Search Results (899)

Highly pathogenic avian influenza H5N1 virus (H5N1 HPAIV) has long circulated in wild waterfowl and poultry and continues to cross species barriers, posing sustained threats to livestock productivity and public health. The 2024 outbreak of H5N1 HPAIV in dairy cattle across multiple U.S. states represents the first recognized large-scale transmission event of this virus in ruminants, reshaping our understanding of its host range and transmission ecology. This review summarizes recent advances in the epidemiology, virological characteristics, transmission, pathogenesis, surveillance, and control of dairy cattle-associated H5N1. The initial multistate outbreak was dominated by clade 2.3.4.4b genotype B3.13, whereas subsequent independent introductions of genotype D1.1 demonstrated that repeated avian-to-cattle spillover also contributes to the evolving outbreak ecology. A defining feature is efficient replication in bovine mammary epithelial cells, resulting in high viral titers in milk, supporting milk-associated exposure and milking-related contamination as plausible components of transmission, although the relative contribution of different routes remains unresolved. Infected cattle typically show reduced feed intake, a marked decline in milk production, abnormal milk, and mild systemic signs, whereas severe respiratory disease and mortality are uncommon. Mutations such as PB2-M631L, PA-K497R, and changes in NP and NS1 may promote replication and immune evasion in bovine cells, while genotype-specific PB2 adaptations highlight the potential for further mammalian adaptation. Human infections reported to date have been predominantly mild and associated with occupational exposure, with no evidence of sustained human-to-human transmission. Experimental vaccine studies have begun to demonstrate immunogenicity in cattle, although protection against mammary infection, viral shedding, and transmission remains to be established. Future priorities include clarifying mammary tropism and transmission dynamics, while strengthening diagnostics, farm biosecurity, vaccination, evaluating cattle vaccination strategies, and integrated One Health surveillance. Full article

Background: Metabolic dysfunction-associated fatty liver disease (MAFLD) is a prevalent chronic liver disorder with limited pharmacological therapies, highlighting the need for novel multi-target agents from traditional medicinal plants. Artemisia rupestris L. has hepatoprotective effects, but its active constituents and mechanisms against MAFLD remain unexplored. Methods: The chemical profile of an ultrasonically-assisted 70% ethanol extract of Artemisia rupestris L. (ARE) was determined using UPLC-Q-TOF-MS. Network pharmacology, molecular docking, 100-ns molecular dynamics (MD) simulations, and in vitro experiments in oleic acid-induced HepG2 cells were integrated to identify active compounds, core targets, and pathways, and to validate lipid-lowering and hepatoprotective effects. Results: Seventy-eight compounds were identified, mainly flavonoids, phenolic acids, and terpenoids. Network analysis highlighted AKR1B10 and MAPK14 as core targets and isorupestonic acid and rupestonic acid as key components. MD simulations supported the stable binding of isorupestonic acid to AKR1B10 (RMSD ~1.6–3.2 Å), driven by van der Waals and electrostatic interactions. In vitro, ARE dose-dependently reduced intracellular triglycerides, total cholesterol, and LDL-C, decreased lipid droplets, and lowered AST/ALT levels, with a high-dose efficacy comparable to atorvastatin. Conclusions: Through in vitro experiments and network pharmacology analyses, this study preliminarily elucidated the beneficial effects of ARE on lipid deposition in hepatocytes and its potential mechanism, thereby providing an experimental basis and potential avenues for subsequent in vivo studies on pharmacodynamics, pharmacokinetics, and toxicology. Full article

This study compared secondary-metabolite profiles, IC₅₀-based antiproliferative activity, and relative selectivity of four Indonesian cardamom (Amomum compactum Soland. ex Maton) accessions: Bogor white, Bogor red, Sukabumi, and Ciamis. Ethanol and ethyl acetate extracts were evaluated in MCF-7, MCM-B2, and MCA-B1 tumor-derived cells, with Vero cells as a non-tumor-derived reference and profiled by untargeted LC-MS/MS. Putatively annotated metabolites were further assessed using chemometrics, terpenoid-focused network pharmacology, molecular docking, and 50 ns molecular dynamics simulation. Extract IC₅₀ values ranged from 0.959 to 2.210 mg mL⁻¹ in MCF-7, 0.818 to 2.733 mg mL⁻¹ in MCM-B2, and 1.382 to 3.426 mg mL⁻¹ in MCA-B1 cells, while Vero values ranged from 2.762 to 4.000 mg mL⁻¹. MCF-7 cells generally showed the greatest overall sensitivity, although the lowest individual IC₅₀ occurred with Sukabumi ethyl acetate extract in MCM-B2 cells. Ciamis ethyl acetate extract showed higher relative selectivity toward MCF-7 and MCM-B2 cells. LC-MS/MS putatively annotated 36 metabolites, with terpenoid-related compounds predominating. Kaur-16-ene was computationally prioritized based on predicted interactions with ESR1, EGFR, SCN5A, and CCND1 and short-timescale ESR1-complex stability. These findings demonstrate accession- and solvent-associated differences and support kaur-16-ene for further experimental validation. Full article

H9N2 avian influenza viruses pose a persistent zoonotic risk owing to their broad host adaptability. Between 2024 and 2025, two H9N2 isolates (ZHY1022B2 and ZHY0417A11) were recovered from quail flocks in Hebei, China. Phylogenetic analysis clustered their HA genes within the dominant B4.7.2 subclade, but diverged into two subgroups (B4.7.2.2 and B4.7.2.1). Of particular interest was ZHY0417A11, which displayed a multigenic reassortment pattern—its PB1 originated from an H3N8 virus, PA and PB2 from H3N3, NS from H10N3, and the M gene was nearly identical (99.32%) to a human H3N8 isolate, whereas the remaining HA, NA and NP segments maintained the H9N2 backbone. Despite the presence of the HA mammalian-adaptive markers (H191N, A198V and Q234L), both strains showed marked antigenic drift from the vaccine strain SS (BJ/94-like; R < 0.5), while retaining reactivity with currently circulating field strains. These findings argue for heightened vigilance in quail populations, given their role as mixing vessels, and highlight the limitations of current vaccine matching in light of ongoing H9N2 evolution. Full article

Nelumbo nucifera Gaertn. is a rich source of bioactive compounds, and has been extensively investigated for its antioxidant and anti-inflammatory properties. However, despite its broad pharmacological potential, detailed molecular-level insights into the interactions of its phenolic and flavonoid constituents with key skin-aging-related enzymes remain limited. Therefore, this study employed an integrated in silico and in vitro approach to systematically evaluate the anti-aging potential of phenolic and flavonoid compounds derived from N. nucifera. Molecular docking was performed to investigate their binding interactions with tyrosinase, elastase, and collagenase. Compounds exhibiting the most promising inhibitory profiles were further subjected to molecular dynamics (MD) simulations for 100 ns and density functional theory (DFT) analyses at the B3LYP/6-31G(d,p) level of theory to elucidate their structural stability and electronic properties. The computational results revealed favorable binding affinities, stable complex formation, and electrostatic features supporting hydrogen bond interactions. In vitro enzyme inhibition assays demonstrated that hesperidin exhibited inhibitory activity against tyrosinase, collagenase, and elastase, with IC₅₀ values of 3.43 mM, 2.26 mM, and 0.55 mM, respectively. Kojic acid was used as a positive control for tyrosinase, while epigallocatechin gallate (EGCG) was used as a positive control for both collagenase and elastase. Additionally, hesperidin exhibited enzyme-inhibitory activity and preliminary intrinsic UV absorption. This compound warrants further investigation, specifically regarding its stability and performance within standardized cosmeceutical formulations. Full article

Japanese encephalitis virus (JEV) is a zoonotic pathogen transmitted primarily by Culex mosquitoes and causes severe neurological diseases in humans and animals. The main endemic areas are the Western Pacific and Southeast Asia, and its geographical distribution has expanded in recent years. The development of a diagnosis for orthoflavivirus infections is hampered by two main problems: the short duration of viremia, resulting in a narrow detection window, and severe cross-reactivity. NS1, a secreted nonstructural protein of orthoflavivirus, holds promise as a new target for overcoming these limitations. In this study, we established a highly specific and sensitive capture ELISA for the JEV NS1 protein. First, the JEV NS1 protein was successfully expressed in mammalian cells and purified using affinity chromatography. Seven mAbs recognizing JEV NS1 were generated, and the mAb 20B6 exhibited the strongest binding affinity. Based on 20B6, a capture ELISA was developed with an optimal coating concentration of 3 μg/mL and an optimal detection antibody working concentration of 0.432 μg/mL. No cross-reactivity was observed with other orthoflaviviruses (including WNV, KUNV, USUV, MVEV, SLEV, and ZIKV) or common porcine viruses. The method could effectively detect NS1 protein in cell culture medium, cell lysates, mouse tissues, and porcine serum samples from JEV-infected subjects. This study provides a solid foundation that may be further developed into an efficient and specific tool for epidemiological surveillance of Japanese encephalitis. Full article

(1) Background: Bovine viral diarrhea virus (BVDV) is an economically important pathogen affecting cattle worldwide. The genetic diversity of BVDV-2 in buffalo remains poorly understood. This study aimed to elucidate the genomic and antigenic features of GX24, the first BVDV-2b isolate identified from dairy buffalo in China. (2) Methods: The virus was isolated from a rectal swab through three blind passages in MDBK cells and identified through immunofluorescence and RT-PCR. The near-complete genome was sequenced using the Illumina platform and subjected to phylogenetic and recombination analysis. B-cell epitopes and glycosylation sites were predicted using BepiPred-3.0, Epitope1D, DiscoTope-3.0, NetNGlyc-1.0, and NetO-Glyc-4.0. (3) Results: The 12,267-nt GX24 genome (GenBank: PX682047) was classified as BVDV-2b. Recombination analysis detected a putative recombination signal within the NS5A gene involving Chinese BVDV-2b and Italian BVDV-2a strains. Integrative analyses predicted conserved linear and conformational epitope clusters in the N- and C-terminal regions of the E2 protein. Spatial analysis indicated that several epitope residues may be masked by glycan shielding. (4) Conclusions: This study provides the first genomic evidence of a BVDV-2b isolate from dairy buffalo in China, suggesting that BVDV-2b may be present in this population. The putative recombination signal and antigenic characteristics offer valuable insights for the development of diagnostics and vaccine design in the future. Full article

This paper presents a dual-quaternion (DQ) framework for the rigid-kinematic modeling and validation of rigid hexagonal Kresling origami robots executing a prescribed FOLD–TWIST–FOLD motion. Triangular panels are modeled as rigid bodies, and crease lines are modeled as fixed revolute axes. Exact spatial 4R closure is decomposed into a primal orientation closure and a dual transported-translation closure. Under explicit non-degeneracy, paired-normal transport, and branch assumptions, the projection of the dual closure recovers a Bennett-type axis ratio. For Kresling, this result is applied only in the intersecting-axis limit d = 0, serving as a local axis-geometry diagnostic rather than a sufficient global closure criterion. The baseline three-cell module (N_s = 6, R = 48 mm, H₀ = 60 mm, initial twist 30°) executes a (−2 mm, +5°, −2 mm) actuation command over 37 states, preserving rigid-edge/panel and DQ consistency within the prescribed tolerances. Comparative benchmarks show that DQ and homogeneous-transform mappings achieve practically identical accuracy, although homogeneous transforms run faster in the tested MATLAB 2025b workload; Direct LM is faster for computing the endpoint solution, whereas DQ-parametric homotopy provides state-by-state path certification. A generalized implementation evaluates 27 configurations (21 accepted, six rejected), with all six controlled actuation-order permutations accepted. The framework serves as a pre-prototyping rigid-kinematic qualification tool that separates local axis compatibility from global rigid-origami feasibility. Full article

Bovine viral diarrhea virus (BVDV) is a pathogen of globally significance in cattle that has two biotypes: non-cytopathogenic (non-cp) and cytopathogenic (cp). The cp biotype arises from the non-cp through genome rearrangements, which frequently involve the insertion of host cellular RNA sequences, often accompanied by viral genome duplications. Here, we performed a systematic analysis of insertions across all complete BVDV genomes available in GenBank. Despite a 10-fold increase in available sequences over the past 10 years (from 59 to 670), the repertoire of known rearrangements associated with the cp phenotype has expanded only modestly, with insertions occurring predominantly at five conserved genomic hotspots. Notably, independent acquisitions of similar insertions—such as DNAJC14 (Jiv), ubiquitin-like sequences, the NS4B-NS5A cleavage site, and the PYPDPQTLG motif—in phylogenetically unrelated virus lineages reflect a limited number of permissive sites. Our analysis further demonstrates that many insertions are mosaic and suggests multiple recombination events that are poorly compatible with template switching by viral polymerase. Mechanistically, we propose that non-replicative recombination provides the most consistent explanation for the observed insertion patterns, particularly common coupling of cellular RNA acquisitions with viral sequence duplications. Both the steric proximity of virus replication to the endoplasmic reticulum and the RNA end requirements (2′-3′ cyclic phosphate and 5′-OH) suggest that non-replicative recombination is likely mediated by the endoplasmic reticulum-associated IRE1 RNase and RtcB ligase, which are responsible for the alternative cytoplasmic splicing of cellular mRNA. Full article

Trust Management Systems (TMSs) have recently emerged as a behavioural complement to identity-based approaches in Industrial IoT (IIoT) cybersecurity by evaluating node trustworthiness. Deep Learning (DL)-based TMSs offer favourable detection over heuristic and Machine Learning (ML) models. The computational density of DL models introduces a trade-off between inference fidelity and deployment feasibility, particularly in Edge-Fog-Cloud (EFC) IIoT architectures where latency and resources are constrained. This work proposes an EFC architectural framework that relocates DL inference to the Fog layer, reducing Cloud communication latency and Edge resource exhaustion. A lightweight Long Short-Term Memory (LSTM)-based model derives continuous trust scores from header-derived, flow-aggregated features, with inference latency bounded through fixed-size sliding windows and stateless execution. The system is trained and evaluated on CIC-IoT-2023 across Denial-of-Service (DoS), Distributed DoS (DDoS), Mirai, and benign scenarios. System scalability is assessed through ns-3 network simulation under benign conditions, with full-system behaviour further evaluated under benign, DoS, and Mirai scenarios. Offline evaluation achieves F1-score 0.9996, accuracy 0.9997, ROC-AUC 0.9999, and PR-AUC 0.9997. Architectural evaluation yields a mean inference latency of 0.049 ms, a maximum enforcement latency of 0.120 ms, and a 302 kB deployment footprint. System simulation confirms a benign False Positive Rate (FPR) 0.07% and a maximum detection latency of 0.22 ms. DoS achieves recall 0.99999 and FPR 0.00186, and Mirai achieves recall 0.99997 with FPR 0. This demonstrates that DL-based trust inference is achievable on resource-constrained Fog nodes, establishing the work as a viable solution for trust evaluation in EFC IIoT deployments. Full article

Cancer remains one of the leading causes of morbidity and mortality worldwide, highlighting the need for safe and effective therapeutic strategies targeting multiple oncogenic pathways. Cinnamon (Cinnamomum spp.) contains several bioactive phytochemicals with reported antioxidant and anticancer properties; however, their potential interactions with key cancer-associated signaling proteins have not been comprehensively investigated. In this study, an integrated computational and preliminary experimental approach was employed to evaluate four major cinnamon phytochemicals, namely e-cinnamaldehyde, eugenol, p-cymene, and cinnamic acid. Consensus molecular docking was performed using AutoDock Vina (v1.2.7), Smina (v2020.12.10), and GNINA (v1.3.3) against phosphoinositide 3-kinase (PI3K), nuclear factor kappa B (NF-κB), and mammalian target of rapamycin (mTOR). Docking analyses were complemented by protein-ligand interaction profiling, pharmacokinetic and toxicity prediction (ADMET), and a 100 ns molecular dynamics simulation with MM/GBSA binding free-energy analysis of the selected mTOR-p-cymene complex. In addition, the antioxidant activity and cytotoxic effects of a crude methanolic cinnamon bark extract were evaluated using in vitro antioxidant assays and MTT assays against HCT-116 and HT-29 colorectal cancer cell lines. Consensus docking predicted that all four phytochemicals were capable of interacting with the selected protein targets, although the predicted binding profiles varied among the compounds. Eugenol showed comparatively more favorable predicted interactions with PI3K, p-cymene produced the lowest predicted docking score for NF-κB, and cinnamic acid displayed a comparatively consistent predicted multitarget binding profile across PI3K, NF-κB, and mTOR. ADMET analysis suggested that all compounds satisfied major drug-likeness criteria and exhibited predicted oral bioavailability, although potential cytochrome P450 interactions and hepatotoxicity were predicted for some compounds. Molecular dynamics simulation indicated that the selected mTOR-p-cymene complex maintained a stable binding pose throughout the simulation, while MM/GBSA analysis yielded a modest binding free-energy estimate (ΔG_bind = −4.70 ± 8.20 kcal/mol), which should be interpreted cautiously because of the observed energetic variability. The crude methanolic cinnamon bark extract exhibited antioxidant activity and reduced the viability of HCT-116 and HT-29 colorectal cancer cells in a concentration-dependent manner. Collectively, these findings provide computational predictions of potential interactions between selected cinnamon-derived phytochemicals and cancer-associated signaling proteins and are consistent with the preliminary observation that the crude cinnamon extract exhibits antioxidant activity and cytotoxic effects in colorectal cancer cell lines. However, the computational analyses do not establish direct inhibition of the PI3K/NF-κB/mTOR signaling pathway, and the biological assays were performed using a crude extract rather than isolated phytochemicals. Therefore, further studies using purified compounds, biochemical target validation, pathway-specific cellular analyses, and in vivo models are required to determine whether the predicted protein-ligand interactions contribute to the observed biological activity. Full article

Japanese encephalitis virus (JEV), the prototype member of the JEV serogroup within the genus Orthoflavivirus (family Flaviviridae), is closely related to West Nile virus (WNV), Murray Valley encephalitis virus (MVEV), and St. Louis encephalitis virus (SLEV). To characterize antigenic cross-reactivity within this group, we evaluated 15 region-specific rabbit antisera, previously generated against nearly the entire JEV protein-coding region, by immunoblotting whole-cell lysates from BHK-21 cells infected with WNV, MVEV, or SLEV, with JEV included as a reference. Six antisera (α-E^N-term, α-NS2B, α-NS3^N-term, α-NS3^C-term, α-NS5^N-term, and α-NS5^C-term) robustly recognized homologous proteins across all three viruses. The remaining nine antisera displayed lineage-restricted or virus-specific reactivity: (a) α-C cross-reacted strongly with WNV and MVEV but weakly with SLEV; (b) α-M, α-NS1, α-NS4A, and α-NS4B^C-term cross-reacted with WNV and MVEV but not SLEV; (c) α-E^C-term and α-NS1′ cross-reacted only with WNV; (d) α-Pr cross-reacted exclusively with MVEV; and (e) α-NS4B^N-term showed no detectable cross-reactivity under the experimental conditions used. Notably, α-NS1 and α-NS1′ detected heat-labile multimers of NS1 and NS1′. These serological patterns mirror the established phylogeny of the JEV serogroup, with JEV clustering most closely with MVEV, followed by WNV and then SLEV. Together, these findings provide a comprehensive cross-reactivity map of JEV antigen-directed antisera and establish a practical framework for dissecting antigenic relationships among JEV serogroup members. These results enhance our understanding of orthoflavivirus antigenic evolution and support the future development of improved diagnostics, broad-acting vaccines, and experimental reagents for emerging and re-emerging encephalitic orthoflaviviruses. Full article

Introduction: The development of bivalent mucosal vaccines capable of providing protection against both influenza and SARS-CoV-2 is a major public health focus. While most COVID-19 vaccines target the spike (S) protein, the highly conserved nucleocapsid (N) protein represents a strategic target for cross-reactive, cell-mediated immunity. This study evaluates Corfluvec, an intranasal vaccine candidate based on an attenuated NS1-truncated influenza vector expressing a fragment of the SARS-CoV-2 N protein. Methods: Protective efficacy, including viral load and pathomorphological changes in the lungs and vessels, was evaluated in Syrian hamsters challenged with high and low doses of SARS-CoV-2 (lineage B.1.1). Cross-protective efficacy against homologous and heterologous influenza A strains (H1N1pdm09, H3N2, and A/PR/8/1934) was tested in a lethal murine model. Additionally, immunogenicity of Corfluvec applied via human-compatible delivery device was tested in cynomolgus macaques (Macaca fascicularis). Results: In Syrian hamsters, vaccination significantly reduced viral loads in the lungs and nasal turbinates. Histopathological analysis revealed a preservation of lung vascular integrity: vaccinated animals showed stable CD31 expression and controlled Ki-67 proliferative activity, accompanied by a marked reduction in vasculitis and perivascular edema compared to placebo controls. In mice, the vaccine provided 100% protection against homologous and heterologous influenza virus challenges. In macaques, the two-dose intranasal immunization was well-tolerated and induced significant systemic IgG and mucosal sIgA responses, alongside robust N-specific IFNγ+ T-cell activation. Conclusions: Corfluvec is a promising bivalent vaccine candidate that provides dual protection against influenza and COVID-19. Its ability to limit viral shedding from the upper respiratory tract and to mitigate SARS-CoV-2-induced pulmonary pathology, contributing to the preservation of lung vascular integrity, underscores the utility of mucosal immunization with Corfluvec as a valuable intranasal complement to current systemic vaccination strategies. Full article

Neglected arboviruses dengue (DENV), Zika (ZIKV), yellow fever (YFV), Japanese encephalitis (JEV), and chikungunya collectively affect hundreds of millions of people annually, yet no specific antiviral drug has been approved for any of them. Alkaloids, nitrogen-containing specialized metabolites produced by diverse plant families, have emerged as a promising source of broad-spectrum antiviral scaffolds. This review compiles and critically analyzes over 100 alkaloid-virus pairs across several RNA virus families, providing a comparative mechanistic analysis. Lycorine, narciclasine, emetine, and berbamine, among others, exhibit potent activity against phylogenetically distant viruses, with the most potent activities reported against flaviviruses (narciclasine: EC₅₀ 0.02 µM against DENV, ZIKV, YFV, and JEV; pancratistatine: 0.0063 µM against ZIKV). Structure-activity analysis of multiple alkaloid classes identifies key pharmacophoric features, including the phenanthridine nucleus (lycorine derivatives) and the bis-benzylisoquinoline scaffold (tetrandrine, berbamine), as determinants of antiviral potency, selectivity, and broad-spectrum activity. Genetic resistance data in West Nile virus challenge the widely accepted model of lycorine as a direct nucleoside inhibitor, instead pointing toward the involvement of the membrane-associated NS4A-2K-NS4B replication complex, though direct validation remains limited for other flaviviruses. Converging structural, biochemical, and transcriptomic evidence suggests that ribosome-mediated translational stress may represent an additional host-directed antiviral mechanism for isoquinoline-type alkaloids, though the causal chain from ribosome binding to activation of the integrated stress response and to antiviral effect has not been established. The present analysis highlights that in vivo validation remains limited to a few alkaloid-virus pairs. Unbiased target deconvolution and formal testing of the ribosome/integrated stress response hypothesis stand out as essential research priorities. Full article

The reaction of 2,5-dimethylfuran (DMF) with two equivalents of α-nitrosostyrene (NS) leading to a tricyclic 1,2-oxazine nitrone (TON) has been examined theoretically at the ωB97X-D/6-311G(d,p) computational level. In this case, the domino process should be considered: (i) a [4+2] cycloaddition (42CA) between DMF and NS yielding a bicyclic 1,2-oxazine (BO); and (ii) a second formal [3+2] cycloaddition (32CA) reaction between BO and NS yielding the final TON. Analysis of the reactivity indices shows that DMF and BO generated along the first 42CA reaction are strong nucleophiles, while NS is a strong electrophile. The first 42CA reaction takes place according to a one-step mechanism. In turn, the second 32CA proceeds according to a two-step mechanism through the zwitterionic intermediate ZW. It is worth noting that the activation enthalpy of the significant point (TS-1mn) of the 42CA is very low, 2.17 kcal·mol⁻¹, the reaction being strongly exothermic by −37.71 kcal·mol⁻¹. This cycloaddition reaction is completely meta regioselective and endo stereoselective. The first step of the formal 32CA reaction has an activation enthalpy of 8.48 kcal·mol⁻¹ (TS-21), the overall domino process being strongly exothermic by −40.34 kcal·mol⁻¹. Both TS-1mn and TS-21 are associated with highly asynchronous single bond processes. The high global electron density transfer (GEDT) found at both TSs, higher than 0.32e, points out the high polar character of these cycloaddition reactions, classified as reverse electron density flux (REDF). Electron Localization Function (ELF) analysis of TS-1mn and TS-21 shows that the C2-C3 and C3-N5 bonds are not formed at the same time, while in the intermediate ZW we observed the creation of one of them. Full article