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The slow rate of anaerobic microbial dechlorination in natural environments limits the application of polychlorinated biphenyl (PCB) bioremediation. Anaerobic digested sludge (ADS), abundant in nutrients and microorganisms, could be an effective additive to improve microbial dechlorination. This research investigates the influence of ADS on Aroclor 1260 dechlorination performance, microbial community composition, and the abundance of functional genes. Moreover, further enrichment of organohalide-respiring bacteria (OHRB) was examined using tetrachloroethene (PCE) as the electron acceptor, followed by the serial dilution-to-extinction method in conjunction with resuscitation promoting factor (Rpf) supplementation. The results demonstrated that the addition of 5 g/L ADS achieved more extensive and efficient dechlorination of PCBs. ADS enhanced the removal of meta- and para-chlorine without significantly changing the dechlorination pathways. The abundances of dechlorinators, including Dehalobium and Dehalobacter within the Chloroflexi and Firmicutes phyla, as well as non-dechlorinators from the Desulfobacterota, Euryarchaeota, and Bacteroidetes phyla, were significantly increased with ADS amendment. Similarly, an increased abundance of bacteria, OHRB, reductive dehalogenase (RDase) genes, and archaeal 16S rRNA genes was observed. Additionally, obligate OHRB, such as Dehalobacter and Dehalobium, were further enriched. These findings indicate that ADS effectively enhances microbial reductive dechlorination and highlight the potential for enriching and isolating OHRB with Rpf., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Adenosine triphosphate (ATP) can be released into the extracellular milieu from various types of cells in response to a wide range of physical or chemical stresses. In the respiratory tract, extracellular ATP is recognized as an important signal molecule and trigger of airway inflammation. Chlorine (Cl 2 ), sulfur dioxide (SO 2 ), and ammonia (NH 3 ) are potent irritant gases and common industrial air pollutants due to their widespread uses as chemical agents. This study was carried out to determine if acute inhalation challenges of these irritant gases, at the concentration and duration simulating the accidental exposures to these chemical gases in industrial operations, triggered the release of ATP in the rat respiratory tract; and if so, whether the level of ATP in bronchoalveolar lavage fluid (BALF) evoked by inhalation challenge of a given irritant gas was elevated by chronic allergic airway inflammation. Our results showed: 1 ) inhalation of these irritant gases caused significant increases in the ATP level in BALF, and the magnitude of evoked ATP release was in the order of Cl 2 > SO 2 > NH 3 . 2 ) Chronic airway inflammation induced by ovalbumin-sensitization markedly elevated the ATP level in BALF during baseline (breathing room air) but did not potentiate the release of ATP in the lung triggered by inhalation challenges of these irritant gases. These findings suggested a possible involvement of the ATP release in the lung in the regulation of overall airway responses to acute inhalation of irritant gases and the pathogenesis of chronic allergic airway inflammation. NEW & NOTEWORTHY Extracellular adenosine triphosphate (ATP) is a contributing factor and signaling molecule of airway inflammation. This study demonstrated for the first time that the ATP release in the lung was markedly elevated after acute inhalation challenges of three common industrial air pollutants; the order of the response magnitude was chlorine > sulfur dioxide > ammonia. These findings provided new information and improved our understanding of the adverse pulmonary effects caused by accidental inhalation exposures to these irritant gases.

The coupling of thermal remediation with microbial reductive dechlorination (MRD) has shown promising potential for the cleanup of chlorinated solvent contaminated sites. In this study, thermal treatment and bioaugmentation were applied in series, where prior higher thermal remediation temperature led to improved TCE dechlorination performance with both better organohalide-respiring bacteria (OHRB) colonization and electron donor availability. The 60 °C was found to be a key temperature point where the promotion effect became obvious. Amplicon sequencing and co-occurrence network analysis demonstrated that temperature was a more dominating factor than bioaugmentation that impacted microbial community structure. Higher temperature of prior thermal treatment resulted in the decrease of richness, diversity of indigenous microbial communities, and simplified the network structure, which benefited the build-up of newcoming microorganisms during bioaugmentation. Thus, the abundance of Desulfitobacterium increased from 0.11 % (25 °C) to 3.10 % (90 °C). Meanwhile, released volatile fatty acids (VFAs) during thermal remediation functioned as electron donors and boosted MRD. Our results provided temperature-specific information on synergistic effect of sequential thermal remediation and bioaugmentation, which contributed to better implementation of the coupled technologies in chloroethene-impacted sites., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Carbon and chlorine isotope effects for biotransformation of chloroform by different microbes show significant variability. Reductive dehalogenases (RDase) enzymes contain different cobamides, affecting substrate preferences, growth yields, and dechlorination rates and extent. We investigate the role of cobamide type on carbon and chlorine isotopic signals observed during reductive dechlorination of chloroform by the RDase CfrA. Microcosm experiments with two subcultures of a Dehalobacter-containing culture expressing CfrA-one with exogenous cobamide (Vitamin B 12 , B12 + ) and one without (to drive native cobamide production)-resulted in a markedly smaller carbon isotope enrichment factor (ε C, bulk ) for B12 - (-22.1 ± 1.9‰) compared to B12 + (-26.8 ± 3.2‰). Both cultures exhibited significant chlorine isotope fractionation, and although a lower ε Cl, bulk was observed for B12 - (-6.17 ± 0.72‰) compared to B12 + (-6.86 ± 0.77‰) cultures, these values are not statistically different. Importantly, dual-isotope plots produced identical slopes of Λ Cl/C (Λ Cl/C, B12+  = 3.41 ± 0.15, Λ Cl/C, B12 - = 3.39 ± 0.15), suggesting the same reaction mechanism is involved in both experiments, independent of the lower cobamide bases. A nonisotopically fractionating masking effect may explain the smaller fractionations observed for the B12 - containing culture., (© 2024 The Author(s). MicrobiologyOpen published by John Wiley & Sons Ltd.)

1,2-dichloropropane (1,2-DCP) and 1,2,3-trichloropropane (1,2,3-TCP) are hazardous chemicals frequently detected in groundwater near agricultural zones due to their historical use in chlorinated fumigant formulations. In this study, we show that the organohalide-respiring bacterium Dehalogenimonas alkenigignens strain BRE15 M can grow during the dihaloelimination of 1,2-DCP and 1,2,3-TCP to propene and allyl chloride, respectively. Our work also provides the first application of dual isotope approach to investigate the anaerobic reductive dechlorination of 1,2-DCP and 1,2,3-TCP. Stable carbon and chlorine isotope fractionation values for 1,2-DCP (Ɛ C  = -13.6 ± 1.4 ‰ and Ɛ Cl  = -27.4 ± 5.2 ‰) and 1,2,3-TCP (Ɛ C  = -3.8 ± 0.6 ‰ and Ɛ Cl  = -0.8 ± 0.5 ‰) were obtained resulting in distinct dual isotope slopes (Λ 12DCP  = 0.5 ± 0.1, Λ 123TCP  = 4 ± 2). However direct comparison of Λ C-Cl among different substrates is not possible and investigation of the C and Cl apparent kinetic isotope effects lead to the hypothesis that concerted dichloroelimination mechanism is more likely for both compounds. In fact, whole cell activity assays using cells suspensions of the Dehalogenimonas-containing culture grown with 1,2-DCP and methyl viologen as electron donor suggest that the same set of reductive dehalogenases was involved in the transformation of 1,2-DCP and 1,2,3-TCP. This study opens the door to the application of isotope techniques for evaluating biodegradation of 1,2-DCP and 1,2,3-TCP, which often co-occur in groundwaters near agricultural fields., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Anaerobic microbial transformation is a key pathway in the natural attenuation of polychlorinated biphenyls (PCBs). Much less is known about the transformation behaviors induced by pure organohalide-respiring bacteria, especially kinetic isotope effects. Therefore, the kinetics, pathways, enantioselectivity, and carbon and chlorine isotope fractionation of PCBs transformation by Dehalococcoides mccartyi CG1 were comprehensively explored. The results indicated that the PCBs were mainly dechlorinated via removing their double-flanked meta-chlorine, with their first-order kinetic constants following the order of PCB132 > PCB174 > PCB85 > PCB183 > PCB138. However, PCBs occurred great loss of stoichiometric mass balance during microbial transformation, suggesting the generation of other non-dehalogenation products and/or stable intermediates. The preferential transformation of (-)-atropisomers and generation of (+)-atropisomers were observed during PCB132 and PCB174 biotransformation with the enantiomeric enrichment factors of -0.8609 ± 0.1077 and -0.4503 ± 0.1334 (first half incubation times)/-0.1888 ± 0.1354 (second half incubation times), respectively, whereas no enantioselectivity occurred during PCB183 biotransformation. More importantly, although there was no carbon and chlorine isotope fractionation occurring for studied substrates, the δ 13 C values of dechlorination products, including PCB47 (-28.15 ± 0.35‰ ∼ -27.77 ± 0.20‰), PCB91 (-36.36 ± 0.09‰ ∼ -34.71 ± 0.49‰), and PCB149 (-28.08 ± 0.26‰ ∼ -26.83 ± 0.10‰), were all significantly different from those of their corresponding substrates (PCB85: -30.81 ± 0.02‰ ∼ -30.22 ± 0.21‰, PCB132: -33.57 ± 0.15‰ ∼ -33.13 ± 0.14‰, and PCB174: -26.30 ± 0.09‰ ∼ -26.01 ± 0.07‰), which further supported the generation of other non-dehalogenation products and/or stable intermediates with enrichment or depletion of 13 C. These findings provide deeper insights into the anaerobic microbial transformation behaviors of PCBs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

We assessed the mechanisms by which nonencapsulated heme, released in the plasma of mice after exposure to chlorine (Cl 2 ) gas, resulted in the initiation and propagation of acute lung injury. We exposed adult male and female C57BL/6 mice to Cl 2 (500 ppm for 30 min), returned them to room air, and injected them intramuscularly with either human hemopexin (hHPX; 5 µg/g BW in 50-µL saline) or vehicle at 1 h post-exposure. Upon return to room air, Cl 2 -exposed mice, injected with vehicle, developed respiratory acidosis, increased concentrations of plasma proteins in the alveolar space, lung mitochondrial DNA injury, increased levels of free plasma heme, and major alterations of their lung proteome. hHPX injection mice mitigated the onset and development of lung and mitochondrial injury and the increase of plasma heme, reversed the Cl 2 -induced changes in 83 of 237 proteins in the lung proteome at 24 h post-exposure, and improved survival at 15 days post-exposure. Systems biology analysis of the lung global proteomics data showed that hHPX reversed changes in a number of key pathways including elF2 signaling, verified by Western blotting measurements. Recombinant human hemopexin, generated in tobacco plants, injected at 1 h post-Cl 2 exposure, was equally effective in reversing acute lung and mtDNA injury. The results of this study offer new insights as to the mechanisms by which exposure to Cl 2 results in acute lung injury and the therapeutic effects of hemopexin. NEW & NOTEWORTHY Herein, we demonstrate that exposure of mice to chlorine gas causes significant changes in the lung proteome 24 h post-exposure. Systems biology analysis of the proteomic data is consistent with damage to mitochondria and activation of eIF2, the master regulator of transcription and protein translation. Post-exposure injection of hemopexin, which scavenges free heme, attenuated mtDNA injury, eIF2α phosphorylation, decreased lung injury, and increased survival.

Chlorine gas (Cl 2 ) has been repeatedly used as a chemical weapon, first in World War I and most recently in Syria. Life-threatening Cl 2 exposures frequently occur in domestic and occupational environments, and in transportation accidents. Modeling the human etiology of Cl 2 -induced acute lung injury (ALI), forensic biomarkers, and targeted countermeasures development have been hampered by inadequate large animal models. The objective of this study was to develop a translational model of Cl 2 -induced ALI in swine to understand toxico-pathophysiology and evaluate whether it is suitable for screening potential medical countermeasures and to identify biomarkers useful for forensic analysis. Specific pathogen-free Yorkshire swine (30-40 kg) of either sex were exposed to Cl 2 (≤240 ppm for 1 h) or filtered air under anesthesia and controlled mechanical ventilation. Exposure to Cl 2 resulted in severe hypoxia and hypoxemia, increased airway resistance and peak inspiratory pressure, and decreased dynamic lung compliance. Cl 2 exposure resulted in increased total leucocyte and neutrophil counts in bronchoalveolar lavage fluid, vascular leakage, and pulmonary edema compared with the air-exposed group. The model recapitulated all three key histopathological features of human ALI, such as neutrophilic alveolitis, deposition of hyaline membranes, and formation of microthrombi. Free and lipid-bound 2-chlorofatty acids and chlorotyrosine-modified proteins (3-chloro-l-tyrosine and 3,5-dichloro-l-tyrosine) were detected in plasma and lung tissue after Cl 2 exposure. In this study, we developed a translational swine model that recapitulates key features of human Cl 2 inhalation injury and is suitable for testing medical countermeasures, and validated chlorinated fatty acids and protein adducts as biomarkers of Cl 2 inhalation. NEW & NOTEWORTHY We established a swine model of chlorine gas-induced acute lung injury that exhibits several features of human acute lung injury and is suitable for screening potential medical countermeasures. We validated chlorinated fatty acids and protein adducts in plasma and lung samples as forensic biomarkers of chlorine inhalation.

Dechlorination is one of the main processes for the natural degradation of polychlorinated biphenyls (PCBs) in an anaerobic environment. However, PCB dechlorination pathways and products vary with PCB congeners, types of functional dechlorinating bacteria, and environmental conditions. The present study develops a novel model for determining dechlorination pathways and fluxes by tracking redox potential variability, transforming the complex dechlorination process into a stepwise sequence. The redox potential is calculated via the Gibbs free energy of formation, PCB concentrations in reactants and products, and environmental conditions. Thus, the continuous change in the PCB congener composition can be tracked during dechlorination processes. The new model is assessed against four measurements from several published studies on PCB dechlorination. The simulation errors in all four measurements are calculated between 2.67 and 35.1% under minimum ( n = 0) and maximum ( n = 34) numbers of co-eluters, respectively. The dechlorination fluxes for para- dechlorination pathways dominate PCB dechlorination in all measurements. Furthermore, the model also considers multiple-step dechlorination pathways containing intermediate PCB congeners absent in both the reactants and the products. The present study indicates that redox potential might be an appropriate indicator for predicting PCB dechlorination pathways and fluxes even without prior knowledge of the functional dechlorinating bacteria.

Substantial natural chlorination processes are a growing concern in diverse terrestrial ecosystems, occurring through abiotic redox reactions or biological enzymatic reactions. Among these, exoenzymatically mediated chlorination is suggested to be an important pathway for producing organochlorines and converting chloride ions (Cl - ) to reactive chlorine species (RCS) in the presence of reactive oxygen species like hydrogen peroxide (H 2 O 2 ). However, the role of natural enzymatic chlorination in antibacterial activity occurring in soil microenvironments remains unexplored. Here, we conceptualized that heme-containing chloroperoxidase (CPO)-catalyzed chlorination functions as a naturally occurring disinfection process in soils. Combining antimicrobial experiments and microfluidic chip-based fluorescence imaging, we showed that the enzymatic chlorination process exhibited significantly enhanced antibacterial activity against Escherichia coli and Bacillus subtilis compared to H 2 O 2 . This enhancement was primarily attributed to in situ-formed RCS. Based on semiquantitative imaging of RCS distribution using a fluorescence probe, the effective distance of this antibacterial effect was estimated to be approximately 2 mm. Ultrahigh-resolution mass spectrometry analysis showed over 97% similarity between chlorine-containing formulas from CPO-catalyzed chlorination and abiotic chlorination (by sodium hypochlorite) of model dissolved organic matter, indicating a natural source of disinfection byproduct analogues. Our findings unveil a novel natural disinfection process in soils mediated by indigenous enzymes, which effectively links chlorine-carbon interactions and reactive species dynamics.

Pharmaceutical transformation products (TPs) generated during wastewater treatment have become an environmental concern. However, there is limited understanding regarding the TPs produced from pharmaceuticals during wastewater treatment. In this study, chloroquine (CQ), which was extensively used for treating coronavirus disease-19 (COVID-19) infections during the pandemic, was selected for research. We identified and fractionated the main TP produced from CQ during chlorine disinfection and investigated the neurotoxic effects of CQ and its main TP on zebrafish (Danio rerio) embryos. Halogenated TP353 was observed as one of the main TPs produced from CQ during chlorine disinfection. Zebrafish embryos test revealed that TP353 caused higher neurotoxicity in zebrafish larvae, as compared to the CQ, and that was accompanied by significantly decreased expression levels of the genes related to central nervous system development (e.g., gfap, syn2a, and elavl3), inhibited activity of acetylcholinesterase (AChE), reduced GFP fluorescence intensity of motor neuron axons in transgenic larvae (hb9-GFP), and reduced total swimming distance and swimming velocity of larvae during light-dark transition stimulation. The results of this study can potentially be utilized as a theoretical reference for future evaluations of environmental risks associated with CQ and its related TPs. This work presents a methodology for assessing the environmental hazards linked to the discharge of pharmaceutical TPs after wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Residual chlorine and biofilm coexistence is inevitable in drinking water transmission and distribution networks. Understanding the microbial response and its mediated effects on disinfection byproducts under different categories of residual chlorine stress is essential to ensure water safety. The aim of our study was to determine the response of pipe wall biofilms to residual chlorine pressure in chlorine and chloramine systems and to understand the microbially mediated effects on the formation and migration of haloacetonitriles (HANs), typical nitrogenous disinfection byproducts. According to the experimental results, the biofilm response changes under pressure, with significant differences noted in morphological characteristics, the extracellular polymeric substances (EPS) spatial structure, bacterial diversity, and functional abundance potential. Upon incubation with residual chlorine (1.0 ± 0.2 mg/L), the biofilm biomass per unit area, EPS, community abundance, and diversity increased in the chloramine group, and the percentage of viable bacteria increased, potentially indicating that the chloramine group provides a richer variety of organic matter precursors. Compared with the chloramine group, the chlorination group exhibited increased haloacetonitrile formation potential (HANFP), with Rhodococcus (43.2%) dominating the system, whereas the prediction abundance of metabolic functions was advantageous, especially with regard to amino acid metabolism, carbohydrate metabolism, and the biodegradation and metabolism of foreign chemicals. Under chlorine stress, pipe wall biofilms play a stronger role in mediating HAN production. It is inferred that chlorine may stimulates microbial interactions, and more metabolites (e.g., EPS) consume chlorine to protect microbial survival. EPS dominates in biofilms, in which proteins exhibit greater HANFP than polysaccharides., Competing Interests: Declaration of competing interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled, “Response mechanisms of pipe wall biofilms in water supply networks under different disinfection strategy pressures and the effect of mediating halogenated acetonitrile formation”., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Microbial dechlorination of polychlorinated biphenyls (PCBs) is limited by the slow growth rate and low activity of dechlorinators. Resuscitation promoting factor (Rpf) of Micrococcus luteus, has been demonstrated to accelerate the enrichment of highly active PCB-dechlorinating cultures. However, it remains unclear whether the addition of Rpf can further improve the dechlorination performance of anaerobic dechlorination cultures. In this study, the effect of Rpf on the performance of TG4, an enriched PCB-dechlorinating culture obtained by Rpf amendment, for reductive dechlorination of four typical PCB congeners (PCBs 101, 118, 138, 180) was evaluated. The results indicated that Rpf significantly enhanced the dechlorination of the four PCB congeners, with residual mole percentages of PCBs 101, 118, 138 and 180 in Rpf-amended cultures being 16.2-29.31 %, 13.3-20.1 %, 11.9-14.4 % and 9.4-17.3 % lower than those in the corresponding cultures without Rpf amendment after 18 days of incubation. Different models were identified as appropriate for elucidating the dechlorination kinetics of distinct PCB congeners, and it was observed that the dechlorination rate constant is significantly influenced by the PCB concentration. The supplementing Rpf did not obviously change dechlorination metabolites, and the removal of chlorines occurred mainly at para- and meta- positions. Analysis of microbial community and functional gene abundance suggested that Rpf-amended cultures exhibited a significant enrichment of Dehalococcoides, Dehalogenimonas and Desulfitobacterium, as well as non-dechlorinators belonging to Desulfobacterota and Bacteroidetes. These findings highlight the potential of Rpf as an effective additive for enhancing PCB dechlorination, providing new insights into the survival of functional microorganisms involved in anaerobic reductive dechlorination., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Cyanobacteria and their metabolites are one of the primary precursors of disinfection by-products (DBPs) in natural water environments. However, few studies have investigated whether the production of DBPs by cyanobacteria changes under complex environmental conditions and possible mechanisms underlying these changes. Therefore, we investigated the effects of algal growth phase, water temperature, pH, illumination and nutrients on the production of trihalomethane formation potential (THMFPs) by Microcystis aeruginosa in four algal metabolic fractions, that is, hydrophilic extracellular organic matter (HPI-EOM), hydrophobic EOM (HPO-EOM), hydrophilic intracellular organic matter (HPI-IOM) and hydrophobic IOM (HPO-IOM). Additionally, correlations between THMFPs and some typical algal metabolite surrogates were analyzed. The results showed that the productivity of THMFPs by M. aeruginosa in EOM could be affected significantly by the algal growth phase and incubation conditions, while the IOM productivity varied insignificantly. M. aeruginosa in the death phase could secrete more EOM and have a higher THMFP productivity than those in the exponential or stationary phases. Cyanobacteria grown under harsh conditions could have increased THMFP productivity in EOM by increasing the reactivity of algal metabolites with chlorine, for example, under low pH conditions, and secreting more metabolites in EOM, for example, under low temperature or nutrient limitation conditions. Polysaccharides were responsible for the enhanced THMFP productivity in HPI-EOM fraction, and a significant linear correlation was found between the concentration of polysaccharides and THMFPs (r = 0.8307). However, THMFPs in HPO-EOM did not correlate with dissolved organic carbon (DOC), ultraviolet absorbance at 254 nm (UV 254 ), specific UV absorbance (SUVA) and cell density. Thus, we could not specify the kind of algal metabolites that contribute to the increased THMFPs in the HPO-EOM fraction under harsh growth conditions. Compared with the case in EOM, the THMFPs in IOM were more stable and correlated with the cell density and total amount of IOM. The results implied that the THMFPs in the EOM were sensitive to growth conditions and were independent of algal density. Considering the fact that traditional water treatment plants cannot remove dissolved organics as efficiently as algal cells, the increased THMFP productivity in EOM by M. aeruginosa under harsh growth conditions could be a potentially serious threat to the safety of the water supply., Competing Interests: Declaration of competing interest I would like to declare on behalf of my co-authors that the work described was original research that has not been published previously, and not under consideration for any other journals, in whole or in part. All authors are aware of, and accept responsibility for the manuscript., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study evaluated the body condition score (BCS) at calving and breed (B) effects on milk composition, yield, performance, physiological parameters, hemogram, blood metabolites, and urinary metabolites in the transition and early lactation periods of Mediterranean (MED) and Murrah (MUR) buffaloes. Twenty MED and fifteen MUR buffaloes were distributed into four experimental treatments, in a completely randomized design, considering their racial groups and BCS (LBCS = low; HBCS = high): LBCS MED (N = 9); HBCS MED (N = 11); LBCS MUR (N = 8); HBCS MUR (N = 7). Animals were monitored during the last 21 days of gestation and first 56 days postpartum and kept under the same management and feeding conditions. During data collection, milk composition, yield, performance, physiological parameters, hemogram, blood metabolites, and urinary metabolites were evaluated. Higher milk production and fat-corrected milk were observed in MED than MUR buffaloes. Breed effects were observed on body weight, rectal temperature, glucose, urea, calcium (Ca) concentrations, and BCS effects on total protein, albumin, urea, and Ca. There were BCS effects on hematocrit, neutrophils, eosinophils, and interactions between B × BCS for lymphocytes and platelets. There were breed effects on urinary concentrations of chlorine, uric acid, and interactions between weight (W) × B on chlorine and urea. The MED buffaloes can be considered the most prepared to undergo physiological changes, including the BCS value at calving, indicating higher physiological health. Besides, this study demonstrates more considerable preparation for the calving, regardless of the body condition score at calving., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Polychlorinated biphenyls (PCBs) and microplastics (MPs) commonly co-exist in various environments. MPs inevitably start aging once they enter environment. In this study, the effect of photo-aged polystyrene MPs on microbial PCB dechlorination was investigated. After a UV aging treatment, the proportion of oxygen-containing groups in MPs increased. Photo-aging promoted the inhibitory effect of MPs on microbial reductive dechlorination of PCBs, mainly attributed to the inhibition of meta-chlorine removal. The inhibitory effects on hydrogenase and adenosine triphosphatase activity by MPs increased with increasing aging degree, which may be attributed to electron transfer chain inhibition. PERMANOVA showed significant differences in microbial community structure between culturing systems with and without MPs (p < 0.05). Co-occurrence network showed a simpler structure and higher proportion of negative correlation in the presence of MPs, especially for biofilms, resulting in increased potential for competition among bacteria. MP addition altered microbial community diversity, structure, interactions, and assembly processes, which was more deterministic in biofilms than in suspension cultures, especially regarding the bins of Dehalococcoides. This study sheds light on the microbial reductive dechlorination metabolisms and mechanisms where PCBs and MPs co-exist and provides theoretical guidance for in situ application of PCB bioremediation technology., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Many disease resistance genes in wheat (Triticum aestivum L.) confer strong resistance to specific pathogen races or strains, and only a small number of genes confer multipathogen resistance. The Leaf rust resistance 67 (Lr67) gene fits into the latter category as it confers partial resistance to multiple biotrophic fungal pathogens in wheat and encodes a Sugar Transport Protein 13 (STP13) family hexose-proton symporter variant. Two mutations (G144R, V387L) in the resistant variant, Lr67res, differentiate it from the susceptible Lr67sus variant. The molecular function of the Lr67res protein is not understood, and this study aimed to broaden our knowledge on this topic. Biophysical analysis of the wheat Lr67sus and Lr67res protein variants was performed using Xenopus laevis oocytes as a heterologous expression system. Oocytes injected with Lr67sus displayed properties typically associated with proton-coupled sugar transport proteins-glucose-dependent inward currents, a Km of 110 ± 10 µM glucose, and a substrate selectivity permitting the transport of pentoses and hexoses. By contrast, Lr67res induced much larger sugar-independent inward currents in oocytes, implicating an alternative function. Since Lr67res is a mutated hexose-proton symporter, the possibility of protons underlying these currents was investigated but rejected. Instead, currents in Lr67res oocytes appeared to be dominated by anions. This conclusion was supported by electrophysiology and 36Cl- uptake studies and the similarities with oocytes expressing the known chloride channel from Torpedo marmorata, TmClC-0. This study provides insights into the function of an important disease resistance gene in wheat, which can be used to determine how this gene variant underpins disease resistance in planta., Competing Interests: Conflict of interest statement. All authors declare no conflict of interest., (© The Author(s) 2023. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Cardiovascular diseases, the leading life-threatening conditions, involve cardiac arrhythmia, coronary artery disease, myocardial infarction, heart failure, cardiomyopathy, and heart valve disease that are associated with the altered functioning of cation-chloride cotransporters. The decreased number of cation-chloride cotransporters leads to reduced reactivity to adrenergic stimulation. The KCC family is crucial for numerous physiological processes including cell proliferation and invasion, regulation of membrane trafficking, maintaining ionic and osmotic homeostasis, erythrocyte swelling, dendritic spine formation, maturation of postsynaptic GABAergic inhibition, and inhibitory/excitatory signaling in neural tracts. KCC2 maintains intracellular chlorine homeostasis and opposes β-adrenergic stimulation-induced Cl- influx to prevent arrhythmogenesis. KCC3-inactivated cardiac tissue shows increased vascular resistance, aortic distensibility, heart size and weight (i.e. hypertrophic cardiomyopathy). Due to KCC4's high affinity for K+, it plays a vital role in cardiac ischemia with increased extracellular K+. The NKCC and NCC families play a vital role in the regulation of saliva volume, establishing the potassium-rich endolymph in the cochlea, sodium uptake in astrocytes, inhibiting myogenic response in microcirculatory beds, regulation of smooth muscle tone in resistance vessels, and blood pressure. NKCC1 regulates chlorine homeostasis and knocking it out impairs cardiomyocyte depolarization and cardiac contractility as well as impairs depolarization and contractility of vascular smooth muscle rings in the aorta. The activation of NCC in vascular cells promotes the formation of the abdominal aortic aneurysm. This narrative review provides a deep insight into the structure and function of KCCs, NKCCs, and NCC in human physiology and cardiac pathobiology. Also, it provides cell-specific (21 cell types) and region-specific (6 regions) expression of KCC1, KCC2, KCC3, KCC4, NKCC1, NKCC2, and NCC in heart., Competing Interests: Declaration of Competing Interest The authors have no competing interests to declare., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

As an important part of China's chemical industry, chlor-alkali enterprises were featured by heavy consumption of natural resource and large amount of pollutants. Circular economy in chlor-alkali industry should be developed in China to achieve a balance between economic growth and resource consumption. This study took three chlor-alkali enterprises in Shanghai Chemical Industry Park as a case, and three scenarios were established for chlorine metabolism: Scenario 1 (Chlorine flows independently), Scenario 2 (Chlorine flows from Enterprise A to Enterprise B and C), Scenario 3 (Closed loop of Chlorine Gas). An integrated methodology based upon Substance Flow Analysis (SFA), Life Cycle Assessment (LCA), Life Cycle Cost (LCC), and Data Envelopment Analysis (DEA) was set up and used to evaluate the Chlor-Alkali Industrial Symbiosis. The results showed that the resource efficiency, production and conversion rate of chlorine was significantly improved with the establishment and optimization of symbiosis scenarios, which also showed very positive environmental and economic benefits according to LCA-LCC analysis. DEA was performed to analyze the Chlorine's resource efficiency in its metabolism, and the results showed that the efficiency of Scenario 1, Scenario 2 and Scenario 3 were 0.8548, 0.9490 and 1, respectively. Especially, in Scenario 3, a new chemical technology, catalytic oxidation of hydrogen chloride, can convert the by-product hydrogen chloride into chlorine, which can be reused and finally made a closed-loop in this industrial symbiosis. The results of this study can provide a new way to optimize the resource recycle, pollutants discharge and carbon emissions of chlor-alkali industry. [ABSTRACT FROM AUTHOR]

Polyvinyl chloride (PVC), a carbon backbone synthetic plastic containing chlorine element, is one of six widely used plastics accounting for 10% global plastics production. PVC wastes are recalcitrant to be broken down in the environment but release harmful chlorinated compounds, causing damage to the ecosystem. Although biodegradation represents a sustainable approach for PVC reduction, virtually no efficient bacterial degraders for additive-free PVC have been reported. In addition, PVC depolymerization by Tenebrio molitor larvae was suggested to be gut microbe-dependent, but to date no additive-free PVC degraders have been isolated from insect guts. In this study, a bacterial consortium designated EF1 was newly enriched from the gut of Tenebrio molitor larvae, which was capable of utilizing additive-free PVC for its growth with the PVC-mass reduction and dechlorination of PVC. PVC films inoculated with consortium EF1 for 30 d were analyzed by diverse polymer characterization methods including atomic force microscopy, scanning electron microscope, water contact angle, time-of-flight secondary ion mass spectrometry, Fourier transform infrared spectroscopy, differential scanning calorimetry, thermogravimetric analysis technique, and ion chromatography. It was found that bio-treated PVC films were covered with tight biofilms with increased -OH and -CC- groups and decreased chlorine contents, and erosions and cracks were present on their surfaces. Meanwhile, the hydrophilicity of bio-treated films increased, but their thermal stability declined. Furthermore, M w , M n and M z values were reduced by 17.0%, 28.5% and 16.1% using gel permeation chromatography, respectively. In addition, three medium-chain aliphatic primary alcohols and their corresponding fatty acids were identified as PVC degradation intermediates by gas chromatography-mass spectrometry. Combing all above results, it is clear that consortium EF1 is capable of efficiently degrading PVC polymer, providing a unique example for PVC degradation by gut microbiota of insects and a feasibility for the removal of PVC wastes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Sensor histidine kinase HprS, an oxidative stress sensor of Escherichia coli, senses reactive oxygen species (ROS) and reactive chlorine species (RCS), and is involved in the induction of oxidatively damaged protein repair periplasmic enzymes. We reinvestigated the roles of six methionine and four cysteine residues of HprS in the response to HClO, an RCS. The results of site-directed mutagenesis revealed that methionine residues in periplasmic and cytoplasmic regions (Met225) are involved in HprS activation. Interestingly, the Cys165Ser substitution reduced HprS activity, which was recovered by an additional Glu22Cys substitution. Our results demonstrate that the position of the inner membrane cysteine residues influences the extent of HprS activation in HClO sensing., (© 2023 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

The anaerobic bioremediation of polychlorinated biphenyls (PCBs) is largely impeded by difficulties in massively enriching PCB dechlorinators in short periods of time. Tetrachloroethene (PCE) is often utilized as an alternative electron acceptor to preenrich PCB-dechlorinating bacteria. In this study, resuscitation promoting factor (Rpf) was used as an additive to enhance the enrichment of the microbial communities involved in PCE/PCBs dechlorination. The results indicated that Rpf accelerates PCE dechlorination 3.8 to 5.4 times faster than control cultures. In Aroclor 1260-fed cultures, the amendment of Rpf enables significantly more rapid and extensive dechlorination of PCBs. The residual high-chlorinated PCB congeners (≥5 Cl atoms) accounted for 36.7% and 59.8% in the Rpf-amended cultures and in the corresponding controls, respectively. This improvement was mainly attributed to the enhanced activity of the removal of meta-chlorines (47.7 mol % versus 14.7 mol %), which did not appear to affect dechlorination pathways. The dechlorinators, including Dehalococcoides in Chloroflexi and Desulfitobacterium in Firmicutes , were greatly enriched via Rpf amendment. The abundance of nondechlorinating populations, including Methanosarcina , Desulfovibrio , and Bacteroides , was also greatly enhanced via Rpf amendment. These results suggest that Rpf serves as an effective additive for the rapid enrichment of active dechlorinating cultures so as to provide a new approach by which to massively cultivate bioinoculants for accelerated in situ anaerobic bioremediation. IMPORTANCE The resuscitation promoting factor (Rpf) of Micrococcus luteus has been reported to resuscitate and stimulate the growth of functional microorganisms that are involved in the aerobic degradation of polychlorinated biphenyls (PCBs). However, few studies have been conducted to investigate the role of Rpf on anaerobic microbial populations. In this study, the enhancement of Rpf on the anaerobic microbial dechlorination of PCE/PCBs was discovered. Additionally, the Rpf-responsive populations underlying the enhanced dechlorination were uncovered. This report reveals the rapid enrichment of active dechlorinating cultures via Rpf amendment, and this sheds light on massively enriching PCB dechlorinators in short periods of time. The enhanced in situ anaerobic bioremediation of PCBs could be expected by supplementing Rpf.

Chlorine is abundant in cells and biomolecules, yet the biology of chlorine oxidation and reduction is poorly understood. Some bacteria encode the enzyme chlorite dismutase (Cld), which detoxifies chlorite (ClO 2 - ) by converting it to chloride (Cl - ) and molecular oxygen (O 2 ). Cld is highly specific for chlorite and aside from low hydrogen peroxide activity has no known alternative substrate. Here, we reasoned that because chlorite is an intermediate oxidation state of chlorine, Cld can be used as a biomarker for oxidized chlorine species. Cld was abundant in metagenomes from various terrestrial habitats. About 5% of bacterial and archaeal genera contain a microorganism encoding Cld in its genome, and within some genera Cld is highly conserved. Cld has been subjected to extensive horizontal gene transfer. Genes found to have a genetic association with Cld include known genes for responding to reactive chlorine species and uncharacterized genes for transporters, regulatory elements, and putative oxidoreductases that present targets for future research. Cld was repeatedly co-located in genomes with genes for enzymes that can inadvertently reduce perchlorate (ClO 4 - ) or chlorate (ClO 3 - ), indicating that in situ (per)chlorate reduction does not only occur through specialized anaerobic respiratory metabolisms. The presence of Cld in genomes of obligate aerobes without such enzymes suggested that chlorite, like hypochlorous acid (HOCl), might be formed by oxidative processes within natural habitats. In summary, the comparative genomics of Cld has provided an atlas for a deeper understanding of chlorine oxidation and reduction reactions that are an underrecognized feature of biology., (© 2022. The Author(s).)

Nutrient imbalance (i.e., deficiency and toxicity) of microelements is an outstanding environmental issue that influences each aspect of ecosystems. Although the crucial roles of microelements in entire lifecycle of plants have been widely acknowledged, the effective control of microelements is still neglected due to the narrow safe margins. Selenium (Se) is an essential element for humans and animals. Although it is not believed to be indispensable for plants, many literatures have reported the significance of Se in terms of the uptake, accumulation, and detoxification of essential microelements in plants. However, most papers only concerned on the antagonistic effect of Se on metal elements in plants and ignored the underlying mechanisms. There is still a lack of systematic review articles to summarize the comprehensive knowledge on the connections between Se and microelements in plants. In this review, we conclude the bidirectional effects of Se on micronutrients in plants, including iron, zinc, copper, manganese, nickel, molybdenum, sodium, chlorine, and boron. The regulatory mechanisms of Se on these micronutrients are also analyzed. Moreover, we further emphasize the role of Se in alleviating element toxicity and adjusting the concentration of micronutrients in plants by altering the soil conditions (e.g., adsorption, pH, and organic matter), promoting microbial activity, participating in vital physiological and metabolic processes, generating element competition, stimulating metal chelation, organelle compartmentalization, and sequestration, improving the antioxidant defense system, and controlling related genes involved in transportation and tolerance. Based on the current understanding of the interaction between Se and these essential elements, future directions for research are suggested., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Chiral polychlorinated biphenyls (PCB) are environmentally relevant developmental neurotoxicants. Because their hydroxylated metabolites (OH-PCBs) are also neurotoxic, it is necessary to determine how PCB metabolism affects the developing brain, for example, in mouse models. Because the cytochrome P450 isoforms involved in the metabolism of chiral PCBs remain unexplored, we investigated the metabolism of PCB 91 (2,2',3,4',6-pentachlorobiphenyl), PCB 95 (2,2',3,5',6-pentachlorobiphenyl), PCB 132 (2,2',3,3',4,6'-hexachlorobiphenyl), and PCB 136 (2,2',3,3',6,6'-hexachlorobiphenyl) using liver microsomes from male and female Cyp2a(4/5)bgs -null, Cyp2f2 -null, and wild-type mice. Microsomes, pooled by sex, were incubated with 50 μM PCB for 30 min, and the levels and enantiomeric fractions of the OH-PCBs were determined gas chromatographically. All four PCB congeners appear to be atropselectively metabolized by CYP2A(4/5)BGS and CYP2F2 enzymes in a congener- and sex-dependent manner. The OH-PCB metabolite profiles of PCB 91 and PCB 132, PCB congeners with one para -chlorine substituent, differed between null and wild-type mice. No differences in the metabolite profiles were observed for PCB 95 and PCB 136, PCB congeners without a para -chlorine group. These findings suggest that Cyp2a(4/5)bgs -null and Cyp2f2 -null mice can be used to study how a loss of a specific metabolic function (e.g., deletion of Cyp2a(4/5)bgs or Cyp2f2 ) affects the toxicity of chiral PCB congeners.

The ability to overcome stressful environments is critical for pathogen survival in the host. One challenge for bacteria is the exposure to reactive chlorine species (RCS), which are generated by innate immune cells as a critical part of the oxidative burst. Hypochlorous acid (HOCl) is the most potent antimicrobial RCS and is associated with extensive macromolecular damage in the phagocytized pathogen. However, bacteria have evolved defense strategies to alleviate the effects of HOCl-mediated damage. Among these are RCS-sensing transcriptional regulators that control the expression of HOCl-protective genes under non-stress and HOCl stress. Uropathogenic Escherichia coli (UPEC), the major causative agent of urinary tract infections (UTIs), is particularly exposed to infiltrating neutrophils during pathogenesis; however, their responses to and defenses from HOCl are still completely unexplored. Here, we present evidence that UPEC strains tolerate higher levels of HOCl and are better protected from neutrophil-mediated killing compared with other E. coli. Transcriptomic analysis of HOCl-stressed UPEC revealed the upregulation of an operon consisting of three genes, one of which encodes the transcriptional regulator RcrR. We identified RcrR as a HOCl-responsive transcriptional repressor, which, under non-stress conditions, is bound to the operator and represses the expression of its target genes. During HOCl exposure, however, the repressor forms reversible intermolecular disulfide bonds and dissociates from the DNA resulting in the derepression of the operon. Deletion of one of the target genes renders UPEC significantly more susceptible to HOCl and phagocytosis indicating that the HOCl-mediated induction of the regulon plays a major role for UPEC's HOCl resistance. IMPORTANCE How do pathogens deal with antimicrobial oxidants produced by the innate immune system during infection? Uropathogenic Escherichia coli (UPEC), the most common etiological agent of urinary tract infections (UTIs), is particularly exposed to infiltrating neutrophils and, therefore, must counter elevated levels of the antimicrobial oxidant HOCl to establish infection. Our study provides fundamentally new insights into a defense mechanism that enables UPEC to fend off the toxic effects of HOCl stress. Intriguingly, the defense system is predominantly found in UPEC and absent in noninvasive enteropathogenic E. coli. Our data suggest expression of the target gene rcrB is exclusively responsible for UPEC's increased HOCl tolerance in culture and contributes to UPEC's survival during phagocytosis. Thus, this novel HOCl stress defense system could potentially serve as an attractive drug target to increase the body's own capacity to fight UTIs.

Plant with a great diversity shows several responses towards the biotic and abiotic stresses. Among these abiotic stresses, salinity is the main damaging factor as it reduces the yield of wheat plant with moderate salt tolerance. For its survival, plant undergoes through some genetic, biochemical and physiological changes to tackle the stress. This review mainly describes the conditions where various ions present in the soil, especially sodium and chlorine, enter into the plant and the genes or proteins involved with survival mechanism against the damage in plants. Salt stress causes alteration in enzymatic activity and Photosynthesis, oxidative stress, damage of cellular structure and components and ionic imbalance. Ion toxicity stress occur due to accumulation of excessive sodium ion and chloride ion. Transcriptional factors TaPIMP, TaSRG and TaMYBsdu 1 play key role in gene expression mechanism to overcome the stress. High affinity potassium transporter gene family is responsible for salt tolerance in wheat plant. HKT1;4 and HKT1;5 genes are responsible for Na exclusion in Triticum monococcum. Forty QTLs were found with the marker assisted selection in bread wheat for salinity tolerance and some morphological traits, 5 QTLs were related to sodium ion exclusion. In bread wheat, salt stress tolerance mechanism is mainly an exclusion of Na + ions but also include K + ion concentration. The salinity tolerant germplasm MW#293 provides an opportunity for the development of future salinity tolerant bread wheat., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Repairing oxidative-targeted macromolecules is a central mechanism necessary for living organisms to adapt to oxidative stress. Reactive oxygen and chlorine species preferentially oxidize sulfur-containing amino acids in proteins. Among these amino acids, methionine can be converted into methionine sulfoxide. This post-translational oxidation can be reversed by methionine sulfoxide reductases, Msr enzymes. In Gram-negative bacteria, the antioxidant MsrPQ system is involved in the repair of periplasmic oxidized proteins. Surprisingly, in this study, we observed in Escherichia coli that msrPQ was highly expressed in the absence of oxygen. We have demonstrated that the anaerobic induction of msrPQ was due to chlorate (ClO 3 - ) contamination of the Casamino Acids. Molecular investigation led us to determine that the reduction of chlorate to the toxic oxidizing agent chlorite (ClO 2 - ) by the three nitrate reductases (NarA, NarZ, and Nap) led to methionine oxidation of periplasmic proteins. In response to this stress, the E. coli HprSR two-component system was activated, leading to the over-production of MsrPQ. This study, therefore, supports the idea that methionine oxidation in proteins is part of chlorate toxicity, and that MsrPQ can be considered as an anti-chlorate/chlorite defense system in bacteria. Finally, this study challenges the traditional view of the absence of Met-oxidation during anaerobiosis., (© 2022 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.)

Polychlorinated biphenyls (PCBs) as typical halogenated persistent organic pollutants are widely distributed in natural environments, and can be enriched and magnified in organisms via food webs. It is consequently urgent and necessary to develop techniques to completely remove these persistent organohalides. In this study, we developed a process (Bio-RD-PAOP) by integrating microbial reductive dechlorination (Bio-RD) with subsequent persulfate activation and oxidation process (PAOP) for effective remediation of PCBs. Results showed the synergistic combination of advantages of Bio-RD and PAOP in dechlorination of higher-chlorinated PCBs and of PAOP in degradation/mineralization of lower-chlorinated PCBs, respectively. For the PAOP, both experimental evidences and theoretical calculations suggested that degradation rate and efficiency decreased with the increased PCB chlorine numbers. Relative to the Bio-RD and PAOP, Bio-RD-PAOP had significantly higher PCB removal efficiencies, of which values were PCB congener-specific. For example, removal efficiency of Bio-RD-PAOP in removing PCB88 is 2.50 and 1.86 times of that of Bio-RD and PAOP, respectively. In contrast, the efficiency is 1.66 and 3.35 times of Bio-RD and PAOP, respectively, for PCB180 removal. The PAOP-derived oxidizing species (mainly sulfate free radical) significantly decreased microbial abundance, particularly of the organohalide-respiring Dehalococcoides. Notably, co-existence of other microorganisms alleviated the inhibitive effect of oxidizing species on the Dehalococcoides, possibly due to formation of microbial flocs or biofilm. This study provided a promising strategy for extensive remediation of organohalide-contaminated sites, as well as new insight into impact of PAOP-derived oxidizing species on the organohalide-respiring community., (Copyright © 2022 Elsevier B.V. All rights reserved.)

SARS-CoV-2, the culprit pathogen of COVID-19, elicits prominent immune responses and cytokine storms. Intracellular Cl - is a crucial regulator of host defense, whereas the role of Cl - signaling pathway in modulating pulmonary inflammation associated with SARS-CoV-2 infection remains unclear. By using human respiratory epithelial cell lines, primary cultured human airway epithelial cells, and murine models of viral structural protein stimulation and SARS-CoV-2 direct challenge, we demonstrated that SARS-CoV-2 nucleocapsid (N) protein could interact with Smad3, which downregulated cystic fibrosis transmembrane conductance regulator (CFTR) expression via microRNA-145. The intracellular Cl - concentration ([Cl - ] i ) was raised, resulting in phosphorylation of serum glucocorticoid regulated kinase 1 (SGK1) and robust inflammatory responses. Inhibition or knockout of SGK1 abrogated the N protein-elicited airway inflammation. Moreover, N protein promoted a sustained elevation of [Cl - ] i by depleting intracellular cAMP via upregulation of phosphodiesterase 4 (PDE4). Rolipram, a selective PDE4 inhibitor, countered airway inflammation by reducing [Cl - ] i . Our findings suggested that Cl - acted as the crucial pathological second messenger mediating the inflammatory responses after SARS-CoV-2 infection. Targeting the Cl - signaling pathway might be a novel therapeutic strategy for COVID-19., (© 2022. The Author(s).)

Microplastics (MPs) and polychlorinated biphenyls (PCBs) generally coexist in the environment, posing risks to public health and the environment. This study investigated the effect of different MPs on the microbial anaerobic reductive dechlorination of Aroclor 1260, a commercial PCB mixture. MP exposure inhibited microbial reductive dechlorination of PCBs, with inhibition rates of 39.43%, 23.97%, and 17.53% by polyethylene (PE), polypropylene (PP), and polystyrene (PS), respectively. The dechlorination rate decreased from 1.63 μM Cl - d -1 to 0.99-1.34 μM Cl - d -1 after MP amendment. Chlorine removal in the meta-position of PCBs was primarily inhibited by MPs, with no changes in the final PCB dechlorination metabolites. The microbial community compositions in MP biofilms were not significantly different (P > 0.05) from those in suspension culture, although possessing greater Dehalococcoides abundance (0.52-0.81% in MP biofilms; 0.03-0.12% in suspension culture). The co-occurrence network analysis revealed that the presence of MPs attenuated microbial synergistic interactions in the dechlorinating culture systems, which may contribute to the inhibitory effect on microbial PCB dechlorination. These findings are important for comprehensively understanding microbial dechlorination behavior and the environmental fate of PCBs in environments with co-existing PCBs and MPs and for guiding the application of in situ PCB bioremediation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The effect of chlorine on Escherichia coli biofilm O157:H7 are well established; however, the effect on biofilm adhesion to food as well as the six emerging E. coli serotypes ("big six") have not been fully understood. Chlorine sanitization with 1-min 100 mg/L was applied against seven pathogenic E. coli (O111, O121:H19, O45:H2, O26:H11, O103:H11, O145, and O157:H7) biofilms on high-density polyethylene (HDPE) and stainless steel (SS) coupons, respectively. Using sponge cake as a food model, the adhesion behavior was evaluated by comparison of bacteria transfer rate before and after treatment. Besides, the metabolic profiles of biofilms were analyzed by nuclear magnetic resonance (NMR) spectrometer. A significant decrease in transfer rate (79% decline on SS and 33% decline on HDPE) was recorded as well as the distinctive pattern between SS and HDPE coupons was also noticed, with a low population (6-7 log CFU/coupon) attached and low survivals (0-3 log CFU/coupon) upon chlorine on SS, while high population (7-8 log CFU/coupon) attached and high survivals (5-7 log CFU/coupon) on HDPE. Moreover, O121:H19 and O26:H11 demonstrated the highest resistance to chlorine with the least metabolic status and pathways affected. O103:H11, O145, and O111 followed similar metabolic patterns on both surfaces. Distinct metabolic patterns were found in O45:H2 and O157:H7, where the former had more affected metabolic status and pathways on SS but less on HDPE, whereas the latter showed an opposite trend. Overall, a potential contamination source of STEC infection in flour products was demonstrated and metabolic changes induced by chlorine were revealed by NMR-based metabolomics, which provides insights to avoid "big six" biofilms contamination in food., (Copyright © 2022. Published by Elsevier Ltd.)

Bacteria capable of dehalogenation via reductive or hydrolytic pathways are ubiquitous. Little is known, however, about the prevalence of bacterial dechlorination in deep terrestrial environments with a limited carbon supply. In this study we analyzed published genomes from three deep terrestrial subsurface sites: a deep aquifer in Western Siberia, the Sanford Underground Research Facility in South Dakota, USA, and the Soudan Underground Iron Mine (SUIM) in Minnesota, USA to determine if there was evidence to suggest that microbial dehalogenation was possible in these environments. Diverse dehalogenase genes were present in all analyzed metagenomes, with reductive dehalogenase and haloalkane dehalogenase genes the most common. Taxonomic analysis of both hydrolytic and reductive dehalogenase genes was performed to explore their affiliation; this analysis indicated that at the SUIM site, hydrolytic dehalogenase genes were taxonomically affiliated with Marinobacter species. Because of this affiliation, experiments were also performed with Marinobacter subterrani strain JG233 ('JG233'), an organism containing three predicted hydrolytic dehalogenase genes and isolated from the SUIM site, to determine whether hydrolytic dehalogenation was an active process and involved in growth on a chlorocarboxylic acid. Presence of these genes in genome appears to be functional, as JG233 was capable of chloroacetate dechlorination with simultaneous chloride release. Stable isotope experiments combined with confocal Raman microspectroscopy demonstrated that JG233 incorporated carbon from 13C-chloroacetate into its biomass. These experiments suggest that organisms present in these extreme and often low-carbon environments are capable of reductive and hydrolytic dechlorination and, based on laboratory experiments, may use this capability as a competitive advantage by utilizing chlorinated organic compounds for growth, either directly or after dechlorination., (© The Author(s) 2022. Published by Oxford University Press on behalf of FEMS.)

The pharmacological actions exerted by benzodiazepines are dependent on the discrete α protein subunits of the γ-aminobutyric acid type A receptor (GABA A R). Recent developments via a cryo-EM structure of the α1β3γ2L GABA A R ion channel provide crucial insights into ligand efficacy and binding affinity at this subtype. We investigated the molecular interactions of diazepam and alprazolam bound GABA A R structures (6HUP and 6HUO) to determine key binding interaction domains. A halogen bond between the chlorine atoms of diazepam and alprazolam with the group on the backbone of the α1 histidine amino acid 102 is important to the positive allosteric modulatory actions of diazepam and alprazolam in the α1β3γ2L GABA A R ion channel. In order to gain insight into α subtype selectivity we designed and synthesized close structural analogs of diazepam and alprazolam. These compounds were then docked into the recently publish cryo-EM structures of GABA A Rs (6HUP and 6HUO). This modeling along with radio-ligand binding data resulted in the conclusion that the non-classical bioisosteric replacement of the chlorine atom at C7 with an ethinyl group (compound 5) resulted in an 11-fold gain in α5 binding selectivity over the α1 subtype. Moreover, the potency of compound 5 resulted in a ligand with less sedation than diazepam, while still maintaining the same anxiolytic potency. These modeling data extend our understanding of the structural requirements for α-subtype-selective compounds that can be utilized to achieve improved medical treatments. It is clear that the ethinyl group in place of a halogen atom decreases the affinity and efficacy of benzodiazepines and imidazodiazepines at α1 subtypes, which results in less sedation and ataxia., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

H. boliviensis strain kknpp38 is a dense exopolysaccharide (EPS) producing bacterium, isolated from the early-stage (72-h-old) of marine biofilm. Laboratory experiments demonstrated that this isolate forms a potent biofilm on various artificial substrata viz. polystyrene, stainless steel as well as titanium and possesses high tolerance to chlorine disinfection. To determine the genes and biosynthetic pathways involved in the EPS production, whole-genome sequencing was performed using high-throughput Illumina tag sequencing. The high-quality reads were first de novo assembled using Unicycler genome assembler (version 0.4.9b) and then annotated using Prokka (version 1.13). The complete genome comes from one circular chromosome containing 4.96 Mbp DNA with G + C content of 55%, and encompasses genes encoding 4476 proteins, 2 rRNAs, and 57 tRNAs. Intriguingly, genomic analysis revealed the existence of genes involved in ATP-binding cassette (ABC) transporter-dependent EPS biosynthesis pathways (ugd, ugd2, galU). In addition, we identified genes involved in ectoine (ectA, ectB, ectC, ectD) and polyhydroxyalkanoates (PHAs; fabA, fabB, fabD, fabF, fabH, fabV, fabZ, phaC, phaD, phaG, phaR, phaZ1) production, which are known to involve in bacterial adaptation in saline environment. The outcomes of this study expand scientific understanding on the genes and pathways involved in EPS biosynthesis by marine bacteria., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The chemical and biological mechanisms of electrochemical elimination of Microcystis aeruginosa (M. aeruginosa) using boron-doped diamond (BDD) anode were comparatively explored in three different electrolytes (chloride, sulfate, and phosphate solutions). The most efficient elimination of M. aeruginosa was observed in chloride solution, which was attributed to the greatest total long-lived oxidants from the favorable formation of active chlorine. Moreover, the high permeability of active chlorine resulted in profound intracellular damages to chlorophyll-a, microcystin-LR (MC-LR), superoxide dismutase (SOD) enzyme, and DNA in the chloride system. The change of membrane permeability and degradation of the released MC-LR induced by active chlorine were further confirmed by the increase of extracellular MC-LR in the initial 5 min and a complete decay in the subsequent 15 min, while the change in morphology of algae cells was insignificant from SEM images. In sulfate and phosphate electrolytes, membrane damages were much more pronounced based on lipid peroxidation observation, although changes in cell morphology was found more significant in phosphate system. The higher concentrations of oxidants (·OH, O 3 , H 2 O 2 , S 2 O 8 2- ) generated in sulfate than in phosphate solution explained the greater efficiency of electrochemical elimination of M. aeruginosa in the sulfate electrolyte in terms of changes of cell density, OD 680 , chlorophyll-a, MC-LR, lipids, SOD enzyme, and DNA., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Two-component systems (TCS) are signaling pathways that allow bacterial cells to sense, respond to, and adapt to fluctuating environments. Among the classical TCS of Escherichia coli, HprSR has recently been shown to be involved in the regulation of msrPQ , which encodes the periplasmic methionine sulfoxide reductase system. In this study, we demonstrated that hypochlorous acid (HOCl) induces the expression of msrPQ in an HprSR-dependent manner, whereas H 2 O 2 , NO, and paraquat (a superoxide generator) do not. Therefore, HprS appears to be an HOCl-sensing histidine kinase. Using a directed mutagenesis approach, we showed that Met residues located in the periplasmic loop of HprS are important for its activity: we provide evidence that as HOCl preferentially oxidizes Met residues, HprS could be activated via the reversible oxidation of its methionine residues, meaning that MsrPQ plays a role in switching HprSR off. We propose that the activation of HprS by HOCl could occur through a Met redox switch. HprSR appears to be the first characterized TCS able to detect reactive chlorine species (RCS) in E. coli. This study represents an important step toward understanding the mechanisms of RCS resistance in prokaryotes. IMPORTANCE Understanding how bacteria respond to oxidative stress at the molecular level is crucial in the fight against pathogens. HOCl is one of the most potent industrial and physiological microbicidal oxidants. Therefore, bacteria have developed counterstrategies to survive HOCl-induced stress. Over the last decade, important insights into these bacterial protection factors have been obtained. Our work establishes HprSR as a reactive chlorine species-sensing, two-component system in Escherichia coli MG1655, which regulates the expression of msrPQ , two genes encoding, a repair system for HOCl-oxidized proteins. Moreover, we provide evidence suggesting that HOCl could activate HprS through a methionine redox switch.

The comparison of chemical and histopathological data obtained from the analysis of excised tumor fragments oral squamous cell carcinoma (OSCC) with the demographic and clinical evolution data is an effective strategy scarcely explored in OSCC studies. The aim was to analyze OSCC tissues for protein expression of enzymes related to oxidative stress and DNA repair and trace elements as candidates as markers of tumor aggressiveness and prognosis. Tumor fragments from 78 OSCC patients that had undergone ablative surgery were qualitatively analyzed by synchrotron micro-X-ray fluorescence for trace elements. Protein expression of SOD-1, Trx, Ref-1 and OGG1/2 was performed by immunohistochemistry. Sociodemographic, clinical, and histopathological data were obtained from 4-year follow-up records. Disease relapse was highest in patients with the presence of chlorine and chromium and lowest in those with tumors with high OGG1/2 expression. High expression of SOD-1, Trx, and Ref-1 was determinant of the larger tumor. Presence of trace elements can be markers of disease prognosis. High expression of enzymes related to oxidative stress or to DNA repair can be either harmful by stimulating tumor growth or beneficial by diminishing relapse rates. Interference on these players may bring novel strategies for the therapeutic management of OSCC patients., (© 2021. The Author(s).)

Background: Saliva provides a primary defense mechanism against several infectious diseases through its numerous immunological and non-immunological factors. Alteration in the composition of saliva often compromises its defense mechanisms, predisposing the oral cavity to disease entities. HIV patients under antiretroviral therapy (ART) have shown to exhibit altered salivary composition. These changes are postulated to be a result of the effect of ART on the salivary protein and electrolytes levels., Objectives: The present study aims to assess the potential difference in the salivary total protein and electrolyte levels in HIV patients with and without ART., Methods: Patients were divided into 3 groups- Group A (HIV-1 positive patient under ART for at least 6 months)-66, Group B (HIV-1 positive patient not started on ART)-66, Group C (HIV negative patients)-66. Saliva samples were collected and evaluated for total salivary protein and electrolyte levels in all the 3 groups., Results: There was a statistically significant difference in the salivary protein (p = 0.000) and electrolyte (Sodium, p = 0.000; Potassium, p = 0.039; chlorine, p = 0.027; ionized calcium, p = 0.002) levels among the three groups., Conclusion: HIV positive individuals with and without ART have alteration in the salivary composition. Some of these alterations (total protein and iCa levels) are due to the HIV infection, while others (Na, K, Cl) could be due to ART or a combined effect of both. Salivary changes in HIV positive individuals could predispose them to oral diseases. Thus, regular oral examination and prophylactic regimen must be formulated to maintain their oral hygiene and quality of life., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest, (Copyright © 2021. Published by Elsevier Inc.)

Soil salinity is a global concern and often the primary factor contributing to land degradation, limiting crop growth and production. Alfalfa (Medicago sativa L.) is a low input high value forage legume with a wide adaptation. Examining the tissue-specific responses to salt stress will be important to understanding physiological changes of alfalfa. The responses of two alfalfa cultivars (salt tolerant 'Halo', salt intolerant 'Vernal') were studied for 12 weeks in five gradients of salt stress in a sand based hydroponic system in the greenhouse. The accumulation and localization of elements and organic compounds in different tissues of alfalfa under salt stress were evaluated using synchrotron beamlines. The pattern of chlorine accumulation for 'Halo' was: root > stem ~ leaf at 8 dSm -1 , and root ~ leaf > stem at 12 dSm -1 , potentially preventing toxic ion accumulation in leaf tissues. In contrast, for 'Vernal', it was leaf > stem ~ root at 8 dSm -1 and leaf > root ~ stem at 12 dSm -1 . The distribution of chlorine in 'Halo' was relatively uniform in the leaf surface and vascular bundles of the stem. Amide concentration in the leaf and stem tissues was greater for 'Halo' than 'Vernal' at all salt gradients. This study determined that low ion accumulation in the shoot was a common strategy in salt tolerant alfalfa up to 8 dSm -1 of salt stress, which was then replaced by shoot tissue tolerance at 12 dSm -1 ., (Copyright © 2021 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Metal-oxide nanoparticles (MO-NPs), such as the highly bioreactive copper-based nanoparticles (CuO-NPs), are widely used in manufacturing of hundreds of commercial products. Epidemiological studies correlated levels of nanoparticles in ambient air with a significant increase in lung disease. CuO-NPs, specifically, were among the most potent in a set of metal-oxides and carbons studied in parallel regarding DNA damage and cytotoxicity. Despite advances in nanotoxicology research and the characterization of their toxicity, the exact mechanism(s) of toxicity are yet to be defined. We identified chlorination toxicity as a damaging consequence of inflammation and myeloperoxidase (MPO) activation, resulting in macromolecular damage and cell damage/death. We hypothesized that the inhalation of CuO-NPs elicits an inflammatory response resulting in chlorination damage in cells and lung tissues. We further tested the protective action of LGM2605, a synthetic small molecule with known scavenging properties for reactive oxygen species (ROS), but most importantly, for active chlorine species (ACS) and an inhibitor of MPO. CuO-NPs (15 µg/bolus) were instilled intranasally in mice and the kinetics of the inflammatory response in lungs was evaluated 1, 3, and 7 days later. Evaluation of the protective action of LGM2605 was performed at 24 h post-challenge, which was selected as the peak acute inflammatory response to CuO-NP. LGM2605 was given daily via gavage to mice starting 2 days prior to the time of the insult (100 mg/kg). CuO-NPs induced a significant inflammatory influx, inflammasome-relevant cytokine release, and chlorination damage in mouse lungs, which was mitigated by the action of LGM2605. Preventive action of LGM2605 ameliorated the adverse effects of CuO-NP in lung.

Aims: Toxaphene is a persistent organic pollutant, composed of approximately 1000 highly chlorinated bicyclic terpenes. The purpose of this study was to evaluate if camphor, a structural analogue of toxaphene, could stimulate aerobic biotransformation of weathered toxaphene., Methods and Results: Two enrichment cultures that degrade camphor as the sole carbon source were established from contaminated soil and biosolids. These cultures were used to evaluate aerobic transformation of weathered toxaphene. Only the biosolids culture could transform compounds of technical toxaphene (CTTs) in the presence of camphor, while no transformation was observed in the presence of glucose or with toxaphene as a sole carbon source. The transformed toxaphene had lower concentration of CTTs with longer retention times, and higher concentration of compounds with lower retention times. Gas chromatography with electron capture negative ion mass spectrometry (GC/ECNI-MS) showed that aerobic biotransformation mainly occurred with Cl 8 - and Cl 9 -CTTs compounds. The patterns of Cl 6 - and Cl 7 -CTTs were also simplified albeit to a much lesser extent. Seven camphor-degrading bacteria were isolated from the enrichment culture but none of them could degrade toxaphene., Conclusion: Camphor degrading culture can aerobically transform CCTs via reductive pathway probably by co-metabolism using camphor as a co-substrate., Significance and Impact of the Study: Since camphor is naturally produced by different plants, this study suggests that stimulation of aerobic transformation of toxaphene may occur in nature. Moreover plants, which produce camphor or similar compounds, might be used in bioremediation of contaminated soils., (© 2020 The Society for Applied Microbiology.)

Halogenated natural products number in the thousands, but only in rare cases are the evolutionary advantages conferred by the halogens understood. We set out to investigate the lissoclimide family of cytotoxins, which includes several chlorinated members, because of our long-standing interest in the synthesis of chlorinated secondary metabolites.Our initial success in this endeavor was a semisynthesis of chlorolissoclimide (CL) from the commercially available sesquiterpenoid sclareolide. Featuring a highly selective and efficient-and plausibly biomimetic-C-H chlorination, we were able to access enough CL for collaborative studies, including X-ray cocrystallography with the eukaryotic ribosome. Through this experiment, we learned that CL's chlorine atom engages in a novel halogen-π dispersion interaction with a neighboring nucleobase in the ribosome E-site.Owing to the limitations of our semisynthesis approach, we established an analogue-oriented approach to access numerous lissoclimide compounds to both improve our understanding of structure-activity relationships and to learn more about the halogen-π interaction. In the course of these studies, we made over a dozen lissoclimide-like compounds, the most interesting of which contained chlorine-bearing carbons with unnatural configurations. Rationalizing the retained potency of these compounds that appeared to be a poor fit for the lissoclimide binding pocket, we came to realize that the chlorine atoms would engage in these same halogen-π interactions even at the expense of a chair to twist-boat conformational change, which also permitted the compounds to fit in the binding site.Finally, because neither of the first two approaches could easily access the most potent natural lissoclimides, we designed a synthesis that took advantage of rarely used terminal epoxides to initiate polyene cyclizations. In this case, the chlorine atom was incorporated early and helped control the stereochemical outcome of the key step.Over the course of this project, three different synthesis approaches were designed and executed, and our ability to access numerous lissoclimides fueled a range of collaborative biological studies. Further, chlorine played impactful roles throughout various aspects of both synthesis and biology. We remain inspired to learn more about the mechanism of action of these compounds and to deeply investigate the potentially valuable halogen-π dispersion interaction in the context of small molecule/nucleic acid binding. In that context, our work offers an instance wherein we might have gained a rudimentary understanding of the evolutionary importance of the halogen in a halogenated natural product.

Aim: To explore the different consequences of acute and chronic exposure to chlorine gas (Cl 2 ) on the functional and histological parameters of health mice., Main Methods: Firstly, male BALB/c mice were acute exposed to 3.3 or 33.3 or 70.5 mg/m 3 Cl 2 . We analyzed the lung function, the inflammatory cells in the bronchoalveolar lavage, cell influx in the peribrochoalveolar space and mucus production. In a second phase, mice were chronic exposed to 70.5 mg/m 3 Cl 2 . Besides the first phase analyses, we also evaluated the epithelial cells thickness, collagen deposition in the airways, immunohistochemistry stain for IL-1β, iNOS, IL-17 and ROCK-2 and the levels of IL-5, IL-13, IL-17, IL-1β and TNF-α in lung homogenate., Key Findings: Acute exposure to chlorine impaired the lung function, increased the number of inflammatory cells in the BALF and in the airways, also increased the mucus production. Furthermore, when chlorine was exposed chronically, increased the airway remodeling with collagen deposition and epithelial cells thickness, positive cells for IL-1β, iNOS, IL-17 in the airways and in the alveolar walls and ROCK-2 in the alveolar walls, lung inflammation with increased levels of IL-5, IL-13, IL-1β and TNF-α in the lung homogenate, and also, induced the acid mucus production by the nasal epithelium., Significance: Acute and chronic exposure to low dose of chlorine gas worsens lung function, induces oxidative stress activation and mucus production and contributes to augmenting inflammation in health mice., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Recent work revealed an active biological chlorine cycle in coastal Arctic tundra of northern Alaska. This raised the question of whether chlorine cycling was restricted to coastal areas or if these processes extended to inland tundra. The anaerobic process of organohalide respiration, carried out by specialized bacteria like Dehalococcoides , consumes hydrogen gas and acetate using halogenated organic compounds as terminal electron acceptors, potentially competing with methanogens that produce the greenhouse gas methane. We measured microbial community composition and soil chemistry along an ∼262-km coastal-inland transect to test for the potential of organohalide respiration across the Arctic Coastal Plain and studied the microbial community associated with Dehalococcoides to explore the ecology of this group and its potential to impact C cycling in the Arctic. Concentrations of brominated organic compounds declined sharply with distance from the coast, but the decrease in organic chlorine pools was more subtle. The relative abundances of Dehalococcoides were similar across the transect, except for being lower at the most inland site. Dehalococcoides correlated with other strictly anaerobic genera, plus some facultative ones, that had the genetic potential to provide essential resources (hydrogen, acetate, corrinoids, or organic chlorine). This community included iron reducers, sulfate reducers, syntrophic bacteria, acetogens, and methanogens, some of which might also compete with Dehalococcoides for hydrogen and acetate. Throughout the Arctic Coastal Plain, Dehalococcoides is associated with the dominant anaerobes that control fluxes of hydrogen, acetate, methane, and carbon dioxide. Depending on seasonal electron acceptor availability, organohalide-respiring bacteria could impact carbon cycling in Arctic wet tundra soils. IMPORTANCE Once considered relevant only in contaminated sites, it is now recognized that biological chlorine cycling is widespread in natural environments. However, linkages between chlorine cycling and other ecosystem processes are not well established. Species in the genus Dehalococcoides are highly specialized, using hydrogen, acetate, vitamin B 12 -like compounds, and organic chlorine produced by the surrounding community. We studied which neighbors might produce these essential resources for Dehalococcoides species. We found that Dehalococcoides species are ubiquitous across the Arctic Coastal Plain and are closely associated with a network of microbes that produce or consume hydrogen or acetate, including the most abundant anaerobic bacteria and methanogenic archaea. We also found organic chlorine and microbes that can produce these compounds throughout the study area. Therefore, Dehalococcoides could control the balance between carbon dioxide and methane (a more potent greenhouse gas) when suitable organic chlorine compounds are available to drive hydrogen and acetate uptake., (Copyright © 2021 American Society for Microbiology.)

Neutrophils generate hypochlorous acid (HOCl) and related reactive chlorine species as part of their defence against invading microorganisms. In isolation, bacteria respond to reactive chlorine species by upregulating responses that provide defence against oxidative challenge. Key questions are whether these responses are induced when bacteria are phagocytosed by neutrophils, and whether this provides them with a survival advantage. We investigated RclR, a transcriptional activator of the rclABC operon in Escherichia coli that has been shown to be specifically activated by reactive chlorine species. We first measured induction by individual reactive chlorine species, and showed that HOCl itself activates the response, as do chloramines (products of HOCl reacting with amines) provided they are cell permeable. Strong RclR activation was seen in E. coli following phagocytosis by neutrophils, beginning within 5 min and persisting for 40 min. RclR activation was suppressed by inhibitors of NOX2 and myeloperoxidase, providing strong evidence that it was due to HOCl production in the phagosome. RclR activation demonstrates that HOCl, or a derived chloramine, enters phagocytosed bacteria in sufficient amount to induce this response. Although RclR was induced in wild-type bacteria following phagocytosis, we detected no greater sensitivity to neutrophil killing of mutants lacking genes in the rclABC operon., (© The Author(s) 2020. Published by Oxford University Press on behalf of FEMS.)

Novel bacterial type II topoisomerase inhibitors (NBTIs) stabilize single-strand DNA cleavage breaks by DNA gyrase but their exact mechanism of action has remained hypothetical until now. We have designed a small library of NBTIs with an improved DNA gyrase-binding moiety resulting in low nanomolar inhibition and very potent antibacterial activity. They stabilize single-stranded cleavage complexes and, importantly, we have obtained the crystal structure where an NBTI binds gyrase-DNA in a single conformation lacking apparent static disorder. This directly proves the previously postulated NBTI mechanism of action and shows that they stabilize single-strand cleavage through asymmetric intercalation with a shift of the scissile phosphate. This crystal stucture shows that the chlorine forms a halogen bond with the backbone carbonyls of the two symmetry-related Ala68 residues. To the best of our knowledge, such a so-called symmetrical bifurcated halogen bond has not been identified in a biological system until now.

Objectives: To investigate the role of sodium in intravesical absorption of water in the bladder and the sodium pathway in the urothelium., Methods: Adult female Sprague-Dawley rats received either saline or a 5% glucose solution injection into their bladders. The changes in intravesical fluid volume; concentrations of sodium and chlorine and osmolality; and expression of aquaporin-2, epithelial sodium channel, and claudins were compared after 3 hours., Results: Intravesical volume decreased significantly in the saline group compared to that in the 5% glucose solution group. The expression of claudin-3 and -6 was higher in the saline group than in the glucose group. There was a significant correlation between changes in the intravesical saline volume and the concentration of sodium and chlorine. Intravesical administration of amiloride did not affect changes in the fluid volume and concentration of sodium., Conclusions: The presence of sodium is important for the absorption of intravesical fluid through aquaporin-2 in the urinary bladders of rats. Claudin-3 and -6 may be associated with the transport of sodium through the bladder urothelium., (© 2020 John Wiley & Sons Australia, Ltd.)

This study tested the hypothesis that exposure to chlorine-sterilised pool water would impair oral nitrate reduction (ONR). ONR was assessed in elite swimmers before and after morning and afternoon pool-based training. Nonswimmers were only assessed in the morning. ONR was similar in swimmers and nonswimmers ( P  = 1.000) and unchanged before and after morning and afternoon training ( P  ≥ 0.341). Therefore, exposure to chlorinated pool water does not interfere with ONR. Novelty Exposure to chlorine-sterilised pool water does not impair oral nitrate reduction in elite swimmers.

The measurement of the potassium concentration (K + ) in vitreous humor (VH) has been a well-known adjunct for the estimation of the post-mortem interval (PMI) since the early 1960s. For years, however, many authors have been using other biochemical markers in an attempt to improve predictions. In this paper we confirm the role of K + in the determination of the PMI adopting a linear regression model and we investigate whether other biochemical markers could improve the model through a multiple regression analysis. Additionally, the research aims to confirm the data of the analytes of interest among different techniques and instrumentations. We deemed this as an important issue because a primary concern in the literature is that automated analytical methods are often calibrated and, for the most part, validated for serum or urine analysis. Our results confirmed the well-established role of K + as well as highlighted Albumin as a novel marker to be considered for further improvement of prediction models, especially since 72 h after death., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)