Surface browning is a significant challenge that hinders high-quality development of the fresh-cut produce industry. Vanillic acid (VA), an intermediate in the phenylpropanoid pathway, presents a potential solution, but its ability to reduce browning in fresh-cut produce was unknown. This study investigated the effects of pre-treating fresh-cut taro slices with varying concentrations of VA, followed by cold storage. The results showed that VA application at concentration of 0.1 g/L and higher reduced browning development in fresh-cut taro. Further investigations, using 0.1 g/L VA treatment, evaluated its mitigation at the metabolic, transcriptional and physiological levels. Metabolomics analysis revealed that VA treatment decreased the abundance of many specific flavonoids and lipids while promoting glutathione metabolism. Transcriptomic analysis, coupled with weighted gene co-expression network analysis and gene set enrichment analysis, indicated that genes down-regulated by VA treatment were predominantly enriched in flavonoid biosynthesis and lipid metabolism pathways. Conversely, VA treatment up-regulated the expression of genes in glutathione metabolism pathway, particularly at 6 d. Molecular docking analysis validated that VA could directly bind to the active sites of four key proteins involved in the flavonoid biosynthesis and lipid metabolism pathways. Physiological measurements showed that VA treatment reduced the activities of POD and PPO, lowered total phenolics, MDA, H2O2, and GSH levels, while increasing GSSG concentration. The multi-omics data, together with physiological results, demonstrate that VA treatment effectively prevents browning in taro. Given its natural origin, VA treatment holds promise as a safe and effective method for mitigating browning in the fresh-cut industry.

Feline calicivirus (FCV) is a major pathogen of upper respiratory tract diseases in cats, posing a significant threat to feline health. While current FCV preventive measures rely primarily on traditional vaccines, messenger RNA (mRNA) vaccines have emerged as a promising alternative, offering high efficacy, safety, rapid clinical development, and potential for fast, cost-efficient production. In this study, we designed a modified nucleotide sequence with a 124-amino acid deletion at a position in the region of the FCV-VP1 protein as an immunogen. The plasmid encoding the codon-optimized VP1 sequence was constructed, and VP1-mRNA was generated by in vitro transcription (IVT) and capping. After transfection into BHK-21 cells, immunofluorescence assay (IFA) and WB confirmed successful FCV-VP1 expression. Subsequently, the mRNA was encapsulated into lipid nanoparticles (LNPs) to prepare the LNP-VP1-mRNA vaccine. Characterization analysis revealed a uniform particle size distribution (polydispersity index [PDI] = 0.169) and a stable surface charge (zeta potential = -1.67 mV). A prime-boost immunization strategy was employed, which involved two intramuscular injections to immunize BALB/c mice or cats with the LNP-VP1-mRNA vaccine. ELISA analysis demonstrated that the vaccine elicited elevated levels of anti-FCV IgG and neutralizing antibodies in a dose-dependent manner, accompanied by the secretion of cytokines including IFN-β, IFN-γ, IL-4, and IL-6. Importantly, the VP1 mRNA vaccine provided complete protection against FCV challenge in cats, without the typical clinical signs and with a 100% survival rate. Our results indicate that the LNP-VP1-mRNA vaccine is a promising candidate for combating FCV infection.

Porcine deltacoronavirus (PDCoV) is an emerging enteropathogenic coronavirus that poses a significant threat to the swine industry. In this study, a novel PDCoV strain, designated PDCoV-ZJHZ2024, was identified from fecal samples of diarrheic pigs in China. Metagenomic analysis revealed co-detection of PDCoV and swine acute diarrhea syndrome coronavirus (SADS-CoV), with the microbial community predominantly composed of bacteria and characterized by abnormal enrichment of Bacillus cereus and pronounced gut microbiota dysbiosis. Genomic analyses demonstrated that PDCoV-ZJHZ2024 has undergone independent recombination events involving the ORF1b region and the spike (S) gene, accompanied by cross-regional genetic exchange, highlighting the critical role of recombination in PDCoV evolution and diversification. Codon usage analysis further indicated that codon preferences in this strain are primarily shaped by natural selection, potentially conferring enhanced translational efficiency in the host. Collectively, these findings underscore the evolutionary adaptability and transmission potential of PDCoV-ZJHZ2024 and provide new insights into PDCoV evolutionary dynamics, thereby informing future surveillance efforts and prevention strategies in swine populations.

Background:Young pigs are often coinfected with porcine circovirus 2 (PCV2) and porcine epidemic diarrhea virus (PEDV). This study aimed to determine the impact of coinfection of pigs with PCV2 and PEDV. Forty 16-day-old crossbred mixed-sex piglets were assigned to four groups (n = 10/group, NEG-CONTROL, PCV2-CONTROL, PCV2+PEDV, and PEDV-CONTROL). At day postinoculation (dpi) 0, NEG-CONTROL pigs were inoculated with saline, PCV2-CONTROL pigs were inoculated with PCV2, PCV2+PEDV pigs were inoculated with PCV2 and PEDV, and PEDV-CONTROL pigs were inoculated with PEDV. Results:No clinical signs were observed in the NEG-CONTROL and PCV2-CONTROL group pigs throughout termination of the study at dpi 21. Other than mild to moderate diarrhea, which lasted for about 7 days, no other clinical signs associated with PEDV or PCV2 infection were observed in the PEDV-CONTROL and PEDV+PCV2 groups. Conclusion:Coinfection of PEDV and PCV2 had no effect on virus shedding, serum antibody profile, and macroscopic or microscopic lesions.

Abrin is a type II ribosome-inactivating protein extracted from the seeds of Abrus precatorius. Due to its extreme toxicity and ease of acquisition, it is classified as a Category B bioterrorism agent, and no effective antidote is currently available. In this study, a fully human antibody against abrin, designated A140, was generated via screening of a fully human phage-display antibody library and subsequent expression. The A140 antibody demonstrates high affinity while maintaining specificity for its target antigen. It exhibited potent neutralizing efficacy against abrin toxin in both in vitro and in vivo assays. Intriguingly, the A140 antibody, which enters cells via clathrin-dependent and macropinocytic pathways and is subsequently trafficked to lysosomes, does not prevent the cellular entry of abrin, suggesting that its intracellular neutralization activity may function within the lysosomal compartment. Molecular docking studies based on AlphaFold 3 (AF3) indicate that A140 primarily recognizes residues Ser14, Arg169, and Asn173 of the alpha chain of abrin-a. Transcriptomic analysis based on Vero E6 cells showed that A140 antibody treatment was able to reverse almost 80% of the transcriptomic changes induced by abrin. Preliminary toxicity analysis shows that a high dose of the A140 antibody has no impact on the heart, liver, spleen, lungs, kidneys, and other major organs in mice. Our findings indicate that the A140 antibody, as a novel fully human antibody, holds promise as a key therapeutic agent for the treatment of abrin intoxication.

Pancreatic cancer is highly refractory and aggressive, with cancer stem cells (CSCs) being primarily responsible for its metastasis and chemoresistance. Deregulated cellular bioenergetics is a hallmark of cancer cells. However, the influence of bioenergetics on the maintenance of pancreatic CSC stemness and its underlying mechanisms have not been fully elucidated. In this study, pancreatic CSCs, isolated either by sorting ALDH+ subpopulation or enriching serially passaged tumorspheres from pancreatic cancer cells and PDX model, exhibited active mitochondrial complex I activity and increased oxidative phosphorylation. Complex I maintains stemness and tumorigenicity through its core subunit, NDUFS1. NDUFS1-mediated pancreatic CSC stemness is reinforced by high expression of CD147, which promotes pSTAT3Tyr705-mediated NDUFS1 transcription. To promote stemness, CD147-NDUFS1 initiates SIRT1-DNMT1 metaboloepigenetic signaling, decreasing promoter hypomethylation and increasing the mRNA expression of the stem cell transcript factor PAX2. Moreover, NDUFS1 and CD147 expressions were highly correlated in pancreatic cancer tissues, and their co-expression was significantly associated with poor patient survival. Taken together, our study provides evidence that mitochondrial complex I functions as a key player in CSC stemness maintenance through NDUFS1-mediated retrograde metaboloepigenetic signaling. Blocking a key regulator of mitonuclear communication by targeting CD147 may be a novel therapy for pancreatic cancer.

ABSTRACT Liquid–liquid phase separation (LLPS) has emerged as a fundamental physicochemical principle that organizes macromolecules into dynamic, membraneless condensates. These assemblies are increasingly recognized as critical regulators of diverse cellular processes. Notably, both viruses and their hosts exploit LLPS to optimize their respective strategies for replication and defense, forming a dynamic interplay centered around phase separation. However, a comprehensive mechanistic understanding of how LLPS modulates the dynamic viral–host battle, and how this knowledge can be leveraged for therapeutic development, remains an active area of investigation. This review systematically explores the dual roles of LLPS in viral infection and antiviral immunity. We detail how viruses hijack LLPS to form replication factories and inclusion bodies that enhance entry, replication, and immune evasion. Conversely, we explore how host cells leverage LLPS to assemble potent immune signaling hubs, such as those nucleated by cGAS–STING, NLRP6 inflammasomes, and T/B‐cell receptor microdomains, to amplify antiviral responses. Furthermore, we critically evaluate emerging therapeutic strategies that target these phase separation interfaces. By integrating recent advances across virology, immunology, and biophysics, this review establishes a unified framework for understanding and targeting LLPS in viral infectious diseases, offering new perspectives for future basic research and clinical intervention.

Porcine epidemic diarrhea virus (PEDV), an enteropathogenic coronavirus causing high mortality in neonatal piglets, continues to threaten global swine industries. Frequent mutations in the spike (S) protein of PEDV, particularly in emerging variants, have substantially compromised commercial vaccine efficacy. Despite the emergence of G2c variants dominating recent epidemics, comprehensive studies integrating viral isolation, phylogenetics, structural modeling, cross-neutralizing antibody response, and pathogenicity assessment remain insufficient. In this study, we successfully isolated a G2c strain (AHCZ02) and obtained 69 S gene sequences from nine provinces during 2021-2024. Phylogenetic analysis identified G2c variants as predominant (69.57%, 48/69) in current outbreaks. Structural comparisons revealed four G2c-specific substitutions (N139D, I287M, F345L, and L998M) inducing conformational changes in critical S domains compared to G2a/G2b strains, potentially disrupting immune recognition. The results of serum neutralizing antibody (nAb) test using the AHCZ02 strain showed that G2c-based feedback exposure strategies elicited 3.9-fold higher geometric mean titers (GMTs) than S-INDEL-based approaches. Furthermore, feedback exposure strategies of G2c (GMT = 480-1893) showed 12- to 189.3-fold higher neutralizing activity versus conventional vaccines (GMT = 10-40). Pathogenicity assessment in neonatal piglets revealed 100% mortality within 66 h post-AHCZ02 inoculation, accompanied by hallmark clinical manifestations including profuse watery diarrhea, rapid weight loss, and severe jejunal villus atrophy. Collectively, these findings provide evidence that G2c variants have developed S protein modifications associated with diminished vaccine efficacy, underscoring the need for next-generation vaccines incorporating G2c-specific antigenic determinants, and strengthened virological surveillance systems to effectively monitor and respond to PEDV evolutionary dynamics.

Prenatal hypoxia (PH) is a common pregnancy complication that can lead to cognitive impairment in the offspring, but the underlying mechanisms remain unclear. In this study, we established a model of PH by exposing C57 mice to hypoxia (10.5% oxygen) environment from gestational day (GD) 12.5-17.5. We found that PH resulted in cognitive impairment and reduced hippocampal neurogenesis in male offspring compared to control offspring. Mechanistically, PH is a form of prenatal stress that promotes placental transfer of maternal glucocorticoids (GC), which induces hyperactivity of the fetal hypothalamic-pituitary-adrenal (HPA) axis, leading to downregulation of the hippocampal glucocorticoid receptor (GR) in the offspring. In addition, PH promotes increased nuclear translocation of the GR and histone deacetylase 3 (HDAC3) complex, which represses the expression of immediate-early gene Npas4. By acting as a GR receptor antagonist, Mifepristone (MIF) mitigates ameliorated neurogenesis and cognitive impairment in the hippocampus of PH male offspring through the GR/HDAC3-Npas4 pathway. Thus, our study reveals that the GR/HDAC3-Npas4 signaling pathway is implicated in reduced hippocampal neurogenesis and cognitive impairment in PH male offspring. This research provides support for the pathogenesis of fetal cognitive impairment caused by PH.

Immune-mediated premature ovarian failure (IPOF) represents a significant pathology within reproductive health, necessitating the advancement of high-precision diagnostic instruments. To meet this clinical exigency, we engineered HD-GSH, a novel hemicyanine-based fluorogenic sensor specifically optimized for the real-time quantification of glutathione (GSH)-a critical redox biomarker inextricably linked to the molecular etiology of IPOF. The sensing mechanism utilizes a GSH-mediated specific transformation that induces a significant amplification of the fluorescence profile, achieving high linearity and a superior limit of detection (LOD) is 0.1 mu M. This analytical framework enables the continuous longitudinal tracking of GSH flux, facilitating early-phase diagnostics and the systematic assessment of IPOF progression. In pathological murine models, HD-GSH elicited markedly higher fluorescence intensities relative to healthy controls, validating its capacity for disease-specific metabolic recognition. These molecular findings were further substantiated by histopathological H&E staining, which documented profound structural degradation in ovarian tissues within the IPOF cohorts. Given its exceptional chemo-specificity and operational resilience under physiological conditions, HD-GSH serves as a robust analytical platform for non-invasive clinical diagnostics and biomedical research, offering transformative potential for real-time disease surveillance and the expedited implementation of targeted therapeutic interventions for IPOF.

Porcine deltacoronavirus (PDCoV) is a significant pathogen of swine with a global distribution, leading to severe gastrointestinal disease and substantial economic losses. Furthermore, PDCoV poses a potential threat to human health, as evidenced by the recent identification of three cases of infection in Haitian children. This study aimed to investigate the effects of PDCoV infection on host intestinal microbiota and bile acid metabolism, as well as the antiviral effects of lithocholic acid (LCA) in vitro and in vivo. Our results revealed that PDCoV infection caused microbiota dysbiosis in piglets, significantly reducing the intestinal abundance of Bacteroides fragilis (B. fragilis), a reduction that correlated with disruptions in bile acid metabolism. Colonization with bile salt hydrolase (BSH)-producing B. fragilis increased the levels of unconjugated bile acids and inhibited PDCoV infection, highlighting the role of microbiota-associated bile acid metabolism in viral pathogenesis. LCA, a prominent unconjugated bile acid, was shown to effectively inhibit PDCoV infection in porcine small intestinal epithelial cells and porcine intestinal enteroids. Notably, LCA inhibited PDCoV replication independently of bile acid receptor signaling and innate immune modulation. Mechanistic studies indicated that LCA prevents PDCoV infection by disrupting the viral entry process, specifically inhibiting the binding between the PDCoV spike protein and its cellular receptor, aminopeptidase N. In vivo experiments further confirmed that LCA significantly inhibited PDCoV infection in piglets. These results collectively highlight the potential of LCA as a therapeutic agent against PDCoV by targeting and disrupting the viral entry process, providing a novel strategy to control zoonotic PDCoV infections.

Porcine deltacoronavirus (PDCoV) is a globally distributed swine enteropathogenic virus that emerged in the last decade. A recent report of PDCoV infection in Haitian children also highlights potential public health implications. In this study, two monoclonal antibodies (mAbs), 1C2 and 5H5, were generated and showed high specificity for the PDCoV S protein. Both mAbs displayed high-titer neutralizing capabilities, suggesting their potential for passive immunotherapy. Epitope mapping revealed that the mAbs likely recognized conformational epitopes in the S1 subunit domains A and B of the native S protein, thereby blocking the interaction between the S1 receptor-binding domain and the cellular receptor, which could inhibit viral entry into host cells. This study offers new biological tools for PDCoV detection and lays the groundwork for the future development of porcine-specific antibodies for the prevention and treatment of PDCoV in piglets.

Prenatal hypoxia (PH) is a common complication of pregnancy, and it is strongly associated with psychiatric disorders such as depression and anxiety in the offspring. However, how prenatal hypoxia contributes to psychiatric disorders in the offspring is unclear. In this study, we established a model of prenatally hypoxic mice, where pregnant females were treated with hypoxia (10.5% O2) during gestational days 12.5-17.5, while controls (CON) were kept in a normoxic (21% O2) environment. Compared to CON offspring, PH male offspring exhibited depression-like behaviors. Prenatal hypoxia resulted in significantly higher protein level of the oxygen-sensitive subunit of hypoxia-inducible factor (Hif-1α) and lower levels of Ten-eleven translocated methylcytosine dioxygenase 1 (Tet1), β-catenin, and downstream Dicer1-miRNAs pathway associated with depressive behavior. Mechanistically, prenatal hypoxia leads to Hif-1α binding to Tet1, which inhibits β-catenin binding to Tet1, leading to an increase in ubiquitination-dependent degradation of β-catenin and down-regulation of the β-catenin-Dicer1-miRNAs pathway. In addition, administration of the β-catenin-specific agonist SKL2001 or overexpressing virus ameliorated the down-regulation of β-catenin-Dicer1-miRNAs signaling and depression-like behavior in PH male offspring. These findings suggest that Hif-1α and β-catenin competition for Tet1 binding is involved in depression-like behaviors in PH offspring, and this study provides important data on the molecular mechanisms by which prenatal hypoxia might be involved in adult psychiatric disorders of fetal origin.

Meplazumab, a humanized CD147 antibody, showed favorable safety and clinical benefits in phase 1 and phase 2/3 seamless clinical studies. Further evaluation of its therapeutic efficacy in patients with severe COVID-19 is needed. In this phase 3 add-on study, we randomized patients with severe COVID-19 in a 1:1 ratio to receive 0.2 mg/kg meplazumab or placebo via intravenous injection, and evaluated efficacy and safety within 56 days. Between February 2023 and November 2023, 108 patients with severe COVID-19 were randomized to two groups, with their baseline characteristics generally balanced. The primary endpoint, 28-day all-cause mortality was 1.96% in the meplazumab group vs 7.69% in the placebo group (P = 0.1703). Supplementary analysis using composite strategy indicated a significant reduction of 28-day all-cause mortality in meplazumab compared to placebo (3.92% vs 15.38%, P = 0.044). Meplazumab also significantly reduced the mortality in smoking subjects on day 28 (P = 0.047) compared to placebo in supplementary analysis. The secondary endpoint, 56-day all-cause mortality, was 1.96% in the meplazumab group and 11.54% in the placebo group (P = 0.048), which was 3.92% and 15.38%, respectively (P = 0.044) by supplementary analysis. Additional secondary endpoints showed potential benefits, including increased hospital discharge rates, improved clinical outcomes, and improved viral nucleotide conversion rate. Meplazumab demonstrated good safety and tolerability, with no grade ≥ 3 TEAEs observed. These promising results indicate that meplazumab reduces mortality and enhances clinical benefits in severe COVID-19 patients with a good safety profile, providing effective and specific therapeutics for severe COVID-19 (the trial was registered at ClinicalTrials.gov (NCT05679479)).

Infections of swine enteric coronavirus (SECoV), including porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and swine acute diarrhea syndrome coronavirus (SADS-CoV), cause severe diarrhea in piglets and result in substantial losses to the pig industry. The intestinal microbiota plays a crucial role in SECoV disease progression and outcomes, yet current research largely focuses on specific age groups or intestinal segments. This review provides a comprehensive analysis of the dynamic microbiota changes in piglets after SECoV infections across different ages and intestinal regions. It discusses differential microbiota analyses, functional changes, metabolic products, alongside their effects on immune responses. Additionally, we explore fecal bacterial transplantation as a potential intervention and highlight the role of the microbiota in either promoting or inhibiting SECoV infections. The development of advanced research tools, including culturomics, sequencing technologies, and multi-omics approaches, is pivotal in understanding the intricate relationship between the porcine intestinal microbiota and SECoV infections, offering potential strategies for preventing and controlling SECoV-related diseases.

Porcine deltacoronavirus (PDCoV), also known as HKU15, is a swine enteropathogenic virus that is believed to have originated in birds. PDCoV belongs to the genus Deltacoronavirus (DCoV), the members of which have mostly been identified in diverse avian species. We recently reported that chicken or porcine aminopeptidase N (APN), the major cellular receptor for PDCoV, can mediate cellular entry via three pseudotyped retroviruses displaying spike proteins from three avian DCoVs (HKU11, HKU13, and HKU17). In the present work, to better understand how avian-origin CoVs may be transmitted to pigs, we investigated the unknown DCoV entry pathway in avian cells. We show that clathrin-mediated endocytosis is involved in the entry of these DCoV pseudoviruses into chicken-origin DF-1 cells. Pseudovirus entry was suppressed by means of pharmacological inhibitors, dominant-negative mutants, and siRNAs targeting various cellular proteins and signalling molecules, suggesting that PDCoV and avian DCoV pseudovirus entry into DF-1 cells depends on clathrin, dynamin-2, cathepsins and a low-pH environment but is independent of caveolae and macropinocytosis. Furthermore, we found that DCoV pseudovirus entry was linked to Rab5- and Rab7-dependent pathways. This is the first report demonstrating that these DCoVs utilize clathrin-mediated endocytosis pathways to enter avian-origin cells, providing new insights into interspecies transmission of DCoVs.

Coronaviruses are a class of RNA viruses that cause disease in animals and humans. Zoonotic coronaviruses, such as severe acute respiratory syndrome coronavirus (SARS-CoV) and SARS-CoV-2, can cross species barriers and cause high mortality rates in humans. Coronavirus infection can cause different forms of cell death, among which apoptosis is the earliest and most systematic cell death. PANoptosis is a newly discovered inflammatory programmed cell death regulated by the supramolecular complex known as the PANoptosome. The cell death not only eliminates virus-infected cells but also further promotes innate and adaptive immune processes. Recent studies have shown that the important signaling pathways of apoptosis and PANoptosis induced by coronavirus infection are closely cross-linked, which plays a key role in regulating the replication and pathogenicity of coronavirus. In this review, we first revisit the molecular biology and the replication cycle of coronaviruses, summarize the definitions and characteristics of apoptosis and PANoptosis, and then focus on the current mechanisms of coronavirus-induced apoptosis and PANoptosis, as well as the regulatory mechanisms of these pathways for coronaviruses. We are exploring the detailed molecular mechanism between multiple programmed cell death pathways to provide new ideas for the pathogenic mechanism and drug development of coronavirus.

Mannanase specifically degrades mannan, increases the efficiency of energy utilization, and improves intestinal health in broilers. The interaction effects between mannanase and soybean meal (SBM) have not been extensively explored. Therefore, the present study aimed to determine effects of adding mannanase to diets with different SBM content on broilers, and to explore interaction effects between mannanase and SBM. This study was conducted on Arbor Acres broilers. Under low-energy conditions (metabolizable energy reduced by 50 kcal/kg), a 3 × 2 factorial design was used with three SBM content diets (control group, 50%, or 25% of the SBM content of control) and with two levels of mannanase (0 or 100 mg/kg) respectively. In experiment 1, growth performance and intestinal health were determined. Experiment 2 measured energy metabolism in broilers by respiratory calorimetry, while feces were collected to determine nutrient digestibility. Results indicated that low SBM diets supplemented with degossypolled cottonseed protein and corn gluten meal significantly reduced broiler growth performance during d 0−42. However, mannanase supplementation in diets containing 35.66% and 17.83% SBM significantly improved growth performance from d 0 to 21, reduced respiratory quotient at 21 d, and improved intestinal health (p < 0.05). In the 35.66% SBM diet, mannanase also enhanced energy metabolism by improving nitrogen retention and protein energy utilization (p < 0.05). However, mannanase showed limited efficacy when SBM content was reduced to 8.92%. Microbiological analyses showed that mannanase significantly reduced Escherichia coli and promoted 2-Oxocarboxylic acid metabolism in cecal microbes. In conclusion, there was a reciprocal relationship between mannanase and SBM content, with mannanase still exerting the above beneficial effects at the SBM level of 17.83%.