Newcastle disease virus (NDV) represents a major threat to the worldwide poultry industry, and the absence of approved antiviral therapeutics highlights the need for alternative intervention strategies. In this study, an integrated computational and experimental methodology to assess the interaction of selected plant-derived flavonoids against NDV. Molecular docking was conducted to evaluate the interactions of karanjin, mammeigin, and 3, 5-dihydroxy-4′,7-dimethoxyflavone (DHDM) with multiple NDV proteins, comprising hemagglutinin neuraminidase (HN), fusion protein (F), matrix protein (M), nucleoprotein (N), and phosphoprotein (P). Based on docking predictions, the NDV phosphoprotein was selected for experimental validation of biophysical interaction studies. Among the tested compounds, karanjin consistently exhibited the strongest binding affinities, with particularly favorable interactions observed for the phosphoprotein, a key regulator of viral transcription and replication. In silico pharmacokinetic analysis further supported the drug-likeness and oral absorption potential of karanjin. Based on docking predictions, the NDV phosphoprotein was selected for experimental validation. Recombinant phosphoprotein was expressed and purified, and flavonoid interactions were examined using UV–Vis absorption spectroscopy, fluorescence quenching, and isothermal titration calorimetry analysis. Karanjin displayed a clear and specific binding profile characterized by moderate-to-strong affinity and an enthalpy-driven interaction, whereas mammeigin and DHDM showed weak or nonspecific interactions. Spectroscopic analyses corroborated these findings, indicating stable complex formation without major structural perturbation. Collectively, these results identify karanjin as a promising flavonoid candidate targeting the NDV phosphoprotein.

Newcastle disease affects many avian species, predominantly chickens. It is known to cause a substantial economic burden to the poultry industry. The velogenic strain of Newcastle disease virus (NDV) can cause mortality up to 100 %. Real-time PCR and ELISA are currently used to diagnose NDV; however, these techniques have some limitations. To address the limitations, we developed an aptamer-probed paper-based analytical device (ePAD) to detect NDV. The NDV HN protein-specific aptamer was identified following 10 rounds of SELEX. The identified aptamer was investigated for its binding and stability with the HN protein with the help of docking, molecular dynamics simulations, and CD spectroscopy. ZnO nanorods were used to enhance the conductivity of the fabricated ePAD aptasensor. The aptamer was then immobilized on the working electrode to detect NDV. Various concentrations of NDV were used to optimize the fabricated aptasensor. All the electrochemical changes were analysed with the help of cyclic voltammetry and impedance. Our data showed the detection of a very low concentration of NDV using the designed ePAD biosensor. The specificity of the aptasensor was studied using a non-related virus from the samples as a negative control. The developed aptasensor could offer a cost-effective, user-friendly, and specific alternative to detect NDV from clinical samples.

Oncolytic virotherapy emerged as a revolutionary approach to immunotherapy in cancer treatment. Newcastle disease virus (NDV), a promising oncolytic agent of the Paramyxoviridae family, has gained considerable attention as an immunotherapeutic agent against cancer mainly due to its inherent tumor selectivity, broad tropism, convenient propagation, and lack of pre-existing immunity in humans. Its selective tropism is primarily mediated by an impaired type I interferon signalling pathway in cancer cells. Apart from the direct lysis, NDV induces immunogenic cell death (ICD), releasing tumor-associated antigens (TAAs) and damage-associated molecular patterns (DAMPs), which in turn lead to potent activation of innate and acquired immunity. The efficiency of the wild-type NDV strains has been improved through genetic engineering. Recombinant strains capable of expressing immuno-stimulatory cytokines and enzymes to remodel the immuno-suppressive tumor microenvironment (TME) are the most promising. This review discusses the history of early events that shaped NDV into an 'oncolytic agent'. It reviews its characteristics and mechanisms of anti-tumor activity, which can directly or indirectly form part of the cancer immunity cycle. Further, limitations and issues surrounding the unmet translational delay of NDV as an immuno-therapeutic agent in personalized and combination immunotherapy strategies are also discussed.

African swine fever (ASF) is a viral disease of domestic pigs and wild boars with a high fatality rate. The causative agent of this disease is the African swine fever virus (ASFV), which continues to represent a major threat to the global swine industry, for which therapeutic options remain scarce. Among ASFV's structural proteins, p54 plays a crucial role in host-virus interactions, although its involvement in host cell stress responses and apoptosis remains incompletely elucidated. In the present study, we examined the functional characteristics of the ASFV p54 protein and assessed a nucleic acid aptamer-based approach to mitigate its pathogenic effects. The ASFV p54 gene was cloned into a mammalian expression vector (pcDNA 3.1-p54) and transfected into the porcine macrophage cell line 3D4/31 to assess its effects on host cells. p54 expression strongly induced apoptosis, accompanied by increased intracellular and mitochondrial ROS levels, which are indicative of cell death and mitochondrial stress. To corroborate these results in a viral context, a recombinant Newcastle disease virus (NDV) expressing ASFV p54 (rNDV-p54) was constructed and employed to infect porcine macrophages. The rNDV-p54 infection mirrored the aforementioned cellular responses, affirming pro-apoptotic and oxidative stress-inducing properties of ASFV p54 protein. Subsequently, for therapeutic purposes, the ASFV p54 protein-targeted aptamer was screened and evaluated. The selected aptamer inhibited p54-induced apoptosis in 3D4/31 cells, accompanied by reduced ROS levels. Collectively, our findings identify ASFV p54 protein as a major inducer of oxidative stress-dependent apoptosis in host cells and show that targeted nucleic acid aptamers can potently neutralize these impacts. The findings highlight an aptamer-based approach with high binding affinity for the ASFV p54 protein, underscoring its potential for diagnostic applications and as a platform for subsequent antiviral research. Nevertheless, additional research is essential to assess its antiviral effectiveness in ASF infections via appropriate in vitro and in vivo models.

The hemagglutinin-neuraminidase (HN) of Newcastle disease virus (NDV) is a multifunctional protein that mediates virus attachment to host sialic acid-containing receptors, exhibits neuraminidase activity, and promotes membrane fusion. Notably, purified HN protein also targets sialic acids on malaria-parasitised red blood cells (PRBCs), thereby reducing parasite viability, and is thus a potential therapeutic molecule against the malaria parasite. To gain further insights into the structural and functional basis of its sialic acid recognition, we performed comparative analyses of the sialic acid-binding pocket across different viral lectins. It revealed a Type-VI sialic acid-binding module in HN, characterised by fewer interactions with the functional groups of sialic acid compared to Type-I influenza neuraminidases (NA). Introduction of a rationale-based specific mutation (I175Y) in the central binding pocket of HN transformed its sialic acid-binding module from Type-VI to Type-I. It resulted in significantly higher affinity for sialic acid, accompanied by a substantial loss in its neuraminidase activity. This disruption in the delicate functional balance between receptor-binding and enzymatic cleavage highlights a trade-off between optimising one activity at the cost of another. Biologically, the mutant HN exhibited enhanced hemadsorption, or cell-binding activity, and thus showed greater ability to block NDV replication in both cell monolayers and in-ovo systems. The mutant protein also demonstrated an increased capacity to target PRBCs and showed potent anti-plasmodial activity, compared to the wild-type. This enhanced targeting capability highlights the mutant as a promising candidate for developing precision drug delivery systems for malaria.

Newcastle disease virus (NDV) remains a substantial threat to the worldwide poultry industry. Targeting ion homeostasis is studied as a potential antiviral strategy to prevent viral infection and treat channelopathies. Quinoline derivatives, such as 8-aminoquinolines (AQ), are well-established scaffolds in medicinal chemistry with known antiviral properties. The study introduces a novel series of AQ derivatives and evaluates their transmembrane zinc (Zn²⁺) transport activity. Our results suggested that a potent Zn²⁺ transporter, AQ-CN, effectively suppressed NDV replication post-infection, both in vitro and in ovo, with reduced hemolytic activity and potent inhibitory concentration. Additionally, elevated Zn²⁺ levels affected host epigenetic regulators, downregulating Class I histone deacetylases (HDAC1, HDAC2, HDAC3, HDAC8) and reducing global lysine acetylation. Furthermore, in the presence of Zn2 +, AQ-CN increased type I interferon signaling and upregulated antiviral cytokine genes. Pharmacological inhibition with valproic acid, a class I HDAC inhibitor, largely restored the antiviral phenotype, indicating that HDAC-dependent mechanisms are involved in the observed effect. Overall, our findings point to AQ-CN-mediated transmembrane transport as a viable anti-NDV strategy and support a model in which zinc-regulated host Class I HDAC pathways help combat NDV replication.

Tau306-311, also known as Ac-PHF6 (CH3CO-VQIVYK-NH2), is a short peptide that forms a viscous solution in water but causes instant gelation of PBS and cell culture media. Ac-PHF6 analogs wherein tyrosine’s phenolic group is substituted with phenyl or electron-deficient aromatic rings, also form hydrogels. These data indicate that the aromatic residues in these peptides may not contribute to self-assembly through their ring electronic effects, but through their hydrophobicity. We test this hypothesis by investigating Ac-PHF6 analogs wherein Tyr is substituted with several aliphatic amino acids with different side-chain hydrophobicities, viz. Lys, Ala, Met, Val, and Ile. Ac-PHF6* (tau275-280: CH3CO-VQIINK-NH2), an aliphatic tau hexapeptide motif similar to Ac-PHF6, was also investigated. The peptides were synthesized via solid-phase peptide synthesis using Fmoc chemistry. Peptide stock solutions were prepared in water and diluted in PBS. The resulting PBS hydrogels were characterized using oscillatory rheology, ThT fluorescence, CD, and IR spectroscopy, while their supramolecular architectures were examined using TEM. The cytocompatibility of the peptides was evaluated using HEK-293 cells. All peptides, except for the Y310K analog, caused instant gelation of PBS. The Y310K analog formed a gel after approximately 2 h of incubation. Additionally, the Y310K hydrogel exhibited significantly lower stiffness compared to the other hydrogels. The other peptides formed hydrogels with stiffness higher than that of Ac-PHF6 hydrogel. Except for Ac-VQIVKK-am, all peptides formed amyloid-like fibrils, as revealed by ThT fluorescence spectroscopy and congo red spectral shift assay. All peptides were found to be cytocompatible with HEK-293 cells. Tyr residue is not essential for Ac-PHF6 hydrogelation. Tyrosine’s contribution to Ac-PHF6 self-assembly and hydrogelation is through its hydrophobicity rather than aromaticity.

AbstractAnatid herpesvirus 1 (AnHV-1) is still a significant limiting factor in the waterfowl industry and there is a clear demand for field-applicable serodiagnostic assays that are robust, sensitive and reproducible. In this report, we designed and conducted a comprehensive comparative evaluation of whole virus or recombinant glycoprotein C (gC), glycoprotein D (gD) alone or a mixture of gC-gD ELISA, to determine which one had the better diagnostic performance. The recombinant proteins were produced, purified, and optimized for use in indirect ELISA, and subsequent validation was carried out using a set of 120 field serum samples. Receiver operating characteristic (ROC) curve analysis demonstrated that the combined gC-gD ELISA exhibited an almost perfect diagnostic performance (AUC = 0.9925), and high sensitivity (93%) and specificity (95%) surpassed those of two antigen-based assays and whole virus assay. Although both gC ELISA (high sensitivity 95% but moderate specificity 80%) and gD ELISA (outstanding sensitivity 99%, but lower overall accuracy: AUC = 0.9150) performed well, the sensitivity of the whole virus ELISA was lower (87%) with marked variation. In particular, intra-assay precision analysis showed a significant decrease in coefficient of variation for the gC-gD ELISA (∼4%) compared with the gD (∼15%), gC (∼20%) and whole virus (∼40%) formats, indicating the highest reproducibility of the gC-gD ELISA. Taken together these results demonstrate that the gC-gD ELISA provides a novel and robust tool for high-throughput AnHV-1 serosurveillance which represents a major breakthrough in terms of economy, when compared to traditional whole virus based ELISAs for large field applications.

Anatid alphaherpesvirus 1 (AnHV-1) is an emerging threat that induces severe disease and death in both domestic and wild waterfowl. Birds have an elevated basal body temperature, and suffer at times from environmental heat stress, yet little are known about the mechanisms involved in avian virus replication at higher temperatures. In this study, we examined the effects of hyperthermia on replication dynamics and host cellular responses to AnHV-1 in chicken embryonic fibroblast cultures and an in ovo model. Hyperthermic conditions dramatically increased AnHV-1 replication, viral gene expression and protein accumulation in vitro and in ovo. Elevated temperature was associated with coordinated changes in metabolic and mitochondrial parameters indicative of elevated glucose uptake, modulated expression of key glycolytic enzymes, elevated ATP production, and altered intracellular NADPH levels consistent with increased redox demand. Hyperthermia was associated with increased mitochondrial membrane potential, ATP production and reactive oxygen species generation while reducing the expression of selected antiviral immune-response genes. These changes were suggestive that thermal stress remodels intracellular environment to promote viral replication. Pharmacological inhibition of these pathways reduced viral replication, supporting their potential contribution to enhanced AnHV-1 replication under elevated temperature.

Insulin aggregation compromises formulation stability and patient safety. Surfactin-inspired peptides Pep 7 and Pep 13 inhibit insulin fibrillation, restore native structure, and preserve bioactivity, depicted by ThT, CD, ITC, TEM, MD, and cell culture.

Background: Salmonella Typhimurium is a major pathogen causing non-typhoidal salmonellosis in humans. Poultry is a major reservoir of S. Typhimurium. Currently available vaccines against S. Typhimurium are not very effective. Therefore, the search for novel adjuvants to improve vaccine efficacy is a priority for developing effective and efficient vaccines. Method: In this study, next-generation adjuvants, such as calcium phosphate nanoparticles, are being evaluated. Our objective was to assess the potential of calcium phosphate nanoparticles, using outer membrane proteins of Salmonella Typhimurium as antigens, for immune-potential testing in poultry, with Montanide as a control. The toxicity of the prepared vaccine formulation was evaluated in rats. Results: CaP-Omp-Nps in the 30-45 nm size range showed a protein entrapment efficiency of 42.5% and a loading capacity of 50.3%. Both vaccinated groups, calcium phosphate outer membrane protein nanoparticles (CaP-Omp-Nps) and Montanide, induced an efficient humoral immune response, with mean titers of 3.48 + 0.0245 and 4.9 + 0.0142 on the 15th day, 3.5 + 0.0118 and 4.79 + 0.009 on the 30th day, and 4.48 + 0.427 and 5.31 + 0.154 on the 45th day post vaccination, respectively, indicating an improvement (CaP-Omp-Nps group) or stability (Montanide group) over the study period. Further, the CaP-Omp-Nps group revealed a better cell-mediated immune response than the Montanide-Omp group. The toxicity study in rats showed no significant differences in serum biomarkers and blood chemistry parameters, indicating that the nano-vaccine formulation is non-toxic and safe. Outer membrane proteins of Salmonella Typhimurium, when used with a few conventional adjuvants, could not produce a balanced Th1 and Th2 immune response against Salmonella Typhimurium. Conclusions: In this study, we developed a novel nano-vaccine formulation composed of outer membrane proteins of Salmonella Typhimurium and calcium phosphate nanoparticles. The vaccine formulation was found to be safe and could elicit the desired Th1 and Th2 immune responses, as evidenced by humoral, cell-mediated, and protective immunity produced by the nano vaccine in poultry. Therefore, the present findings suggest that the CaP-Omp-Nps vaccine may be an efficient, safe, and cost-effective vaccine against Salmonella Typhimurium.

The safe disposal of pathogenic waste from H1N1 outbreaks poses challenges for municipal solid waste (MSW) landfill operations. Needle-punched (NP) geosynthetic clay liners (GCLs) containing granular bentonite (GB) are widely used to contain various contaminants. However, GB particle size distribution, needle-punching reinforcement, and exposure to saline leachates can adversely influence hydraulic performance. Understanding the sorption, hydraulic, and diffusion characteristics of H1N1 under field conditions is essential. Current research on pathogenic waste behavior in MSW landfill environments, particularly the impact of GB size on osmotic efficiency in the presence of viral contamination, remains limited. This study employed H1N1 to evaluate the sorption capacity, hydraulic behavior, and diffusion parameters of GBs with varying grain sizes, alongside powdered bentonite (PB). Additionally, the study assessed the role of NP fibers in GCLs on H1N1 permeation under chemo-mechanical loading. Results show robust sorption across all GBs, with the finest GB and PB achieving optimal diffusion and retardation characteristics; in contrast, coarser GBs significantly increase fluid permeation rates. The GCL exhibited approximately one order of magnitude higher permeation under identical chemo-mechanical loading due to preferential flow through the NP fibre. These findings provide mechanistic insights into bentonite–virus interactions and serve as a preliminary basis for designing barrier systems to manage pandemic-related wastes safely.

Wild waterfowl constitute the primary natural reservoir of influenza A viruses, and wetlands at the convergence of major migratory flyways serve as critical hubs for viral genetic exchange. Baikal Siberia, situated at the intersection of the East African-West Asian, Central Asian, and East Asian-Australasian flyways, represents a unique yet understudied region in this context. Here we report the results of long-term virological surveillance of wild birds in the Lake Baikal basin conducted between 2018 and 2024. A total of 1036 cloacal swab samples from 28 bird species were screened, yielding 42 influenza A virus isolates belonging to 12 HA/NA subtype combinations: H1N1, H3N1, H3N2, H3N5, H3N6, H3N8, H4N6, H6N1, H6N2, H6N3, H6N8, and H12N5. Among the detected subtypes, H6 viruses-identified with four distinct neuraminidase combinations (N1, N2, N3, N8)-are of particular public health relevance owing to their documented capacity for dual-receptor binding and potential for zoonotic transmission to mammals, including humans. Full-genome sequencing followed by cluster analysis of internal gene segments identified 16 distinct segment constellations, indicating extensive reassortment. BLAST searches against the GISAID database revealed closest genetic relatives in Mongolia, South Korea, Japan, China, and Western Siberia, with more distant links to Bangladesh, Europe, and a possible intercontinental connection via the Pacific flyway. Maximum-likelihood phylogenetic analysis of the HA and NA segments confirmed that all isolates belong to the Eurasian genetic lineage, yet they are distributed across multiple clades rather than forming a single monophyletic group, reflecting the role of Buryatia as a mixing zone for genetically diverse viral populations. These findings substantially expand the understanding of influenza A virus ecology in the Lake Baikal basin and underscore the importance of continued surveillance at this key migratory crossroads in Northern Asia.

Arginylation is a well-known, evolutionarily conserved, and recognizable post-translational modification (PTM). In this modification, an arginine residue is enzymatically attached to the target residue, such as aspartic acid (D), glutamic acid (E), and oxidized cysteine (Cys-SO₂H, Cys-SO₃H) by arginyltransferase 1 (ATE1) utilizing arginyl-tRNA. This phenomenon occurs both at the N-terminal and mid-chain of a protein. Arginylation is required for various biological activities like overall cellular development, cytoskeletal organization, and cellular stress adaptation. For predicting arginylation likelihood, only laboratory-based experimental procedures are currently available. To solve this, we developed the web server ARGpredict to predict arginylation events in protein sequences. The designed web server analyzes the arginylation target sites and assigns specific scores. To evaluate its effectiveness, this web server was tested against various known and experimentally validated datasets. ARGpredict yielded promising prediction results, which were also verified by statistical methods. The web server is available at https://iitg.ac.in/sachinku/ARGpredict/.

Viral co-infections are emerging as major contributors to disease outcomes; however, their biological implications for avian hosts remain poorly described. Birds often encounter more than one pathogen simultaneously, thereby promoting the possibility of complex viral interactions. In this report, we studied the impactof co-infection with two widely spread avian pathogens Anatid herpesvirus-1 (AnHV-1) and Newcastle disease virus (NDV) using an in vitro and in ovo models. Co-infection dramatically altered the dynamics of viral replication and host immune responses compared with single-virus infections. Most importantly, co-infected cells displayed disrupted expression of major pro-inflammatory cytokines and immune modulatorssuch as IL-1β, NLRP3, IL-18, TNF-α, NF-κB, IFN-α and -β together with CH25H implying increased inflammatory signalling and tissue injury. These results reveal that AnHV-1-NDV co-infection aggravates immune disturbance and potentially exacerbates the severity of disease, implicating the importance of accountingfor viral co-infections in avian disease monitoring, diagnosis and control.

Efforts for developing biocompatible and efficient photosensitizers (PSs) that enable Reactive Oxygen Species (ROS) (Type-I and Type-II) generation have received increased attention due to their comprehensive therapeutic utility. This work presents six novel metal-free ionic PSs based on peri-functionalized cationic naphthalimides (RNICs) viz. R = -H (HNIC), -Ph (PNIC), -PhNH2 (ANIC), -PhN(CH3)(2) (NNIC), -PhN(Ph)(2) (TNIC), -PhN(BiPh)(2) (BNIC), engineered by modulating donor units to achieve diverse photophysical properties and specific ROS generation in cancer cells. They exhibited large Stokes shifts (113-245 nm), solid-state white light emission capabilities, and rare photoredox catalytic efficiency. Among them, HNIC exhibits solid-state white light emission (CIE: 0.34, 0.33), while PNIC achieved a photoluminescence quantum yield (Phi(PL)) of 0.99 in aqueous media, far surpassing typical aggregation-induced emission (AIE/AIEE) luminogens. HNIC also introduces the previously unexplored excimer-driven phenomena for new PS design due to its unique excimer formation. Although ANIC demonstrates negligible Phi(Delta) (0.006), NNIC displays the highest singlet oxygen quantum yield (Phi(Delta)) of 0.95 in aqueous media. TNIC/BNIC demonstrated high photoredox catalytic efficiency in cancer cells (IC50: 700 nM/1.67 mu M). Thus, careful tweaking of the functional units, anion-pi(+) interactions, and the imidazole unit, are very crucial for obtaining superoxide (O-2(center dot-)) generating photoredox catalysts and anti-cancer therapeutics, facilitated by rare triplet-ground state splitting energy (type-I PS design).

The Japanese encephalitis virus (JEV) remains a major cause of viral encephalitis in Asia, with significant morbidity and mortality. This review offers a comprehensive overview of the current landscape of JEV research, focusing on its genomic structure, protein composition, and global epidemiology. We highlight the complexity of JEV transmission and pathogenesis, examining the interplay of demographic factors and geographic spread. In particular, we assess the evolution of diagnostic methodologies from traditional molecular and serological techniques to emerging biosensor-based approaches, emphasizing advancements in sensitivity and rapidity. The application of CRISPR/Cas systems for JEV detection marks a promising frontier in molecular diagnostics. Additionally, we review the current status of JEV vaccines, discussing recent innovations in vaccine development aimed at enhancing immunogenicity and accessibility. Beyond prevention, a spectrum of antiviral strategies-including direct-acting antivirals, entry inhibitors, host-directed modulators, neuroprotective agents, and steroidal/synthetic compounds-has demonstrated potent in vitro and in vivo efficacy, targeting viral enzymes, structural proteins, and host pathways. This review underscores the critical role of advanced detection strategies and vaccines in controlling JEV, offering insights into ongoing efforts to mitigate its impact in endemic regions.