Search Results (4,994)

Environmental heat stress disrupts cellular homeostasis through coordinated molecular responses involving protein quality control, oxidative stress regulation, inflammatory signaling, and water homeostasis. Although saliva plays an essential role in maintaining oral homeostasis, the molecular pathways underlying salivary adaptation to heat stress remain insufficiently characterized. We hypothesized that integrating the human salivary proteome with systems biology approaches would identify a reproducible heat-stress-responsive molecular signature. Human salivary proteomic datasets retrieved from the Human Salivary Proteome Wiki were integrated into a non-redundant dataset comprising 15,594 protein accessions corresponding to 2540 distinct proteins. Following deduplication, functional annotation, and biological curation, proteins associated with heat stress response, oxidative stress regulation, inflammatory signaling, water homeostasis, salivary secretion, and mucosal protection were assembled into a Salivary Climate Stress Panel (SCSP). Protein–protein interaction and functional enrichment analyses were performed, and the biological relevance of the identified proteins was independently evaluated using a human heat stress transcriptomic meta-analysis comprising 322 comparisons and the GEO dataset GDS3004. Functional filtering identified 2050 heat-stress-associated salivary proteins, from which a curated SCSP of 35 proteins was established. Network analysis identified HSP90AB1, HSP90AA1, and IL1B as the principal hub proteins linking heat stress, oxidative stress, inflammatory signaling, and water homeostasis pathways, while transcriptomic validation confirmed consistent activation of representative SCSP genes under heat stress conditions. These findings provide a reproducible systems-level framework for investigating heat-stress-responsive molecular pathways in the human salivary proteome and support future studies on salivary biomarkers of environmental heat exposure. Full article

Peste des petits ruminants virus (PPRV) causes a highly fatal disease that severely impacts small ruminant production and global food security. This study aimed to develop a rapid, user-friendly colloidal gold immunochromatographic test strip for detecting PPRV-specific antibodies using a double-antigen sandwich format. Bioinformatic analysis using DNASTAR Protean was performed to predict candidate antigenic regions in the PPRV H and N proteins. Three predicted candidate regions from each protein were selected and incorporated into the design of the recombinant fusion antigen PPRV-H₃N₃EP. The recombinant antigen was expressed in E. coli, purified, and refolded to obtain a final concentration of 8.52 mg/mL. The strip was assembled with colloidal gold-labeled fusion protein as the detection probe and unlabeled protein coated on the test line, plus an independent mouse IgG/goat anti-mouse IgG control system. Performance evaluation showed that the results were readable within 10–15 min. The strip showed satisfactory analytical sensitivity and cross-reactivity performance, consistent qualitative results in within-batch repeatability testing, and preliminary short-term storage stability. In a comparative evaluation using sheep serum samples and a commercial competitive enzyme-linked immunosorbent assay (ELISA) kit, the overall agreement reached 97.9%. Collectively, the constructed PPRV-H₃N₃EP antigen enabled a simple, rapid, and reliable strip assay suitable for field detection of PPRV antibodies and post-vaccination monitoring, while also providing a methodological reference for developing antibody tests for other pathogens. Full article

Euonymus alatus is a species of medicinal and ornamental value, yet high-quality chloroplast genome resources for this taxon remain scarce. The complete chloroplast genome of E. alatus was assembled and compared with 15 congeneric species to investigate genomic structure and evolutionary dynamics. The genome is 157,416 bp with a GC content of 37.3%, containing 131 genes and 156 repeats (67 tandem repeats, 49 dispersed repeats, and 40 SSRs). Divergence was concentrated in non-coding regions, with 14 hypervariable regions identified as potential markers. IR expansion occurred independently in E. fortunei and E. japonicus, while no genome-wide inversions were detected. 11 protein-coding genes were identified under positive selection, among which clpP exhibited the strongest signal, suggesting a possible role in adaptive evolution. Codon usage bias analysis revealed that both mutation pressure and natural selection shape codon usage patterns, with the latter playing a relatively prominent role; 17 optimal codons were identified. Phylogenetic analysis strongly supported E. alatus as sister to E. phellomanus. This study provides a valuable genomic resource for species authentication, phylogenetic revision, and breeding in this genus. Full article

Osteoarthritis is an irreversible degenerative joint disease driven by sustained NF-κB-mediated inflammatory responses, and conventional intra-articular hyaluronic acid or small-molecule NF-κB inhibitors cannot achieve targeted on-demand treatment due to poor solubility, rapid clearance and lack of lesion microenvironment responsiveness. OA with inherent anti-chondrolytic activity can self-assemble into nanocarriers in water, yet it lacks stimuli-responsive capacity. Herein, we rationally designed and synthesized an OA-Der by covalently conjugating o-phenylenediamine fragments to the OA backbone. ¹H NMR and HRESI-MS spectra fully verified the accurate chemical structures of intermediate and final OA-Der. Blank OA-Der micelles exhibited uniform spherical core–shell nanostructures (50–150 nm) under TEM and AFM, while pathological high NO triggered complete disassembly of micellar assemblies. We further co-assembled OA-Der with NF-κB inhibitor BAY 11-7082 to construct NO-responsive BAY@OA-Der supramolecular micelles. In vitro experiments using human C28/I2 chondrocytes with LPS-induced inflammatory injury demonstrated that BAY@OA-Der significantly improved cell viability and reduced apoptotic chondrocyte proportion. At mRNA and protein levels, the supramolecular micelle formulation remarkably suppressed NF-κB p65 phosphorylation, downregulated cartilage-degrading ADAMTS5, and upregulated ACAN compared with free BAY or blank OA-Der. Collectively, this natural bioactive self-assembled NO-responsive delivery platform achieves synergistic anti-inflammatory and matrix-protective effects by precisely releasing drugs at NO-overexpressed osteoarthritis inflammatory sites and offers an in vitro design strategy for osteoarthritis responsive delivery systems. Full article

Deep eutectic systems (DESs) have evolved from sustainable solvent alternatives to promising bioactive platforms with reported antimicrobial, anti-inflammatory, regenerative, cryoprotective, and cytoprotective properties. However, despite the growing number of biological studies, the mechanistic basis of these effects remains poorly understood because biological activity is still interpreted predominantly from the chemical identity of the hydrogen-bond donor and acceptor, rather than from the supramolecular organization of the eutectic system itself. This review is intended to provide anyone interested in the biomedical and pharmaceutical applications of DESs with a conceptual framework for understanding how supramolecular organization may influence the biological performance of DES-based systems, without requiring extensive expertise in physical chemistry. It critically analyzes the current evidence linking DES structure with biological function. The literature reveals that many reported biological responses cannot be fully explained by the properties of the individual constituents alone, supporting the existence of emergent physicochemical behavior associated with eutectic formation. Current evidence further demonstrates that DESs are dynamic supramolecular systems characterized by hydrogen-bond networks, nanoscale heterogeneity, hydration-dependent structural rearrangement, and persistent local organization under biologically relevant conditions. These structural features generate localized physicochemical microenvironments capable of modulating membrane organization, protein hydration, osmotic balance, and biomolecular interactions, providing a plausible mechanistic basis for the diverse biological effects reported to date. Our analysis also highlights a fundamental disconnect between the extensive physicochemical characterization of DESs and the predominantly composition-based interpretation of their biological activity. While conventional Quantitative Structure–Activity Relationship (QSAR) approaches rely on molecular descriptors of individual components, they fail to capture the higher levels of organization that characterize these dynamic multicomponent systems. Based on concepts established in supramolecular chemistry, self-assembled biomaterials, colloidal science, and soft matter, we propose a Hierarchical Structure–Activity Relationship (H-SAR) framework in which biological activity emerges from successive levels of organization extending from molecular composition and hydrogen-bond networks to nanostructural organization, hydration-dependent restructuring, localized physicochemical microenvironments, and biological interfaces. This framework provides a mechanistic basis for interpreting DES bioactivity and could offer a conceptual roadmap for the rational design, predictive modeling, and biomedical translation of next-generation bioactive deep eutectic systems. Full article

Background: Clematis tangutica (Maxim.) Korsh. is a Tibetan medicinal plant, but its chloroplast genome and plastid gene evolution remain unexplored. Here, we assembled the complete chloroplast genome of C. tangutica and investigated the adaptive evolution of the ycf2 gene. Methods: Chloroplast DNA was extracted from fresh leaves and sequenced on the Illumina HiSeq PE150 platform. We then performed comprehensive genomic analyses, including genome structure characterization, repeat and SSR identification, comparative genomics, and positive selection analysis of the ycf2 gene. Results: The genome is 159,584 bp with a typical quadripartite structure, containing 134 genes and 23 SSRs. Comparative analyses revealed that ycf2 is the most variable gene among the 13 divergent loci identified. Positive selection analysis detected 12 significant sites in ycf2, all clustered in the middle region of the protein. Conclusions: This study provides the first complete chloroplast genome resource for C. tangutica, offers new insights into the adaptive evolution of ycf2 in Ranunculaceae, and yields molecular markers applicable to population genetics, phylogenetic studies, and conservation planning for this species of medicinal importance. Full article

Hydrophobins are small, cysteine-rich amphipathic proteins predominantly produced by filamentous fungi, known for their ability to self-assemble at hydrophobic–hydrophilic interfaces. These proteins are essential for fungal development, surface interactions, pathogenicity, and environmental adaptation, and they have attracted growing interest for biotechnological applications. In this work, we provide a narrative review of fungal hydrophobins, based on a systematic literature search and manual curation of eligible studies integrated with protein database records from Ascomycota and Basidiomycota. Information was compiled from peer-reviewed publications selected according to predefined eligibility criteria and complemented with UniProt records, covering taxonomic distribution, functional and biophysical properties, and physiological and pathogenic roles. Significant diversity in molecular features and physicochemical profiles was observed, indicating functional specialization across different ecological niches and lifestyles. Additionally, the compiled data highlight various biotechnological applications, such as surface modification, enzyme immobilization, drug delivery systems, biomaterial development, and environmentally sustainable technologies. By consolidating molecular, functional, and applied information across a wide range of fungal species, this review provides a comprehensive reference framework for hydrophobin research and biotechnological innovation. Full article

Protein aggregation strongly depends on the local environment, yet investigating it in compartmentalized, high-surface-area volumes remains challenging. Here, we present an optically controlled platform for inducing and monitoring protein aggregation in microliter droplets. Local pH changes are generated in situ via photoconversion of 2-nitrobenzaldehyde, enabling noninvasive environmental control without mechanical mixing or direct perturbation of the sample. By combining fluorescein-based pH measurements with Raster Image Correlation Spectroscopy (RICS), pH variations and protein diffusion were monitored. As a model system, we investigated β-lactoglobulin, whose association state strongly depends on pH. Light-induced acidification progressively shifted the protein toward its isoelectric region, resulting in a measurable decrease in the diffusion coefficient consistent with the formation of larger supramolecular species. Furthermore, modulation of electrostatic interactions through NaCl addition revealed the sensitivity of the system to environmental factors governing the balance between electrostatic repulsion and short-range attractive interactions. Under these conditions, the platform enabled the observation of aggregation behaviors that were significantly less pronounced in corresponding bulk conditions. Overall, this droplet-based approach allows for the precise manipulation of local chemistry and quantitative analysis of biomolecular self-assembly, making it readily extendable to various noninvasive, time-resolved monitoring applications. Full article

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) selectively triggers apoptosis in malignant, infected, or stressed cells while sparing normal tissues, making it an attractive therapeutic candidate. However, many tumors exhibit intrinsic or acquired resistance to TRAIL, driven by reduced DR4/DR5 surface expression, elevated decoy receptor levels, dysregulated DISC assembly, overexpression of c-FLIP and anti-apoptotic Bcl-2 family proteins, or activation of survival pathways such as NF-κB, PI3K/Akt, and MAPK. Moreover, TRAIL receptors can initiate non-canonical signaling pathways that promote migration, invasion, and metastasis in specific oncogenic contexts, thereby further limiting therapeutic efficacy. We aimed to integrate mechanistic insights into TRAIL biology with current therapeutic advances, providing a comprehensive framework for understanding resistance and for designing rational TRAIL-based combination strategies. We summarized the structural and signaling features of TRAIL receptors, outlined the major determinants of TRAIL sensitivity, and evaluated predictive biomarkers that may guide patient selection. In addition, we examined next-generation TRAIL agonists and targeted delivery systems developed to enhance receptor clustering, pharmacokinetics, and tumor specificity. Together, these insights highlight the therapeutic promise of mechanistically informed TRAIL combinations. A deeper understanding of resistance pathways and biomarker-guided stratification will be essential for restoring apoptotic competence and improving clinical outcomes. Full article

Feeds rich in protein are the dominant nitrogen input in intensive aquaculture, yet only part of dietary nitrogen is retained as animal biomass; the remainder enters water and sediment through uneaten feed, feces, dissolved wastes, mucus, sloughed tissue, and branchial ammonia excretion. This review aims to integrate nutritional, physiological, microbial, and disease-related evidence into an evidence-graded framework that positions ammonia nitrogen as a reactive node linking feed, host, water, sediment, and pathogen risk. To assemble this evidence, we conducted a structured narrative search of Web of Science and PubMed for records in English or Chinese published from 2006 to July 2026, with no restriction on publication type, and classified evidence as direct, indirect, or conceptual. The strongest evidence shows that dietary protein supply, amino acid balance, digestibility, and feeding regime regulate nitrogen retention and ammonia output, while microbial ammonification, nitrification, denitrification, dissimilatory nitrate reduction to ammonium, anammox, and assimilation determine whether reactive nitrogen is regenerated, retained, or removed. Experimental studies further show that ammonia impairs oxidative balance, mucosal barriers, immunity, and disease resistance. In contrast, evidence that pathogen infection quantitatively alters nitrogen retention, ammonia excretion, organic nitrogen release, and sedimentary ammonium regeneration remains limited and largely indirect. Accordingly, ammonia nitrogen is framed as a measurable reactive node rather than a unique source or a universally validated causal loop. Practical management should combine precision nutrition with water, biofloc, sediment, and disease surveillance, while future factorial studies and isotope tracer studies should quantify the complete nitrogen budget under pathogen challenge. Full article

The present study was performed to determine the antibiotic resistance genes (ARGs), virulence determinants, and mobile genetic elements in multidrug-resistant Escherichia coli CUK-76 isolated from wastewater in Himachal Pradesh, India. Whole genome sequencing was performed using the Illumina Miseq system, and the draft genome sequence was assembled by Unicycler v0.5.1 and annotated by the NCBI Prokaryotic Genome Annotation Pipeline (PGAP v6.10). The bioinformatics-based prediction analysis was performed using ResFinder v4.7.2 and CARD v4.0.1 (antibiotic resistance genes), VirulenceFinder v2.0, VFDB and MGEFinder v1.0.3 (virulence determinants), PlasmidFinder v2.0.1 (plasmid sequences), MLST v2.0 (sequence type), ISFinder and TnCentral v2.0 (insertion sequences and transposons), PathogenFinder2 v0.6.0 (pathogenicity), RAST (subsystems category) and Phigaro (prophage sequences). The draft genome of E. coli CUK-76 strain comprised 4,607,136 bp with a GC content of 51%. Genome annotation revealed 4498 genes of which 4289 were protein-coding genes, 78 RNA genes, and 131 pseudogenes. It was related to sequence type ST949 and its predicted resistome consisted of bla_CTX-M-15, bla_TEM-1B (class A β-lactamase genes), bla_EC-14 (class C β-lactamase gene), aph(6)-Id, aph(3″)-Ib, qnrS1, sul2, tet(A), and dfrA14 genes. Additionally, multiple virulence genes, two plasmid sequences viz. IncFIB(K) and IncFIB(AP001918), insertion sequences, transposons and prophage sequences were detected. The genomic dataset of this strain will be a valuable resource for comparative genomic studies on E. coli. Full article

This study investigated the RNA virome associated with the phlebotomine sand fly Pintomyia fischeri using a metatranscriptomic approach applied to the Meta29 library, composed of 21 specimens collected at the São Paulo Zoo, Brazil. Read-based taxonomic analysis revealed a diverse viral community composed of viruses associated with the families Iflaviridae, Narnaviridae, Partitiviridae, Reoviridae, Solemoviridae, Tombusviridae, Totiviridae, and Tymoviridae, in addition to highly abundant unclassified RNA viruses related to the ShiM 2016 group. Assembly and annotation analyses enabled the characterization of four viral RNA genomes associated with the families Partitiviridae (PfPartitiV-1a-SP, PfPartitiV-1b-SP, and PfPartitiV-2-SP) and Narnaviridae (PfNarnaV-1-SP). Sequence comparisons revealed low amino acid identity relative to currently available reference viral sequences, supporting the classification of these sequences as highly divergent and potentially novel viral lineages. Structural modeling of the RNA-dependent RNA polymerase (RdRp) proteins identified the viral polymerase-specific catalytic motifs A, B, and C. Phylogenetic analyses further supported the evolutionary divergence of the identified viruses relative to currently described taxa. Overall, these findings expand current knowledge of viral RNA diversity associated with Neotropical sand flies and highlight the value of metatranscriptomic approaches for the detection and characterization of previously undescribed viruses associated with medically important insect vectors. Full article

Splicing factors, as core determinants of splice-site selection and dynamic spliceosome assembly, play pivotal roles in stress responses. This review systematically categorizes splicing factors involved in plant abiotic stress responses according to their functions as major spliceosomal components, dividing them into small nuclear ribonucleoproteins (snRNPs) and associated components, spliceosome assembly and disassembly factors, splicing regulatory factors, and proteins related to non-canonical RNA splicing. On this basis, we summarize their regulatory mechanisms of these factors under salt, drought, abscisic acid (ABA) signaling, temperature, and oxidative stresses. Through analyses across multiple species—including Arabidopsis thaliana, rice, maize, soybean, and wheat—we reveal both the evolutionary conservation and species-specific divergence of splicing-factor-mediated regulation. Currently, a large amount of research is still mainly at the transcriptome analysis or single phenotype validation stages, lacking in-depth analysis of direct targets, splicing isomer functions, and molecular mechanisms. Furthermore, current research is heavily concentrated on Arabidopsis, with relatively insufficient functional validation and breeding applications in crops such as maize and wheat. Despite substantial progress, several bottlenecks remain for translational applications in breeding, such as functional redundancy among splicing factor family members, growth penalties associated with overexpression, and tissue-specific and developmental-stage-dependent effects. To address these challenges, we discuss promising strategies, including CRISPR/Cas9-mediated splice-site editing, the use of inducible or tissue-specific promoters, and targeted modulation of upstream kinases, although extensive field trials and rigorous evaluations remain necessary. Collectively, this review provides a theoretical framework for understanding the roles of splicing factors in RNA-level regulation of plant stress adaptation and highlights their potential for breeding improvement. Full article

Human papillomavirus type 16 (HPV16) is a major etiological agent of cervical and other epithelial cancers, yet the mechanisms underlying host–pathogen interactions remain incompletely understood. In this study, we investigated the responses of human peripheral blood leukocytes to engineered HPV16 L1/L2 chimeric virus-like particles (VLPs), produced in suspension by HEK 293-F cells. These VLPs were designed to mimic native viral structures while incorporating chimeric features that enhance stability and immunogenicity. Through experimental assays, we characterized leukocyte engagement, primarily involving leukocyte phenotyping, VLP internalization, confocal colocalization, and endocytic pathway analyses. We demonstrated that recombinant L1/L2 proteins assembled into structured VLPs capable of interacting with mononuclear cells, including lymphocytes and monocytes, but not with polymorphonuclear cells, such as neutrophils, eosinophils and basophils. Uptake occurred via the CD71 transferrin receptor-mediated pathway, in addition to other endocytic routes analyzed, as confirmed by blockage assays using chlorpromazine, rCTB, filipin, nystatin, liquemine, and sodium azide. Confocal colocalization and endocytic pathway analyses further supported receptor-mediated uptake. These findings demonstrate that HPV16 L1/L2 chimeric VLPs interact with and are internalized by human peripheral blood mononuclear cells through CD71-associated and other endocytic pathways. The study provides new insights into HPV16 VLP–leukocyte interactions and contributes to a better understanding of the cellular mechanisms involved in VLP uptake, which may be relevant for future studies on HPV biology and VLP-based vaccine development. Full article

Background: Aztreonam–avibactam (ATM-AVI) is an important therapeutic option for infections caused by metallo-β-lactamase-producing Enterobacterales. However, the emergence of resistance may limit its clinical effectiveness. Here, we report two high-level ATM-AVI-resistant Escherichia coli isolates recovered from critically ill patients in a tertiary hospital in Guangzhou, both of whom had prior exposure to broad-spectrum antimicrobial agents. Methods: Antimicrobial susceptibility testing was performed by broth microdilution. Whole-genome sequencing (WGS) was conducted using the Illumina NovaSeq and Oxford Nanopore Technologies platforms for hybrid assembly. Resistome analysis, multilocus sequence typing (MLST), plasmid replicon typing, and phylogenomic analysis were performed. Comparative genomics with global ST410 isolates was used to investigate the evolutionary origin. Results: Antimicrobial susceptibility testing revealed high-level resistance to ATM-AVI. Hybrid whole-genome sequencing showed that both isolates belonged to the globally disseminated high-risk ST410 lineage and carried bla_NDM-5, CTX-M-type extended-spectrum β-lactamase genes, and an identical four-amino-acid insertion in penicillin-binding protein 3 (ftsI_I334IYRIK). Both isolates exhibited highly conserved chromosomal backbones, differed by only 29 core-genome single-nucleotide polymorphisms, and clustered within a China-associated ST410 clade. Notably, their key resistance determinants were located on distinct plasmid backgrounds: bla_NDM-5 was found on a predicted conjugative IncFIB/IncFIC plasmid in one isolate and on an IncX1 element with potential mobilization in the other. Conclusions: These findings provide important clinical evidence of high-level ATM-AVI resistance in Escherichia coli ST410 isolates in China and highlight the emergence of ATM-AVI resistance-associated determinants within a high-risk genomic background. Active phenotypic and genomic surveillance is warranted as ATM-AVI enters broader clinical use. Full article