Search Results (12,138)

Background: Transforming growth factor beta 1 (TGF-β1) is a master regulator of fibrogenesis and extracellular matrix (ECM) remodeling. Among molecules involved in profibrotic signaling, periostin (POSTN) and the Snail family transcriptional repressor 2 (SLUG, SNAI2) have emerged as potential regulators of tissue remodeling and fibroblast activation. However, the temporal relationship between TGF-β1-induced expression of SLUG and periostin in human dermal fibroblasts remains poorly understood. Objectives: This study aimed to evaluate the effect of TGF-β1 stimulation on SNAI2/SLUG and POSTN/periostin expression dynamics in normal human dermal fibroblasts (NHDFs) and characterize their association with profibrotic responses. Methods: NHDFs were stimulated with recombinant human TGF-β1 (20 ng/mL) for 24, 48, and 72 h. SNAI2 and POSTN mRNA expression was quantified by RT-qPCR, intracellular SLUG protein levels were assessed by Western blotting, and periostin accumulation in the culture medium was measured by enzyme-linked immunosorbent assay (ELISA). The RT-qPCR and ELISA data were analyzed using two-way ANOVA with Bonferroni correction; because of the small number of replicates (n = 2), the Western blot data are presented descriptively. Results: TGF-β1 significantly increased POSTN mRNA expression after 24 and 48 h (both p < 0.001); at 72 h, POSTN mRNA levels remained numerically elevated relative to the control, although this difference did not reach statistical significance. Periostin accumulation in the culture medium increased progressively and was significantly higher than in the control group after 48 and 72 h (p < 0.001). In contrast, SNAI2/SLUG exhibited transient expression kinetics, with increased SNAI2 mRNA at 48 h (p < 0.05) and the highest SLUG protein levels also observed after 48 h of stimulation, followed by a decline at 72 h. Conclusions: TGF-β1 induced time-dependent changes in SNAI2/SLUG and POSTN/periostin expression in human dermal fibroblasts. As fibroblast activation and ECM remodeling were not directly assessed, the present conclusions are restricted to the expression changes in the two investigated molecules, whose concurrent yet temporally distinct regulation may indicate potential relevance in the fibroblast response to profibrotic signaling. Further studies are required to determine whether they participate in common regulatory mechanisms or independently contribute to fibrotic processes. Full article

Hypervirulent Klebsiella pneumoniae (hvKP) causes invasive infections, including bloodstream infections and liver abscesses, and dysregulated host inflammation may exacerbate liver injury. We investigated whether pretreatment with Humanin, a mitochondria-derived anti-inflammatory peptide, could attenuate hvKP-associated liver injury. We measured serum Humanin in 20 patients with hvKP-associated bloodstream infection and liver abscess and 20 matched healthy controls. C57BL/6 mice received intraperitoneal Humanin (2.5 or 5 mg/kg/day) for five days before intravenous hvKP challenge. We assessed liver injury, inflammatory responses, hepatic transcriptomes, NF-κB activation, and survival. Serum Humanin was higher in patients than in the controls (p < 0.001). In infected mice, Humanin pretreatment was associated with less prominent histopathological liver injury and lower serum ALT, AST, and PCT, hepatic myeloperoxidase activity, F4/80 immunoreactivity, CD86 fluorescence, and pro-inflammatory gene expression. The effects were generally greater at 5 mg/kg than at 2.5 mg/kg. Transcriptomic analysis showed attenuation of inflammatory pathways, including NF-κB signaling (NES = −1.86, p = 0.002, FDR = 0.012). Humanin also reduced p-p65/p65 and p-IκBα/IκBα ratios in liver tissue and hvKP-stimulated bone-marrow-derived macrophages; 5 mg/kg prolonged survival (p < 0.05). Humanin pretreatment may therefore limit inflammatory liver injury during systemic hvKP infection. This effect was associated with reduced macrophage-associated inflammation and NF-κB activation, although its molecular targets remain unknown. Full article

Transcranial magnetic stimulation (TMS) enables non-invasive, focal modulation of cortical circuits by inducing electric currents in the brain through electromagnetic induction, thereby influencing neuronal excitability and synaptic plasticity. High inter- and intra-individual variability has led, however, to moderate efficacy and reproducibility of stimulation and treatment protocols, motivating a shift toward brain-state-dependent stimulation. Over the past decade, real-time phase-triggered EEG-TMS has established the oscillatory phase—particularly focusing on the sensorimotor mu rhythm—as a key determinant of cortical excitability and plasticity modulation. The field, however, remains largely confined to univariate, sensor-space analyses of local mu-rhythm phase, missing large-scale network dynamics. Recent advances in online EEG source reconstruction and multivariate machine and deep learning (ML/DL) approaches have begun to move beyond local phase toward whole-brain, network-level state estimation, achieving encouraging preliminary accuracies in predicting trial-by-trial cortical excitability, with promising applications in network-dysregulation conditions such as chronic pain. Integrating source-space reconstruction and individual biological variability, and adaptive ML/DL pipelines into closed-loop frameworks promises to move beyond generic stimulation protocols toward selective, network-targeted neuromodulation tailored to the individual’s dynamic brain state. Against this background, this review provides a critical overview of current achievements and limitations, while highlighting emerging methodological directions toward fully brain-state-adaptive and network-targeted EEG-TMS. We further present an illustrative use case of adaptive EEG-TMS for pain modulation, where treatment responses remain heterogeneous and the relevant dynamics are distributed across networks, and which therefore stands to gain most from individualized, network-targeted protocols. Full article

Acute inflammatory challenge can impair intestinal barrier function and bile acid homeostasis in weaned piglets. Glutamate serves as an important energy substrate and functional amino acid for intestinal epithelial cells, yet the epitranscriptomic mechanisms underlying its potential protective effects remain poorly defined. This study investigated whether dietary glutamate supplementation is associated with m⁶A-dependent regulation of SLC10A2 and bile acid signaling in weaned piglets subjected to an acute LPS challenge, with supporting validation in IPEC-J2 cells. LPS challenge (100 μg/kg body weight) impaired ileal morphological structure, activated the TLR4/NF-κB signaling pathway, and aggravated intestinal inflammatory responses in piglets. In contrast, glutamate supplementation was associated with restored ileal mucosal morphology, up-regulated expression of intestinal tight junction proteins, and suppressed TLR4/NF-κB pathway activation. Further metabolomic analysis indicated that LPS stimulation and glutamate intervention altered ileal bile acid metabolism: LPS decreased total bile acid (TBA) content in ileal tissue while increasing TBA accumulation in ileal chyme, whereas glutamate treatment partially reversed this pattern, by elevating ileal tissue TBA and reducing chyme TBA, suggesting a facilitatory effect of glutamate on intestinal bile acid reabsorption. Glutamate supplementation was also associated with increased ileal ASBT (SLC10A2) and FXR (NR1H4) expression. The SLC10A2 transcript exhibited detectable m⁶A modification; glutamate supplementation rescued the LPS-induced downregulation of m⁶A demethylases FTO and ALKBH5, which was associated with reduced global and SLC10A2-specific m⁶A modification. In vitro experiments in IPEC-J2 cells suggested that inhibition of FTO or glutamate dehydrogenase attenuated the effects of glutamate on m⁶A modification and SLC10A2 expression, and that YTHDF2 may contribute to the degradation of hyper-m⁶A-modified SLC10A2 mRNA. Glutamate-derived α-KG may serve as a cofactor supporting demethylase expression and activity after LPS stimulation. Collectively, these findings suggest that glutamate may alleviate LPS-induced intestinal inflammation in weaned piglets through a pathway involving α-KG-dependent maintenance of m⁶A demethylase expression, reduced m⁶A modification of SLC10A2, and preserved ASBT/FXR signaling. This study provides evidence for a nutritional–epitranscriptomic axis that may contribute to intestinal protection under acute inflammatory stress. Full article

Helminth infections continue to present a substantial public health challenge worldwide, particularly in tropical and developing areas. Despite the widespread use of anthelmintic drugs for treatment and control, their effects on host cells remain poorly understood. In this study, the three commonly used anthelmintics albendazole, ivermectin, and praziquantel were investigated by transcriptome analysis for their effects on gene activity in the human intestinal cell line FHs 74 Int after 24-h exposure. The drug concentrations used were substantially higher than in the treatment of helminthiases but did not affect cell viability after 24 h and less than 50% after 72 h. Praziquantel had negligible toxic effects on the intestinal cells at the highest achievable concentration of 1 mM whereas albendazole and ivermectin were already effective at low micromolar concentrations and were tested at 1 µM and 15 µM concentrations. Consistent with the observed cytotoxicity, transcriptome analysis showed no transcriptional changes for 1 mM praziquantel at low cutoff criteria of adjusted p-value (padj) ≤ 0.05, log₂ fold change ≥ 0. At the same padj but with log₂ fold change ≥ 2, 17 upregulated and 2 downregulated genes were found after treatment with 1 µM albendazole and 106 upregulated and 65 downregulated genes with 15 µM ivermectin. Analysis of these genes showed that albendazole caused a modest upregulation of cell cycle related processes, especially mitotic spindle formation, and chromosome segregation. The cells showed a stronger response to ivermectin and the data suggested inhibition of cell proliferation through downregulated transcription factors and nuclear receptors and stimulation of cell growth and cell survival mechanisms through upregulated metabolic enzymes. Full article

NF-κB p65 (RelA) is a key regulator of inflammation, immunity, and stress responses. Recent evidence indicates that alternative p65 isoforms may diversify NF-κB signaling, but their functions remain largely unexplored. We previously identified a novel splice variant, p65 iso5, which contains an additional upstream exon and displays distinct molecular properties, including the ability to interact with dexamethasone in a glucocorticoid receptor-dependent manner. Here, we define the transcriptional programs regulated by p65 iso5 using RNA-seq analysis of HeLa cells expressing either p65 iso5 or canonical p65, with or without dexamethasone treatment. Under basal conditions, p65 iso5 expression was associated with reduced expression of genes involved in type I interferon and antiviral pathways as compared to the effects of canonical p65, while genes involved in translation, ribosome biogenesis, and metabolic activity were not reduced as with p65. Upon dexamethasone stimulation, p65 iso5 expression was associated with extensive transcriptome remodeling characterized by suppression of biosynthetic and proliferative programs and activation of metabolic and stress-adaptive pathways. Comparative analyses reveal that glucocorticoid responses are strongly isoform-dependent, with p65 iso5 expression being associated with distinct gene expression networks supported by distinct protein–protein interaction hubs. p65 iso5 is overexpressed in cirrhotic and hepatocellular carcinoma tissues as well as in high-grade colon tumors, suggesting clinical relevance. Our findings identify p65 iso5 as a context-dependent NF-κB modulator with unique transcriptional and metabolic functions and potential relevance in inflammation-associated diseases and cancer. Full article

Ferroptosis is a regulated form of cell death caused by iron-dependent membrane lipid peroxidation. Iron metabolism, membrane lipid composition, cellular metabolic pathways, and antioxidant defense systems regulate ferroptosis. Recently, ferroptosis has attracted interest as a target in cancer therapy, in particular cancer cells have developed several biological adaptative mechanisms to evade ferroptosis, thus enhancing tumor progression, metastatic dissemination, stemness, and resistance to conventional therapies. Interestingly, ferroptosis is regulated by the tumor microenvironment, where hypoxia, immune cells, and stromal components can either stimulate or inhibit ferroptotic cell death. It has been demonstrated that targeting ferroptosis, alone or in combination with chemotherapy, radiotherapy and immunotherapy, has anticancer effects in preclinical models and may contribute to avoid treatment resistance. However, the role of ferroptosis in pediatric cancer remains incompletely understood since these kinds of tumors show different developmental, genomic, and metabolic features that may contribute to create different ferroptosis vulnerabilities. Emerging evidence in neuroblastoma, B-cell acute lymphoblastic leukemia, osteosarcoma, and medulloblastoma supports the involvement of ferroptosis-related pathways in tumor biology and treatment response. Preclinical studies also indicate that ferroptosis induction may represent a therapeutic strategy in selected pediatric cancer models, although tumor heterogeneity, drug delivery, and potential toxicity to developing tissues remain important translational challenges. This review summarizes the molecular mechanisms underlining the relationship between ferroptosis and cancer biology, tumor progression, tumor microenvironment, and therapy resistance, with particular interest in its emerging relevance and therapeutic potential in pediatric oncology, while critically considering the current evidence and major challenges for clinical translation. Full article

More than one type of policy is necessary to advance the sustainability transition and stimulate innovation. However, limited empirical evidence exists regarding the synergistic effects of environmental and innovation policies on corporate green innovation. Using data on Chinese listed companies in heavy-pollution industries from 2007 to 2022 as samples, this study employs a multi-phase difference-in-differences (DID) approach to examine the synergistic effect of China’s Innovative City Pilot Policy (ICPP) and Low-Carbon City Pilot Policy (LCPP) on corporate green innovation. The results show that the ICPP-LCPP synergy promotes green innovation among enterprises in heavily polluting industries. Further analyses reveal that this effect is concentrated among leading enterprises, with large state-owned leading enterprises (LSLEs) exhibiting a particularly pronounced response. Meanwhile, environmental, social, and governance (ESG) performance positively moderates the relationship between the ICPP–LCPP synergy and green innovation among leading enterprises. Green innovation by leading enterprises has a positive effect on follower enterprises’ green patent grants, particularly green invention patent grants. This study provides important insights into how policy synergy can foster corporate green innovation and facilitate a collaborative sustainability transition involving both leading and follower enterprises. Full article

Objective. The main objective of this study was to examine the effects of transcutaneous auricular vagus nerve stimulation (taVNS) on psychological, sleep, and performance outcomes in powerlifters. Previous research has primarily focused on taVNS in clinical or healthy samples and showed improvements in psychological functioning, recovery, and sleep. Evidence for beneficial effects of taVNS in healthy athletes, however, is missing so far. Methods. Over the course of 4 weeks, 10 healthy adult powerlifters self-administered taVNS five times a week for 30 min after strength training or before bedtime, in an uncontrolled, single-arm pilot design without a sham or control condition. Measurements were taken at three time points: before the intervention phase (pre), immediately after the 4-week intervention phase (post) and 1 month after the post-assessment (follow-up). Pre, post, and follow-up measurements included various subjective psychological domains (recovery, stress, perceived stress, affect, memory, attention, anxiety, and well-being), objective sleep domains (nocturnal HRV, sleep score, sleep duration, and subjective sleep quality), and submaximal strength performance outcomes in squat, bench press, and deadlift. The small sample size required non-parametric testing. Additionally, a responder analysis was conducted to explore individual differences in outcome-specific responses. Results. Results showed nominally significant pre-post improvements (uncorrected for multiple comparisons) in positive affect and self-reported attentional difficulties, both revealing large effect sizes; however, given the absence of a control group and the lack of correction for multiple comparisons across multiple outcomes tested, these findings should be interpreted as exploratory and hypothesis-generating rather than confirmatory. The remaining psychological domains and sleep outcomes did not show meaningful changes. Regarding performance, only two of the ten participants provided complete baseline training data, so no reliable conclusions about a beneficial effect of taVNS on strength performance could be drawn. The responder analysis was consistent with the two nominally significant findings but did not reveal a clear overall pattern of taVNS effectiveness. Conclusions. The current study is the first to implement a 4-week taVNS intervention in strength training and demonstrate differences in domain-specific responsiveness to taVNS in athletes for psychological health and self-reported cognitive performance. These findings offer important insights into the feasibility and practical application of taVNS in sports settings. Further studies with larger, sham-controlled samples, a pre-registered primary outcome and a longer intervention period are needed to determine whether these preliminary, hypothesis-generating findings reflect genuine effects of taVNS. Full article

T helper cells (Th) are central to mucosal IgA induction and key targets for modulation by vaccine adjuvants. To improve understanding of cellular mechanisms underlying mucosal vaccine-induced immunity, we analyzed antigen-specific peripheral blood Th responses elicited by the oral enterotoxigenic Escherichia coli (ETEC) vaccine ETVAX, administered with or without the double mutant heat-labile toxin (dmLT) adjuvant. ETVAX, consisting of inactivated E. coli overexpressing colonization factors CFA/I, CS3, CS5, and CS6 with a heat-labile toxin B-subunit toxoid, was given orally in two doses to adult volunteers, either alone or with 10 or 25 µg dmLT. Antigen-specific Th-associated cytokine responses were assessed in stimulated peripheral blood mononuclear cells isolated from 15 to 18 individuals/group using ELISA and electrochemiluminescence assays. ETVAX predominantly induced Th1 (IFN-γ) and Th17 (IL-17A) responses, with minimal Th2-associated cytokines. Responses were markedly reduced after CD4+ T-cell depletion, supporting a Th cell origin. The strongest responses targeted LTB and CS3, with IFN-γ responses detected in 60–80% and IL-17A in 40–60% across all vaccinees. Responses to CFA/I, CS5 and CS6 were generally weaker. Exploratory comparisons suggested broader IFN-γ responses and more consistent IFN-γ and IL-17A responses to lower-dose antigens, particularly CS6, in recipients receiving vaccine plus 10 µg dmLT. These trends paralleled IgA antibody-secreting cell response patterns, with significantly enhanced IgA responses to CS6 in the vaccine plus 10 µg dmLT group. In conclusion, ETVAX induces antigen-specific Th1- and Th17-type responses in peripheral blood, supporting a role for cellular immunity in mucosal responses to oral ETEC vaccines. Full article

Flavonoids are natural compounds that have shown promise against chronic inflammation. Despite sharing a common core scaffold, flavonoids exhibit substantial structural heterogeneity, and comparisons of biological activities across subclasses remain limited. These studies are often constrained by the cost and sensitivity limitations of conventional analytical approaches. The main objective of this study was to validate a cell-based reporter system, HEK-Blue IL-1R cells, as an alternative to immunoassays for assessing flavonoid structure–activity relationships on the modulation of inflammasome-driven IL-1β secretion by macrophages. Under mild inflammatory stimulation with lipopolysaccharide (LPS), the reporter displayed sensitive and efficient detection of low-level IL-1β signaling. In a comparative analysis of three flavonoids differing in polarity and glycosylation, quercetin, a polar aglycone, and nobiletin, a methoxylated aglycone, significantly inhibited reporter responses relative to LPS-stimulated controls, with a trend toward greater inhibition detected for quercetin. In contrast, no inhibitory effect was detected for rutin, a glycosylated derivative of quercetin. These findings were consistent with the expression of NLRP3 inflammasome proteins responsible for IL-1β maturation and secretion. Collectively, these results support the potential of flavonoid aglycones against inflammasome-associated inflammation and highlight the HEK-Blue IL-1R reporter system as a sensitive and cost-effective platform for screening IL-1β signaling under conditions of low-grade inflammation. Full article

This review proposes that selected micronutrients may contribute to mitohormetic readiness, defined here as the pre-existing biochemical and regulatory capacity of a cell, tissue, or organism to sense, buffer, and transduce a bounded mitochondrial stressor, mount a proportionate adaptive response, and recover toward baseline, or a more capable post-challenge state, without persistent functional loss. This nutrient-centered construct extends the Energy Allocation System (EAS), in which resilience is shaped by how organisms allocate finite metabolic resources under stress and conservation-dominant phenotypes reflect coordinated energy-conserving responses rather than isolated dysfunction. The proposed relevance of micronutrients lies less in direct mitochondrial stimulation than in preserving the endocrine, redox, enzymatic, and one-carbon conditions required for beneficial adaptation. Thyroid physiology is used as the principal nutrient-centered exemplar, particularly through the roles of iodine, selenium, and iron, before the analysis is extended to methylfolate, cobalamin, riboflavin, magnesium, and niacin. The review does not present mitohormetic readiness as an established physiological principle or validated clinical construct, but as a hypothesis-generating extension of the EAS intended to refine how resilience, metabolic flexibility, and biochemical sufficiency are interpreted in clinical bioenergetics. Full article

Background: Glioblastoma (GBM) is a primary malignant tumor of the central nervous system and has a high lethal rate despite therapeutic advances. Although immunotherapies have achieved great success in solid tumors, the highly immunosuppressive tumor microenvironment and the blood–brain barrier (BBB) obstruction hinder the development of immunotherapies for GBM. In this study, we innovatively developed a nanohydrogel composite vaccine (nanoCOM-GEL) for GBM immunotherapy via intranasal immunization. Methods: The nanoCOM-GEL used GelMA as the hydrogel matrix and was co-formulated with antigenic peptides, the BBB-penetrating peptide (peptide 22), as well as immune cell stimulants and chemokines. The efficiency of this vaccine in bypassing the BBB and its capacity to induce anti-GBM immune responses were evaluated in vitro and in vivo. Results: The nanoCOM-GEL vaccine demonstrated superior BBB-bypassing and BBTB-penetrating capabilities, potent immunostimulatory activity, and effective GBM-targeting efficacy, as validated in both cellular and animal models. In an orthotopic GBM mouse model (n = 8 per group), intranasal immunization with nanoCOM-GEL significantly extended median survival from 21 days (control group) to more than 60 days (nanoCOM-GEL group), representing a 2.8-fold increase (p < 0.001). The vaccine markedly inhibited tumor growth, as evidenced by an 82.5% reduction in tumor tissue at day 21 compared to controls (p < 0.01). Mechanistically, nanoCOM-GEL increased intratumoral CD8⁺ T cell infiltration by 9.5-fold and upregulated DCs by 6.4-fold, while simultaneously reducing intratumoral M2-type tumor-associated macrophages by 79.3% (p < 0.001), effectively reshaping the immunosuppressive tumor microenvironment. Conclusions: This innovative nanoCOM-GEL vaccine achieved potent anti-GBM therapeutic efficacy and provided a promising strategy for effective GBM immunotherapies. Full article

Erythropoiesis-stimulating agent (ESA) hyporesponsiveness is a major challenge in the management of anemia in chronic kidney disease (CKD). This study evaluated the association between serum 1,25-dihydroxyvitamin D [1,25(OH)₂D] levels and ESA responsiveness in patients with pre-dialysis CKD. This retrospective cohort study included 363 adult patients with anemia and pre-dialysis CKD who received continuous erythropoietin receptor activator therapy for at least three months. ESA responsiveness was assessed using the erythropoietin resistance index (ERI). The highest ERI tertile was used to define ERI-based relative ESA hyporesponsiveness. Among 363 patients, 121 (33.3%) were classified as having relative ESA hyporesponsiveness. Serum 1,25(OH)₂D levels were significantly lower in the relative ESA-hyporesponsive group. In multivariable analyses, lower serum 1,25(OH)₂D levels were independently associated with relative ESA hyporesponsiveness and higher log-transformed ERI. Exploratory ROC analysis showed modest discriminative ability for identifying patients in the highest ERI tertile. Lower serum 1,25(OH)₂D levels were independently associated with ERI-based relative ESA hyporesponsiveness in pre-dialysis CKD. These findings suggest that active vitamin D status may be linked to ESA responsiveness, as assessed by ERI. Full article

Pseudomonas aeruginosa ST235 isolates drive a distinct macrophage inflammatory program characterized by IL-1β axis dominance and suppression of canonical NF-κB-associated cytokine responses. In THP-1-derived macrophages, ST235 isolates induce robust inflammasome activation, resulting in elevated IL-1β production and altered cell death responses, while eliciting markedly lower TNFα, IL-6, IL-8, and IL-10 responses than non-ST235 isolates. In contrast, nonST235 isolates preferentially stimulate NF-κB-dependent cytokine production with weaker inflammasome activation. Mechanistically, ST235 infection suppresses NF-κB signaling and reduces caspase-1 expression, whereas caspase-8 remains unchanged, indicating that increased IL-1β production can be due to alternative inflammasome pathways or posttranscriptional modifications. Rapid macrophage death further restricts overall cytokine production, reinforcing an IL-1β-dominant inflammatory profile. ST235 isolates also selectively reprogram M2-polarized macrophages toward an M1 phenotype, as demonstrated by increased CD86 expression, while both isolate groups promote M1 polarization in unpolarized macrophages. Importantly, pyoverdine purified from ST235 isolates reproduced aspects of these responses by inducing IL-1β production and cytotoxicity more effectively than pyoverdine from non-ST235 isolates, supporting a potential contribution of ST235-derived pyoverdine to the IL-1β-dominant inflammatory phenotype. Collectively, these findings suggest that ST235 isolates modulate macrophage immune responses through suppression of NF-κB-associated responses and promotion of IL-1β-dominant inflammation, providing insights into the virulence of this high-risk clone and highlighting potential therapeutic targets. Significant statement: ST235 isolates of P. aeruginosa uniquely modulate macrophage immune responses by robustly activating inflammasome signaling, inducing IL-1β expression while suppressing other cytokines, and driving macrophage polarization toward a pro-inflammatory M1 phenotype. These findings highlight distinct pathogenic strategies of ST235 isolates that may inform targeted therapeutic interventions. Full article