Search Results (684)

Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) poses a serious threat to the swine industry in China. As a major pig-producing province, Shandong requires continuous epidemiological monitoring of PRRSV. To elucidate the molecular epidemiology of the virus, 1621 clinical samples were collected from suspected cases across different regions of Shandong Province between 2023 and 2025, primarily from Tai’an, Linyi, Jining, and Liaocheng. RT-qPCR detection showed that the positive rate for PRRSV-2 was 20.05% (325/1621). Genetic analysis based on ORF5 and NSP2 genes indicated that Sublineage L1C (NADC30-like) was the dominant strain for 38.38% of ORF5 gene and 72.73% of NSP2 sequencing results. This was followed by Sublineage L8E and L1A and L5A strains. Key virulence-related mutations were identified at residues R13 and R151 in the GP5 protein, which are associated with enhanced pathogenicity. Additionally, variations in neutralizing epitope and the number of N-glycosylation sites (ranging from 2 to 5 per strain) suggested potential immune evasion. Notably, 26.79% (15/56) of sequenced samples showed discordant ORF5 and NSP2 genotyping results, indicating widespread recombination among PRRSV strains in Shandong Province. These finding demonstrated that the genetic diversity, high recombination frequency, and key amino acid variations in circulating PRRSV strains collectively undermine vaccine effectiveness. This study highlights the need to optimize vaccination strategies, enhance biosecurity measures, and implement effective disease control and elimination programs to reduce the impact of PRRSV in Shandong Province. Full article

Background/Objectives: Coronaviruses are a family of positive-sense RNA viruses that cause respiratory and gastrointestinal disease in mammals and birds. Their zoonotic nature and high mutability make them a pandemic threat. UNICOR-v is a pre-pandemic, pan-coronavirus vaccine composed of an adjuvanted mix of twelve synthetic peptides originating from conserved regions within Nsp12 and M coronavirus proteins containing clusters of predicted T-cell epitopes. Here, we evaluate the immunogenicity of UNICOR-v and its efficacy against Middle East Respiratory Syndrome-related coronavirus (MERS). Methods: Animals were vaccinated with an adjuvanted equimolar mix of UNICOR-v. Humoral and cellular immunogenicity were assessed 28 days later through ELISA and FLUOROSpot. Vaccine efficacy was assessed in a DPP4 knock-in (HDPP4-KI) mouse model where mice were challenged post-vaccination with a lethal or non-lethal dose of MERS-CoV-MA. Results: Vaccination with UNICOR-v induced high IgG titers in both mice and rabbits and cellular secretion of pro-inflammatory cytokines. Vaccination with UNICOR-v, or passive serum transfer, significantly reduced viral lung titers 4 days post-infection compared to placebo. Vaccination induced lower immune cell infiltration in the alveolar space and increased repair of the cells lining the major airways in vaccinated mice, translating to increased survival rate compared to placebo. Conclusions: These data demonstrate the ability of conserved T-cell epitopes to protect against MERS-CoV infection, supporting further characterization of the breadth of protection of UNICOR-v against other coronaviruses that affect humans and livestock, following a One Health approach to control this highly zoonotic family of viruses. Full article

This work presents the synthesis, characterization, and application of zirconium oxide (ZrO₂)-based catalysts, modified with macro (silica nanospheres, NSP-SiO₂) and mesopore templates (Pluronic 123), impregnated with tungstophosphoric acid (TPA), in the catalytic pyrolysis of tomato agro-industrial residues. The NSP-SiO₂ (SXX) and P123 (PYY) amount mainly influences the ZrO₂SXXPYY-specific surface area (S_BET) and average pore diameter (D_p). ³¹P MAS NMR and FT-IR characterization results show that TPA (H₃PW₁₂O₄₀) was partially transformed into [P₂W₂₁O₇₁]⁶⁻ and [PW₁₁O₃₉]⁷⁻ during the synthesis steps. The acidic properties of ZrO₂SXXPYY samples containing 25 and 50 wt% of TPA (ZrO₂SXXPYYT25 and ZrO₂SXXPYYT50, respectively) are dependent on both the TPA content and the support nature. Bio-oil composition and product selectivity were strongly influenced by the textural and acid-based properties of the catalysts. Notably, non-catalytic pyrolysis favored pathways leading to C2 compounds, with a high content of acetic acid and hydroxyacetone. In contrast, the use of catalysts promoted the formation of higher molecular weight oxygenated compounds (C5–C6), specifically furans, aldehydes, and ketones. Full article

The SARS-CoV-2 public health challenges have highlighted the urgent need for coronavirus-targeting life-saving therapeutics. Given the emergence of drug-resistant strains, the development of antivirals against viral proteins beyond the commonly targeted main protease or RNA-dependent RNA polymerase is critical. The SARS-CoV-2 nonstructural protein 13 (nsp13) is a highly conserved RNA helicase and an essential component of the viral replication–transcription complex (RTC). It unwinds double-stranded RNA to facilitate viral transcription and replication, making it a strong target for drug development. To identify nsp13 inhibitors, we used an ultra-high-throughput nucleic acid unwinding assay to screen a library of FDA-approved drugs and bioactive compounds. We identified forty inhibitors with IC₅₀ values ranging from 1.4 to 10 μM. Ten were further selected for biochemical and biophysical characterization. Four of these are bound to nsp13 without interacting with the nucleic acid substrate and without inhibiting the ATPase activity of nsp13. Hydrogen–deuterium exchange coupled with Mass Spectrometry (HDX-MS) studies show compound binding causes differential exchange in two regions of nsp13. Furthermore, these compounds have antiviral activity against infectious SARS-CoV-2 in multiple cell lines, with cytotoxicity affecting, in some cases, the apparent antiviral effect. Future optimization efforts could help develop therapeutics against SARS-CoV-2 and other potential coronavirus threats. Full article

This study assesses the stability, in vitro bioaccessibility and potential bioavailability, and in vivo genotoxicity and toxicity of polyethylene glycol–soy lecithin (PEGSL-ACG-NSps) or β-cyclodextrin–soy lecithin (βCDSL-ACG-NSps) nanosuspensions (NSps). Both formulations exhibited initial particle sizes below 130 nm and PDI values below 0.3. Under simulated gastrointestinal conditions, PEGSL-ACG-NSps preserved structural integrity, with only a moderate size increase (~239 nm) in the intestinal phase and controlled release of acetogenins (ACGs); in contrast, βCDSL-ACG-NSps destabilized considerably (size > 500 nm) and released ACGs rapidly. Consistently, βCDSL-ACG-NSps achieved higher in vitro bioaccessibility and a potential bioavailability (up to 95% from post-digestion recovery). In contrast, PEGSL-ACG-NSps displayed a more gradual release profile (up to 55%). In vivo toxicity tests in mice showed no significant genotoxic or cytotoxic effects for either formulation, even at high doses. These findings suggest that selecting appropriate food-grade stabilizing polymers is crucial for optimizing NSps for the oral delivery of ACGs as therapeutic agents. Full article

Flesh mealiness, a textural disorder in apples, reduces storage quality and consumer acceptance. The ‘Delicious’ and ‘Fuji’, prominent apple cultivars in China, exhibit contrasting susceptibility to mealiness, though the underlying mechanisms remain unclear. This study compared cytological, physiological and cell wall metabolic changes between mealy ‘Oregon Spur II Delicious’ and non-mealy ‘Miyazaki Spur Fuji’ during ambient storage. Toluidine blue staining and scanning electron microscopy revealed that ‘Delicious’ exhibited larger intercellular spaces and cell separation in contrast to ‘Fuji’. This observation aligns with the earlier onset of mealiness in ‘Delicious’: its mealiness degree increased from 3.06% at harvest to 19.62% after 28 d of storage (a 6.4-fold rise), whereas that of ‘Fuji’ only increased from 2.13% to 3.90% (1.8-fold). This pronounced increase in ‘Delicious’ was accompanied by a significant increase in air space volume and a reduction in expressible juice. Furthermore, the occurrence of mealiness in ‘Delicious’ involved a sharp increase in respiration rate and ethylene production, alongside rapid declines in firmness and starch content. Notably, there was a substantial accumulation of water-soluble pectin (WSP) and chelator-soluble pectin (CSP) in ‘Delicious’, whereas the content of Na₂CO₃-soluble pectin (NSP) remained consistently lower. Monosaccharide composition analysis confirmed significantly reduced arabinose and galactose levels across pectin fractions (WSP, CSP, and NSP) in ‘Delicious’. Correspondingly, immunofluorescence labeling showed a pronounced degradation of arabinan and galactan within the side chains of rhamnogalacturonan-I (RG-I). In addition, the activities of pectin methylesterase, α-L-Arabinofuranosidase, and β-D-Galactosidase remained significantly elevated in ‘Delicious’. Collectively, these findings demonstrate that cultivar differences in flesh mealiness are attributable to divergent physiological senescence and cell wall disassembly processes. Full article

With the ongoing emergence of SARS-CoV-2 variants, continued surveillance of antiviral susceptibility remains critical for detecting resistance that could compromise treatment efficacy. This study evaluated the activity of 2 SARS-CoV-2 RNA-dependent RNA polymerase (Nsp12) inhibitors against emerging Omicron variants: remdesivir (RDV), an approved antiviral for the treatment of COVID-19, and obeldesivir (ODV), an oral prodrug that shares the same parent nucleoside as RDV. Both RDV and ODV were shown to retain antiviral activity against the Omicron subvariants BA.2.86.1, JN.1.7, KP.2, KP.3.1.1, KP.3.3, LP.8.1, NB.1.8.1, XBB.2, XEC, and XFG compared with wild-type reference strains. Only 1 new lineage-defining Nsp12 substitution, D284Y (detected in NB.1.8.1), was observed. Phenotypic analysis demonstrated that a replicon containing this substitution remained susceptible to both RDV and ODV. These findings are consistent with previous studies showing that RDV and ODV retain potent activity against previously identified Omicron variants, support the continued clinical use of RDV against circulating SARS-CoV-2 variants, and reinforce the potential of ODV as an oral antiviral therapeutic. Full article

The detection rate of Porcine Reproductive and Respiratory Syndrome Virus type 1 (PRRSV-1) in China has been increasing, with its growing genetic diversity and evolving pathogenicity posing significant challenges to disease control. In this study, a novel PRRSV-1 strain, designated GD18-2, was identified from a pig farm in Guangdong Province that experienced an outbreak despite vaccination with a PRRSV-2 vaccine. Whole-genome sequencing indicated that the GD18-2 strain possesses a genome length of 14,932 bp and exhibits 81.4% to 83.9% nucleotide identity with classical PRRSV-1 strains. Phylogenetic analyses based on both the complete genome and the ORF5 gene indicated that GD18-2 belongs to a distinct, new lineage. A unique amino acid deletion (positions 306–357) was identified in the non-structural protein Nsp2, along with specific mutations within the hypervariable regions of the structural proteins GP3 and GP4. Pathogenicity assessment demonstrated that GD18-2 induced fever, respiratory symptoms, and mortality in piglets. In pregnant sows, it caused reproductive failure (abortion, stillbirth, weak piglets) and was capable of vertical transmission via the placenta. This study highlights the emergence of a PRRSV-1 strain with a unique genetic background and high pathogenicity in southern China, underscoring the necessity for enhanced molecular epidemiological surveillance and updated control strategies. Recombination analysis using RDP4 revealed no significant recombination events in GD18-2. Full article

G-quadruplexes (G4s) are emerging as potential antiviral targets. SARS-CoV-2 genomic RNA contains 42 G-rich regions harboring putative G-quadruplex-forming sequences (PQSs). Here, we performed a systematic genomic and structural analysis of SARS-CoV-2 PQSs. It was proposed that non-G-tetrads or different triads may stabilize most G4s in this RNA. Many G4s may include the most stable U·A-U triad. Several G-quadruplexes may be significantly stabilized by 3′ U-tetrad. Large-scale mutational analysis of RNA structural elements containing PQSs showed that most PQSs are highly conserved, while persistent G4-destroying mutations were observed only for one PQS and were transient for two others. Based on G4 position and structural context, we propose that: (i) G4 370 in nsp1 may contribute to cap-independent translation initiation; (ii) certain putative G4s in different genes may assist in co-translational folding of viral proteins; (iii) G4 13385, located upstream of the frameshift stimulation element, may promote formation of a pseudoknot competent for −1 frameshifting. For putative G4s at positions 3467, 13385 and 28903, we analyzed binding to 13 compounds by molecular docking and selected four candidates for molecular dynamics simulations. The ligand EKM emerged as a promising antiviral candidate due to its specific binding to G4 3467. Full article

Serine proteases are a class of enzymes that orchestrate an immune response. These proteases can be hijacked by viruses to facilitate entry and spread, while simultaneously supporting the innate immune system in neutralizing pathogens. This review highlights the dual roles of exogenous serine proteases, emphasizing neutrophil serine proteases (NSPs) that facilitate viral entry and promote disease progression while also contributing to antiviral defense by degrading viral glycoproteins. Additionally, the potential to modulate serine protease activity to boost host defenses will be discussed, offering both significant challenges and new opportunities for therapeutic intervention. Full article

Coronaviruses pose a global pandemic threat, making development of a pan-coronavirus inhibitor crucial for preparedness and containment in the event of a new coronavirus outbreak. The 3C-like protease (3CL^pro) is a key target for antiviral development, as it is essential for viral replication and conserved across human coronaviruses. We previously developed an assay to monitor SARS-CoV-2 3CL^pro activity in cells. This assay uses a single vector that coexpresses the 3CL^pro enzyme and the reporter, which consists of two luciferase fragments linked by a 3CL^pro cleavage site. Cleavage of this site by 3CL^pro decreases luciferase activity, whereas inhibition of 3CL^pro increases the luciferase activity. Here, we adapted this assay to examine 3CL^pro activity from six other human coronaviruses: SARS-CoV, MERS-CoV, HCoV-NL63, HCoV-229E, HCoV-OC43, and HCoV-HKU1. We further determined the effects of different cleavage sites to improve the signal-to-background ratio. The Nsp4-Nsp5 site and super-active substrate (SAS) resulted in the largest dynamic range for most coronaviruses in our assay. Using the broad-spectrum 3CL^pro inhibitor GC376, we observed increased reporter activity, indicating the assay’s efficacy for identifying inhibitors across multiple coronaviruses. The adaptation and improvement of the assay can facilitate the development of inhibitors against 3CL^pro from multiple or novel coronaviruses. Full article

3-Amino-4-hydroxybenzoic acid (3,4-AHBA) is a non-proteinogenic aromatic compound that functions as a key biosynthetic precursor for diverse secondary metabolites with pharmaceutical and industrial value. Microbial production of 3,4-AHBA offers a sustainable alternative to petroleum-based chemical synthesis; however, metabolic complexity and trade-offs between growth and product formation constrain rational strain design. Here, genome-scale metabolic (GSM) modeling and flux balance analysis (FBA) were integrated with targeted genetic engineering to elucidate and enhance 3,4-AHBA production in Streptomyces thermoviolaceus. A genome-scale metabolic model was constructed and expanded by incorporating the nspH–nspI gene operon, which encodes the 3,4-AHBA biosynthetic pathway. In silico FBA predicted substantial rewiring of central carbon metabolism, with carbon flux redirected from glycolysis and the tricarboxylic acid cycle toward aspartate-derived intermediates and 3,4-AHBA synthesis, accompanied by reduced biomass-associated flux. Guided by these predictions, an engineered strain (St::NspHI) was developed and experimentally evaluated. Consistent with model predictions, the engineered strain exhibited lower growth rates and glucose uptake than the wild type, reflecting a metabolic burden. Nevertheless, 3,4-AHBA production was achieved exclusively in the engineered strain. Comparison of simulated and experimental fluxes revealed overestimation by FBA, likely due to secondary metabolism and incomplete genome annotation. Overall, GSM-guided design enables optimization of precursor production. Full article

Non-starch polysaccharides, the primary structural component of dietary fiber, play critical roles in metabolic and digestive health through multiple physiological mechanisms, yet their composition in Mediterranean aromatic plants remains poorly characterized, limiting the development of novel functional food ingredients. This study provides the first comprehensive NSP profiling of 22 populations across three Tunisian Satureja species (S. nervosa, S. graeca, and endemic S. barceloi), using enzymatic analysis, gas chromatography, and multivariate statistics. Total non-starch polysaccharides reached exceptional levels (21.5 ± 3.0 g/100 g dry weight (DW)), with several populations exhibiting unprecedented soluble fiber proportions exceeding 50%, including population SG4 achieving 79.7%. Monosaccharide analysis revealed uronic acid dominance (42.9–52.5% of total NSP), indicating pectin-rich cell walls with distinct functional properties. Principal component analysis (explaining 61.5–84.9% of variance) demonstrated that populations cluster by fiber chemotype rather than taxonomic classification. Hierarchical and K-means clustering identified three distinct clusters in the soluble and total fiber fractions, with uronic acid-dominated populations (SG4, SB, SG18, SN8) and arabinose–xylose enriched populations (SN13, SN12, SN22, SG21) as extreme chemotypes. Intraspecific variation (coefficient of variation, CV: 14.0–50.0%) substantially exceeded interspecific differences. These findings establish Tunisian Satureja as an exceptional functional fiber source and demonstrate that population-level chemical screening outperforms taxonomic classification for developing nutraceuticals targeting cholesterol reduction, glycemic control, and gut microbiome modulation. Full article

Highland barley (HB), a nutrient-rich grain, is limited in bread applications due to its weak gluten network and high content of non-starch polysaccharides (NSPs) such as β-glucan and arabinoxylan. This study aimed to improve the dough properties and bread quality of a composite flour containing 40% whole-grain highland barley through synergistic use of xylanase and β-glucanase. Rheological analysis revealed that dual-enzyme treatment significantly reduced dough rigidity (G′ decreased by ~40%) and increased extensibility (tan δ raised by ~25%), shifting the network from a brittle NSP-dominated gel toward an elastic gluten-based structure. Low-field NMR showed that enzymes promoted redistribution of water from tightly bound states with NSPs to protein phases, enhancing gluten hydration. Microstructural observations confirmed a more continuous and uniform gluten network with finely embedded starch granules. Consequently, enzyme-treated bread exhibited a 35% higher specific volume, reduced hardness (~50% lower), improved springiness and cohesiveness, and superior sensory scores in texture, taste, and overall acceptability compared to the untreated composite. Single-enzyme treatments yielded partial improvements, highlighting the necessity of synergistic action. These results demonstrate that combined xylanase and β-glucanase treatment effectively mitigates the negative impact of NSPs, enabling the production of high-quality, sensorially appealing HB-enriched bread with optimized structural and textural properties. Full article

Papain-like protease (PLpro), a crucial functional domain of the SARS-CoV-2 non-structural protein 3 (nsp3), plays a dual role in both hydrolyzing viral polyprotein precursors and modulating host immune responses. These critical functions position PLpro as a key target in the ongoing development of antiviral therapies for SARS-CoV-2. This review analyzes more than 100 PLpro-ligand co-crystal structures and summarizes the major binding modes between these ligands and PLpro. Most of these ligands bind to sites analogous to those targeted by the classical non-covalent inhibitor GRL0617, primarily involving the P3 and P4 subsites and the BL2 loop. Based on these structural insights, optimized inhibitors have expanded targeting beyond the canonical binding site to auxiliary regions such as the BL2 groove and the Val70 site, and in some cases toward the catalytic Cys111 buried within a narrow pocket. Certain ligands identified through various screening approaches bind to non-canonical or allosteric regions, such as the S1 and S2 sites or the zinc-finger domain, engaging PLpro through distinct interaction modes and thereby offering additional opportunities for PLpro inhibitor design. The review also discusses potential strategies for future PLpro inhibitor development informed by recent structural advances. Taken together, these structural and functional insights support ongoing efforts in the structure-guided design and optimization of PLpro inhibitors. Full article