Comparative impact of pristine and aged microplastics with triclosan on lipid metabolism in larval zebrafish: Unveiling the regulatory role of miR-217.

The prevalence of microplastics (MPs), especially aged particles, interacting with contaminants like triclosan (TCS), raises concerns about their toxicological effects on aquatic life. This study focused on the impact of aged polyamide (APA) MPs and TCS on zebrafish lipid metabolism. APA MPs, with rougher surfaces and lower hydrophobicity, exhibited reduced TCS adsorption than unaged polyamide (PA) MPs. Co-exposure to PA/APA MPs and TCS resulted in higher TCS accumulation in zebrafish larvae, notably more with PA than APA. Larvae exposed to PA + TCS exhibited greater oxidative stress, disrupted lipid metabolism, and altered insulin pathway genes than those exposed to TCS. However, these negative effects were lessened in the APA + TCS group. Through miRNA-seq and miR-217 microinjection, it was revealed that PA + TCS co-exposure upregulated miR-217, linked to lipid metabolic disorders in zebrafish. Moreover, molecular docking showed stable interactions formed between PA, TCS, and the insulin signaling protein Pik3r2. This study demonstrated that PA and TCS co-exposure significantly inhibited the insulin signaling in zebrafish, triggering lipid metabolism dysregulation mediated by miR-217 upregulation, while APA and TCS co-exposure alleviated these disruptions. This research underscored the ecological and toxicological risks of aged MPs and pollutants in aquatic environments, providing crucial insights into the wider implications of MPs pollution.

Mpox virus Clade IIb infected Cynomolgus macaques via mimic natural infection routes closely resembled human mpox infection.

Generating an infectious non-human primate (NHP) model using a prevalent monkeypox virus (MPXV) strain has emerged as a crucial strategy for assessing the efficacy of vaccines and antiviral drugs against human MPXV infection. Here, we established an animal model by infecting cynomolgus macaques with the prevalent MPXV strain, WIBP-MPXV-001, and simulating its natural routes of infection. A comprehensive analysis and evaluation were conducted on three animals, including monitoring clinical symptoms, collecting hematology data, measuring viral loads, evaluating cellular and humoral immune responses, and examining histopathology. Our findings revealed that initial skin lesions appeared at the inoculation sites and subsequently spread to the limbs and back, and all infected animals exhibited bilateral inguinal lymphadenopathy, eventually leading to a self-limiting disease course. Viral DNA was detected in post-infection blood, nasal, throat, rectal and blister fluid swabs. These observations indicate that the NHP model accurately reflects critical clinical features observed in human MPXV infection. Notably, the animals displayed clinical symptoms and disease progression similar to those of humans, rather than a lethal outcome as observed in previous studies. Historically, MPXV was utilized as a surrogate model for smallpox. However, our study contributes to a better understanding of the dynamics of current MPXV infections while providing a potential infectious NHP model for further evaluation of vaccines and antiviral drugs against mpox infection. Furthermore, the challenge model closely mimics the primary natural routes of transmission for human MPXV infections. This approach enhances our understanding of the precise mechanisms underlying the interhuman transmission of MPXV.

Toxic effects of triclosan on hepatic and intestinal lipid accumulation in zebrafish via regulation of m6A-RNA methylation.

Triclosan (TCS), recognized as an endocrine disruptor, has raised significant concerns due to its widespread use and potential health risks. To explore the impact of TCS on lipid metabolism, both larval and adult zebrafish were subjected to acute and chronic exposure to TCS. Through analyzes of biochemical and physiological markers, as well as Oil Red O (ORO) and hematoxylin and eosin (H&E) staining, our investigation revealed that TCS exposure induced hepatic and intestinal lipid accumulation in larval and adult zebrafish, leading to structural damage and inflammatory responses in these tissues. The strong affinity of TCS with PPARγ and subsequent pathway activation indicate that PPARγ pathway plays a crucial role in TCS-induced lipid buildup. Furthermore, we observed a decrease in m6A-RNA methylation levels in the TCS-treated group, which attributed to the increased activity of the demethylase FTO and concurrent suppression of the methyltransferase METTL3 gene expression by TCS. The alteration in methylation dynamics is identified as a potential underlying mechanism behind TCS-induced lipid accumulation. To address this concern, we explored the impact of folic acid-a methyl donor for m6A-RNA methylation-on lipid accumulation in zebrafish. Remarkably, folic acid administration partially alleviated lipid accumulation by restoring m6A-RNA methylation. This restoration, in turn, contributed to a reduction in inflammatory damage observed in both the liver and intestines. Additionally, folic acid partially mitigates the up-regulation of PPARγ and related genes induced by TCS. These findings carry substantial implications for understanding the adverse effects of environmental pollutants such as TCS. They also emphasize the promising potential of folic acid as a therapeutic intervention to alleviate disturbances in lipid metabolism induced by environmental pollutants.

Chlorpromazine inhibits EAG1 channels by altering the coupling between the PAS, CNBH and pore domains.

EAG1 depolarization-activated potassium selective channels are important targets for treatment of cancer and neurological disorders. EAG1 channels are formed by a tetrameric subunit assembly with each subunit containing an N-terminal Per-Arnt-Sim (PAS) domain and C-terminal cyclic nucleotide-binding homology (CNBH) domain. The PAS and CNBH domains from adjacent subunits interact and form an intracellular tetrameric ring that regulates the EAG1 channel gating, including the movement of the voltage sensor domain (VSD) from closed to open states. Small molecule ligands can inhibit EAG1 channels by binding to their PAS domains. However, the allosteric pathways of this inhibition are not known. Here we show that chlorpromazine, a PAS domain small molecule binder, alters interactions between the PAS and CNBH domains and decreases the coupling between the intracellular tetrameric ring and the pore of the channel, while having little effect on the coupling between the PAS and VSD domains. In addition, chlorpromazine binding to the PAS domain did not alter Cole-Moore shift characteristic of EAG1 channels, further indicating that chlorpromazine has no effect on VSD movement from the deep closed to opened states. Our study provides a framework for understanding global pathways of EAG1 channel regulation by small molecule PAS domain binders.

Acute/chronic triclosan exposure induces downregulation of m6A-RNA methylation modification via mettl3 suppression and elicits developmental and immune toxicity to zebrafish.

Triclosan (TCS), a prevalent contaminant in aquatic ecosystems, has been identified as a potential threat to both aquatic biota and human health. Despite its widespread presence, research into the immunotoxic effects of TCS on aquatic organisms is limited, and the underlying mechanisms driving these effects remain largely unexplored. Herein, we investigated the developmental and immune toxicities of environmentally relevant concentrations of TCS in zebrafish, characterized by morphological anomalies, histopathological impairments, and fluctuations in cytological differentiation and biomarkers following both acute (from 6 to 72/120 hpf) and chronic exposure periods (from 30 to 100 dpf). Specifically, acute exposure to TCS resulted in a significant increase in innate immune cells, contrasted by a marked decrease in T cells. Furthermore, we observed that TCS exposure elicited oxidative stress and a reduction in global m6A levels, alongside abnormal expressions within the m6A modification enzyme system in zebrafish larvae. Molecular docking studies suggested that mettl3 might be a target molecule for TCS interaction. Intriguingly, the knock-down of mettl3 mirrored the effects of TCS exposure, adversely impacting the growth and development of zebrafish, as well as the differentiation of innate immune cells. These results provide insights into the molecular basis of TCS-induced immunotoxicity through m6A-RNA epigenetic modification and aid in assessing its ecological risks, informing strategies for disease prevention linked to environmental contaminants.

Pest recognition in microstates state: an improvement of YOLOv7 based on Spatial and Channel Reconstruction Convolution for feature redundancy and vision transformer with Bi-Level Routing Attention.

Introduction: In order to solve the problem of precise identification and counting of tea pests, this study has proposed a novel tea pest identification method based on improved YOLOv7 network. Methods: This method used MPDIoU to optimize the original loss function, which improved the convergence speed of the model and simplifies the calculation process. Replace part of the network structure of the original model using Spatial and Channel reconstruction Convolution to reduce redundant features, lower the complexity of the model, and reduce computational costs. The Vision Transformer with Bi-Level Routing Attention has been incorporated to enhance the flexibility of model calculation allocation and content perception. Results: The experimental results revealed that the enhanced YOLOv7 model significantly boosted Precision, Recall, F1, and mAP by 5.68%, 5.14%, 5.41%, and 2.58% respectively, compared to the original YOLOv7. Furthermore, when compared to deep learning networks such as SSD, Faster Region-based Convolutional Neural Network (RCNN), and the original YOLOv7, this method proves to be superior while being externally validated. It exhibited a noticeable improvement in the FPS rates, with increments of 5.75 HZ, 34.42 HZ, and 25.44 HZ respectively. Moreover, the mAP for actual detection experiences significant enhancements, with respective increases of 2.49%, 12.26%, and 7.26%. Additionally, the parameter size is reduced by 1.39 G relative to the original model. Discussion: The improved model can not only identify and count tea pests efficiently and accurately, but also has the characteristics of high recognition rate, low parameters and high detection speed. It is of great significance to achieve realize the intelligent and precise prevention and control of tea pests.

A screening study on the detection strain of Coxsackievirus A6: the key to evaluating neutralizing antibodies in vaccines.

The increasing incidence of diseases caused by Coxsackievirus A6 (CV-A6) and the presence of various mutants in the population present significant public health challenges. Given the concurrent development of multiple vaccines in China, it is challenging to objectively and accurately evaluate the level of neutralizing antibody response to different vaccines. The choice of the detection strain is a crucial factor that influences the detection of neutralizing antibodies. In this study, the National Institutes for Food and Drug Control collected a prototype strain (Gdula), one subgenotype D1, as well as 13 CV-A6 candidate vaccine strains and candidate detection strains (subgenotype D3) from various institutions and manufacturers involved in research and development. We evaluated cross-neutralization activity using plasma from naturally infected adults (n = 30) and serum from rats immunized with the aforementioned CV-A6 strains. Although there were differences between the geometric mean titer (GMT) ranges of human plasma and murine sera, the overall trends were similar. A significant effect of each strain on the neutralizing antibody test (MAX/MIN 48.0 ∼16410.3) was observed. Among all strains, neutralization of the S112 strain by 15 different sera resulted in higher neutralizing antibody titers (GMTS112 = 132.0) and more consistent responses across different genotypic immune sera (MAX/MIN = 48.0). Therefore, S112 may serve as a detection strain for NtAb testing in various vaccines, minimizing bias and making it suitable for evaluating the immunogenicity of the CV-A6 vaccine.

Integrated assessment of the pollution and risk of heavy metals in soils near chemical industry parks along the middle Yangtze River.

Industrialization in riparian areas of critical rivers has caused significant environmental and health impacts. Taking eight industrial parks along the middle Yangtze River as examples, this study proposes a multiple-criteria approach to investigate soil heavy metal pollution and associated ecological and health risks posed by industrial activities. Aiming at seven heavy metals, the results show that nickel (Ni), cadmium (Cd), and copper (Cu) exhibited the most significant accumulation above background levels. The comprehensive findings from Pearson correlation analysis, cluster analysis, principal component analysis, and industrial investigation uncover the primary sources of Cd, arsenic (As), mercury (Hg), and lead (Pb) to be chemical processing, while Ni and chromium (Cr) are predominantly derived from mechanical and electrical equipment manufacturing. In contrast, Cu exhibits a broad range of origins across various industrial processes. Soil heavy metals can cause serious ecological and carcinogenic health risks, of which Cd and Hg contribute to >70 % of the total ecological risk, and As contributes over 80 % of the total health risk. This study highlights the importance of employing multiple mathematical and statistical models in determining and evaluating environmental hazards, and may aid in planning the environmental remediation engineering and optimizing the industry standards.

Neurotoxic effects of 2-ethylhexyl diphenyl phosphate exposure on zebrafish larvae: Insight into inflammation-driven changes in early motor behavior.

The extensive utilization and potential adverse impacts of the replacement flame-retardant 2-Ethylhexyl Diphenyl Phosphate (EHDPP) have raised concerns. Currently, there is limited knowledge regarding the developmental, neurological, and immunotoxic consequences of EHDPP exposure, as well as its potential behavioral outcomes. In this study, we undertook a comprehensive examination and characterization of the toxic effects over the EHDPP concentration range of 14-1400 nM. Our findings unveiled that EHDPP, even at an environmentally relevant concentration of 14 nM, exhibited excitatory neurotoxicity, eliciting a 13.5 % increase in the swimming speed of zebrafish larvae. This effect might be attributed to the potential influence of EHDPP on the release of neurotransmitters like serotonin and dopamine, which, in turn, mediated anxiety-like behavior in the zebrafish larvae. Conversely, sublethal dose EHDPP (1400 nM) exposure significantly suppressed the swimming vigor of zebrafish larvae, accompanied by morphological changes, abnormal behaviors, and alterations in intracerebral molecules. Transcriptomics revealed the underlying mechanism. The utilization of pathway inhibitors reshaped the inflammatory homeostasis and alleviated the toxicity induced by EHDPP exposure, anchoring the pivotal role played by the TLR4/NF-κB signaling pathway in EHDPP-induced adverse changes in zebrafish behavior and neurophysiology. This study observed the detrimental effects of EHDPP on fish sustainability at environmentally relevant concentrations, highlighting the practical significance for EHDPP risk management. Elucidating the toxic mechanisms of EHDPP will contribute to a deeper comprehension of how environmental pollutants can intricately influence human health.

Evaluation of Stability, Inactivation, and Disinfection Effectiveness of Mpox Virus.

BACKGROUND: Mpox virus (MPXV) infections have increased in many countries since May 2022, increasing demand for diagnostic tests and research on the virus. To ensure personnel safety, appropriate and reliable measures are needed to disinfect and inactivate infectious samples; Methods: We evaluated the stability of infectious MPXV cultures stored at different temperatures and through freeze-thaw cycles. Heat physical treatment (56 °C, 70 °C, 95 °C), chemical treatment (beta-propiolactone (BPL)) and two commercialized disinfectants (Micro-Chem Plus (MCP) and ethanol) were tested against infectious MPXV cultures; Results: The results indicated that MPXV stability increases with lower temperatures. The MPXV titer was stable within three freeze-thaw cycles and only decreased by 1.04 log10 (lg) 50% cell culture infective dose (CCID50) per milliliter (12.44%) after twelve cycles. MPXV could be effectively inactivated at 56 °C for 40 min, 70 °C for 10 min, and 95 °C for 5 min. For BPL inactivation, a 1:1000 volume ratio (BPL:virus) could also effectively inactivate MPXV. A total of 2% or 5% MCP and 75% ethanol treated with MPXV for at least 1 min could reduce >4.25 lg; Conclusions: MPXV shows high stability to temperature and freeze-thaw. Heat and BPL treatments are effective for the inactivation of MPXV, while MCP and ethanol are effective for disinfection, which could help laboratory staff operate the MPXV under safer conditions and improve operational protocols.

Cerebral Lactate Participates in Hypoxia-induced Anapyrexia Through its Receptor G Protein-coupled Receptor 81.

Hypoxia-induced anapyrexia is thought to be a regulated decrease in body core temperature (Tcore), but the underlying mechanism remains unclear. Recent evidence suggests that lactate, a glycolysis product, could modulate neuronal excitability through the G protein-coupled receptor 81 (GPR81). The present study aims to elucidate the role of central lactate and GPR81 in a rat model of hypoxia-induced anapyrexia. The findings revealed that hypoxia (11.1% O2, 2 h) led to an increase in lactate in cerebrospinal fluid (CSF) and a decrease in Tcore. Injection of dichloroacetate (DCA, 5 mg/kg, 1 µL), a lactate production inhibitor, to the third ventricle (3 V), alleviated the increase in CSF lactate and the decrease in Tcore under hypoxia. Immunofluorescence staining showed GPR81 was expressed in the preoptic area of hypothalamus (PO/AH), the physiological thermoregulation integration center. Under normoxia, injection of GPR81 agonist 3-chloro-5-hydroxybenzoic acid (CHBA, 0.05 mg/kg, 1 µL) to the 3 V, reduced Tcore significantly. In addition, hypoxia led to a dramatic increase in tail skin temperature and a decrease in interscapular brown adipose tissue skin temperature. The number of c-Fos+ cells in the PO/AH increased after exposure to 11.1% O2 for 2 h, but administration of DCA to the 3 V blunted this response. Injection of CHBA to the 3 V also increased the number of c-Fos+ cells in the PO/AH under normoxia. In light of these, our research has uncovered the pivotal role of central lactate-GPR81 signaling in anapyrexia, thereby providing novel insights into the mechanism of hypoxia-induced anapyrexia.

The sulfur microbial diet and risk of nonalcoholic fatty liver disease: a prospective gene-diet study from the UK Biobank.

BACKGROUND: The gut microbiota is closely related to liver diseases. The dietary pattern associated with sulfur-metabolizing bacteria in stool has been found to influence intestinal health. OBJECTIVE: We aimed to investigate whether consuming the sulfur microbial diet is associated with nonalcoholic fatty liver disease (NAFLD). METHODS: We included 143,918 participants of European descent from the UK Biobank. Information on serving sizes used per diet component was recorded by an online 24-h dietary assessment tool (Oxford WebQ). The total sulfur microbial diet score was constructed by summing the product of ß-coefficients and corresponding serving sizes. NAFLD was ascertained using hospital inpatient and death records. Cox proportional hazard models were used to estimate the adjusted hazard ratio (HR) and 95% confidence interval (CI). Mediation analyses were used to investigate underlying mediators including body mass index, waist circumference, glucose, triglyceride, urate, and C-reactive protein. A polygenic risk score for NAFLD was constructed and stratified to assess whether the association is modified by genetic predisposition. RESULTS: After a median follow-up of 11.7 y (interquartile range: 11.3-12.5 y), we documented 1540 incident cases of NAFLD. After adjustment for covariates, we observed an overall J-shaped relationship between the sulfur microbial diet and risk of NAFLD. Those in the highest quartile of sulfur microbial diet score had a 46% increased risk of NAFLD [HRQ4vsQ1 (95% CI): 1.46 (1.26, 1.69)]. We also found that this association is partly mediated by metabolic disorders and systemic inflammation. In addition, the positive association was stronger among individuals at higher genetic risk for NAFLD (Pinteraction = 0.044). CONCLUSIONS: The sulfur microbial diet had adverse associations with incident NAFLD, particularly in those at a higher genetic risk. Our study may provide evidence on the role of sulfur-metabolizing bacteria in the diet-NAFLD association.

Normalizing Flow-Based Distribution Estimation of Pharmacokinetic Parameters in Dynamic Contrast-Enhanced Magnetic Resonance Imaging.

OBJECTIVE: The pharmacokinetic (PK) parameters estimated from dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) provide valuable information for clinical research and diagnosis. However, these estimated PK parameters suffer from many sources of variability. Thus, the estimation of the posterior distributions of these PK parameters could provide a way to simultaneously quantify the values and uncertainties of the PK parameters. Our objective is to develop an efficient and flexible method to more closely approximate and estimate the underlying posterior distributions of the PK parameters. METHODS: The normalizing flow model-based parameters distribution estimation neural network (FPDEN) is proposed to adaptively learn and estimate the posterior distributions of the PK parameters. The maximum likelihood estimation (MLE) loss is directly constructed based on the parameter distributions learned by the normalizing flow model, rather than pre-defined distributions. RESULTS: Experimental analysis shows that the proposed method can improve parameter estimation accuracy. Moreover, the uncertainty derived from the parameter distribution constitutes an effective indicator to exclude unreliable parametric results. A successful demonstration is the improved classification performance of the glioma World Health Organization (WHO) grading task, specifically in terms of distinguishing between low and high grades, as well as between Grade III and Grade IV. CONCLUSION: The FPDEN method offers improved accuracy for estimation of PK parameters and boosts the performance of the glioma grading task. SIGNIFICANCE: By enhancing the precision and reliability of DCE-MRI, the proposed method promotes its further applications in clinical practice.

Interference of gut-brain-gonad axis originating from triclocarban exposure to parent zebrafish induces offspring embryonic development abnormality by up-regulation of maternal circSGOL1.

Triclocarban (TCC) is a widely used antibacterial ingredient possessing acute toxicity effects; however, its chronic toxicity and underlying molecular mechanisms remain uncertain. Herein, we demonstrated that chronic TCC exposure affects the growth and development of adult zebrafish through inducing an intestinal flora disorder in the gut. The imbalance of intestinal flora caused functional barriers within the intestinal-brain-gonadal axis. This resulted in a series of anomalous nerve and motor behaviors, and reproductive toxicity as reflected in pathological damage to parental gonads and F1-larval developmental malformations. Abnormal development of F1 larvae was attributed to apoptosis induced by the up-regulation of circSGOL1. This up-regulation affected the activity and localization of the hnRNP A1 protein, which then promoted overexpression of pro-apoptotic related genes that ultimately lead to apoptosis during early embryonic development. Overall, these novel findings systematically elucidated the TCC toxicity mechanism in parent-offspring dyads, and provide important theoretical guidance for early risk warning and control of chronic TCC toxicity.

Deciphering the molecular mediators of triclosan-induced lipid accumulation: Intervention via short-chain fatty acids and miR-101a.

As a potential environmental obesogen, triclosan (TCS) carries inherent risks of inducing obesity and metabolic disorders. However, the underlying molecular mechanisms behind the lipid metabolism disorder induced by TCS have remained elusive. Through a fusion of transcriptomics and microRNA target prediction, we hypothesize that miR-101a as a responsive miRNA to TCS exposure in zebrafish, playing a central role in disturbing lipid homeostasis. As an evidence, TCS exposure triggers a reduction in miR-10a expression that accompanied by elevation of genes linked to regulation of lipid homeostasis. Through precision-controlled interventions involving miRNA expression modulation, we discovered that inhibition of miR-101a enhanced expression of its target genes implicated in lipid homeostasis, subsequently triggering excessive fat accumulation. Meanwhile, the overexpression of miR-101a acts as a protective mechanism, counteracting the lipid metabolism disorder induced by TCS in the larvae. Notably, the combination of short-chain fatty acids (SCFAs) emerged as a potential remedy to alleviate TCS-induced lipid accumulation partially by counteracting the decline in miR-101a expression induced by TCS. These revelations provide insight into a prospective molecular framework underlying TCS-triggered lipid metabolism disorders, thereby paving the way for pre-emptive strategies in combating the ramifications of TCS pollution.

Virological Characteristics of Five SARS-CoV-2 Variants, Including Beta, Delta and Omicron BA.1, BA.2, BA.5.

SARS-CoV-2 variants of concern (VOCs) show increasing transmissibility and infectivity and induce substantial injuries to human health and the ecology. Therefore, it is vital to understand the related features for controlling infection. In this study, SARS-CoV-2 WIV04 (prototype) and five VOCs (Beta, Delta, Omicron BA.1, BA.2 and BA.5 variants) were inoculated in Vero cells to observe their growth activities. Apart from evaluating the environmental stability at different temperatures, residual virus titers and infectivity at different temperatures (4 °C, room temperature (RT) and 37 °C) were measured over 7 days. The experiment also assessed the infectivity for different incubation durations. The growth capacity assay suggested that the WIV04, Beta and Delta variants replicated efficiently in Vero cells compared with Omicron Variants, and BA.2 replicated more efficiently in Vero cells than BA.1 and BA.5. In addition, all variants exhibited longer survivals at 4 °C and could remain infectious after 7 days, compared to RT' survival after 5 days and at 37 °C after 1 day. The virus infection assay indicated that the Omicron variant had a weaker ability to infect cells compared to the WIV04, Beta and Delta strains, and a longer infection time was required for these strains, except for BA.2.

Human monoclonal antibody F61 nasal spray effectively protected high-risk populations from SARS-CoV-2 variants during the COVID-19 pandemic from late 2022 to early 2023 in China.

Following the national dynamic zero-COVID strategy adjustment, the utilization of broad-spectrum nasal neutralizing antibodies may offer alternative approach to controlling the outbreak of Omicron variants between late 2022 and early 2023 in China. This study involved an investigator-initiated trial (IIT) to assess the pharmacokinetic, safety and efficacy of the F61 nasal spray. A total of 2,008 participants were randomly assigned to receive F61 nasal spray (24 mg/0.8 mL/dose) or normal saline (0.8 mL/dose) and 1,336 completed the follow-up in the IIT. Minimal absorption of F61 antibody into the bloodstream was detected in individuals receiving F61 nasal spray for seven consecutive days. No treatment-emergent adverse reactions of grade 3 severity or higher were reported. In the one-dose cohort, the 7-day cumulative SARS-CoV-2 infection rate was 79.0% in the F61 group and 82.6% in the placebo group, whereas, in the multiple-dose (once daily for 7 consecutive days) cohort, the rates were 6.55% in the F61 group and 23.83% in the placebo group. The laboratory-confirmed efficacy of F61 was 3.78% (-3.74%-10.75%) in the one-dose cohort and 72.19% (57.33%-81.87%) in the multiple-dose cohort. In the real-world study, 60,225 volunteers in four different regions were administered the F61 nasal spray based on subject's wishes, over 90% efficacy rate was observed against different Omicron variants. The F61 nasal spray, with its favorable safety profile, could be a promising prophylactic monoclonal antibody against SARS-CoV-2 VOCs.

Molecular mechanism of EAG1 channel inhibition by imipramine binding to the PAS domain.

Ether-a-go-go (EAG) channels are key regulators of neuronal excitability and tumorigenesis. EAG channels contain an N-terminal Per-Arnt-Sim (PAS) domain that can regulate currents from EAG channels by binding small molecules. The molecular mechanism of this regulation is not clear. Using surface plasmon resonance and electrophysiology we show that a small molecule ligand imipramine can bind to the PAS domain of EAG1 channels and inhibit EAG1 currents via this binding. We further used a combination of molecular dynamics (MD) simulations, electrophysiology, and mutagenesis to investigate the molecular mechanism of EAG1 current inhibition by imipramine binding to the PAS domain. We found that Tyr71, located at the entrance to the PAS domain cavity, serves as a "gatekeeper" limiting access of imipramine to the cavity. MD simulations indicate that the hydrophobic electrostatic profile of the cavity facilitates imipramine binding and in silico mutations of hydrophobic cavity-lining residues to negatively charged glutamates decreased imipramine binding. Probing the PAS domain cavity-lining residues with site-directed mutagenesis, guided by MD simulations, identified D39 and R84 as residues essential for the EAG1 channel inhibition by imipramine binding to the PAS domain. Taken together, our study identified specific residues in the PAS domain that could increase or decrease EAG1 current inhibition by imipramine binding to the PAS domain. These findings should further the understanding of molecular mechanisms of EAG1 channel regulation by ligands and facilitate the development of therapeutic agents targeting these channels.

Pore-scale investigation of surfactant-enhanced DNAPL mobilization and solubilization.

Surfactant-enhanced aquifer remediation has been proved successful to remove dense non-aqueous phase liquids (DNAPLs) from contaminated sites. However, the underlying mechanisms of the DNAPL mobilization and solubilization at the pore scale remains to be addressed for efficient application to the field remediation system. In this work, the emerging microfluidic and imaging technologies are applied to investigate the dynamics of DNAPL remediation. Visualized experiments of the evolution of DNAPL remediation are performed to study the role of surfactant type, concentration and injection rate. The DNAPL remediation is dominated by mobilization followed by solubilization for most surfactants. Mobilization occurs as soon as surfactants and DNAPL are in contact until forming a new stable phase structure, and the solubilization continues until the end of injection. We observe the breakup behavior of long droplets and ganglia during the mobilization, which is attributed to the surfactant-reduced interfacial tension and thus expedites DNAPL mobilization and redistribution. During the solubilization, the formation of micelles incorporating DNAPL fractions increases the DNAPL concentration gradient and thus enhances the mass transfer, but the rate-limited diffusion of micelles reduces the mass transfer rate coefficient. Increasing the surfactant content and decreasing the injection rate can promote mobilization and solubilization. The DNAPL mobilization ability of the surfactants SDS and SDBS is stronger than SAOS and Tween 80 regardless of the injection rates. Tween 80 may be considered an ideal surfactant of only solubilization but not mobilization is desired. This work elucidates the pore-scale mechanisms during surfactant-enhanced DNAPL remediation, which are beneficial for upscaling studies, predictive modeling, and operation optimization of DNAPL remediation in the field.

METTL3 exacerbates insulin resistance in hepatocytes by regulating m6A modification of cytochrome P450 2B6.

BACKGROUND: Insulin resistance (IR) in hepatocytes endangers human health, and frequently results in the development of non-alcoholic fatty liver disease (NAFLD). Research on m6A methylation of RNA molecules has gained popularity in recent years; however, the molecular mechanisms regulating the processes of m6A modification and IR are not known. The cytochrome P450 (CYP450) enzyme system, which is mainly found in the liver, is associated with the pathogenesis of NAFLD. However, few studies have been conducted on CYP450 related m6A methylation. Here, we investigated the role of the methyltransferase METTL3 in exacerbating IR in hepatocytes, mainly focusing on the regulation of m6A modifications in CYP2B6. METHODS AND RESULTS: Analysis using dot blot and epitranscriptomic chips revealed that the m6A modification pattern of the transcriptome in high-fat diet (HFD)-induced fatty liver and free fatty acid (FFA)-induced fatty hepatocytes showed significant changes. CYP450 family members, especially Cyp2b10, whose homolog in humans is CYP2B6, led to a noticeable increase in m6A levels in HFD-induced mice livers. Application of the METTL3 methyltransferase inhibitor, STM2457, increased the level of insulin sensitivity in hepatocytes. We then analyzed the role of METTL3 in regulating m6A modification of CYP2B6 in hepatocytes. METTL3 regulated the m6A modification of CYP2B6, and a positive correlation was found between the levels of CYP2B6 translation and m6A modifications. Furthermore, interference with METTL3 expression and exposure to STM2457 inhibited METTL3 activity, which in turn interfered with the phosphorylated insulin receptor substrate (pIRS)-glucose transporter 2 (GLUT2) insulin signaling pathway; overexpression of CYP2B6 hindered IRS phosphorylation and translocation of GLUT2 to membranes, which ultimately exacerbated IR. CONCLUSION: These findings offer unique insights into the role that METTL3-mediated m6A modifications of CYP2B6 play in regulating insulin sensitivity in hepatocytes and provide key information for the development of strategies to induce m6A modifications for the clinical treatment of NAFLD.