important roles. In this study, experiments using aortic arch–aortic nerve preparations isolated from rats revealed that both ASIC2 and TRPV1 are functionally necessary, as blocking either abrogated nearly all pressure-dependent neural discharge. However, whether ASIC2 and TRPV1 work in coordination remained unclear. So we carried out cell-attached patch-clamp recordings in HEK293T cells co-expressing ASIC2 and TRPV1 and found that inhibition of ASIC2 completely blocked stretch-activated currents while inhibition of TRPV1 only partially blocked these currents. Immunofluorescence staining of aortic arch–aortic adventitia from rats showed that ASIC2 and TRPV1 are co-localized in the aortic nerve endings, and co-immunoprecipitation assays confirmed that the two proteins form a compact complex in HEK293T cells and in baroreceptors. Moreover, protein modeling analysis, exogenous co-immunoprecipitation assays, and biotin pull-down assays indicated that ASIC2 and TRPV1 interact directly. In summary, our research suggests that ASIC2 and TRPV1 form a compact complex and function synergistically in the mechano-electrical transduction of arterial baroreceptors. The model of synergism between MSCs may have important biological significance beyond ASIC2 and TRPV1. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12264-021-00737-1.

Biotin is an essential micro-nutrient across the three domains of life. The paradigm earlier step of biotin synthesis denotes “BioC-BioH” pathway in Escherichia coli. Here we report that BioZ bypasses the canonical route to begin biotin synthesis. In addition to its origin of Rhizobiales, protein phylogeny infers that BioZ is domesticated to gain an atypical role of β-ketoacyl-ACP synthase III. Genetic and biochemical characterization demonstrates that BioZ catalyzes the condensation of glutaryl-CoA (or ACP) with malonyl-ACP to give 5’-keto-pimeloyl ACP. This intermediate proceeds via type II fatty acid synthesis (FAS II) pathway, to initiate the formation of pimeloyl-ACP, a precursor of biotin synthesis. To further explore molecular basis of BioZ activity, we determine the crystal structure of Agrobacterium tumefaciens BioZ at 1.99 Å, of which the catalytic triad and the substrate-loading tunnel are functionally defined. In particular, we localize that three residues (S84, R147, and S287) at the distant bottom of the tunnel might neutralize the charge of free C-carboxyl group of the primer glutaryl-CoA. Taken together, this study provides molecular insights into the BioZ biotin synthesis pathway., Biotin is an essential enzyme cofactor and two pathways for the generation of the biotin precursor pimeloyl-ACP are known. Here, the authors identify and characterize a third pathway for biotin precursor synthesis involving BioZ and they also present the Agrobacterium tumefaciens BioZ crystal structure.

Accurate nationwide spatiotemporal distribution of ambient ozone product is critical for environment & health departments and for researches to investigate the influence of ozone for epidemiological studies. Our hybrid method, the novel CAMS (The Copernicus Atmosphere Monitoring Service) ozone improvement (CAO3_I) method, is the first attempt to predict ambient ozone by improving CAMS ozone (CAO3) products. For this novel framework, the SVM (Support Vector Machine) has been adopted for classification through the most significant regional ozone patterns which were extracted through the REOF (Rotated Empirical Orthogonal Function) technique. For each classified region, meteorological data, geographical data, CAMS ozone and ground-sites ozone are fed into random forest for regional regulation training and prediction. The CAO3_I method has shown its great feasibility in daily ozone surface distribution prediction. Based on daily averaged ozone concentrations for each station (STO3), the performance of CAO3 products (R2 = 0.35, RMSE = 25.77 μg/m3, MAPE = 42.06) have significantly improved to CAO3_I (R2 = 0.81, RMSE = 14.10 μg/m3, MAPE = 22.37), which shows above 97.4% R2 and RMSE have been improved. Our model is also capable to predict high level ozone concentrations in summer where the R2 has improved from 0.37 (for CAO3) to 0.81. In comparison with ground monitoring stations, the CAMS ozone improvement results can excellently reflect the distribution of daily ground-level ozone concentration and outperform previous statistical models in predicting ambient O3 concentrations. Therefore, the prediction results and our proposed model can be used for future epidemiological studies and air pollution controlling programs.

In this study, the effect of chondroitin sulphate nano-selenium (CS@Se) on Alzheimer's disease (AD) in mice was investigated. CS@Se alleviated anxiety and improved the spatial learning and memory impairment in AD mice. CS@Se significantly reduced cell oedema and pyknosis, protected the mitochondria, and improved abnormal changes in the ultrastructure of hippocampal neuron synapses of AD mice. Moreover, CS@Se significantly increased the levels of superoxide dismutase(SOD), glutathione peroxidase (GSH-Px), Na+/K+-ATPase assay (Na+/K+-ATPase) and acetyltransferase (ChAT), and decreased the levels of malondialdehyde (MDA) and acetylcholinesterase (ChAE) in AD mice. Western blot results showed that CS@Se can attenuate excessive phosphorylation of tau (Ser396/Ser404) by regulating the expression of glycogen synthase kinase-3 beta (GSK-3β). In addition, CS@Se can activate the extracellular signal-regulated kinase 1/2 (ERK 1/2) and p38 mitogen-activated protein kinase (p38 MAPK) signalling pathways to inhibit nuclear transcription factor kappa B (NF-κB) nuclear translocation, thereby regulating the expression of pro-inflammatory cytokines. In summary, CS@Se can reduce oxidative stress damage, inhibit excessive tau phosphorylation, reduce inflammation to delay AD development, and increase the learning and memory capacities of AD mice.

The reduction of nitrate using internal circulation micro-electrolysis technology (ICE) was investigated. The effect of the reaction time, initial pH, Fe/C ratio, and aeration rate on the nitrate reduction was investigated using a single factor experiment. Based on the results of the single factor experiment, a response surface methodology (RSM) was applied to optimize the N2 generation selectivity. The effects and interactions of three independent variables were estimated using a Box–Behnken design. Using the RSM analysis, a quadratic polynomial model with optimal conditions at pH = 8.8, Fe/C = 1:1, and an aeration rate of 30 L·min−1 was developed by means of the regression analysis of the experimental data. Using the RSM optimization, the optimal conditions yielded a N2 generation selectivity of 72.0%, which is in good agreement with experimental result (73.2% ± 0.5%) and falls within the 95% confidence interval (IC: 66.8%–77.3%) of the model results. This indicates that the model obtained in this study effectively predicts the N2 generation selectivity for nitrate reduction by the ICE process, thus providing a theoretical basis for process design.

Summary Biotin is an indispensable cofactor in the three domains of life. The unusual virulence factor BioJ of Francisella catalyzes the formation of pimeloyl-ACP, an intermediate in biotin synthesis. Here, we report the 1.58 Å crystal structure of BioJ, the enzymatic activity of which is determined with the in vitro reconstituted reaction and biotin bioassay in vivo. Unlike the paradigm BioH, BioJ displays an atypical α/β-hydrolase fold. A structurally conserved catalytic triad (S151, D248, and H278) of BioJ is functionally defined. A proposed model for BioJ catalysis involves two basic residues-rich cavities, of which cavity-1, rather than cavity-2, binds to the ACP moiety of its physiological substrate, pimeloyl-ACP methyl ester. In summary, this finding provides molecular insights into the BioJ gatekeeper of biotin synthesis., Graphical Abstract, Highlights • BioJ is a distinct gatekeeper of biotin synthesis • We report structural and functional definition of BioJ • We propose a working model for BioJ-substrate binding in addition to catalytic triad • It furthers our understanding how the substrate is gatekept by BioJ in biotin synthesis, Biological Sciences; Microbiology; Structural Biology

The large conductance Ca2+-activated potassium (BKCa) channel is activated by stretch. The stress-regulated exon (STREX) in α-subunits is known to affect the mechanosensitivity of BKCa channels. However, in human colonic smooth muscle cells (HCoSMCs), we found that the α-subunits without STREX (ZERO-BK) and β1-subunits could be detected yet the cells were mechanosensitive. Whether the β1-subunit is involved in the regulation of BKCa mechanosensitivity is unclear. In the present study, ZERO-BK and β1-subunits were individually expressed or coexpressed in HEK293 cells and cell-attached patch-clamp techniques were used to measure BKCa currents defining mechanosensitivity. Single-channel patch-clamp recordings from HEK293 cells cotransfected with ZERO-BK and β1-subunits showed stretch sensitivity, but there was no mechanosensitivity in HEK293 cells transfected only with ZERO-BK. We also showed that expression of the β1-subunit could increase mechanosensitivity of the STREX-type α-subunits (STREX-BK). To identify the domain in β1-subunits responsible for affecting stretch sensitivity, we expressed β1- LoopDel (chimeric β1-subunits without the extracellular loop) or β1- C TermDel (chimeric β1-subunits without COOH terminus) with ZERO-BK in HEK293 cells. The data showed that deletion of the extracellular loop but not the COOH terminus of β1-subunits virtually abolished the mechanosensitivity. These results suggest that the extracellular loop of the β1-subunit is involved in the regulation of BKCa channel mechanosensitivity and that role is independent of STREX.

Stretch-induced relaxation has not been clearly identified in gastrointestinal tract. The present study is to explore the role of large conductance calcium-activated potassium channels (BKCa) in stretch-induced relaxation of colon. The expression and currents of BKCawere detected and the basal muscle tone and contraction amplitude of colonic smooth muscle strips were measured. The expression of BKCain colon is higher than other GI segments (P< 0.05). The density of BKCacurrents was very high in colonic smooth muscle cells (SMCs). BKCain rat colonic SMCs were sensitive to stretch. The relaxation response of colonic SM strips to stretch was attenuated by charybdotoxin (ChTX), a nonspecific BKCablocker (P< 0.05). After blocking enteric nervous activities by tetrodotoxin (TTX), the stretch-induced relaxation did not change (P> 0.05). Still, ChTX and iberiotoxin (IbTX, a specific BKCablocker) attenuated the relaxation of the colonic muscle strips enduring stretch (P< 0.05). These results suggest stretch-activation of BKCain SMCs was involved in the stretch-induced relaxation of colon. Our study highlights the role of mechanosensitive ion channels in SMCs in colon motility regulation and their physiological and pathophysiological significance is worth further study.