Search Results (27,222)

Background/Objectives: Gamma-oryzanol (ORZ), a bioactive compound extracted from rice bran oil, has health-promoting properties but limited therapeutic use due to poor stability and bioavailability. This study aimed to synthesize gamma-oryzanol-encapsulated nanoparticles (ORZ-NPs) and investigate their anti-inflammatory effects in lipopolysaccharide-stimulated RAW 264.7 macrophages. Methods: ORZ-NPs were synthesized via nanoprecipitation and characterized by dynamic light scattering and transmission electron microscopy. ORZ content was assessed using high performance liquid chromatography. In vitro release was determined using a dialysis method. Inducible nitric oxide synthase (iNOS) was assessed by Western blotting, nitric oxide (NO) by Griess assay, and tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) by enzyme-linked immunosorbent assay. Results: ORZ-NPs exhibited spherical morphology with a mean particle size of 93.320 ± 2.027 nm, polydispersity index 0.149 ± 0.025, and zeta potential −22.400 ± 0.252 mV. ORZ remained stable for 90 days. In vitro release reached 70% at 24 h in PBS (pH 7.4). At 50 μg mL⁻¹, ORZ-NPs significantly decreased iNOS and NO production (approximately 65% of control, p < 0.01), without affecting TNF-α or IL-6. Conclusions: ORZ-NPs demonstrate selective anti-inflammatory activities by suppressing iNOS and NO production while pro-inflammatory cytokines remain unaffected. These findings suggest a partial modulatory effect on the inflammatory signaling pathway. Full article

Inflammatory bowel disease (IBD) is triggered by genetic predisposition and chronic inflammation, with aberrant activation of the innate immune complex NLRP3 inflammasome playing a pivotal role in its pathogenesis. In this study, we investigated the effects of a hot water extract from the brown alga Endarachne binghamiae (EB-WE) on the inhibition of NLRP3 inflammasome activation, with a focus on its antioxidant properties, in various inflammation models. In bone marrow-derived macrophages (BMDMs), NLRP3 inflammasome activation was induced using LPS and ATP, and EB-WE pretreatment (100, 200 µg/mL) significantly reduced the secretion of IL-1β and IL-18. Confocal immunofluorescence analysis further confirmed that EB-WE suppressed the formation of the NLRP3-ASC/caspase-1 complex. Furthermore, the in vivo anti-IBD efficacy of EB-WE was assessed using a DSS-induced mouse model, in which colonic inflammation and NLRP3-mediated responses were prominent. Oral administration of EB-WE (2 or 5 mg/day) markedly ameliorated clinical symptoms, such as weight loss, diarrhea, and rectal bleeding, and significantly reduced the disease activity index (DAI). EB-WE also decreased serum pro-inflammatory cytokine levels and the expression of NLRP3 inflammasome-related molecules in colon tissue at both the gene and protein levels. In both BMDMs and the IBD mouse model, we further analyzed the upstream regulatory pathway involving NOX2-iNOS. EB-WE efficiently inhibited the activation of the NOX-iNOS axis and NF-κB phosphorylation, thereby alleviating inflammasome activation associated with DSS-induced oxidative stress and neutrophil/macrophage infiltration. Collectively, these results demonstrate that EB-WE effectively suppresses the formation and activation of the NLRP3 inflammasome by modulating the NOX-iNOS axis and the NF-κB pathway via antioxidant mechanisms. These findings suggest that EB-WE holds promise as a novel marine-derived natural therapeutic agent for the treatment of chronic inflammatory diseases. Full article

This study evaluated Apis mellifera brood fat extracts as a sustainable alternative to beeswax for anti-inflammatory topical delivery, including their formulation into nanostructured lipid carriers (NLCs). Brood fat was extracted using acetone, ethyl acetate (EA), and hexane, and the resulting extracts were characterized for fatty acid composition and physicochemical properties. Safety was assessed using the hen’s egg chorioallantoic membrane test and cytotoxicity testing in RAW 264.7 macrophages. Anti-inflammatory activity was assessed by inhibition of lipopolysaccharide-induced interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) production. The most suitable extract was formulated into NLCs using sugar squalane as liquid lipid, and the effects of lipid ratio and preparation method were investigated. The results showed that the ethyl acetate extract had the highest yield. Compared with beeswax, all fat extracts exhibited a favorable oleic acid–rich fatty acid profile with comparable crystallinity and thermal behavior, while showing significantly enhanced anti-inflammatory activity (p < 0.05). All extracts and their NLCs were non-irritating and non-cytotoxic. Ethyl acetate extract-based NLCs exhibited favorable particle sizes (72.1 ± 0.3 nm) and narrow polydispersity (0.14 ± 0.00), with high-pressure homogenization producing smaller particles compared to probe sonication without affecting IL-6 or TNF-α inhibition. Therefore, A. mellifera brood fat extract is a sustainable anti-inflammatory lipid source with strong potential as an alternative to beeswax in topical nano-formulations. Full article

Natural killer (NK) cells, key effectors of innate immunity, are classically categorized into CD56^dim and CD56^bright subsets in humans. While murine NK cell heterogeneity has become increasingly recognized, the classification of mature NK cell subsets remains incompletely defined. Here, we comprehensively characterized CD127⁺ NK cells in mice and identified them as a distinct, mature subset, developing independently of the thymus and interleukin (IL)-15 signaling. Flow cytometric analyses revealed that CD127⁺ NK cells are broadly distributed across lymphoid and non-lymphoid tissues—including in C57BL/6 wild-type and athymic Foxn1^−/− mice—and exhibit a surface phenotype distinct from CD127⁻ NK and thymus-derived CD127⁺ NK cells. Functional assays demonstrated that CD127⁺ NK cells produce interferon-γ and exert cytotoxic activity, despite expressing markers typically associated with immature NK cells. CD127⁺ NK cells were absent in IL-7Rα^−/− mice but present in IL-15^−/− and IL-15Rα^−/− mice, indicating a selective dependence on IL-7 signaling. IL-7 promoted their proliferation and activation both in vitro and in vivo. These findings revise current models of NK cell development by identifying a novel, IL-7-responsive, IL-15-independent, thymus-independent, and functionally competent CD127⁺ NK cell subset that is phenotypically distinct from helper-like innate lymphoid cells (ILCs). This study provides a framework for future investigations on NK cell heterogeneity, tissue specialization, and cytokine-mediated regulation. Full article

Staphylococcus aureus is a major cause of skin and soft tissue infections, necessitating the development of new topical agents with rapid bactericidal activity and low resistance potential. Here, we evaluated the antibacterial activity of a pyrazolone copper complex (P-FAH-Cu-phen) against S. aureus, investigated its in vitro mode of action, and its assessed therapeutic efficacy in a murine model of S. aureus-infected skin trauma. P-FAH-Cu-phen exhibited potent bactericidal activity (minimum inhibitory concentration [MIC] 1.4 μg/mL; minimum bactericidal concentration [MBC] 2.8 μg/mL) and rapid killing (>91% eradication within 2.5 min), with no detectable MIC increase under the tested serial passaging conditions. Cell-envelope dysfunction was evidenced by increased supernatant alkaline phosphatase activity, elevated leakage of nucleic acids and proteins, and reduced membrane-associated Na⁺/K⁺- and Ca²⁺/Mg²⁺-ATPase activities. At sub-inhibitory concentrations, P-FAH-Cu-phen reduced haemolytic and coagulase activities, modulated virulence gene expression (sea, hla, agrA), and inhibited biofilm formation and biofilm-associated metabolic activity. In vivo, topical treatment accelerated wound closure and histopathological repair, increased hydroxyproline content, reduced bacterial burden, and lowered TNF-α and IL-10 levels in wound tissues. Collectively, P-FAH-Cu-phen shows multi-faceted anti-infective activity and exhibits further development as a topical candidate for S. aureus-infected skin wounds. Full article

Background/Objectives: The strategy of targeting endotoxins and circulating histones to alleviate excessive inflammation and tissue damage has been proposed as an important immunoregulatory strategy against sepsis. However, the development of a multifunctional hemoperfusion adsorber that simultaneously removes endotoxins and histones remains an unmet clinical need in sepsis management. Methods: We synthesized chitosan-cellulose composite (CSCE) microspheres utilizing phase inversion technology, while heparin-grafted chitosan-cellulose composite (CSCEHEP) microspheres were developed by grafting heparin onto CSCE microspheres through the carbodiimide coupling method. In our experimental design, we allocated healthy New Zealand rabbits to four distinct groups: a healthy control group, a lipopolysaccharides (LPS) group, a CSCE group, and a CSCEHEP group. Following the administration of LPS for 12 h, septic rabbits underwent extracorporeal hemoperfusion with either CSCE or CSCEHEP microspheres for a duration of 6 h, notably without the inclusion of heparin in the blood circuits. Post-hemoperfusion, we conducted an analysis of thrombus formation and total protein adsorption on the column. Concurrently, blood samples were collected from the venous side to evaluate inflammatory cytokine concentrations, liver and kidney function levels, LPS levels, the histone presence, and to perform histopathological assessments of liver and kidney injury. Results: Our in vivo experiments demonstrated that CSCEHEP microspheres for extracorporeal circulation could achieve a 6 h hemoperfusion session in septic rabbits without the need for continuous anticoagulation with heparin. A CSCEHEP column turns into a very light-red color (almost the original white) and light contamination or clotting was observed after the 6 h hemoperfusion. Moreover, CSCEHEP microspheres effectively reduced the concentration levels of leukocyte, serum IL-6 and TNF-α, mitigated pathological damage to the liver and kidneys, and removed over 56.7% of LPS and nearly 58.6% of histone H3 from the blood of septic rabbits during hemoperfusion. Conclusions: Hemoperfusion utilizing CSCEHEP microspheres exhibits excellent self-anticoagulation capabilities, remarkable anti-inflammatory performance, efficient endotoxin adsorption and histone antagonism properties, rendering it both effective and safe for use in septic rabbits. Full article

The mycotoxin deoxynivalenol (DON) is a common contaminant found in swine diets, causing decreased growth performance and poor health. Additionally, F18 enterotoxigenic E. coli is a leading cause of post-weaning diarrhea. Nursery pigs are often exposed to each of them after weaning; however, it is unknown what impact the combination of these stressors has on gastrointestinal health. Therefore, the objective of this study was to investigate the effect of pre-exposure to DON on the response of intestinal porcine epithelial cells (IPEC-J2) to challenge with enterotoxigenic F18 E. coli. Four groups were compared: Control (untreated cells), DON (cells treated with 0.5 μM DON for 24 h), F18 E. coli (multiplicity of infection 5:1, varied duration) and DON + E. coli (DON treatment with subsequent E. coli infection). Gene expression of IL-8, IL-6 and TNFα was significantly increased in cells infected with E. coli for 3 h vs. uninfected cells (p < 0.0001, p < 0.0001 and p < 0.0001, respectively). There was an interactive effect between DON and E. coli on IL-8 gene expression; cells pretreated with DON before E. coli infection had a higher expression of IL-8 than those not pretreated (p < 0.05). The concentration of IL-8 protein was significantly increased by E. coli (p < 0.0001). Claudin 1 and Occludin protein abundance were reduced by E. coli as measured by Western blot. Cytotoxicity was increased by E. coli vs. Control (p < 0.05). Pretreatment with DON increased the amount of E. coli that adhered to IPEC-J2 cells (p < 0.01) 30 min post-infection. FITC-dextran passage was increased in the DON + E. coli treatment vs. E. coli alone (p < 0.0001). Transepithelial electrical resistance (TEER) was decreased by DON when compared to untreated cells at 0 h (p < 0.0001). Similarly, DON + E. coli exhibited lower TEER vs. E. coli alone at 2 h post-infection (p < 0.0001). Taken together, these results indicate that DON pre-exposure increased the severity of E. coli infection on endpoints such as barrier permeability and E. coli adhesion. Full article

Platelet-rich gel (PRG) is a fibrin-based biobased biomaterial generated by activating platelet-rich plasma (PRP), yet its biological characterization has commonly relied on univariate measurements of isolated mediators. This study aimed to define the multivariate biological organization of PRG and related hemocomponents (PRP, chemically induced platelet lysate (CIPL), and plasma) in a canine model under single exposure to non-steroidal anti-inflammatory drugs (NSAIDs). In a randomized crossover design (n = 6 dogs), hemocomponents were produced at baseline (0 h) and 6 h after administration of carprofen or firocoxib. Platelet and white blood cell (WBC) counts, growth factors (platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor beta-1 (TGF-β1)), and cytokines (tumor necrosis factor alpha (TNF-α), interleukin-1 beta, and interleukin-10) were integrated using linear mixed-effects modeling, principal component analysis (PCA), and hierarchical clustering. PRG was derived from a leukocyte-poor PRP precursor with moderate platelet enrichment (~1.6-fold vs. whole blood) and a marked WBC reduction (~8–9-fold). In mixed-effects modeling, hemocomponent type significantly influenced the PDGF-BB:TNF-α log-ratio, with PRG (estimate −1.12; 95% CI −1.34 to −0.90) and plasma (−2.06; 95% CI −2.28 to −1.84) lower than PRP, while CIPL did not differ. Time and NSAID effects were not supported. PCA identified two orthogonal axes explaining 61.3% of total variance (PC1 = 43.7%, PC2 = 18.6%), separating a platelet/trophic dimension (log(PDGF-BB), log(TGF-β1), platelet count, PDGF-BB:TNF-α log-ratio) from an inflammatory dimension (log(TNF-α), log(IL-1β)). Overall, hemocomponent composition emerged as the primary determinant of mediator organization, supporting the interpretation of PRG as a structured, biomaterial defined by coordinated mediator networks. Full article

Purpose: Due to the lack of effective local therapeutic strategies for oral squamous cell carcinoma (OSCC), this study aimed to develop a novel gelatin/lignin hydrogel loaded with mesenchymal stem cell (MSC)-derived exosomes enriched in microRNA-185 (miR-185 EV) for intraoral delivery, followed by systematic evaluation of its therapeutic efficacy and underlying molecular mechanisms. Materials and Methods: The gelatin/lignin hydrogel was prepared and subsequently loaded with miR-185 EV. The physicochemical properties of the hydrogel, including microstructure, swelling behavior, chemical composition, and rheological characteristics, were systematically evaluated. Next, the stability, viscosity, biocompatibility, and exosome release kinetics of the hydrogel were further assessed. A 4-nitroquinoline-1-oxide (4NQO)-induced mouse tongue carcinogenesis model was established to assess the in vivo antitumor activity of the hydrogel via intraoral administration. Moreover, a proteomic analysis was conducted to investigate the molecular mechanisms of miR-185 EV on OSCC. Results: The miR-185 EV-loaded gelatin/lignin hydrogel exhibited favorable physicochemical properties, stability, and biocompatibility while prolonging the tissue retention time of miR-185 EV. In vivo antitumor efficacy experiments showed that the miR-185 EV-loaded hydrogel significantly inhibited tumor occurrence and alleviated epithelial dysplasia. Immunohistochemical analyses revealed significant suppression of tumor proliferation and epithelial–mesenchymal transition (EMT) of the hydrogel. Proteomic analysis indicated that miR-185 EV suppressed OSCC progression by downregulating interleukin-1β (IL-1β), consequently inhibiting the NF-κB signaling pathway. Conclusion: The findings demonstrate the successful development of the miR-185 EV-loaded gelatin/lignin hydrogel that represents an effective nanomedicine platform for intraoral drug delivery, providing a promising strategy for the clinical treatment of OSCC. Full article

Background/Objectives: Chlorpyrifos (CPF), a widely used organophosphate pesticide, has been associated with oxidative stress-mediated renal injury. Prenatal exposure may pose a risk for developmental nephrotoxicity; however, data regarding protective natural agents remain limited. This study evaluated the protective effects of Ganoderma lucidum (GNL) against CPF-induced renal alterations in rat offspring. Methods: Pregnant rats received CPF (5 mg/kg) and/or GNL (400 mg/kg) orally throughout gestation. On postnatal day 28, blood and kidney tissues from male offspring were collected for biochemical, ELISA, histopathological, immunohistochemical, and stereological analyses. Results: Prenatal CPF exposure significantly elevated serum urea and creatinine levels and induced oxidative stress, evidenced by increased malondialdehyde (MDA) and nitric oxide (NO) levels and decreased antioxidant enzyme activities (Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH)) (all p < 0.05). Renal TNF-α and IL-6 levels were significantly increased, indicating inflammatory activation. Apoptotic signaling was enhanced, demonstrated by elevated cleaved caspase-3 levels and an altered Bax/Bcl-2 ratio. Tubular injury biomarkers, kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL), were markedly increased. Histopathological findings revealed tubular degeneration, while stereological analysis confirmed significant increases in cortical and glomerular volumes. GNL co-treatment attenuated oxidative stress, suppressed inflammatory cytokines, reduced caspase-3 activation, lowered KIM-1 and NGAL levels, and preserved renal structure. Conclusions: Prenatal CPF exposure induces developmental nephrotoxicity through interconnected oxidative, inflammatory, and apoptotic mechanisms. Ganoderma lucidum mitigates these alterations by restoring antioxidant defense systems, modulating the Bax/Bcl-2 apoptotic balance, suppressing pro-inflammatory cytokine production, reducing tubular injury markers, and normalizing stereologically detected renal structural changes. Full article

Background/Objectives: Patients with pre-existing heart failure (HF) represent a clinically vulnerable population with increased susceptibility to adverse outcomes during acute systemic illnesses, including coronavirus disease 2019 (COVID-19). Systemic inflammation is increasingly recognized as a central pathophysiological mechanism linking cardiovascular vulnerability with infection-related organ dysfunction. However, the prognostic role of inflammatory biomarkers in hospitalized COVID-19 patients with pre-existing HF remains incompletely defined. This study aimed to evaluate the association between inflammatory biomarkers and clinical outcomes in this high-risk population. Methods: This retrospective single-center cohort study included 395 consecutive adult patients hospitalized with confirmed COVID-19 between March 2020 and December 2024 at a tertiary referral center. Pre-existing HF was documented in 143 patients (36.2%). Inflammatory biomarkers, including C-reactive protein (CRP), interleukin-6 (IL-6), procalcitonin, and D-dimer, were measured at admission. The primary outcomes were development of sepsis and in-hospital mortality. Multivariable logistic regression models were constructed to identify independent predictors of adverse outcomes after adjustment for demographic characteristics, comorbidities, disease severity, and cardiac biomarkers. Results: Patients with pre-existing HF had significantly higher in-hospital mortality compared with those without HF (11.9% vs. 4.8%, p = 0.016) and showed a trend toward increased intensive care unit admission. HF patients exhibited higher admission IL-6 levels, indicating enhanced inflammatory activation. In univariable analysis, HF was associated with mortality (OR 2.67, 95% CI 1.22–5.83, p = 0.014). After multivariable adjustment, the association between HF and mortality was attenuated, whereas IL-6 remained an independent predictor of mortality (adjusted OR 1.38, 95% CI 1.04–1.82, p = 0.021). Elevated IL-6 and procalcitonin levels were also independently associated with sepsis development. Conclusions: Pre-existing heart failure identifies a population at increased risk of adverse outcomes in hospitalized COVID-19 patients, and this excess risk appears to be partly mediated by systemic inflammatory activation. Interleukin-6 emerged as a key biomarker linking cardiovascular vulnerability, immune dysregulation, and clinical deterioration. These findings support the potential role of inflammation-based risk stratification to improve prognostic assessment and guide personalized management in high-risk patients with underlying cardiovascular disease. Full article

Energy management systems operating under dynamic electricity pricing require fast and cost-optimal control strategies for flexible loads. Mixed-integer linear programming (MILP) can compute theoretically optimal control trajectories but is computationally expensive and typically relies on accurate load forecasts, limiting its practical real-time applicability. This paper proposes a supervised imitation learning (IL) framework that learns optimal setpoint trajectories for a conventional proportional (P) controller directly from electricity price signals and temporal features, thereby eliminating the need for explicit load forecasting. The learned model predicts setpoint trajectories in an open-loop manner, while a lower-level P controller ensures stable closed-loop operation within a two-stage control architecture. The approach is validated in an industrial case study involving load shifting of a refrigeration system under dynamic electricity pricing and benchmarked against MILP optimization, reinforcement learning (RL), heuristic strategies, and various machine learning models. The MILP solution achieves a cost reduction of 21.07% and represents a theoretical upper bound under perfect information. The proposed Transformer model closely approximates this optimum, achieving 19.33% cost reduction while enabling real-time inference. Overall, the results demonstrate that the proposed supervised IL approach can achieve near-optimal control performance with substantially reduced computational effort for real-time energy management applications. Full article

Hypertension-induced erectile dysfunction is associated with endothelial dysfunction in the corpus cavernosum. Membrane depolarization activates the NLRP3 inflammasome, with downregulation of endothelial Ca²⁺-activated K⁺ channels type 2.3 (K_Ca 2.3) and upregulation of endothelin-1 (ET-1) linked to erectile dysfunction. However, underlying mechanisms remain incompletely understood. We hypothesized that activating K_Ca 2.2/2.3 channels reverses erectile dysfunction and ET-1-induced NLRP3 activation in hypertensive DOCA/salt mice. Hypertension was induced in mice using a DOCA/salt model, with unilaterally nephrectomized mice as controls. We measured blood pressure, intracavernous pressure (ICP), and corpus cavernosum (CC) contractility, and performed immunoblots for K_Ca 2.3, caspase-1, and interleukin-1β (IL-1β). DOCA/salt mice showed impaired erectile function and increased IL-1β activity and reduced K_Ca 2.3 expression. Treatment with the endothelin receptor antagonist bosentan or the K_Ca 2.2/2.3 channel opener NS13001 reversed these dysfunctions and reduced ET-1-induced NLRP3 activation. NS13001 also restored decreased currents in endothelial cells exposed to ET-1. These findings establish that hypertension-induced erectile dysfunction involves an ET-1/membrane depolarization/NLRP3 inflammasome axis in corpus cavernosum endothelial cells, and that targeting endothelial K_Ca 2.2/2.3 channels represents a promising therapeutic strategy to counteract erectile dysfunction. Full article

Background/Objectives: Acute myeloid leukemia (AML) comprises a heterogeneous group of diseases, with some subtypes displaying promyelocytic features and altered differentiation programs. Aberrant cytokine receptor signaling has been implicated in leukemogenesis, and IL-5Rα has recently emerged as a potential marker in selected AML subsets, including promyelocytic variants. Benralizumab is a monoclonal antibody directed against the alpha chain of the interleukin-5 receptor (CD125), which blocks IL-5Rα–mediated signaling. This proof-of-concept study aimed to explore the effects of the anti-IL-5Rα monoclonal antibody Benralizumab in an in vitro AML cell model with promyelocytic characteristics. Methods: Public transcriptomic datasets were analyzed to evaluate IL-5Rα expression in AML subtypes. HL-60 cells, an AML cell line expressing IL-5Rα, were treated with Benralizumab and analyzed for cell cycle distribution and modulation of key signaling and apoptotic pathways by flow cytometry and Western blotting. Results: IL-5Rα was highly expressed in AML, particularly in M2 and M3 subtypes. Benralizumab treatment reduced STAT3 expression, activated ERK and NF-κB signaling, induced p21 and p27 expression, altered cell cycle distribution, and induced caspase-8 cleavage, suggesting activation of extrinsic apoptotic signaling. Conclusions: These findings provide preliminary proof-of-concept evidence that IL-5Rα targeting by Benralizumab may directly affect cell survival and cell cycle regulation in AML cells with promyelocytic characteristics. When interpreted together with the in silico analyses performed on AML patient datasets, these results support the rationale for future validation in APL-oriented models carrying the PML::RARα fusion, the disease-defining oncogenic driver generated by the t(15;17) translocation that blocks myeloid differentiation. However, the in silico and in vitro datasets were not formally integrated at the patient level, and these functional results should be considered exploratory. Full article

Chlorinated polyfluoroalkyl ether sulfonate (F-53B), a common substitute for perfluorooctane sulfonate (PFOS), exhibits similar environmental persistence and bioaccumulation potential, raising concerns about its ecological and health impacts. However, comprehensive toxicological data remain limited for adequate environmental risk assessment. In this study, we evaluated the developmental toxicity of F-53B using embryos/larvae of a commercially important benthic fish, blotched snakehead (Channa maculata). Embryos (<1 h post-fertilization, hpf) were exposed to various concentrations of F-53B (0.002, 0.02, 0.2, and 2 mg/L) for 120 h. Exposure resulted in concentration-dependent adverse effects, including reduced hatching success, increased mortality, and morphological malformations (yolk sac edema, spinal curvature). Histopathological analysis revealed substantial hepatic injury (vacuolization, nuclear pyknosis) and intestinal damage (villi atrophy) at higher concentrations (0.2 and 2 mg/L). Mechanistically, F-53B induced oxidative stress through inhibition of superoxide dismutase (SOD) and catalase (CAT), depletion of glutathione (GSH), and elevated malondialdehyde (MDA). Additionally, the observed immune dysregulation was characterized by the up-regulation of pro-inflammatory cytokines, including interleukin 1β (IL-1β), interleukin 8 (IL-8), and tumor necrosis factor-α (TNF-α), consistent with activation of the TLR-MAPK signaling pathway, and coincided with a shift from metabolic adaptation to pronounced inflammation. These integrated findings indicate that F-53B impairs early development in C. maculata through pathways involving oxidative damage, tissue injury, and immune disruption. This underscores the ecological risk F-53B poses to aquatic organisms and highlights the need for more comprehensive environmental risk assessment. Full article