The integrity of the blood–brain barrier (BBB) is crucial for maintaining the function and homeostasis of the central nervous system (CNS), with astrocytes playing a key role in this process. Our study found that infection with the Japanese encephalitis virus (JEV) promoted the translocation of high-mobility group box 1 (HMGB1) from the nucleus to the extracellular space of astrocytes, a process directly associated with BBB disruption. Through bioinformatics analysis, we identified potential targets of encephalitis and constructed a protein–protein interaction (PPI) network. Subsequent functional enrichment analyses, including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, highlighted the calcium signaling pathway as an important regulatory mechanism. Evidence from our in vitro and in vivo model experiments showed that HMGB1 can induce the increase of calcium ions (Ca²+) in astrocytes, thereby activating the calcium signaling pathway and promoting the translocation of aquaporin-4 (AQP4) to the plasma membrane, ultimately leading to BBB disruption. We also performed molecular docking and molecular dynamics simulations to determine the binding affinity between trifluoperazine (TFP) and calmodulin (CaM). TFP binds to CaM and blocks the translocation of AQP4 to the plasma membrane, thereby alleviating HMGB1-mediated BBB disruption. Overall, our data indicate that TFP protects BBB integrity through the CaM–CaMKII–AQP4 axis and identifies this pathway as a promising therapeutic target for the clinical treatment of Japanese encephalitis and other central nervous system diseases.

African swine fever virus (ASFV), a highly lethal pathogen, poses a catastrophic threat to global pork production. While some commercial vaccines exist, their availability remains regional limitations. This review investigates the mechanistic interplay between ASFV and antigen-presenting cells (APCs), with a particular focus on viral evasion strategies that disrupt antigen presentation pathways. Although current understanding of ASFV immunology has largely focused on monocyte/macrophage infection, we provide a systematic analysis of ASFV-mediated immunosuppression across various APCs, including dendritic cells (DCs), B lymphocytes, and γδ T cells, and compare these mechanisms with those employed by other viral pathogens. Key objectives include: (1) synthesizing ASFV-induced dysregulation of antigen presentation, (2) identifying critical gaps in APC-virus interactions, and (3) guiding vaccine development priorities. By highlighting understudied DC-ASFV interplay and potential molecular targets, this work provides a strategic framework for next-generation vaccine design based on host-pathogen conflict at the antigen presentation interface.

African swine fever (ASF) is an acute and highly infectious disease caused by African swine fever virus (ASFV) that has dealt a massive blow to the development of the pig industry in China. Macrophages, the primary target cells of ASFV, exhibit high plasticity. However, their phenotypic changes during infection remain poorly understood. In this study, we observed a significant increase in M2 monocytes within the peripheral blood of ASFV-infected pigs. In vitro experiments demonstrated that ASFV drives macrophage polarization toward the M2 phenotype through early phosphorylation of STAT3. STAT3 inhibition with STATTIC not only blocked M2 polarization but also suppressed ASFV replication. While M2 macrophages do not impede viral attachment or internalization, they display reduced killing capacity compared to M1 macrophages. Furthermore, in a mixed lymphocyte reaction (MLR) system, CD4+ T cells cocultured with ASFV-infected M2-polarized macrophages presented suppressed early activation, marked by downregulated CD25 expression, ultimately impairing adaptive immunity. These findings reveal a critical immune evasion strategy employed by ASFV and provide key insights into ASF pathogenesis and viral persistence.

Alternative splicing (AS) regulates the diversity and level of the proteome. The specificity in AS is in turn regulated by RNA-binding proteins, but our understanding of how they act is far from complete. Here, we identify the Angel wing (Anw) protein, previously CG10948, as a novel AS regulator. Loss of Anw in Drosophila disrupts splicing in muscle genes and subsequently muscle function. Based on a mini-gene assay in which Anw and its RNA targets are co-expressed in cultured cells, we demonstrated orthologous splicing regulation of the mini-gene transcripts, interaction between Anw and its RNA targets, and a remarkable functional conservation among Anw homologs. Anw forms nuclear foci, and genetic ablation of Anw domains suggests that maintaining distinctive features of these foci is important for its function. The evolution of Anw is dynamic with gene gains and losses, but as others suggested, preserves a cross-phyla "ultra conserved element" as an alternative exon that potentially regulates its own level by nonsense-mediated mRNA decay. As the human anw homolog is a candidate gene for myasthenia gravis, our work suggests a mechanism for cellular dysfunction in this disease.

Sodium hypochlorite (SHC) is the most commonly utilized carcass and equipment disinfectant in the poultry industry. However, prolonged exposure to SHC can result in the development of bacterial tolerance and exert co-selection on antimicrobial resistance. This study investigated the co-resistance to SHC and multiple antimicrobial agents among Salmonella enterica serovar Indiana (S. Indiana), with a specific focus on the co-occurrence of disinfectant resistance gene qacEΔ1 and the antimicrobial resistance genes (ARGs) revealed by whole genome sequencing (WGS). Additionally, the study examined the transcriptional response of qacEΔ1 and its closely associated ARGs under SHC pressure. Moreover, the study determined the optimal SHC concentration for the decontamination of multidrug-resistant (MDR) S. Indiana on chicken. The results indicated that S. Indiana exhibited a resistance rate of 73.31 % to SHC, and varying levels of resistance to 13 antimicrobial agents. Furthermore, the analysis revealed a significant correlation between the qacEΔ1 gene and ARGs, including catB3, sul1, arr-3 and blaOXA-1. The genetic contexts surrounding the qacEΔ1 gene demonstrated a high degree of homology, allowing for the categorization into 11 distinct genetic context types, among which the gene cluster aacA4-blaOXA-1-catB3-arr-3-qacEΔ1-sul1 was the most prevalent. Further analysis of the MDR IndS97 strain using PacBio SMRT sequencing revealed that the qacEΔ1 gene was located on plasmid pLKQY01, with IS26 and ISRle7 positioned at the flanks of the composite transposon aacA4-blaOXA-1-catB3-arr-3-qacEΔ1-sul1. The transcription levels of qacEΔ1, arr-3 and sul1 genes in response to SHC stress initially increased, followed by a decline as SHC concentrations rose. At an SHC concentration of 0.5 MIC, the transcription levels of these genes were notably low, and the results indicated a decontamination efficacy of 86.51 % against Salmonella contamination while relatively preserving the freshness of the chicken. This study enhanced the understanding of disinfectant effects on the antimicrobial resistance of S. Indiana and provided evidence to support the regulated use of disinfectants.

Hydrogen peroxide (H2O2) is an important reactive oxygen species, and its abnormal concentration can cause cell distortion and apoptosis, leading to occurrence of various diseases. Therefore, real-time and sensitive monitoring of H2O2 is essential for early disease diagnosis and pathological research. Herein, a novel nanocomposite of Feporphyrin covalent organic frameworks decorated with molybdenum disulfide and multi-walled carbon nanotubes (Fe-pCOFs/MoS2-MWCNTs), which had glorious enzyme-like activity, was reported for efficient electrochemical detection of H2O2. The prepared Fe-pCOFs nanosheets with a large specific surface area and fully dispersed Fe2+ active sites, could enhance mass transfer and atomic utilization, thereby contributing to excellent catalytic efficiency toward H2O2. The flower-like MoS2 nanoflakes not only could catalyze the reduction reaction of H2O2, but also could serve as co-catalyst and form a catalytic cycle with Fe2+ of Fe-pCOFs, contributing the continuous emergence of catalytic active sites. The highly conductive MWCNTs was employed to promote electron transfer rate. Ultimately, the obtained Fe-pCOFs/MoS2-MWCNTs exhibited excellent electrocatalytic activity and conductivity, significantly amplifying the H2O2 response signal. Under the optimal conditions, the sensor had a wide detection range from 20 to 2650 mu & Mcy; for H2O2 with a detection limit as low as 2.2 mu & Mcy;. Moreover, the sensor successfully recorded trace amounts of H2O2 released from RAW264.7 cells, showing promising application for real-time and sensitive monitoring of H2O2 in clinic or pathological research.

Japanese encephalitis (JE), caused by Japanese encephalitis virus (JEV), is a mosquito-borne zoonotic disease and a leading cause of viral encephalitis worldwide. While JEV has the ability to traverse the blood-brain barrier (BBB), the precise mechanisms by which it inhibits the immune response prior to penetrating the BBB remain unclear, presenting obstacles in the development of efficacious therapeutic interventions. This study investigated the impact of JEV on CD8(+) T cell responses, with a particular focus on the dysfunction of CD8(+) T cells during JEV infection. Our results demonstrated that JEV infection significantly elevated the expression of PD-1 and TIM-3 on CD8(+) T cells, which are markers of T cell exhaustion, leading to inhibited function and impaired differentiation, resulting in a poorer prognosis in mice. Compared with nondiseased mice, symptomatic mice presented a greater proportion of exhaustion-like CD8(+) T cells. In vitro experiments further demonstrated that MDSCs induced an exhaustion-like state in CD8(+) T cells, characterized by significant upregulation of PD-1 and TIM-3 expression. Notably, blocking TIM-3 or depleting MDSCs restored CD8(+) T cell functionality by rescuing the expression of IFN-gamma and TNF-alpha. Furthermore, the depletion of MDSCs not only alleviated T cell exhaustion-like phenotypes but also improved survival rates in JEV-infected mice. These findings suggest that JEV promotes immune evasion through MDSC-induced CD8(+) T cell exhaustion-like states and identify TIM-3 as a promising therapeutic target for JE treatment.

The S2 subunit of infectious bronchitis virus (IBV) is a heavily glycosylated protein that can impact various characteristics of the virus. It is currently known that N-glycosylation modifications are predominantly located on the S2 subunit. However, the exact role of their N-glycosylation modification remains undisclosed. To elucidate the function of these N-glycosylation sites, we identified 14 common sites distributed on the S2 subunit of the 5 genotypes of IBV in present study. Subsequently, we selected 7 sites to generate mutants and assessed their impact on viral virulence, replication ability, and antigenicity. Our finding revealed that only 2 substitutions, N545S and K717N, increased the viral replication titer and antigenicity, and ultimately the pathogenicity in chicks. To delve into the mechanisms underlying this increased pathogenicity, we discovered that K717N can change the structure of antigenic epitopes. The N545S substitution not only influenced antigenic epitope structure, but also enhanced the ability of the virus to enter CEKs during the early stages of viral replication. These results suggest that the enhanced viral pathogenicity associated with N545S and K717N substitutions is multifaceted, with acceleration of the viral membrane fusion process and alterations in epitope structure representing crucial factors in the capability of N-glycosylation modifications to boost viral virulence. These insights provide valuable guidance for the efficient development of live attenuated vaccines.

Pets become vital companions in human life.They offer companionship and emotional support,and contribute significantly to physical and mental health,as well as social activities.Living with pets can stimulate human bodies to release more"affinity hormones"such as ser-otonin,dopamine,and oxytocin,which helps alleviate negative emotions.The human-animal bond can enhance the human body's ability to eliminate toxins,reduce the risk of illnesses,strengthen the immune system,and alle-viate depressive symptoms.Moreover,pet-related social activities can facilitate the making of new friends and the establishment of social connections,serving as a bridge for those who struggle with social interactions.

The ribosomal RNA (rRNA) is one of the most heavily modified RNA species in nature. Although we have advanced knowledge of the sites, functions, and the enzymology of many of the rRNA modifications from all kingdoms of life, we lack basic understanding of many of those that are not universally present. A single N 3 modified uridine base (m 3 U) was identified to be present on the 28 S rRNA from humans and frogs but absent in bacteria or yeast. Here, we show that the equivalent m 3 U is present in Drosophila and that the Ptch/CG12128 enzyme and its human homolog SPOUT1 are both necessary and sufficient for carrying out the modification. The Ptch-modified U is at a functional center of the large ribosomal subunit, and, consistently, ptch -mutant cells suffer loss of ribosomal functions. SPOUT1, suggested to be the most druggable RNA methyltransferases in humans, represents a unique target where ribosomal functions could be specifically compromised in cancer cells.

Blood-brain barrier (BBB) integrity is crucial for maintaining the function and environmental homeostasis of the central nervous system (CNS). Astrocytes play a critical role in the constitution of the BBB. In this study, we found that Japanese encephalitis virus (JEV) infection causes HMGB1 to translocate and release from the nucleus to the extracellular space in astrocytes, as well as elevated HMGB1 levels in the brain, which is related to BBB breakdown. Mechanistically, extracellular HMGB1 induces Ca2+ influx into astrocytes, leading to the overexpression of calmodulin (CaM) and the water channel protein aquaporin-4 (AQP4), triggering the phosphorylation of calmodulin kinase II (CaMKII) and promoting the translocation of AQP4 from the cytoplasm to the cell membrane. Inhibition of CaM, CaMKII, and AQP4 could block the cell membrane translocation of AQP4, thereby alleviating HMGB1-mediated BBB disruption. Extracellular HMGB1 is considered a potential target for mitigating neuroinflammation and BBB disruption. The positive feedback loop of HMGB1 exacerbates the disturbance of the BBB. These findings indicate that the HMGB1-AQP4 axis plays a role in the regulation of BBB integrity, presenting a new therapeutic target for the clinical treatment of JE and other CNS illnesses.

Myeloid-derived suppressor cells (MDSCs) are a group of heterologous populations of immature bone marrow cells consisting of progenitor cells of macrophages, dendritic cells and granulocytes. Recent studies have revealed that the accumulation of MDSCs in the mouse spleen plays a pivotal role in suppressing the immune response following JEV infection. However, the mechanisms by which JEV induces MDSCs are poorly understood. Here, it was found that JEV infection induces mitochondrial damage and the release of mitochondrial DNA (mtDNA), which further leads to the activation of TLR9. TLR9 deficiency decreases the M-MDSCs population and their suppressive function both in vitro and in vivo. Moreover, the increase of MHCII expression on antigen-presenting cells and CD28 expression on T cells in TLR9-/- mice was positively correlated with M-MDSCs reduction. Accordingly, the survival rate of TLR9-/- mice dramatically increased after JEV infection. These findings reveal the connections of mitochondrial damage and TLR9 activation to the induction of M-MDSCs during JEV infection. (c) 2024 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Organophosphorus pesticides (OPs) residues in agricultural products such as vegetables, fruits, and grains pose a threat to food safety and human health. In this work, a novel and highly sensitive electrochemical biosensor was developed for OPs residues detection in real sample, based on the immobilization of acetylcholinesterase (AChE) by electrostatic adsorption on a glassy carbon electrode (GCE) modified with composite nanomaterials of porphyrin-based covalent organic frameworks (p-COFs) loaded by gold nanoparticles (AuNPs) and poly (diallyldimethylammonium chloride) (PDDA). Due to the synergistic effect of p-COFs, AuNPs and PDDA, the fabricated biosensor displayed the remarkable properties with the high electrocataytic activity, and excellent biocompatibility, resulting in a significant signal amplification of as-prepared biosensor. Under optimized conditions, the biosensor showed excellent sensing performances for the detection of methyl parathion (MP), with wide linear ranges from 1.9 x 10- 9-3.8 x 10-5 M and a low detection limit as low as 2.3 x 10-10 M, as well as the satisfactory selectivity, excellent reproducibility, anti-interference properties and good stability. It was also successfully applied to the monitoring of MP content in vegetables and fruits, which may be explore a new opportunity for the potential utilization in analysis field.

The ribosomal RNA (rRNA) is one of the most heavily modified RNA species in nature. Although we have advanced knowledge of the sites, functions and the enzymology of many of the rRNA modifications from all kingdoms, we lack basic understanding on many of those that are not universally present. A single N3 modified Uridine base (m3U) was identified on the 28S rRNA from human and frog over thirty years ago, which is absent in bacteria or yeast. Here we show that the equivalent m3U is present in Drosophila, and that the Ptch enzyme and its human homolog are both necessary and sufficient for carrying out the modification. The Ptch-modified U is at a functional center of the large ribosome, and consistently ptch -mutant cells suffer loss of ribosomal functions. Ptch, proposed to be the most druggable RNA methyltransferases in human, represents a unique target where ribosomal functions could be specifically compromised in cancer cells. ### Competing Interest Statement The authors have declared no competing interest.

Pathogenic African swine fever virus (ASFV) remains a lethal causative agent in the domestic pig industry, which poses a burden on the swine market and causes substantial socioeconomic losses worldwide. Currently, there are no commercially efficacious vaccines or specific treatments available for ASF prevention and control. Unfortunately, little is known about the swine immune response upon ASFV infection. Here, we investigated the host immune response discrepancy induced by the field moderately virulent strain ASFV HB-2208 among healthy, diseased and asymptomatic pigs. In the peripheral blood of diseased swine, lymphopenia is caused by the massive loss of bystander lymphocytes, such as γδ T cells, B cells and CD4+ T cells. Conversely, ASFV has a strong tropism for the mononuclear phagocyte system (MPS) and partial dendritic cells (DCs), whose antigen-presenting ability is impeded by the downregulation of CD80 and MHC I. However, no significant difference in the number of CD8αhigh T cells was detected, whereas the frequencies of NK cells, NKT cells, and regulatory T cells (Tregs) were significantly increased. Additionally, an in vitro model was established with a coculture of primary pulmonary alveolar macrophages (PAMs) and peripheral blood mononuclear cells (PBMCs), which significantly reduced γδ T cells, B cells and CD4+ T cells and increased Tregs. The differentiated immune response might aid in enhancing the understanding of ASFV pathogenesis in suids and provide insights into the mechanism of ASFV-induced lymphopenia for further studies.

Human brain microvascular endothelial cells (hBMECs) are the main component cells of the blood-brain barrier (BBB) and play a crucial role in responding to viral infections to prevent the central nervous system (CNS) from viral invasion. Interferon-inducible transmembrane protein 1 (IFITM1) is a multifunctional membrane protein downstream of type-I interferon. In this study, we discovered that hIFITM1 expression was highly upregulated in hBMECs during Japanese encephalitis virus (JEV) infection. Depletion of hIFITM1 with CRISPR/Cas9 in hBMECs enhanced JEV replication, while overexpression of hIFITM1 restricted the viruses. Additionally, overexpression of hIFITM1 promoted the monolayer formation of hBMECs with a better integrity and a higher transendothelial electrical resistance (TEER), and reduced the penetration of JEV across the BBB. However, the function of hIFITM1 is governed by palmitoylation. Mutations of palmitoylation residues in conserved CD225 domain of hIFITM1 impaired its antiviral capacity. Moreover, mutants retained hIFITM1 in the cytoplasm and lessened its interaction with tight junction protein Occludin. Taken together, palmitoylation of hIFITM1 is essential for its antiviral activity in hBMECs, and more notably, for the maintenance of BBB homeostasis.

Glycosylation of the S1 subunit is a critical post-translational modification in coronavirus antigen proteins and plays a key role in vaccine development. The biological role of highly mutable glycosylation sites in virus evolution, however, remains underexplored. In this study, we identified nine glycosylation sites with high mutation rates on the S1 subunit of Avian Infectious Bronchitis Virus (IBV) through evolutionary analysis. Functional analysis of mutant strains revealed that specific mutations, particularly at position 76 (N → V), significantly enhance the strain's protective efficacy and limit viral replication. These findings provide important insights into the role of glycosylation in viral evolution and offer valuable guidance for the development of more effective IBV vaccines.