Biology, Volume 15, Issue 17 (September-1 2026) – 124 articles

Background: Psoralea corylifolia is a widely used traditional medicinal plant, with its dried mature fruits as the main medicinal part. Flavonoids and coumarins are the primary bioactive compounds of this species. However, the tissue-specific metabolic profiles, gene expression patterns, and potential regulatory factors underlying active compound biosynthesis remain largely uncharacterized across different tissues of P. corylifolia; Methods: In this study, five tissue types (roots, stems, leaves, flowers and fruits) of P. corylifolia were collected as experimental materials. We performed integrated widely targeted metabolomic and transcriptomic analysis, combined with weighted gene co-expression network analysis (WGCNA), to screen co-expression modules and candidate regulatory factors associated with flavonoid and coumarin accumulation; Results: Distinct tissue specificity was observed at both metabolomic and transcriptomic levels among different tissues, with the most remarkable difference between fruits and other tissues. Signature bioactive compounds including isobavachalcone, bavachin and corylin were specifically and highly accumulated in fruits. Differentially expressed genes were mainly enriched in phenylpropanoid biosynthesis, flavonoid biosynthesis and isoflavonoid biosynthesis pathways. WGCNA revealed that the magenta module was significantly positively correlated with fruit tissues and the contents of the above bioactive metabolites. Six candidate transcription factors were identified from this module and classified into three candidate-priority tiers based on a TF–pathway gene co-expression network (344 edges, |r| ≥ 0.8, p < 0.05) and connectivity metrics. The prioritized hubs were Cluster_22013.0 (C3H-type zinc finger transcription factor) and Cluster_21217.8 (NF-YA; Arabidopsis homolog NFYA9/AT3G20910), with Cluster_9299.0 (Rcd1-like) and Cluster_20910.0 (NAC; Arabidopsis homolog NAC002/AT5G04410) as highly connected positively correlated candidates, while Cluster_31326.1 (bZIP) and Cluster_11436.0 (C2H2) were identified as negatively correlated candidates, with all their significant edges representing negative correlations with pathway genes; Conclusions: This study characterizes tissue-specific metabolic and transcriptomic patterns in P. corylifolia, and identifies candidate co-expression modules and transcription factors associated with flavonoid and coumarin accumulation in fruits. The prioritized TF tiers, including candidate hub and negatively correlated TFs, provide a foundation for future functional studies on the regulation of active compound biosynthesis in P. corylifolia. Full article

Kazakh horses are typical seasonal breeders, and ovarian activity is markedly reduced during the non-breeding period. Hormonal stimulation can induce ovarian functional responses during seasonal anestrus; however, the early ovarian transcriptional changes associated with this response remain incompletely understood. In this study, ovarian transcriptome sequencing was performed in 12 Kazakh mares examined during the seasonal anestrous period. The mares were classified into a non-hormonally stimulated seasonal quiescent group (DB, n = 6) and a hormone-induced ovarian activation group (DY, n = 6) following cloprostenol and eCG treatment. RNA sequencing and bioinformatics analyses were conducted to identify differentially expressed genes and enriched biological pathways. Using false-discovery-rate-corrected criteria (FDR < 0.05 and |log2FC| > 1), a total of 2119 differentially expressed mRNAs and 530 differentially expressed lncRNAs were identified between the two groups. FDR-corrected GO and KEGG enrichment analyses revealed significant changes associated with immune regulation, cellular communication, metabolic processes, and reproductive functions. KEGG pathway analysis demonstrated significant enrichment of complement and coagulation cascades, cytokine–cytokine receptor interaction, chemokine signaling pathway, PI3K–Akt signaling pathway, and ovarian steroidogenesis-related pathways (q ≤ 0.05). C1QB, CREB5, and IRS1 were prioritized as transcriptomic candidate genes based on their differential expression and pathway associations, but their functional roles were not experimentally tested. RT-qPCR analysis of selected DEGs showed expression trends concordant with the RNA-seq results and was used as a technical consistency assessment. Because the DY group was pharmacologically induced during seasonal anestrus and tissues were collected shortly after ultrasound confirmation of ovarian activation, the observed transcriptional profile should be interpreted as an early hormone-responsive state rather than a stable estrous state. Furthermore, because untreated naturally cycling mares during the reproductive season were not included, endogenous seasonal effects cannot be distinguished from pharmacological effects of hormonal stimulation. Full article

Cadmium (Cd) pollution threatens global ecosystems and human health owing to its high toxicity, long-term persistence, and dual irreversibility of environmental geochemical behavior and biological toxicological effects. The interaction between phosphate-solubilizing fungi (PSF) and clay minerals governs the biogeochemical transformation, chemical speciation, and migration fate of soil Cd. However, the dose-dependent mechanisms underlying Cd²⁺ removal by Aspergillus niger (A. niger) and montmorillonite remain unclear. In this study, three Cd²⁺ stress levels were simulated, and four A. niger-montmorillonite composite systems were established. Key physicochemical and metabolic indicators, including pH, available phosphorus (AP) concentration, oxalic acid concentration, acid phosphatase (ACP) activity, and Cd²⁺ removal rate, were determined. The results showed that the single A. niger treatment achieved the optimal Cd removal performance among all composite treatment groups. Severe Cd²⁺ stress induced abundant oxalic acid and ACP secretion, which simultaneously mobilized phosphorus and facilitated Cd removal via phosphate precipitation and oxalate complexation. Low-to-medium montmorillonite dosage temporarily alleviated Cd²⁺ toxicity, whereas high montmorillonite dosage restricted fungal metabolic exudates and, together with acid-mediated mineral-Cd interactions, triggered antagonistic rather than synergistic Cd removal. This study clarifies how montmorillonite dosage reshapes the metabolic defense of A. niger, providing theoretical references for optimizing microbe–montmorillonite composite remediation formulations. Full article

Genetic modification and genome editing have become important tools in agriculture, animal production, biotechnology, and human medicine, but their safety and societal acceptance remain subjects of debate. This review examines genetically modified (GM) and gene-edited organisms, distinguishing transgenesis from precision genome editing technologies, including CRISPR/Cas9, base editing, and prime editing. Representative applications in crops, livestock, pharmaceutical production, and xenotransplantation are discussed, together with their regulatory framework and public perception. Current scientific assessments indicate that approved GM foods are not inherently more hazardous to human health than their conventional counterparts when evaluated case by case. Potential benefits include improved nutritional quality, biofortification, disease resistance, increased agricultural efficiency, production of therapeutic proteins, and applications in animal health and medicine. Possible concerns include allergenicity, toxicity, unintended genetic or phenotypic effects, altered nutritional composition, environmental consequences, animal welfare issues, and uncertainties associated with long-term or large-scale deployment. Public acceptance varies substantially according to geographical region, application, cultural and ethical considerations, regulatory environment, scientific literacy, and institutional trust. Overall, GM and gene-edited organisms should not be considered a homogeneous category. Their benefits, risks, and societal acceptability depend on the specific organism, genetic modification, intended trait, and context of use, supporting a balanced, evidence-based, and case-specific approach. Full article

Spermatogenesis in Drosophila melanogaster is a classical model for studying cell polarity establishment, organelle remodeling, and extreme cellular morphogenesis. Spermatid elongation involves a series of highly coordinated dynamic events, including nuclear polarization, axonemal extension, mitochondrial remodeling, and individualization-mediated cytoplasmic clearance. Current studies have largely focused on individual structures, molecules, or pathways, resulting in a fragmented understanding of this process and a lack of an integrated framework that links mitochondrial function, cytoskeletal dynamics, and temporal translational regulation. Based on existing evidence, this review organizes current findings into three regulatory layers involving mitochondrial remodeling, cytoskeletal dynamics, and post-transcriptional translational control. Mitochondrial remodeling provides structural and metabolic support for sperm-tail formation, cytoskeletal processes execute axoneme organization and individualization, and post-transcriptional regulation controls the timing of protein production. Experimental evidence is strongest for mechanisms operating within each layer, whereas direct molecular coupling among the three layers remains incompletely established. We therefore present this model as a working framework rather than an established causal pathway. This framework helps organize current knowledge of extreme cellular differentiation and provides a basis for comparative studies of spermatogenesis and selected mechanisms associated with human male infertility. Full article

Finding signatures of selective sweeps in genomes is a major goal of current population genomics, as it allows estimating the rate of beneficial mutations going to fixation and identifying the genes involved in selection. Models of recurrent selective sweeps traditionally assume that in chromosomal regions of normal recombination rates at most one beneficial allele is on the way to fixation. We review and extend here the theoretical studies on interference between closely linked beneficial mutations suggesting that this assumption may be violated. We show that interference between beneficial mutations may lead to substantially increased fixation times even in chromosomal regions of normal recombination rates. Furthermore, we discuss how interference can be detected in population genomic studies by analyzing genetic footprints of selective sweeps, and search for empirical evidence of interference in published datasets. Full article

The potential of combining Clostridium butyricum (CB) and glucose oxidase (GOD) in poultry production remains to be systematically evaluated. This study evaluated the effects of dietary CB and GOD, supplied alone or in combination, on ileal antioxidant status, jejunal immune responses, gut epithelial barrier function, and cecal short-chain fatty acid (SCFA) composition in broilers, with the aim of comparing their practical effects rather than investigating synergistic or interactive effects. A total of 192 1-day-old broilers (white-feathered) were randomly distributed among 4 experimental groups, each having 6 replicate pens of 8 chicks. The groups had comparable starting weights (43.53 ± 0.51 g) and an equal male-to-female ratio. The control diet (CON) contained no additives, whereas the other three diets were formulated with CB at an inclusion rate of 400 mg/kg, GOD at the same level, or a mixture of both (400 mg/kg each), corresponding to the CB, GOD, and MIX groups, respectively. This experiment spanned 42 days, comprising an early stage (from day 1 to day 21) and a later stage (from day 22 to day 42). At day 21, the GOD group exhibited a significant elevation in ileal SOD activity relative to the CON group (p < 0.05). Jejunal interleukin-6 (IL-6) levels were significantly decreased in the CB and MIX groups (p < 0.05), while the MIX group also showed a significant decrease in IL-2 and a significant increase in interleukin-10 (IL-10) levels (p < 0.05). The GOD and MIX groups showed a significant elevation of sIgA (p < 0.05), while the CB and GOD groups exhibited marked upregulation of Occludin mRNA (p < 0.05). Butyrate concentrations in the cecum were significantly higher in the CB group than in the CON group (p < 0.05). Likewise, acetate levels were elevated in the MIX group compared with CON (p < 0.05). At day 42, the MIX group exhibited a significant reduction in ileal MDA content compared with the control (p < 0.05). In the jejunum of the MIX group, IL-6 was significantly reduced and IL-10 was significantly elevated (p < 0.05). Additionally, ZO-1 mRNA expression was markedly upregulated in the same group (p < 0.05). Ileal GSH-Px activity was significantly higher only in the CB group (p < 0.05). Overall, combined supplementation with CB and GOD exerted beneficial effects on broiler antioxidant status, immune responses, intestinal barrier-related parameters, and cecal SCFA composition, although the magnitude of these effects varied among endpoints and sampling ages. Full article

Freshwater prawns are important components of freshwater ecosystems, yet their diversity and genetic structure in northeastern Thailand remain insufficiently documented, particularly across habitats differing in soil salinity. This study assessed freshwater prawn diversity and phylogenetic relationships in saline and non-saline habitats using an integrative taxonomic approach combining morphological examination with mitochondrial and nuclear DNA barcoding. A total of 225 mature specimens were collected from 15 sampling sites in Maha Sarakham province (saline habitats) and Mukdahan province (non-saline habitats). Representative specimens were examined morphologically and characterized using four barcode loci: cytochrome c oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), 18S ribosomal RNA (18S rRNA), and the internal transcribed spacer (ITS). Morphological assessment identified three taxa, Macrobrachium lanchesteri, Macrobrachium niphanae and Caridina sp. Molecular analyses recovered three principal lineages broadly corresponding to these taxonomic groups, although the resolution varied among loci. COI provided consistent species-level discrimination, while 16S rRNA revealed additional genetic variation within Macrobrachium. The conserved 18S rRNA marker supported separation of the major taxonomic lineages, whereas ITS contributed complementary sequence variation for lineage assessment. The concatenated multilocus dataset provided the most stable phylogenetic reconstruction, with strong support for the principal lineages. Samples from saline and non-saline habitats were not consistently separated, suggesting that salinity was not the primary factor underlying the observed genetic structure. Overall, integrating morphological and multilocus molecular evidence provided a robust framework for assessing freshwater prawn diversity and phylogenetic relationships and establishes a baseline for future taxonomic, phylogeographic, and conservation studies in the Mekong region. Full article

Background: Influenza remains a significant global public health threat, causing substantial morbidity and mortality worldwide. While vaccination serves as the best preventive strategy, considerable interindividual variability in vaccine-induced immune responses persists. The ZBTB46 rs2281929 polymorphism (c.A31G; p.T11A; ACG>GCG), which corresponds to the evolutionarily conserved mouse mutation Zbtb46^T11A (c.A31G; ACT>GCT), has been associated with enhanced antibody responses to influenza vaccination in humans, though its functional mechanisms remain unknown. This study aimed to investigate how this mutation affects influenza vaccine immunogenicity using a knock-in mouse model. Methods: A Zbtb46^T11A knock-in mouse model was generated using CRISPR/Cas9 technology. Homozygous (HO) and wild-type (WT) mice were immunized with a quadrivalent influenza vaccine in a prime-boost regimen. Humoral immune responses were assessed by Enzyme-linked immunosorbent assay, hemagglutination inhibition (HI), and microneutralization (MN) assays. Antibody-secreting cells (ASCs) were quantified by Enzyme-linked immunospot assays. Germinal center B cells, plasma cells, plasmablast cells, conventional dendritic cell (cDC) subsets, and T helper (Th) cells were analyzed by flow cytometry. Statistical comparisons were performed using a two-sample t-test. Results: The Zbtb46^T11A mutation did not alter Zbtb46 protein expression or its abundance in cDCs. Following vaccination, HO mice exhibited significantly enhanced humoral responses, including higher HA-specific IgG titers, HI and MN antibody levels, and increased numbers of ASCs. Flow cytometry revealed elevated proportions of germinal center B cells and plasma cells in HO mice. Furthermore, HO mice showed a selective expansion of type 1 cDCs (cDC1s) and a concomitant increase in Th1 cell frequencies and IFN-γ-secreting cells, while cDC2 proportions and Th2 responses remained unchanged. Conclusions: The Zbtb46^T11A mutation is associated with enhanced influenza vaccine immunogenicity, concomitant with increased cDC1 proportions, Th1 polarization, and germinal center-dependent humoral immunity. These observed associations suggest a candidate mechanism whereby Zbtb46 modulation may shape adaptive immunity, though further functional studies are required to establish causality. These findings provide insights into host genetic variation in vaccine responsiveness and may inform personalized vaccination strategies. Full article

3-Nitropropionic acid (3-NPA) is an irreversible inhibitor of succinate dehydrogenase (SDH; mitochondrial complex II) that is widely used to model mitochondrial metabolic stress. Direct reproductive studies now show that 3-NPA can increase ovarian oxidative stress, granulosa-cell apoptosis, follicular atresia, alter ovarian reserve, impair oocyte maturation, and reduce fertility in experimental models. However, no ovarian study has yet demonstrated the complete succinate–HIF-1α–NLRP3 cascade proposed here. We therefore propose a testable framework in which SDH inhibition causes succinate accumulation and impaired respiratory electron flux, while redox stress provides an additional signal for HIF-1α stabilization and NLRP3 activation. Succinate-mediated inhibition of prolyl hydroxylases provides a mechanistic route to HIF-1α stabilization under normoxic or near-normoxic conditions, whereas NLRP3 activation may integrate mitochondrial danger signals with inflammatory signaling. Importantly, HIF-1α is not intrinsically pathogenic in the ovary: physiological HIF-1α signaling supports angiogenesis, ovulation, granulosa-cell survival, autophagy, and luteal remodeling, whereas persistent or excessive activation may become maladaptive. We further refine the proposed metabolic–inflammatory threshold as a measurable state in which combined succinate/redox burden and inflammasome activation exceed the adaptive capacity of a follicular unit. The model predicts that time-resolved measurements of succinate, SDH activity, HIF-1α stabilization, NLRP3 activation, and follicular outcomes should reveal ordered relationships that can be tested by pharmacological and genetic intervention. This review distinguishes direct ovarian evidence from cross-system mechanistic evidence and identifies the experiments required to establish causality. Full article

Mulberry leaf contains abundant phytochemicals with antioxidant and immunomodulatory activities. However, systematic insight into its dose-dependent regulatory effects on the intestinal health of Amur sturgeon remains limited. In the present study, multi-omics approaches, including 16S rRNA gene sequencing, untargeted metabolomics, transcriptomics, together with RT-qPCR, were applied to investigate graded dietary mulberry leaf supplementation in Acipenser schrenckii. Juvenile sturgeons were fed four experimental diets containing 0%, 2%, 4% and 6% mulberry leaf over a 10-week feeding trial. Dietary mulberry leaf caused no adverse impacts on growth performance or intestinal digestive capacity. Although the overall structure of the intestinal microbiota remained stable, beneficial bacterial taxa were enriched in a dose-dependent manner. Intestinal metabolism underwent hierarchical remodelling: low inclusion levels supported basal nutrient metabolism, medium inclusion strengthened antioxidant capacity, and high inclusion reprogrammed lipid metabolism and immune function. Mulberry leaf reinforced the intestinal chemical barrier by balancing redox homeostasis and reducing mucosal epithelial permeability. Moreover, intestinal immunity was modulated through three sequential phases: initial innate immune priming, B-cell homing, and the establishment of sustained immune tolerance. In conclusion, mulberry leaf maintains intestinal chemical–immune barrier homeostasis in a dosage-tunable manner, supporting its potential application as a functional aquafeed ingredient. Full article

Alpacas (Vicugna pacos) are of increasing economic importance, mainly due to their high-quality fiber. This study aimed to develop and validate molecular tools for sex identification, individual identification and parentage testing, and to characterize genetic diversity and population structure in Huacaya alpacas bred in Poland. Forty-five previously described microsatellites and one newly developed SRY-based marker were organized into three multiplex panels containing sixteen, seventeen and thirteen markers. The SRY marker was validated in 17 reference animals of known sex (5 males, 12 females) and subsequently in 239 study animals, showing complete concordance with recorded sex. The 44 microsatellites, excluding SRY and the monomorphic YWLL02, showed a mean of 11.45 alleles per locus, observed heterozygosity (Ho) of 0.708, expected heterozygosity (He) of 0.755 and mean polymorphism information content (PIC) of 0.727. Combined non-exclusion probabilities were 1.58 × 10⁻¹¹ for a first parent and 1.08 × 10⁻³⁰ for a parent pair. Seven loci showing elevated null allele frequencies were excluded from population structure analyses. Multivariate, phylogenetic and Bayesian analyses revealed no strong genetic subdivision. The developed tools provide practical support for genetic management of alpacas in Poland. Full article

The euryhaline shrimp Exopalaemon carinicauda is a valuable model for studying osmoregulation, yet the developmental establishment and spatial localization of its key ion-transport sites remain unclear. This study investigated the spatiotemporal expression of Na⁺/K⁺-ATPase (EcNKA), cytoplasmic carbonic anhydrase (EcCAc), and glycosylphosphatidylinositol-linked carbonic anhydrase (EcCAg) during early ontogeny and their subcellular responses to salinity and alkalinity stress in juvenile gills of E. carinicauda. Quantitative real-time PCR was used to detect the changes in mRNA expression of the three genes at different developmental stages, and immunohistochemistry was employed to determine the spatial localization of the corresponding three proteins. EcNKA and EcCAg mRNA expression peaked at zoea I, while EcCAc peaked at zoea III–IV. Immunostaining demonstrated a wide distribution of the three proteins across multiple tissues in early larvae, with a later restriction primarily to the gill and intestine. In gills, the EcNKA protein localized to septal cells, whereas EcCAc and EcCAg were located in pillar cells with distinct membrane polarities. Under low-salinity and high-alkalinity stress, transcriptional changes and protein-level responses of EcCAc and EcCAg were discordant. In addition, the protein location of EcCAc exhibited a translocation from the apical to the basolateral membrane, while EcCAg showed apical concentration under stress. These findings provide new insights into the ontogenetic establishment and subcellular plasticity of key ion-transport sites in E. carinicauda and help us to understand how euryhaline shrimp integrate developmental programming and saline–alkaline stress responses at both the mRNA and protein levels. Full article

Rising global temperatures pose a serious threat to wild small mammals’ population persistence. In this study, we investigated the molecular mechanisms underlying heat-induced testicular impairment in Brandt’s vole (Lasiopodomys brandtii), a dominant small mammal species of the Eurasian temperate steppe. Adult males were subjected to short-term heat exposure at 37 °C, 39 °C, and 41 °C. Heat stress at temperatures ≥ 39 °C significantly reduced the testicular index and caused histopathological damage. Integrated transcriptomic and data-independent acquisition proteomic analyses revealed significant enrichment of pathways related to endoplasmic reticulum protein processing and barrier function. Further molecular validation showed significant upregulation of key ER stress-related markers, including total eIF2α, ATF4, ATF6B, and XBP1, suggesting the potential involvement of the unfolded protein response. Together, these results identify endoplasmic reticulum stress as a key mediator of heat-induced testicular injury and highlight that 41 °C leads to marked activation of ER stress markers and severe testicular impairment in Brandt’s voles, even following short-term exposure. Full article

Sterile alpha and Toll/interleukin-1 receptor motif-containing protein 1 (SARM1) is an inducible NAD-consuming enzyme and execution factor in axon degeneration. Rapid ATP collapse after SARM1 activation, however, is not fully explained by NAD depletion alone. We used SARM1-overexpressing HEK293 cells and the cell-permeant activator CZ-48 to examine SARM1-induced non-apoptotic cell death, termed sarmoptosis. CZ-48 induced cell death that was suppressed by HSP90/70-annotated ATP-competitive compounds, especially geldanamycin and VER-155008 (VER), without reducing SARM1 abundance. VER preserved NAD and ATP during SARM1 activation but failed to rescue FK866-mediated NAD starvation, thereby distinguishing CZ-48/SARM1-driven cytotoxicity from generic NAD depletion. In cell-free assays, purified SARM1 reduced ATP levels; this effect was enhanced by SARM1’s activator NMN and attenuated by its pharmacological inhibitors, although the in vitro activity was modest and the reaction products remain to be identified. ATPase-related perturbations, including thapsigargin and bafilomycin A1, also protected cells from CZ-48-induced death, further supporting a central role for ATP collapse in sarmoptosis. iTRAQ proteomics, MitoSOX Red staining, and DiOC6(3) staining revealed that CZ-48 treatment was associated with mitochondrial and metabolic remodeling, mitochondrial ROS accumulation, and mitochondrial depolarization, all of which were mitigated by VER. Collectively, these findings support a convergent ATP-collapse model in which SARM1 activation promotes NAD depletion, directly consumes ATP, and is associated with mitochondrial dysfunction that may amplify ATP-production failure. Full article

Marine cyanobacteria represent a promising but underexplored bioresource for rare earth element (REE) management. This study investigated the effects of lanthanum (La, 10 mg L⁻¹) on four cyanobacterial strains isolated from Black Sea periphyton: Desertifilum tharense AqMaPh-IBSS-CYA-1, Toxifilum mysidocida AqMaPh-IBSS-CYA-10, Salileptolyngbya sp. IBSS-CYA-8, and Cyanobium sp. AqMaPh-IBSS-CYA-15. La induced a delayed hormetic response in Cyanobium sp. Under suboptimal light and temperature conditions, La supplementation supported 100% survival of all strains over 35 days, whereas controls perished. SEM-EDX indicated predominantly intracellular La localization, with surface-bound La detected only sporadically. ICP-MS revealed hyperaccumulation of La (9.5–13.3 mg g⁻¹ dry weight, 110–226-fold enrichment) and co-accumulation of light REEs in filamentous strains. Strain-specific responses suggest distinct protection mechanisms: T. mysidocida may employ metal efflux and polyphosphate granule sequestration, suggesting potential for REE recovery; Salileptolyngbya sp. appears to show broad-spectrum metal binding, potentially useful for bioremediation; D. tharense seems to be the most sensitive under the tested conditions, suggesting potential as a REE bioindicator. These findings are preliminary, but they suggest that La may act not only as a toxicant but also as a metabolic modulator, highlighting the potential of these cyanobacteria as bioresources for sustainable REE management. Full article

Cancer nanomedicine has generated extensive preclinical evidence of improved drug delivery, pharmacokinetics, and tolerability, yet its clinical impact has often remained modest. This narrative review examines the interconnected biological, pharmacokinetic, manufacturing, regulatory, and clinical factors underlying this translational paradox. A structured literature search was conducted primarily in PubMed and Google Scholar, focusing on studies published between 2022 and 2026 while retaining seminal earlier reports. Major biological barriers include protein corona formation, mononuclear phagocyte system clearance, heterogeneous enhanced permeability and retention, complex tumor microenvironments, and intratumoral heterogeneity. These factors limit circulation, tumor accumulation, tissue penetration, drug release, and interpatient reproducibility. Translation is further constrained by off-target accumulation, uncertain long-term toxicity, non-standardized experimental methods, batch-to-batch variability, scale-up challenges, and fragmented regulatory pathways. Clinical experience shows that successful products are dominated by relatively established platforms and reformulations of known anticancer agents, whereas many actively targeted or structurally complex systems have failed to demonstrate sufficient efficacy or safety. Future progress will require mechanism-driven design, human-relevant preclinical models, harmonized characterization, quality-by-design manufacturing, early regulatory integration, biomarker-guided patient selection, and adaptive clinical trials. Aligning nanoparticle engineering with biological and clinical realities is essential for achieving meaningful patient benefit. Full article

Objective: This study established a rat model of lipopolysaccharide (LPS)-induced acute lung injury (ALI) to evaluate pathological damage, collagen deposition, inflammatory cytokine levels, and key gene/protein expression following Hedyotis diffusa water extract (HDWE) intervention. Combined with ultra-high-performance liquid chromatography-quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS), transcriptomic analysis, and molecular simulation, this study identified the bioactive components of HDWE, evaluated their potential interactions with ALI-related targets, and explored the multi-omics-based protective mechanisms of HDWE. Methods: Thirty-six Sprague–Dawley (SD) rats were randomly divided into six groups: Control group, ALI group, DXMS group, HDWE-L group (100 mg/kg), HDWE-M group (200 mg/kg), and HDWE-H group (300 mg/kg). Hematoxylin and eosin (H&E) and Masson’s trichrome staining were used to evaluate lung pathological changes and collagen deposition. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum tumor necrosis factor-α $\mathrm{T}\mathrm{N}\mathrm{F}-\mathsf{\alpha }$ interleukin-1β $\mathrm{I}\mathrm{L}-1\mathsf{\beta }$, erleukin-6 (IL-6), and interleukin-10 (IL-10) levels. Transcriptomic analysis identified differentially expressed genes (DEGs), followed by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), receiver operating characteristic (ROC), and immune infiltration analyses. Quantitative real-time polymerase chain reaction (qRT-PCR) detected the mRNA expression levels of SPHK1, RELA, and NFKBIA. Immunohistochemistry evaluated the expression of eight hub targets, including endothelin-1 (EDN1), sphingosine kinase 1 (SPHK1), intercellular adhesion molecule 1 (ICAM1), interleukin-17 (IL-17), prostaglandin-endoperoxide synthase 2 (PTGS2/COX-2), NF-κB p65 (encoded by RELA), WT1-associated protein (WTAP), and myeloperoxidase (MPO). UHPLC-Q-Orbitrap HRMS characterized HDWE constituents. Molecular docking analysis was performed between 22 compounds and eight hub targets, followed by 100 ns molecular dynamics simulations and molecular mechanics-Poisson–Boltzmann surface area (MM/PBSA) binding free energy calculations for five core targets. Compared with the control group, the ALI group showed increased levels of TNF-α (86%), IL-1β (107%), and IL-6 (66%), accompanied by a 43% reduction in IL-10 and a 300% increase in lung collagen deposition. All HDWE doses alleviated inflammatory responses, with medium-dose HDWE showing the most pronounced effects. Specifically, medium-dose HDWE increased IL-10 levels by 52% and reduced IL-6, TNF-α, and IL-1β levels by 18%, 22%, and 11%, respectively. Transcriptomic analysis identified 2512 DEGs between the control group and ALI groups, 832 exclusive DEGs between the ALI group and HDWE-M groups, and 876 overlapping DEGs enriched in TNF, IL-17, and NF-κB signaling pathways. The eight-hub-gene diagnostic model achieved an area under the curve (AUC) of 0.969. RELA, SPHK1, and four other hub genes showed positive correlations with Th1, Th17, and neutrophil infiltration. In the ALI group, SPHK1, RELA, and NFKBIA mRNA expression levels were 1.30-, 0.96-, and 0.71-fold of those in the control group, respectively. Compared with the ALI group, high-dose HDWE treatment and low-dose HDWE treatment reduced SPHK1 expression to 0.62- and 0.57-fold, respectively, and increased NFKBIA expression to 1.68- and 1.58-fold, respectively. High-dose HDWE treatment reduced RELA expression to 0.43-fold. The expression levels of inflammation-related proteins were increased in the ALI group and were reduced after HDWE treatment. Twenty-two HDWE components were identified, 16 of which met the docking criteria. Asperulosidic acid exhibited favorable predicted binding affinities with all eight targets, with calculated binding free energies of −14.74, −14.92, −17.58, −23.04, and −16.10 kcal/mol for MPO, IL-17, NF-κB p65, PTGS2/COX-2, and SPHK1, respectively. Conclusions: This study provides systematic in vivo pharmacodynamic and in silico component-target evidence regarding the protective effects of HDWE against LPS-induced ALI. HDWE treatment increased NFKBIA expression and reduced SPHK1, RELA, and multiple inflammatory protein levels, suggesting that HDWE may regulate the IL-17/NF-κB-associated inflammatory network, although direct causal relationships require further validation. Asperulosidic acid may represent a key bioactive component with broad target-binding potential. This study was limited by the use of an LPS-induced rat ALI model without gene knockout or target inhibitor validation; therefore, further functional experiments are required to confirm the proposed regulatory mechanisms. Full article

Tobamoviruses (viruses in the genus Tobamovirus, family Virgaviridae) lead to major yield losses in economically important crops around the world. In this review, we go beyond the canonical gene expression framework by integrating recent discoveries of reverse open reading frames (rORFs) on the negative-strand RNA. These rORFs have only been experimentally validated in cucumber green mottle mosaic virus (CGMMV), with predicted sequence-conserved homologs across a subset of the genus, including TMV, ToBRFV, and PMMoV. However, they are not universally present in all tobamoviruses. We systematically dissect the infection cycle—from disassembly and replication to cell-to-cell and systemic movement—with an emphasis on the host factors hijacked at each stage. We synthesize current understanding of plant antiviral immunity, focusing on RNA silencing and NLR receptor-mediated resistance as two pillars of defense, along with the transcription factors and microRNAs that orchestrate these responses. We critically evaluate the experimental evidence for both plant defenses and viral counter-strategies, noting that many mechanistic models derive from limited model systems. We further characterize host genetic resistance and susceptibility factors applicable to crop breeding. These resources include dominant NLR and non-NLR resistance, as well as recessive resistance derived from modified host susceptibility genes. We address how viral mutations, recombination and fitness trade-offs undermine resistance durability. We then evaluate their practical deployment through conventional breeding, the exploitation of quantitative resistance, and genome editing, and outline associated agronomic drawbacks and regulatory constraints. Using ToBRFV as a case study, we analyze its epidemiological traits and assess the current arsenal of surveillance tools, from field diagnostics to remote sensing. Finally, we survey management strategies across a spectrum of maturity. Some approaches, including sanitation protocols and conventionally bred resistant cultivars, have proven effective under field conditions. The first dsRNA-based biopesticide has recently been registered in China, while other biological control agents and low-risk chemical approaches remain largely at the experimental stage. We also discuss the bottlenecks that impede lab-to-field transition and highlight promising solutions such as precision breeding and evolution-oriented cultivar deployment. By bridging molecular virology, epidemiology, and integrated disease management, this review provides a critical, bench-to-field framework for the sustainable control of tobamoviruses. Full article

Fungi play essential roles in ecosystems as pathogens, mutualists, and ubiquitous decomposers. However, like many important microbes, the spatial distribution of species and natural populations remains poorly understood compared to plants and animals. Many fungi are described as global generalists because they occur across wide geographic areas, but it remains unclear how and if these species are constrained by climate or geographic barriers. In this study, we used Species Distribution Models to infer the global climatic suitability of three common and globally distributed fungi: Aspergillus flavus, Penicillium chrysogenum and Aspergillus fumigatus. Models were constructed using global occurrence data from the Global Biodiversity Information Facility and were trained with Bioclimatic variables from the WorldClim dataset. All species’ models showed high prediction fit, with predicted occurrence concentrated in the temperate and subtropical regions and broadly structured patterns. Each species showed distinct predicted distributions, but they displayed considerable spatial overlap on a global scale. Together, these results demonstrate that even apparently globally occurring and generalist fungal species occupy climatically structured niches. This study highlights the utility of SDMs and it provides a framework for future studies integrating ecological, genomics and evolutionary perspectives among the difficult to assess geographically widespread and common fungi. Full article

Marine bacteria represent a vast and largely untapped resource for biotechnological innovation, offering solutions to global challenges in health, sustainability, and environmental conservation. The ocean’s unique conditions have driven marine bacteria to evolve diverse metabolic capabilities, resulting in the production of bioactive compounds, enzymes, and other metabolites with wide-ranging applications. Recent advances in high-throughput sequencing, metagenomics, and analytical chemistry have unlocked new opportunities for leveraging these microorganisms in fields as varied as medicine, agriculture, and bioremediation. This review highlights the role of marine bacteria in the One Health framework, showcasing their contributions to antimicrobial discovery, nutraceutical development, pathogen biocontrol, and environmental cleanup, including microplastic degradation. This review also examines emerging methodologies such as microbiome mining and advanced culturing techniques, which hold the key to realizing the full potential of marine bacteria in a sustainable bioeconomy. By bridging fundamental research with applied sciences, marine biotechnology promises to deliver transformative impacts on human, animal, and environmental health. Full article

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) characterized by neuroinflammation and demyelination. Although treatment options for MS have increased over the past decade, current therapies primarily target the peripheral immune system and are often associated with significant side effects. Moreover, these therapies still do not offer solutions for the resulting neurodegeneration that accompanies the progression of the disease. Findings from research using cellular and animal models and studies in humans highlight the neuroprotective and anti-inflammatory properties of apigenin, a flavonoid present in several commonly consumed plant species, such as cabbage, orange, tea, onion, and chamomile. This review compiles chemical and current evidence on the therapeutic potential of apigenin, primarily on its ability to modulate neuroinflammation and underscore the need for further investigation into its clinical applications as an adjuvant therapy for MS. Full article

The intestinal immune system is essential for insects to defend against pathogenic infections, while the dual oxidase-reactive oxygen species (DUOX-ROS) pathway is a key component. However, the role of the DUOX-ROS pathway in Bombyx mori against Nosema bombycis, a significant pathogen of this species, remains poorly characterized. In this study, four core components of the DUOX-ROS pathway in B. mori (BmGαq, BmPLCβ1, BmPLCβ4, and BmDUOX) were identified using bioinformatics. Pathogen induction experiments showed significant upregulation of BmDUOX and increased ROS production following N. bombycis infection. Immunofluorescence and antibody-blocking assays demonstrated that BmDUOX localizes to the plasma membrane and is essential for inhibiting spore adhesion and invasion. This study provides the first systematic evidence of DUOX-ROS-mediated anti-microsporidian immunity in B. mori, which may provide a theoretical basis for the future development of ROS-based disease control strategies in sericulture. Full article

To quantitatively characterize the pharmacokinetic and pharmacodynamic interactions between Hydroxysafflor Yellow A (HSYA) and Calycosin (CA) in cerebral ischemia–reperfusion injury (CIRI), a factorial experimental design was combined with a hierarchical coupled PK-PD modeling framework. The coupled PK model adequately described the concentration-time profiles of HSYA and CA under co-administration, with R² values of 0.83 and 0.86, respectively, while the PK-PD model showed good fitting performance for Caspase-3 and HIF-1α, with all R² values exceeding 0.97. Structural identifiability analysis showed that the newly introduced PK and PD coupling parameters were globally identifiable, and their Bootstrap 95% confidence intervals excluded zero. After Holm-Bonferroni correction, AUC_0-t, AUC_0-∞ and CL differed significantly between single and combined administration, whereas MRT did not. The model-derived relative contribution weights of HSYA and CA were quantified for both Caspase-3 and HIF-1α, with uncertainty assessed using Bootstrap 95% confidence intervals. Overall, the proposed coupled PK-PD framework provides a quantitative approach for characterizing component interactions and relative pharmacodynamic contributions in multi-component systems. Full article

The LOR (LURP-one related) gene family encodes proteins containing conserved LOR domains; however, its functions in plant abiotic stress responses remain largely unexplored. In this study, we systematically identified and characterized the LOR gene family in the stress-tolerant wild tomato Solanum lycopersicoides using comprehensive bioinformatic analyses and conducted functional validation of the candidate gene ScLOR16. A total of 19 ScLOR members were identified and classified into eight phylogenetic subgroups. Numerous cis-acting elements associated with responses to abscisic acid (ABA), cold, and drought, including ABRE, LTR, and MBS, were detected in the promoter regions, suggesting that the ScLOR family may be broadly involved in ABA-mediated stress signaling pathways. RT-qPCR analysis revealed that ScLOR16 expression was significantly induced by both cold and drought treatments. Subcellular localization assays demonstrated that ScLOR16 is localized in both the nucleus and cytoplasm. Virus-induced gene silencing (VIGS) was subsequently employed to generate ScLOR16-silenced plants. Following 24 h of cold treatment at 4 °C and four days of drought stress, ScLOR16-silenced seedlings exhibited significantly less severe wilting symptoms than empty-vector controls. Physiological analyses showed that silenced plants exhibited enhanced superoxide dismutase (SOD) and peroxidase (POD) activities, increased proline accumulation, and decreased thiobarbituric acid-reactive substances (TBARS) content. Collectively, these results indicate that reduced ScLOR16 transcript levels are associated with enhanced cold and drought tolerance, accompanied by alterations in antioxidant defense and osmoprotection-related physiological markers. This study yields new insights into the evolution and stress-related functions of the ScLOR family. Full article

Canavanine (CAN) is a nonproteinogenic amino acid, synthesised by many legumes (e.g., alfalfa, hairy vetch) and acts as a strong inhibitor of root growth. Thus, CAN may be considered as an allelopathic compound, influencing natural and agroecosystems. CAN is a structural analogue of arginine (Arg), so its toxicity results from its primary mode of action due to interference with Arg-dependent processes, especially incorporation into proteins instead of Arg. The aim of the work was to investigate the impact of CAN (10, 50 µM) treatment (24 or 72 h) on Arg catabolism and content of polyamines (PAs) in the roots of tomato (Solanum lycopersicum L.) seedlings. In the roots of plants cultured in the presence of CAN for 72 h, whosegrowth was completely inhibited, arginase activity and transcript levels of ARG1 and ARG2 were elevated, while ornithine (the product of Arg catabolism by arginase) content was low. CAN application resulted also in decreased total PAs level and increased relative contribution of spermine in the roots of tomato seedlings. CAN-induced alterations in PAs content in the roots were accompanied by changes in transcript levels of genes related to PAs synthesis (SPMS, SPDS) and PAs catabolism (PAO1). Full article

Managers of protected areas are charged with providing public access and enjoyment while minimizing wildlife disturbance, a difficult feat given that wild animals often respond to humans as predators. Evaluating the effects of people on wildlife is critical to inform management actions, but designing methods to assess visitor impacts or test mitigations with free-ranging wildlife is challenging. In Glacier Bay National Park, Alaska, we tested the effects of vessel-based bear viewing on the behavior of brown bears using experimental approaches, and we tested the effects of human presence on wildlife activity at one frontcountry and four backcountry locations during times of normal (2017–2018) vs. low visitation (2020) using remote cameras. We found that vessel approaches within 100 m of brown bears led to increased stress behaviors. Detection rates for five species (black bear, brown bear, wolf, moose, and coyote) varied by site and year. For most species, we did not detect a threshold of human activity at which wildlife activity clearly declined. However, at the four backcountry sites, wolf activity dropped to zero as human activity exceeded 20 camera minutes/week. Furthermore, wolves were not detected at the frontcountry site during all years, suggesting spatial avoidance of a high-human-use area. Additional research is warranted to determine the strength of these associations and whether these observations are generalizable. Our study provides a framework for estimating thresholds of human disturbance to improve management of protected areas. Full article

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by social communication deficits and repetitive behaviors, now affecting approximately 1 in 31 children. While traditionally defined behaviorally, ASD is increasingly understood as a disorder of brain connectivity arising from altered synaptic formation and refinement. This narrative review synthesizes evidence on neuroimmune dysregulation in ASD, focusing on immune-mediated synaptic pruning mechanisms. We conducted a comprehensive literature search in PubMed, Scopus, and Web of Science (2010–2026), prioritizing high-impact peer-reviewed research. Convergent findings suggest that the classical complement cascade (C1q-C3) tags specific synapses for elimination, while microglia participate in the phagocytic removal of tagged connections. Genetic studies have reported associations between ASD and variants in complement-related genes (C1q, C3, CR3, and C4A, although the strongest evidence for C4A-mediated pruning comes from schizophrenia research), as well as in microglial function genes (TREM2, PTEN, SHANK3). Neuroimaging reveals a dynamic pattern of local hyperconnectivity transitioning to long-range hypoconnectivity during development, particularly affecting prefrontal, insular, and cerebellar regions. Systemic inflammation, including gut–brain axis dysbiosis and maternal immune activation, may amplify neuroimmune dysregulation. We conclude that ASD can be understood, in part, as a disorder of synaptic immunology, where disrupted neuroimmune communication during critical developmental windows may contribute to altered connectivity. The complement–microglia axis therefore represents a potential mechanistic target for future therapeutic investigation. Full article

Background: To conserve wild populations and ensure a sustainable supply of musk, China initiated the captive breeding of forest musk deer. The temporal dynamics of gut antibiotic resistance gene (ARG) profiles in captive forest musk deer along a breeding duration gradient remain poorly characterized. Methods: In this study, we employed metagenomic sequencing to systematically characterize the profiles and potential mobility of ARGs. Samples were divided into short-term, medium-term and long-term groups according to breeding durations. Results: A total of 331 ARG subtypes and 71 mobile genetic element (MGE) subtypes were annotated across all samples. ARG Shannon diversity differed overall across groups (Kruskal-Wallis, p = 0.03); Bonferroni-adjusted Dunn’s test showed no significant pairwise differences. PCoA (Bray–Curtis) demonstrated distinct separation of the ST group (p = 0.002), and shared core ARG subtypes gradually increased with extended breeding years. A strong positive correlation between ARG and MGE abundances was identified (r = 0.85, p = 0.0001). In total, 63 contigs carrying co-localized ARG-MGE complexes were recovered. The ST group contained the highest proportion of such contigs. The ST group displayed tight physical ARG-MGE linkage within 1–3 kb genomic intervals. Conclusions: Our results reveal that breeding duration is associated with the gut ARG characteristics of captive forest musk deer. Short-term captivity has higher ARG-MGE co-localization, suggesting a higher possibility of mobilization. Full article