Search Results (805)

Proactive disease transmission pathway analyses break complex transmission routes into simpler steps, making risks and uncertainties easier to identify. This approach is especially valuable for African Swine Fever (ASF), a difficult-to-control disease in low-biosecurity settings or when biosecurity practices are inconsistently applied. To support targeted biosecurity planning, a pathway analysis was conducted that is specific to pork harvest facilities in the United States. The analysis focused on two federally inspected plants that slaughter market hogs and produce primal cuts. Inputs, outputs, and potential transmission pathways were identified through a literature review, site visits, and facility personnel interviews. Because ASF virus remains stable at low temperatures and in many pork products, particular attention was given to pathways involving storage conditions, waste materials, and processing steps such as heating or pH modification. Processing steps were evaluated against existing process control plans and ASF inactivation thresholds to determine mitigation status. Of 42 identified pathways, 39 were classified as unmitigated or of unknown mitigation status. These unmitigated or unknown pathways—broadly involving pigs, people, vehicles, and waste—represent the highest priorities for further risk assessment work and for exploring ways to develop or strengthen biosecurity protocols that reduce ASF transmission. Full article

Background: Systematic infectious screening is recommended before initiation of biologic therapies in chronic inflammatory rheumatic diseases (CIRDs), yet the clinical impact of this strategy in low-prevalence settings remains insufficiently characterized. This study aimed to evaluate the proportion of abnormal findings and their impact on treatment management. Methods: We conducted a retrospective single-center study including adult patients with CIRDs who underwent systematic pre-biologic infectious screening between January 2019 and June 2025. Screening included HIV, hepatitis B virus (HBV), hepatitis C virus (HCV), interferon-γ release assay (IGRA), and chest radiography. The primary outcome was the proportion of abnormal results and their impact on biologic initiation. Results: A total of 418 patients was included (mean age 48.2 ± 14.6 years; 69.1% female). No active HIV, HBV, or HCV infections were detected. Past HBV infection markers were identified in 2.6% of patients, and anti-HCV antibodies in 0.7%, all without detectable viremia. None of these findings required modification of biologic therapy. IGRA positivity was observed in 4.3% of patients and indeterminate results were seen in 3.1%. Preventive antituberculous therapy was initiated in most newly identified IGRA-positive cases, leading to delayed biologic initiation in several patients. Chest radiography yielded limited additional diagnostic value. Conclusions: In this population, systematic pre-biologic infectious screening identified few clinically actionable viral infections, whereas latent tuberculosis screening represented the main determinant of therapeutic modification. These findings support continued emphasis on tuberculosis risk assessment and warrant further prospective studies to evaluate optimized and potentially targeted screening strategies incorporating cost-effectiveness analyses. Full article

Objectives: To investigate the potential mechanisms of maternal pregestational diabetes-induced neural tube defects (NTDs) by integrating proteomic data and histone H4 lysine 5 acetylation (H4K5ac) ChIP-seq data from the mouse model. Methods: The diabetic mouse model was established by intraperitoneal injection of streptozotocin (STZ) into female friend leukemia virus B strain (FVB) mice, with subsequent blood glucose monitoring. Diabetic females were then mated with healthy males, and embryonic tissues were collected on embryonic day 9.5. Among the embryos obtained from diabetic pregnancies, six NTDs embryos and six control embryos were selected for protein expression profiling using tandem mass tag (TMT)-labeled liquid chromatography-tandem mass spectrometry (LC-MS/MS), as well as for assessment of H4K5ac modification by ChIP-seq. Multi-omics integration was performed to identify common differentially expressed genes, followed by functional enrichment analysis. Key genes were validated using RT-qPCR. Results: Proteomic analysis revealed that differentially expressed proteins were significantly enriched in focal adhesion pathway. Protein–protein interaction (PPI) network analysis indicated that these proteins (e.g., Integrin alpha 3 (Itga3), glycogen synthase kinase 3 beta (Gsk3b), mitogen-activated protein kinase 9 (Mapk9)) were associated with focal adhesion and cytoskeletal functions. Integrated multi-omics analysis identified 923 common differentially expressed genes, which were also significantly enriched in focal adhesion pathway. Within this pathway, the protein expression levels of Itga3, Gsk3b, and Mapk9 exhibited a consistent co-variation trend with H4K5ac enrichment. RT-qPCR results confirmed that Itga3 was significantly up-regulated, while Gsk3b was down-regulated in the NTDs group (p < 0.05). Conclusions: Maternal pregestational diabetes may contribute to NTDs by disrupting cytoskeletal reorganization, cell adhesion, and migration processes. This disruption is likely mediated through H4K5ac-regulated expression of key focal adhesion pathway genes such as Itga3 and Gsk3b. Full article

Chronic infection with the hepatitis C virus (HCV) is the most significant risk factor for the development of hepatocellular carcinoma (HCC). It has been shown that the progression of HCV-related liver disease is mediated by both viral and host-specific factors. The HCV replication cycle is a host-dependent process that relies on intracellular signalling pathways within target cells. Thus, intracellular signal transduction plays a pivotal role in the modification of interactions between the host and HCV. These pathways are key regulators of liver diseases, including cirrhosis and HCC. In addition, HCV induces epigenetic modifications in the host genome that inhibit the expression of various tumour-suppressor genes. Some of these changes persist even after successful antiviral treatment and represent a continued risk for HCC development. Despite significant progress in the management of chronic HCV infection, this challenge remains unresolved. In this narrative review, we summarise the mechanisms of HCV-induced disease progression, focusing on the host immune response, the regulatory roles of viral and cellular proteins, and viral survival strategies during chronic infection. We also discuss HCV-induced epigenetic alterations that contribute to hepatocarcinogenesis both during infection and after viral clearance. These insights are important for identifying novel, reliable molecular biomarkers for patient surveillance and for designing new therapeutic approaches. Full article

Background/Objectives: Immunodeficiency can be induced by a variety of factors, such as aging, stress and poor nutrition, and leads to increased susceptibility to infection and disease. The current research was conducted to determine the immunoenhancing potential of yam and its underlying mechanism in a murine model of cyclophosphamide (CTX)-induced immunosuppression. Methods: The gut microbial community and generation of short-chain fatty acids (SCFAs) in response to yam were analyzed by 16S rRNA sequencing and GC-MS. The immune cells in the spleen were analyzed using flow cytometry. GPR41/GPR43/GPR109A triple-knockout mice were used to demonstrate the critical involvement of SCFAs in mediating the protective effect of yam, and RNA-sequencing technology was applied to investigate the potential mechanism by which yam orchestrated the observed metabolic, immune and reparative responses. Results: Yam alleviated the decline in spleen and thymus indices and modulated the frequency of B cells and CD4⁺ and CD8⁺ T cells and promoted the production of IgA, IgG and IgM. Yam increased the secretion of cytokines in the intestine and upregulated the levels of claudin and ZO-1. Yam also increased the content of SCFAs and induced beneficial modifications to the gut microbiota composition. The immune-enhancing activity of yam was confirmed, as evidenced by a notable decrease in viral load in immunosuppressed mice inoculated with influenza virus and its capacity to mitigate inflammatory response in pulmonary tissues. Conclusions: This study suggests that yam enhances immunity by synergistically regulating the gut–immune axis, supporting its development as a functional food intervention in managing immunodeficiency conditions. Full article

The 3,4-dihydropyrimidin-2(1H)-one (DHPM) scaffold possesses diverse biological activities and chemical tunability, allowing structural modifications to modulate antiviral, anticancer, and anti-inflammatory effects. In this study, a series of DHPM derivatives with varied substituents were designed and synthesized, and their structures were characterized by ¹H NMR, ¹³C NMR and HRMS. Their antiviral activities against pseudorabies virus (PRV) and vesicular stomatitis virus (VSV) were evaluated. In vitro assays revealed that several compounds exhibited significant antiviral effects, with 4bf (SI = 243.08 against PRV), 4ce (SI = 196.4 against VSV), and 4be (SI = 124.2 against PRV; SI = 181.1 against VSV) showing the most potent activity. Further studies demonstrated effective inhibition of viral titers, and Western blot and qRT-PCR analyses confirmed downregulation of viral proteins and related genes. Cytotoxicity tests indicated that 4bf, 4ce, and 4be had CC₅₀ values of 270.0, 147.1, and 190.9 μg/mL, respectively, suggesting favorable safety profiles. In vivo experiments showed that 4bf, without affecting normal growth, alleviated PRV-induced pulmonary inflammation and tissue damage, and improved survival in mice. Taken together, DHPM-based compounds demonstrate promising potential as candidate antiviral agents, warranting further development. Full article

The flavor of rabbit meat has always been a major factor hindering the development of the rabbit industry. One of the main factors affecting the flavor of rabbit meat is intramuscular fat. N6-methyladenosine (m⁶A) regulates multiple aspects of the physiology of animals. In this study, qRT-PCR and m⁶A-qPCR were used to identify genes and methylation levels. AAV virus was used as a vector to overexpress genes. To explore the regulatory mechanism of m⁶A on intramuscular fat in rabbits, we first explored the regulation of the LPL gene of rabbits by m⁶A at the cellular level using interfering RNA. Subsequently, we further validated the mechanism and explored the regulation of metabolites by LPL genes in living dorsal muscles. The results demonstrate that METTL3 inhibited LPL expression through m⁶A modification under the recognition of YTHDF2 in adipocytes and muscles. LPL promotes adipocyte differentiation and intramuscular fat deposition. In addition, LPL regulates intramuscular fat deposition through L-Glutamine/multiple pathways and 3-Methyl-L-histidine. This study confirms that m⁶A can affect the expression of the LPL gene in rabbits, thereby regulating the IMF of rabbit meat by L-Glutamine/multiple pathways and 3-Methyl-L-histidine. This study lays the molecular foundation for cultivating high-quality rabbit meat. Full article

Hierarchical functionalisation of the UiO-66(Zr)-NH₂ metal–organic framework with cysteine, poly(ethylene glycol) (PEG), and the SARS-CoV-2 spike receptor-binding domain (RBD) was developed to enable receptor-specific interaction with the angiotensin-converting enzyme 2 receptor (ACE2) in model cells. Post-synthetic modification using cysteine and heterobifunctional PEG linkers allowed controlled bioconjugation of SpyTag-labelled RBD via SpyTag/SpyCatcher chemistry, while preserving the crystallinity, microporosity, and intrinsic optical properties of the UiO-66(Zr)-NH₂ framework. Comprehensive physicochemical characterisation confirmed successful surface functionalisation, tunable aggregation behaviour, and retention of multimodal optical characteristics. Cellular studies in HEK293T and HeLa cells overexpressing EGFP-tagged ACE2 demonstrated enhanced and selective association and uptake of RBD-functionalised nanoparticles compared with non-targeted analogues. Multimodal fluorescence imaging, fluorescence lifetime imaging microscopy, flow-cytometry, and electron microscopy indicated ACE2-dependent endocytic internalisation, with predominant localisation in endosomal and autophagosomal compartments, while both amine- and cysteine-modified formulations exhibited good biocompatibility. Overall, this study establishes a virus-mimetic, ACE2-targeted UiO-66(Zr)-based nanosystem as a proof-of-concept biointerface platform for receptor-specific cellular delivery and imaging, providing a foundation for future MOF-based nanocarriers exploiting ligand–receptor interactions. Full article

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has greatly impacted public health due to high rates of transmissibility and mutation during the COVID-19 pandemic. Macrodomain 1 (Mac1) of non-structural protein 3 remained well conserved across variants and is critical to suppression of host immune response to infection, making Mac1 a promising target for therapeutic development. Mac1 binds and cleaves the post-translational modification ADP-ribose and is hypothesized to have a downstream effect on the host interferon response, but the exact cellular targets of Mac1 are still unknown. Characterizing the substrates of Mac1 ADP-ribosyl hydrolase activity using a catalytically inactive mutant N40D can reveal critical virus–host interactions to identify protein targets of Mac1 and reveal mechanisms of host interferon suppression. Here, we performed affinity enrichment with WT Mac1 and Mac1 N40D in HEK293T and A549 cells and quantified changes in protein interactions by TMT-multiplexed tandem mass spectrometry. We identified interactions between Mac1 and ADP-ribosylated substrates involved in DNA damage response, cytoskeletal components, and cell cycle regulation. Additionally, several members of the TRiC complex involved in protein folding were selectively enriched with mutant Mac1 from A549 cells. These findings suggest a novel role of Mac1 in regulating host protein folding. Full article

Background/Objectives: Epigenetic regulation plays a fundamental role in controlling the spatiotemporal expression of genes in plants under stressful environmental conditions. While LIKE HETEROCHROMATIN PROTEIN 1 (LHP1) is known to be involved in histone modification, its function in regulating the biosynthesis of specialized metabolites, particularly monoterpenoid indole alkaloids (MIAs) in Catharanthus roseus, remains elusive. Methods: CrLHP1 was identified by mining the C. roseus proteome and characterized through sequence alignment, phylogenetic analysis, and conserved domain assessment. Virus-induced gene silencing (VIGS) was employed to suppress CrLHP1 expression, after which the transcript levels of jasmonic acid (JA)-responsive genes and key MIA biosynthetic genes, as well as the accumulation of vindoline and catharanthine, were analyzed. Furthermore, deep learning-based protein structure prediction (AlphaFold3) and yeast two-hybrid (Y2H) assays were conducted to explore protein-protein interactions. Results: CrLHP1 was confirmed as the ortholog of Arabidopsis thaliana LHP1 (AtLHP1). Exposure to 75 μM MeJA upregulated MIA upstream pathway genes while downregulating CrLHP1 transcription. Silencing CrLHP1 significantly upregulated JA-responsive and MIA biosynthetic genes, leading to enhanced catharanthine accumulation. Additionally, the structural prediction and Y2H assays revealed a physical interaction between CrLHP1 and CrJAZ1. Conclusions: These findings suggest that CrLHP1 negatively regulates MIA biosynthesis, potentially by modulating JA signal transduction through interaction with CrJAZ1. This study provides new insights into the possible epigenetic mechanisms governing alkaloid production in C. roseus. Full article

(1) Background: Nitric oxide (NO) serves as a crucial signaling molecule in plant abiotic stress responses. Although its role in enhancing drought resistance in cotton has been recognized, the specific mechanisms underlying this physiological and molecular regulation remain largely unexplored. This study aims to elucidate the multi-layered mechanisms by which NO modulates drought resistance in cotton; (2) Methods: Cotton seedlings were subjected to drought stress with the application of the NO donor sodium nitroprusside (SNP). A combination of confocal laser scanning microscopy, transcriptional expression analysis, biochemical assay of enzyme activity, virus-induced gene silencing (VIGS), and in vitro protein modification assays was applied to characterize the effects of NO on the drought stress response in cotton; (3) Results: Exogenous NO significantly reinforced drought resistance in cotton seedlings by improving leaf water retention capacity and photosynthetic efficiency, eliminating excessive drought-induced reactive oxygen species (ROS), upregulating the transcription and enzymatic activity of antioxidant enzymes, and promoting stomatal closure. Mechanistically, NO triggered S-nitrosylation of the plasma membrane H⁺-ATPase isoform GhHA2, thereby enhancing its protein stability; (4) Conclusions: These findings reveal that exogenous NO orchestrates cotton drought tolerance via multiple interconnected physiological and molecular pathways, in which the activation of the antioxidant defense system and the modulation of stomatal closure serve as central regulatory mechanisms. Full article

Norovirus is the leading cause of acute infectious gastroenteritis in the world and accounts for a significant proportion of outbreaks at the food-borne and person-to-person levels. Due to their low infectious dose, persistence in the environment, and broad genetic diversity, they can quickly spread and reappear in even the most diverse populations. This review integrates current knowledge on the epidemiology of noroviruses, genomic organization, structural biology, virus–host interactions, and replication mechanisms, with a focus on factors that determine virus evolution and strain dominance. Literature has been systematically searched in the PubMed and Scopus databases to incorporate recent experimental and epidemiological findings. Analysis of global surveillance data indicates ongoing genetic diversification of circulating strains, with periodic replacement of major variants, particularly the GII.4 lineage. Variability of the capsid and recognition of histo-blood Group Antigens strongly affects the host’s susceptibility, viral attachment and immune escape. The capsid consists of most of the viral protein complexes. The structural proteins VP1 and VP2 are responsible for determining the contours of the capsid and antigenic specificity. Non-structural proteins are responsible for coordinating the genome replication and the modification of host cell pathways to favor the production of the virus. Eliminating these gaps by means of integrated genomic surveillance and functional studies will provide insight into the evolution of norovirus and help to develop broadly effective vaccines and antiviral strategies. Full article

The hemagglutinin (HA) of influenza A/H3N2 virus evolves rapidly, with glycosylation driving immune evasion. However, how host microenvironmental cues influence this process remains poorly understood. We identified a novel N-linked glycosylation site at position 110 (N110) in contemporary H3N2 viruses (NSS genotype) that enhances viral fitness by increasing receptor-binding signal, HA cleavage, and replication. Remarkably, hypoxia, which mimics the respiratory tract microenvironment, significantly augments N110 glycosylation. Mechanistically, we identified the B4GAT1-B4GALT1 complex as the key mediator of this modification. Hypoxia upregulates their expression and strengthens their interaction with HA. In ferret models, N110-glycosylated viruses exhibit heightened pathogenicity and evade ancestral antibodies. Furthermore, immunization with N110-containing HA confers broad-spectrum protection, whereas reciprocal immunization is ineffective. Our findings reveal hypoxia-driven glycosylation as a previously unrecognized mechanism of H3N2 adaptation, providing critical insights for vaccine efficacy and highlighting the importance of integrating microenvironmental factors into future antiviral strategies. Full article

Although RNA cytosine-5 methylation (m⁵C) is an important post-transcriptional regulatory mechanism, its contribution to plant antiviral immunity remains unclear. In this study, we identified Thiamine thiazole synthase 2 (TaTHI2) as a host mRNA target of the wheat m⁵C methyltransferase TaNSUN2 during infection by Chinese wheat mosaic virus (CWMV), a soil-borne virus that poses a major threat to wheat production. TaNSUN2 contributes to the m⁵C modification of TaTHI2 transcripts, enhancing mRNA stability and sustaining TaTHI2 accumulation. The disruption of a key m⁵C site markedly reduced methylation, weakened TaNSUN2–RNA binding, and accelerated transcript decay, leading to the compromised production of reactive oxygen species (ROS) and increased viral infection. Mechanistically, the TaNSUN2-dependent m⁵C modification stabilized TaTHI2 mRNA, thereby promoting ROS-mediated antiviral defense. Collectively, our results establish the m⁵C modification of TaTHI2 mRNA as a critical post-transcriptional control point in CWMV resistance and highlight TaNSUN2-dependent RNA methylation as an integral component of host antiviral immunity. Full article

Background: Translational regulation constitutes a critical layer of gene expression control in plants, yet the contribution of endogenous 5′ untranslated regions (5′ UTRs) to translational efficiency remains incompletely defined. While viral and synthetic leader sequences have been widely used to enhance protein production, comparatively few native plant 5′ UTRs have been systematically characterised. The objective of this study was to identify and functionally evaluate endogenous plant 5′ UTR elements that promote translation through post-transcriptional mechanisms. Methods: A 79-nucleotide fragment (CYSTM α) derived from the 5′ UTR of Arabidopsis thaliana CYSTM1 (AT1G05340) was cloned upstream of reporter genes and assessed using dual-luciferase assays in transient expression systems (Nicotiana benthamiana and A. thaliana) and in stable transgenic Arabidopsis lines. Translational activity was further evaluated in monocot wheat germ extract and in Escherichia coli. Transcript abundance was quantified by qRT-PCR. Publicly available ribosome profiling and m⁶A datasets were analysed to assess translational efficiency and RNA modification status. Results: In N. benthamiana and A. thaliana, CYSTM α increases reporter protein production 3–7 fold relative to the control and 30–130% above the benchmark Tobacco Mosaic Virus (TMV) Ω leader, without altering mRNA abundance. The CYSTM α sequence also enhances luciferase translation in monocot wheat germ extract and elevates translation 5-fold in E. coli. CYSTM α contains three motifs that may promote translation, namely three CAA repeats that are associated with translation initiation, an AMAYAA motif that is associated with eIF3 binding, and two N6-adenosine DRACH sites that are associated with cap-independent translation. Additionally, ribosome profiling revealed high translational efficiency (TE = 3.25) of native CYSTM1. Conclusions: CYSTM α represents a compact endogenous 5′ UTR element that enhances translation across multiple experimental systems. These findings expand the repertoire of plant-derived translational enhancers and provide insight into sequence features associated with efficient mRNA translation in plants. Full article