Search Results (2,804)

Soil salinity disrupts redox homeostasis and limits plant growth and development. Although catalase (CAT) and superoxide dismutase (SOD) are key enzymatic antioxidants, the CAT and SOD gene families have not been characterized in Artemisia argyi (A. argyi), a species of medicinal and ecological importance. While SOD and CAT serve as the primary enzymatic scavengers for reactive oxygen species (ROS) detoxification, their genomic architecture and stress-responsive regulatory networks in A. argyi have remained uncharacterized. In this study, we conducted the first comprehensive genome-wide analysis of these gene families in A. argyi, identifying 22 structurally conserved members (8 AarCATs and 14 AarSODs). Collinearity and synteny analyses revealed strict lineage-specific evolutionary conservation, while tertiary protein modeling and subcellular localization illustrated a highly organized multi-organelle defense compartmentalization. High salinity (up to 200 mM NaCl) reduced the stomatal conductance and net photosynthetic rate. Salt stress reduced growth and increased osmoprotectant and antioxidant accumulation in A. argyi. Furthermore, histochemical staining using nitroblue tetrazolium (NBT) and 3,3′-Diaminobenzidine (DAB) provided comprehensive evidence of significant accumulation of ROS in leaves, which indicates the intense oxidative stress triggered by ionic stress. Tissue-specific analysis revealed that AarCAT1, AarCSD1, and AarFSD2 were 3.9-, 7.9-, and 12.7-fold higher in leaves than in roots, respectively. Under stress, AarCAT6 and AarCSD1 were strongly repressed in leaves by ~50% and ~46–70%, respectively, whereas AarMSD2 and AarMSD3 were significantly induced in roots by ~2.2- and ~1.8-fold. These distinct expression patterns suggest their potential involvement in tissue-specific stress adaptation and ROS homeostasis. These findings uncover the evolutionary and physiological basis of salt tolerance in A. argyi, providing genetic targets for climate-resilient breeding. Full article

This study evaluated the effects of graded fermented grape seed (FGS) supplementation in breeding-pigeon diets on the growth performance, immune responses, and redox status of squabs. Forty-eight healthy pairs of Gray King breeding pigeons were randomly assigned to four treatments, with 12 cages per treatment. Breeding pigeons received the experimental diets beginning 7 d before the expected hatch date. The basal diet was supplemented with 0, 10, 20, or 30 kg FGS per 1000 kg of diet (CON, FGS10, FGS20, and FGS30, respectively), and each pair reared three squabs until 28 d of age. Growth performance, carcass traits, meat quality, serum biochemical parameters, immunoglobulins, inflammatory cytokines, and serum and hepatic redox indices were evaluated. FGS supplementation affected body weight at 14 d of age and average daily gain (ADG) from 14 to 21 d and from 14 to 28 d of age (p < 0.05), but did not affect body weight at 28 d of age. FGS30 reduced ADG from 14 to 21 and 14 to 28 d and increased relative carcass yields, immunoglobulin A, G, and M levels, interleukin-10, tumor necrosis factor-α, aspartate aminotransferase, low-density lipoprotein cholesterol, and serum and hepatic malondialdehyde levels, while decreasing hepatic superoxide dismutase and glutathione peroxidase activities. FGS20 and FGS30 increased breast muscle L* values and press loss. FGS20 showed numerical advantages in several production traits, but increased breast muscle press loss and did not improve hepatic redox status. Overall, FGS supplementation in breeding-pigeon diets resulted in stage-specific and dose-related responses. FGS30 may exceed the appropriate inclusion range. Based on the present data, none of the tested supplementation levels can be considered optimal, and the range of 10–20 kg FGS per 1000 kg basal diet requires further validation. Full article

Uremic conditions are common in end-stage kidney disease (ESKD) patients. Accelerated vascular diseases in uremic patients lead to heart failure, stroke, and hypertension. To investigate the effects of uremia on porcine arterial smooth muscle cells (aSMCs), bulk RNA sequencing was used to identify uremia-induced alterations in signaling pathways of aSMCs that might explain the aggressive cardiovascular diseases seen in patients with chronic kidney disease (CKD) and ESKD. Bulk RNA sequencing was performed on porcine aSMCs cultured with serum from normal or uremic pigs. Differentially expressed gene (DEG) analysis revealed that 295 genes were upregulated and 138 genes were downregulated after uremic serum exposure. Gene Ontology molecular function analysis demonstrated that ATP-dependent activity, translation factor activity, and ATP-dependent protein folding chaperones were predicted to be negatively enriched after uremic serum exposure, while proton transmembrane transporter activity, antioxidant activity, and glutathione peroxidase activity were predicted to be positively enriched. Gene set enrichment analysis indicated that the cell cycle was predicted to be negatively enriched after uremic serum exposure in aSMCs. Overrepresentation analysis found that focal adhesion, protein processing in the endoplasmic reticulum (ER) and cell senescence were predicted to be negatively enriched, while lysosome, phagosome, apoptosis, and autophagy were predicted to be positively enriched after uremic serum exposure. This study suggests that the signaling pathways that regulate cellular redox homeostasis, the cellular waste disposal system, ER stress and autophagy are major signaling pathways involved in aSMCs’ responses to uremic serum exposure. These pathways may contribute to the severe arterial-specific clinical symptoms observed in CKD/ESKD patients, such as arterial stiffness, vascular calcification and cardiovascular disease. Full article

Spinal cord injury (SCI) is a devastating neurological condition characterized by irreversible primary damage followed by a complex secondary injury cascade involving oxidative stress, neuroinflammation, iron dysregulation, and regulated cell death. Among these mechanisms, ferroptosis, a distinct, iron-dependent form of regulated cell death driven by lipid peroxidation and redox imbalance, is increasingly recognized as a critical mediator of neurodegeneration after SCI. The miRNAs play essential roles in neural injury responses by modulating inflammation, oxidative stress, and cell-death pathways. Growing evidence indicates that miRNAs closely regulate ferroptosis-related signaling networks following SCI, influencing key molecular targets including iron metabolism, antioxidant defense systems, and lipid peroxidation pathways. Conversely, ferroptotic stress may alter miRNA expression profiles, suggesting a bidirectional regulatory relationship. In addition, ferritinophagy, a selective autophagy pathway degrading ferritin via nuclear receptor coactivator 4 (NCOA4), has emerged as an important yet underexplored regulator of intracellular iron homeostasis and ferroptosis susceptibility in SCI. This review systematically summarizes current evidence on the molecular mechanisms linking miRNAs and ferroptosis in SCI, highlights how miRNA-mediated regulation of ferroptosis contributes to neuronal death, glial responses, and impaired regeneration, and discusses emerging therapeutic strategies targeting this axis to promote neuroprotection and functional recovery. By integrating recent experimental findings, we aim to provide mechanistic insight and identify translational opportunities for miRNA- and ferroptosis-based interventions in SCI. Full article

Acute liver injury (ALI) is a severe clinical syndrome characterized by systemic inflammation and mitochondrial oxidative stress. This study investigated the hepatoprotective effects of paeonol (PA) in LPS/D-GalN-induced ALI and explored the potential involvement of macrophage mitochondrial function and NDUFS7. Proteomic analysis showed that the expression of mitochondrial respiratory chain-related proteins was suppressed in the liver of ALI mice, while PA treatment was associated with increased expression of several of these proteins. Integrated metabolomic analysis further indicated that PA modulated metabolites associated with energy metabolism and redox processes. In vitro experiments suggested that inflammatory RAW264.7 macrophages contributed to hepatocyte injury, whereas PA attenuated inflammatory responses and reduced macrophage-mediated injury to AML12 cells. In RAW264.7 cells, PA attenuated ROS accumulation and modulated multiple mitochondrial functional parameters, including mitochondrial membrane potential, ATP levels, the NAD⁺/NADH ratio, and complex I activity. Metabolic studies in human liver microsomes and primary human hepatocytes identified three major PA glucuronide metabolite products with five possible structural assignments. Molecular docking and microscale thermophoresis further indicated that several proposed PA metabolite structures exhibited more favorable predicted interactions and stronger binding to recombinant NDUFS7 than the parent PA. Collectively, these findings support the hepatoprotective effects of PA and its modulation of macrophage mitochondrial function, as well as the potential involvement of PA metabolites and NDUFS7, while further functional validation is required to establish their causal roles. Full article

A novel furan-functionalized polyisoprene (Furan-PIP) was synthesized via a one-pot, redox-initiated cascade approach using tert-butyl hydroperoxide (TBHP) and tetraethylenepentamine (TEPA) as the initiating system. The polymerization of isoprene proceeded through radical emulsion polymerization, accompanied by in situ epoxidation of the resulting double bonds, followed by ring-opening and furanization reactions. The chemical structure of Furan-PIP was qualitatively characterized by FT-IR and ¹H NMR spectroscopy. The appearance of characteristic signals at δ 3.97 and 3.37 ppm (furan ring protons) and δ 8.43 ppm (formyloxy proton) in the ¹H NMR spectrum, together with the corresponding FT-IR absorptions at 1725 cm⁻¹ (C=O) and 1015 cm⁻¹ (furan ring), confirmed the successful incorporation of furan and ring-opened moieties. Residual epoxide signals were negligible, indicating near-complete consumption of epoxy groups during the cascade process. Gel permeation chromatography (GPC) revealed a high-molecular-weight polymer ($\overline{{\mathrm{M}}_{\mathrm{n}}}$ = 45,892 g/mol, PDI = 2.362). The Furan-PIP exhibited a glass transition temperature (Tg) of −49.6 °C and a single-stage thermal degradation at 312 °C. Subsequently, a pre-synthesized antibacterial zinc complex Zn(L-Cl) was physically encapsulated into the Furan-PIP matrix to fabricate a composite material. The composite exhibited time-dependent antibacterial activity against Staphylococcus aureus (S. aureus) over 48 h, leveraging the intrinsic antibacterial property of Zn(L-Cl) previously reported by our group. This work presents a simple and efficient strategy for preparing furan-functionalized elastomers with potential applications in antibacterial materials. Full article

Saline irrigation is a major constraint on vegetation establishment in arid coal-mining landscapes. Nitraria tangutorum Bobrov is a xerohalophytic shrub established in the Dananhu mine-restoration area of Hami, Xinjiang, but its physiological responses to long-term saline irrigation remain insufficiently characterized. In this field study, approximately two-year-old plants were maintained for approximately two growing seasons under freshwater drip irrigation or irrigation water containing 8 or 12 g L⁻¹ total dissolved solids. Unlike short-term controlled salinity assays that evaluate individual response pathways, the present field study integrates root and shoot responses across osmolyte accumulation, oxidative injury, redox regulation, nitrogen metabolism, phytohormone signaling, and lipid remodeling under long-term mixed-salt irrigation. Increasing salinity significantly reduced shoot total chlorophyll content, whereas shoot carotenoid content showed a nonsignificant numerical increase. Soluble sugars, proline, and soluble proteins increased, while total free amino acids declined. Superoxide anion (O₂⁻), hydrogen peroxide (H₂O₂), and malondialdehyde (MDA) increased progressively, demonstrating oxidative injury. The protein concentrations of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX) increased, whereas glutathione peroxidase (GSH-PX) and glutathione reductase (GR) declined. Concurrent decreases in ascorbic acid/ascorbate (AsA) and reduced glutathione (GSH), together with increases in dehydroascorbic acid (DHA) and oxidized glutathione (GSSG), indicated progressive oxidation of the cellular redox environment. Nitrate reductase (NR) declined under salinity, while glutamine synthetase (GS) and glutamate synthase (GOGAT) increased at 8 g L⁻¹ but decreased at 12 g L⁻¹. Growth-associated hormones declined, whereas stress-associated hormones increased. Together, these responses reveal a dose-dependent transition from co-occurring biochemical adjustment and oxidative injury at 8 g L⁻¹ to broader redox and metabolic disruption at 12 g L⁻¹ under long-term field irrigation. These variables provide candidate indicators of downstream salinity response, but ion homeostasis, plant water status, growth, survival, and long-term performance must be evaluated before salt tolerance or irrigation thresholds can be established. Full article

Electrochromic (EC) technology has garnered attention for its low energy consumption and aesthetic versatility. Among various EC materials, tungsten trioxide (WO₃) is notable for its high optical contrast, durability, and coloration efficiency. However, its single-color output limits its versatility in applications such as multicolor displays and windows. In addition, conventional complementary EC devices require the pairing of anodic and cathodic materials, restricting independent control of redox reactions on the electrodes. Hence, this study introduced a three-electrode EC energy storage device with various configurations: vanadium oxide/zinc/vanadium oxide (V₂O₅/Zn/V₂O₅) (energy status indicator) and V₂O₅/Zn/WO₃ (energy storage window and multicolor display). This design allowed independent redox control on both electrodes, enhanced multicolor functionality through V₂O₅, and incorporated zinc metal for efficient energy storage and discharge-induced coloration. The study revealed that V₂O₅/Zn/V₂O₅ configuration exhibited higher coloration depth (58.92% for orange, 32% for yellow-green, and 10.65% for blue) across all color states while achieving a high energy storage capacity of 185 mAh/m², making it ideal for energy status indicator applications. Conversely, the V₂O₅/Zn/WO₃ structure achieved a significantly higher optical contrast of 77.86%, excellent coloration efficiency of 46.24 cm²/C, and energy storage capacities of 104.8 mAh/m² (V₂O₅) and 142 mAh/m² (WO₃), making it suitable for multicolor energy storage windows. Additionally, the V₂O₅/Zn/WO₃ configuration demonstrated suitability for multicolor display applications. Overall, these three-electrode EC energy storage architectures address the intrinsic color limitations and redox control constraints of conventional devices, enabling high-capacity, tunable multicolor operation with significant potential for integration into advanced smart windows, energy indicators, and adaptive display technologies. Full article

This work reports the synthesis of the organic–inorganic copper chloride perovskite (C₆H₅CH₂NH₃)₂[CuCl₄] and its application as an electrode material for lithium-ion batteries. Hybrid copper halide perovskites have emerged as environmentally friendly alternatives to lead-based systems due to their structural versatility and chemical stability; however, their electrochemical energy storage properties remain largely unexplored. To the best of our knowledge, this is the first report investigating (C₆H₅CH₂NH₃)₂[CuCl₄] as an electrode material for lithium-ion batteries. The structural and morphological characteristics of the material were investigated by X-ray diffraction (XRD), scanning electron microscopy (SEM), and infrared spectroscopy (IR), confirming the formation of a layered perovskite structure. Electrochemical performance was evaluated by cyclic voltammetry and galvanostatic charge–discharge measurements. The material delivered an initial specific capacity of 114 mAh g⁻¹ at a current density of 20 mA g⁻¹ and maintained a coulombic efficiency around 99% after 100 cycles, indicating good cycling stability. The electrochemical behavior suggests that the reversible redox activity of Cu species contributes significantly to the charge storage mechanism. Postmortem XRD analysis revealed that the original perovskite framework was only partially retained after cycling, as several characteristic diffraction peaks remained while others disappeared, indicating partial structural decomposition. These results highlight the potential of chlorocuprate-based hybrid perovskites as alternative electrode materials for lithium-ion batteries. Full article

Oxidative imbalance can accompany physiologically demanding and environmental transitions in sheep, but its magnitude varies with tissue, production stage, diet, health status, and the biomarkers used. Nuclear factor erythroid 2-related factor 2 (Nrf2; encoded by NFE2L2) coordinates inducible cytoprotective responses through Kelch-like ECH-associated protein 1 (Keap1), antioxidant response elements (AREs), and Keap1-independent regulatory routes. This structured narrative review evaluates whether dietary antioxidants engage Nrf2-related responses in sheep and whether such responses translate into health, productivity, or product-quality benefits. Evidence was classified by model and endpoint. Tier 1 comprised ovine dietary interventions with tissue-level Nrf2-pathway measurements; Tier 2 comprised ovine dietary studies with redox or phenotypic outcomes but no pathway assay; Tiers 3 and 4 comprised other-ruminant and non-ruminant or in vitro mechanistic evidence, respectively. Only rutin in transition-period ewes and a water extract of Artemisia annua in lambs met Tier 1 criteria. Both altered Nrf2-related gene expression, but neither used pathway perturbation, DNA-binding assays, or definitive target-engagement methods; the results therefore indicate association rather than causality. Evidence for tannins, essential oils, vitamins, selenium, carotenoid-rich feeds, and other phytochemicals in sheep is broader for oxidative status and product stability than for Nrf2 activation. Growth, fertility, milk yield, and survival outcomes are heterogeneous and strongly context-dependent. Dietary antioxidants may act through direct radical interception, microbial metabolites, metal chelation, membrane protection, mitochondrial effects, receptor signaling, inflammation control, and, in some settings, Nrf2-related adaptation. Future studies should combine dose–response designs, exposure measurements, multiple redox markers, tissue-specific pathway assays, and causal validation. Accordingly, Nrf2 is a plausible mechanistic framework for sheep nutrition, but current evidence does not support broad causal or productivity claims. Full article

Ketamine has pharmacological actions beyond N-methyl-D-aspartate receptor antagonism, whereas sodium selenite modulates redox-dependent signaling. Transient receptor potential vanilloid 1 (TRPV1) channels regulate Ca²⁺ influx and may contribute to oxidative stress-associated apoptosis. We investigated whether ketamine and sodium selenite, alone and combined, promote apoptosis through TRPV1-associated mechanisms in SH-SY5Y neuroblastoma cells. Cells were assigned to control, ketamine+sodium selenite (Ket+Sel), Ket+Sel+capsazepine (CPZ), ketamine, ketamine+CPZ, sodium selenite, and sodium selenite+CPZ groups. Ketamine (10 µM) and/or sodium selenite (500 nM) were applied for 48 h; CPZ (0.1 mM, 30 min) was used for pharmacological antagonism, and capsaicin (0.1 mM) for stimulation. Intracellular Ca²⁺ responses were assessed using Fura-2-AM, whereas apoptosis, reactive oxygen species production, mitochondrial depolarization, and caspase-3/9 activities were measured spectrophotometrically or fluorometrically. Under capsaicin-stimulated assay conditions, ketamine and sodium selenite increased all endpoints relative to control, and the combined treatment produced the greatest responses for most endpoints. CPZ attenuated Ca²⁺ responses and several downstream oxidative and apoptotic markers. These findings indicate that ketamine and sodium selenite, particularly in combination, enhance apoptosis through a CPZ-sensitive pathway compatible with TRPV1-associated Ca²⁺-ROS-mitochondrial signaling, while not establishing exclusive TRPV1 mediation. Full article

River sediments act not only as important sinks for heavy metal pollution in watersheds, but also as potential secondary sources under changing environmental conditions. Heavy metals can enter river systems through industrial wastewater discharge, agricultural non-point runoff, urban stormwater and sewage inputs, mining and smelting activities, and atmospheric deposition. During adsorption onto suspended particles, sedimentation, and resuspension, metals such as Cd, Pb, Cr, Cu, Zn, Ni, As, and Hg progressively accumulate in sediments. Because heavy metals are persistent, non-degradable, and bioaccumulative, contaminated sediments can record historical watershed pollution while also releasing metals back into overlying water under hydrodynamic disturbance, pH and redox fluctuations, organic matter mineralization, benthic bioturbation, and dredging activities, thereby threatening aquatic ecosystem stability and human health. Using a global methodological framework with particular emphasis on Chinese river basins, this review systematically summarizes key issues in the study of heavy metal pollution in river sediments, including spatial–temporal distribution and operationally defined fractionation, pollution levels and ecological risk assessment, source apportionment, and remediation and management technologies. Current evidence indicates that heavy metal contamination in river sediments exhibits pronounced spatial heterogeneity and watershed-specific characteristics. Its distribution is jointly controlled by geological background, land use patterns, source input intensity, hydrodynamic conditions, sediment particle size composition, and organic matter content. Methodologically, the field has evolved from single total concentration monitoring and exceedance-based evaluation toward integrated assessment systems that combine total concentrations, operationally defined fractionation, bioavailability, ecological risk, health risk, and source contribution. The joint use of BCR sequential extraction, the geoaccumulation index (Igeo), the pollution load index (PLI), the potential ecological risk index (RI), the risk assessment code (RAC), sediment quality guidelines (SQGs), receptor models, isotope tracing, and machine learning has substantially improved pollution identification, risk zoning, and source apportionment. Overall, research on heavy metal pollution in river sediments has shifted from descriptive judgments of whether contamination exists toward mechanistic and management-oriented questions concerning pollution sources, risk evolution, and remediation strategies. However, important gaps remain in compound pollution transformation mechanisms, regional background values and evaluation benchmarks, uncertainty in model parameters, long-term dynamic monitoring, and engineering-scale verification of remediation technologies. Future studies should strengthen multi-media, multi-scale, and long-term monitoring and further integrate fractionation analysis, toxicological effects, source apportionment models, and remediation technologies to provide a scientific basis for watershed ecological security and precision management of contaminated sediments. Full article

Visible light (VL) accounts for approximately 50% of the solar radiation reaching Earth’s surface and has emerged as a major contributor to skin photoaging and pigmentation disorders, particularly in individuals with darker Fitzpatrick skin phototypes. Increasing evidence suggests that VL exposure induces oxidative stress, inflammation, and melanogenesis by generating reactive oxygen species. In this study, we investigated the protective effects of a topical antioxidant formulation (AOX Mix) containing 15% ascorbic acid, 0.5% ferulic acid, and 1% tocopherol against VL exposure in ex vivo skin biopsies from donors with Fitzpatrick skin phototypes IV–V. To preserve physiological tissue tension and closely replicate in vivo skin responses, human skin explants were maintained using the TenSkin™ culture system. Samples were pretreated with AOX Mix for 30 min, then exposed to VL for 8 h, and collected on Days 2 and 7 for histological and molecular analyses. Our results demonstrated that prolonged exposure to VL disrupted redox homeostasis and induced structural alterations in skin explants, accompanied by increased markers of oxidative stress and pigmentation. In contrast, pre-treatment with AOX Mix significantly attenuated these effects, preserving tissue architecture and reducing molecular indicators of photodamage. In addition, we observed the co-localization of 4-hydroxynonenal and collagen type I, suggesting that oxidative post-translational modification of collagen may contribute to its loss. These findings suggest that antioxidant-based interventions, such as AOX Mix, may be a promising strategy for protecting dark skin against VL exposure. Full article

Background: In our companion study, the gingival mesenchymal stem cell (GMSC) secretome suppresses oxidative stress and induces apoptosis in primary oral squamous cell carcinoma (OSCC) cells, where those wet-lab results are themselves reported as preliminary; here we ask whether it also engages a proliferation × migration programme in patient tissue. The two arms differ in read-out type (apoptosis and redox there, transcript abundance here), not in opposed function. Methods: Primary OSCC cells received GMSC-conditioned medium (GMSC-CM) or indirect Transwell co-culture, assayed by RT-qPCR (VEGFA, TGFB1, MMP9, CXCL12, CCND1, PCNA, MYC, EGFR), MTT, and scratch-wound migration. Thirteen computational layers plus a GeoMx spatial verify-and-decide layer were applied to public data: TCGA-HNSC, GEO, CPTAC, single-cell inference, prognostic modelling, DepMap and LINCS L1000. Results: All eight transcripts were raised in all three arms (16 of 24 comparisons significant), indicating a pro-tumour transcriptional shift; metabolic activity was unchanged and wound closure was reduced under conditioned medium (p < 0.01). Of 1358 genes significant in all three modalities, 1219 (89.8%, 95% CI 88.0–91.3%) share direction against 25.0% expected by chance (3.59-fold; exact binomial p below machine precision). The pre-registered oral-cavity signature did not validate externally (GSE41613; C = 0.562), and no ligand–receptor pair survived permutation calibration. Conclusions: GMSC paracrine exposure induces a pro-tumour transcriptional programme in primary OSCC cells that replicates at patient level, without demonstrated functional or therapeutic consequence; gefitinib is nominated only as a hypothesis-generating candidate. The evidence rests on three primary cultures (n = 3), one GMSC donor preparation, one 24 h time point, a single reference gene, no cell-line authentication and no test of gefitinib; the study programme has concluded, and these experiments cannot be performed now. Full article

Bacterial diseases caused by Flavobacterium covae (Fc), Vibrio harveyi (Vh), Vibrio vulnificus (Vv) and Photobacterium damselae (Pd) seriously constrain Asian seabass aquaculture. Here we dissect the mucosal immune mechanisms engaged by a five-month sequential vaccination strategy that combines nanoemulsion immersion priming with multivalent oral hydrogel boosting. Juvenile seabass were vaccinated, then challenged with F. covae by freshwater immersion and with a Vibrio–Photobacterium (Vh/Vv/Pd) mix by immersion or intraperitoneal injection. Gills were sampled after immersion challenges and intestine after injection, and profiled by RNA sequencing and label-free quantitative proteomics, with selected genes validated by RT-qPCR. Principal component analysis showed clear separation of vaccinated and control fish in all tissues and challenges, indicating a strong and coherent transcriptional reprogramming. Vaccination markedly increased the number of upregulated genes, with Gene Ontology enrichment revealing dominant signatures of ribosome biogenesis, RNA processing, lysosomal organization and immune response. KEGG analysis highlighted cytokine receptor interaction; NOD and Toll-like receptor signaling; oxidative phosphorylation; and phagosome, lysosome and cell adhesion molecule pathways, consistent with heightened antimicrobial readiness. Volcano plots and focused heatmaps showed strong induction of interferon-stimulated genes, cytokines and chemokine receptors, complement components, macrophage mannose receptor, epithelial barrier mediators and numerous immunoglobulin transcripts, with tissue- and challenge-specific patterns. Proteomics corroborated these trends, demonstrating a higher abundance of immunoglobulin heavy chains, complement proteins, cathepsins, heat shock and redox chaperones, ribosomal proteins and cytoskeletal and adhesion regulators in vaccinated mucosae. Integrated pathway mapping linked endothelial adhesion molecules and leukocyte integrins with T cell costimulation networks and an intestinal immune network for immunoglobulin production, including enhanced pIgR-mediated transcytosis. Overall, the sequential vaccination regimen was associated with coordinated transcriptomic and proteomic signatures related to epithelial responses, innate immunity, and humoral immune functions across gill and intestinal tissues. These molecular patterns were accompanied by improved survival following bacterial challenge; however, the present data do not directly demonstrate the functional activity of the inferred immune mechanisms in Asian seabass. Full article