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Alcohol consumption is associated with increased breast cancer risk, partly due to the accumulation of toxic aldehydes like acetaldehyde, a carcinogenic byproduct of ethanol metabolism. Acetaldehyde Dehydrogenase 2 (ALDH2), a key mitochondrial enzyme, detoxifies acetaldehyde and other reactive aldehydes that drive oxidative stress, DNA damage, and hormonal dysregulation—processes central to carcinogenesis. Although alcohol consumption has been implicated in breast cancer, the role of ALDH2 deficiency itself, in the absence of exogenous alcohol exposure, in mammary gland biology and cancer susceptibility remains unclear. Genetic variants that impair ALDH2 activity are highly prevalent in East Asian populations, where carriers of inactive ALDH2 alleles exhibit impaired aldehyde detoxification. While such individuals are more susceptible to alcohol-related cancers, the effects of ALDH2 deficiency on mammary gland development and homeostasis without alcohol exposure remain unexplored. To investigate the effects of ALDH2 deficiency on mammary proliferation and development, we utilized a C57BL/6-based ALDH2 knockout (Aldh2−/−) mouse model. Our findings revealed that Aldh2−/− mice displayed hyperproliferative mammary glands with increased epithelial cell density, ductal expansion, and increased numbers of Ki67+ cells. Flow cytometry analysis revealed expansion of luminal and basal epithelial subpopulations, accompanied by enhanced mammary epithelial stemness, as indicated by increased mammosphere formation and colony-forming efficiency. At the molecular level, ALDH2 deficiency activated oxidative stress pathways, reflected by elevated 8-OHdG, p38 MAPK, NF-κB, and Nrf2 signaling, along with DNA damage responses involving p53 and H2A.X. We also identified a novel upregulation of RANK and RANKL in Aldh2−/− mammary glands, identifying the RANK/RANKL upregulation associated with NF-κB/p38 MAPK activation and enhanced mammary stemness. Furthermore, hormonal dysregulation was observed, with a significant increase in ERα and PR expression and phosphorylation. Dysregulated ER signaling correlated with enhanced erbB3 activation and downstream signaling, including the cyclin D1–pRb-E2F1 axis. These findings suggest that ALDH2 deficiency, possibly through accumulated endogenous aldehydes, profoundly alters mammary morphogenesis, epithelial repopulation, and stemness. These effects are associated with activation of oxidative stress and DNA damage pathways, together with upregulation of RANKL, estrogen receptor and receptor tyrosine kinase signaling. This study is the first to identify ALDH2 deficiency as a novel factor associated with mammary epithelial alterations that may create a tissue state that could predispose to oncogenic transformation. Full article

Hybrid rice outyields conventional rice, but the role of rhizosphere microbiome in this yield advantage remains unclear. We conducted a single-site field experiment to compare three hybrid rice cultivars with one conventional variety. We tested whether the hybrid rice cultivars were associated with distinct rhizosphere microbial consortia, and whether these consortia correlated with soil nutrient availability and yield. Our results showed that the hybrid cultivars produced significantly higher yields than the conventional variety. Rhizosphere soil was collected at booting and mature stages. We integrated 16S rRNA and metagenomic sequencing, soil physicochemical and enzyme activity analyses, and structural equation modeling to explore associations among rice genotype, microbial community, nutrient availability, and yield. Hybrid rice showed higher yield and 1000-grain weight (Yongyou-1540: 69.76 g/plant, 26.85 g). Genotype-associated differences in microbiome were strongest at booting: hybrids enriched Pseudomonadota and Chloroflexota but depleted Acidobacteriota; genera Geobacter, Anaeromyxobacter, and Burkholderiaceae were enriched. Hybrid rhizospheres had higher available phosphorus and potassium (Heliangyou-165: +62.8% P and +24.7% K at booting; +18.4% P and +350.8% K at maturity). Metagenomic profiling revealed a higher abundance of genes associated with nitrogen fixation, siderophore biosynthesis, and sulfur cycling pathways. Nitrogen fixation gene abundance correlated with alkali-hydrolyzable N (r = 0.66), siderophore biosynthesis with available P (r = 0.73), and sulfate respiration with available K (r = 0.61). Enzyme activities showed genotype-and stage-dependent patterns. Structural equation modeling supported a sequential association path: genotype → microbial community (β = 0.58) → soil nutrient composite (β = 0.52) → grain yield (β = 0.44). The indirect effect of genotype on yield via microbiome and nutrients was significant (β = 0.13), while the direct effect was not. The yield advantage observed in the tested hybrid cultivars was accompanied by cultivar-associated differences in rhizosphere microbiomes that correlate with enhanced nutrient bioavailability during critical growth stages. These correlative findings suggest a potential belowground ecological pathway linking cultivar identity, rhizosphere microbiome functional potential, nutrient availability, and yield; however, they do not demonstrate a causal role of the microbiome in the yield advantage of hybrid rice. Given the single-site, single-year design and the use of only one conventional control cultivar, these results should be considered preliminary and require validation across broader genotype panels and environments. Experimental validation is required to test the hypothesized microbial associations. Full article

Background: The classification of metabolic risk solely based on body mass index (BMI) fails to acknowledge the heterogeneity inherent in obesity. Soluble urokinase plasminogen activator receptor (suPAR) has been proposed as a stable biomarker of chronic inflammation; however, its behaviour across metabolic obesity phenotypes defined jointly by adiposity and insulin resistance remains uncertain. Methods: In this cross-sectional study, 190 adults aged 18 to 60 years attending internal medicine outpatient clinics were stratified by BMI (≥30 kg/m²) and homeostasis model assessment of insulin resistance (HOMA-IR ≥ 2.5) into four phenotypes: non-obese insulin-sensitive (NOIS, n = 64), non-obese insulin-resistant (NOIR, n = 50), obese insulin-sensitive (OIS, n = 33), and obese insulin-resistant (OIR, n = 43). Serum suPAR, interleukin-6 (IL-6), and C-X-C motif chemokine ligand 10 (CXCL10) were measured using enzyme-linked immunosorbent assays. Group comparisons, rank-based two-factor analyses, correlation analyses, logistic regression, and receiver operating characteristic (ROC) analyses were performed. Results: Serum suPAR did not differ across the four phenotypes (p = 0.19) and was not correlated with HOMA-IR (ρ = −0.09, p = 0.24). IL-6 was lower in the OIR group than in the NOIS group (p = 0.012). CXCL10 was higher in both obese phenotypes than in both non-obese phenotypes (p < 0.001), and rank-based two-factor analysis attributed this difference to obesity (p < 0.001) rather than insulin resistance (p = 0.71). CXCL10 correlated with BMI (ρ = 0.29, p < 0.001) and remained associated with obesity after adjustment for age, sex, and C-reactive protein (odds ratio per 10 ng/L, 1.12; 95% confidence interval, 1.06 to 1.18). CXCL10 discriminated obesity with an area under the ROC curve of 0.74 (95% CI, 0.66 to 0.81). None of the three biomarkers discriminated insulin resistance (areas under the curve, 0.43 to 0.50). The association between CXCL10 and obesity was consistent in both sexes and unchanged with alternative HOMA-IR cutoffs of 2.0 and 3.0. Conclusions: In this cohort, circulating CXCL10 tracked adiposity rather than insulin resistance, whereas suPAR and IL-6 did not separate metabolic obesity phenotypes. These findings do not support suPAR as a marker of insulin resistance independently of body fat. Full article

Supraventricular tachycardias (SVTs), including atrial fibrillation (AF), atrioventricular nodal re-entrant tachycardia (AVNRT), atrioventricular re-entrant tachycardia (AVRT), and focal atrial tachycardias, can possibly arise from the interaction of genetic predisposition and environmental exposures. While genome-wide association studies (GWASs) have identified 525 loci for atrial fibrillation and a small number of loci for AVNRT and accessory pathway-mediated tachycardia, the contribution of air pollution, lifestyle factors and psychosocial stress to epigenetic remodeling of atrial tissue remains insufficiently integrated into current mechanistic models of arrhythmogenesis. This review aims to synthesize evidence on how environmental exposures, including PM_2.5, NO₂, ozone, tobacco smoke, obesity, alcohol use, and physical inactivity, modulate epigenetic pathways relevant to SVT susceptibility, and to evaluate whether AF-derived epigenetic insights can be extrapolated to other SVTs. Thus, a narrative synthesis was conducted across studies examining environmental determinants, epigenetic mechanisms (DNA methylation, histone modifications, non-coding RNAs), and genetic susceptibility in SVTs. Literature from cardiac tissue studies, circulating epigenetic biomarker analyses, and mechanistic AF models was integrated to construct a unified gene–environment–epigenome framework. In atrial fibrillation, environmental exposures are consistently associated with epigenetic alterations affecting atrial electrophysiology, inflammation, oxidative stress and structural remodeling, and air pollutants and lifestyle factors modulate methylation signatures, histone-modifying enzymes and microRNA networks implicated in atrial conduction and re-entry. For AVNRT, AVRT and focal atrial tachycardia the evidential position is different. Large prospective cohort and case-crossover analyses now link air pollution to incident and acute supraventricular tachycardia, and genome-wide association studies have identified susceptibility loci for AVNRT and for accessory-pathway-mediated tachycardia, including one gene encoding a cardiac chromatin-remodeling protein; but no epigenomic profiling of nodal or accessory-pathway tissue has been reported, and no study has measured an environmental exposure, an atrial epigenetic mark and a non-AF SVT endpoint in the same participants. Twin data indicate that approximately 35% of SVT risk is attributable to genetic and 65% to unique environmental factors, which motivates a gene–environment–epigenome framework without validating its mechanistic detail outside AF. Mapping GWAS-identified loci onto environmentally responsive regulatory pathways identifies candidate convergence points between inherited risk and exposure-driven remodeling; for non-AF SVT, these are designated working hypotheses rather than established mechanisms. Air pollution and lifestyle factors are associated with supraventricular arrhythmia across the spectrum, and in atrial fibrillation there is direct evidence that they act, in part, through epigenetic reprogramming of atrial tissue. No epigenetic panel has been prospectively validated for the prediction of any supraventricular arrhythmia, and precision risk stratification therefore remains a research objective rather than a near-term clinical horizon. Integrating environmental exposure data with genetic and epigenomic profiling is nonetheless the most plausible route toward it. Future priorities include exposure-stratified, cell-resolved epigenomic profiling of atrial and nodal tissue, prospective validation of candidate circulating markers against incident arrhythmia, and replication in non-European populations and in both sexes. Full article

Atmospheric nitrogen (N) deposition is increasing N inputs to forests, but consequences for soil carbon (C) status and respiration partitioning remain uncertain. We conducted a five-year field N-addition experiment in an alpine coniferous forest in Xizang, China. We measured post-treatment soil properties, extracellular enzyme activities, estimated microbial C use efficiency (CUE), total soil respiration (Rs), root-exclusion heterotrophic respiration (Rh), and residual autotrophic respiration (Ra = Rs − Rh). Relative to the zero-N control (N0; 0 kg N ha⁻¹ yr⁻¹), N1 and N2 had higher post-treatment soil organic C, whereas all N-addition treatments had >60% lower available phosphorus and 19.12%–64% lower Rs. The Rs contrast was more strongly reflected in estimated Ra (35.93%–87% lower), whereas Rh responses varied among treatments and campaigns. N addition altered enzyme allocation and was associated with 6.70%–16.60% lower estimated CUE. Exploratory random forest and partial least squares path models indicated that Rh covaried more closely with microbial biomass, whereas Rs and estimated Ra covaried with soil physicochemical conditions and phosphorus availability. Thus, higher post-treatment soil C in some N-addition treatments co-occurred with lower Rs. However, the absence of pretreatment soil data and direct root measurements limits causal interpretation. Full article

Diabetes mellitus is a long-term metabolic disease associated with elevated glucose levels in blood and still constitutes one of the major public health issues worldwide. Inhibition of carbohydrate-digesting enzymes like α-amylase and α-glucosidase has been found to be effective in controlling postprandial hyperglycemia. The current study focused on designing, synthesis, characterization, and evaluation of novel 2,4-thiazolidinedione derivatives (D1–D5) as potent antidiabetic drugs utilizing combined in silico, in vitro, and in vivo techniques. Results from drug-likeness and ADME analyses indicated that all synthesized derivatives met Lipinski’s rule of five and had desirable pharmacokinetics properties along with reduced toxicity. Molecular docking against maltase-glucoamylase (human; PDB ID: 3TOP) protein showed good binding affinities of both D1 and D5 derivatives (−7.74 and −7.40 kcal/mol respectively) due to stable interactions with catalytic residues of enzymes. Inhibition of enzymes in vitro showed that D1 and D5 had the highest inhibitory activities of all synthesized derivatives, with IC₅₀ of 33.86 ± 2.1 and 37.55 ± 1.7 μM against α-amylase and 29.81 ± 3.2 and 32.43 ± 1.2 μM against α-glucosidase, respectively. Cytocompatibility tests on L6 myoblast cells proved that the lead compounds were well tolerated. In addition, studies in a model of Drosophila melanogaster induced by a high-sugar diet revealed a significant decrease in the level of glucose concentration depending on the dose, especially for D1 and D5, indicating their antihyperglycemic activity in vivo. Thus, these data confirm that D1 and D5 can be regarded as promising lead compounds for the development of new antidiabetics acting via inhibition of carbohydrate-metabolizing enzymes. Full article

Stress-induced sebaceous hyperactivity is a key driver of acne and seborrheic dermatitis; however, research investigating the pathway-specific mechanisms through which stress exerts its effects and the suppression of sebum production via targeting stress signaling remains relatively limited. In this study, we investigated whether Camellia nitidissima flower extract (CNF) could counteract stress-induced sebaceous dysfunction and its underlying mechanisms. Using a cortisone-stimulated SZ95 human sebocyte model, we evaluated lipid accumulation, cortisol production, signaling pathway activation, and apoptotic markers. CNF treatment dose-dependently suppressed cortisone-induced lipid production, reducing triglyceride, cholesterol, and free fatty acid levels by up to 35.44%, 38.02%, and 46.39%. Mechanistically, CNF inhibited 11β-HSD1 expression (17.17% reduction) and cortisol secretion (32.84% decrease), thereby blocking local cortisol reactivation. This upstream interception subsequently attenuated PI3K/Akt/mTOR hyperphosphorylation, downregulated lipogenic transcription factors (SREBP-1, PPARγ, LXRα, C/EBP-α) and their target enzymes (FAS, ACC, DGAT). Beyond lipid synthesis inhibition, CNF reversed cortisone-induced apoptosis resistance by reducing the Bcl-2/Bax ratio and suppressing PCNA-mediated hyperproliferation, thereby decreasing sebocyte number. These findings demonstrate that CNF exerts dual oil-control effects: reducing lipid production per cell and reducing lipid-producing cell abundance. Collectively, CNF represents a promising multi-target botanical agent for managing stress-related sebaceous disorders and cosmetic sebum regulation. Full article

Soil salinization acts as a major abiotic stress and severely hampers global agricultural productivity. Elucidating the molecular mechanisms underlying plant salt tolerance is of great significance for crop improvement. Remorin (REM) proteins are plant-specific membrane marker proteins that play a key role in stress signaling; however, their function in the salt tolerance response of sugar beet remains unclear. In this study, 14 BvREM gene family members were identified. Phylogenetic analysis revealed that this family can be divided into five subgroups and exhibits significant interspecies collinearity with Arabidopsis. Transcriptomic analysis showed that several BvREM genes were up-regulated under salt stress, with BvREM12 displaying greater expression fold change than most other family members. Using sugar beet line M14 as the gene resource material, we explored the potential function and proposed a working regulatory model for candidate gene BvREM12 in salt stress response. Subcellular localization results indicated that the BvREM12 protein is localized to the cell membrane and nucleus. Overexpression of BvREM12 significantly increased biomass accumulation in Arabidopsis, elevated antioxidant enzyme activities, decreased MDA levels, and increased proline content; a high K⁺/Na⁺ ratio was maintained, whilst Ca²⁺ levels were significantly higher than in the Col-0. Furthermore, qRT-PCR analysis revealed that several key salt-responsive genes were significantly up-regulated under salt stress. In summary, BvREM12 integrates the ‘Ca²⁺-SOS3-SOS2-SOS1’ signaling pathway with the ‘H⁺-ATPase-AKT1’ ion transport system to synergistically regulate Na⁺ efflux and K⁺ influx, thereby maintaining cellular ion homeostasis and redox balance, and significantly enhancing the plant salt tolerance. This study systematically characterized the sugar beet BvREM gene family and functionally dissected BvREM12. Different from previous studies on REM proteins mainly focusing on ROS scavenging at plasma membrane nanodomains, our work reveals that BvREM12, which is dual-localized to the plasma membrane and nucleus, coordinates the Ca²⁺-dependent SOS signaling cascade and H⁺-ATPase-AKT1 ion transport system to jointly maintain ion homeostasis and redox balance under salt stress. This work provides candidate genes and a mechanistic reference for molecular breeding of salt-tolerant sugar beet. Full article

Background and Objectives: Traditional open anterior cruciate ligament transection (ACLT) surgical models performed through arthrotomy may introduce capsular trauma and postoperative inflammatory responses that can interfere with the interpretation of post-traumatic osteoarthritis (OA). This study aimed to evaluate the feasibility of a minimally invasive, arthroscopic ACLT rabbit model performed entirely under arthroscopic visualization, without arthrotomy, using a 2.4-mm arthroscope and 2.5-mm shaver, and to provide preliminary structural, histological, synovial, and systemic biochemical characterization of the model for future therapeutic testing. Materials and Methods: Bilateral arthroscopic ACLT was performed in three skeletally mature New Zealand rabbits, followed by a supervised 12-week exercise protocol. Joint degeneration was evaluated macroscopically and histopathologically using the standardized Pritzker-Osteoarthritis Research Society International (OARSI) grading system. Collagen matrix remodeling was assessed via Picrosirius Red staining. Local intra-articular catabolism was quantified via Enzyme-Linked Immunosorbent Assay (ELISA) for matrix metallopeptidase 13 (MMP-13) and interleukin-1β (IL-1β) from synovial fluid. Systemic oxidative stress, including lipid peroxidation biomarkers, malondialdehyde (MDA), and nitric oxide (NO), was evaluated in serum. Results: The arthroscopic approach induced osteoarthritic changes, with severity varying across the cohort. Histopathology revealed vertical matrix fissures and disruption of the normal columnar chondrocyte organization, yielding a median comprehensive Pritzker-OARSI score of 6 (range 6–10), with horizontal degradation restricted to stage 2. Synovial fluid analysis demonstrated measurable concentrations of MMP-13 (188.77 ± 93.87 ng/mL) and IL-1β (174.93 ± 95.51 pg/mL). Furthermore, serum oxidative stress parameters at the 12-week endpoint included lipid peroxidation (MDA: 1.01 ± 0.03 nmol/mL) and NO levels (82.72 ± 0.32 µmol/L), presented descriptively in the absence of a comparator group. Conclusions: This study establishes the first multifaceted pathological baseline of a minimally invasive arthroscopic ACLT rabbit model before therapeutic intervention. The arthroscopic ACLT model produced structural cartilage degeneration alongside measurable intra-articular catabolic and systemic oxidative stress changes, establishing a baseline phenotype and a starting point for future therapeutic studies. Full article

Catechol 2,3-dioxygenase (C23O) is the rate-limiting enzyme in the meta-cleavage pathway of aromatic compound degradation, yet its functional annotation within the structurally conserved VOC superfamily remains challenging due to high sequence homology. Here, we isolated a catechol-degrading strain from bovine feces and identified it as Bacillus thuringiensis HHY919 via whole-genome sequencing. Genome annotation revealed four VOC genes sharing the same COG annotation (“catechol 2,3-dioxygenase”) but divergent KO annotations (two as glyoxalases and two as C23O), suggesting functional divergence. Using homology modeling and molecular docking, we compared their binding affinities toward catechol and 11 derivatives. All four proteins showed typical meta-cleavage binding energies (−4.9 to −5.4 kcal/mol), with slightly more favorable binding than an ortho-cleavage control. Notably, gene4224 exhibited the broadest and strongest predicted affinities in virtual screening against 212 compounds, particularly for trichlorophenol (−6.0 kcal/mol) and complex natural products (qvina_score ≤ −7.6 kcal/mol). Phylogenetic analysis and docking results jointly identified gene3355 and gene4224 as computationally prioritized C23O candidates, with gene4224 recommended as the top candidate for future enzyme engineering and bioremediation studies. This study provides a computational workflow for resolving functional ambiguity in VOC family enzymes and generating testable hypotheses for experimental validation. Full article

Alheira is a traditionally smoked, fermented, non-ready-to-eat meat sausage produced in the Trás-os-Montes region of Portugal, where lactic acid bacteria (LAB) are major determinants of product safety and quality. This study used whole-genome sequencing to characterize 59 LAB isolates collected from artisanal alheira produced in six municipalities. Eight species belonging to six genera were identified, and average nucleotide identity analysis resolved 24 non-redundant strain groups. Functional annotation revealed broad repertoires of carbohydrate-active enzymes, proteases, lipases, transport systems, and genes associated with tolerance to acid, oxidative, bile, heat, and salt stress. Genotype–phenotype inference was performed with a subset of 22 strain groups represented by unflagged genome assemblies. After false-discovery-rate correction, carbon-metabolism, transport, energy-metabolism, total functional-gene, acid-stress, bile-stress, and total-stress counts were positively associated with total pH decline. A mixed-effects model showed a positive association between standardized total functional-gene count and acidification ($b=0.137\pm 0.059$, $p=0.049$; marginal $R^2=0.266$, conditional $R^2=0.582$), but within–between decomposition indicated that the association was detectable between species rather than within species. Regularized regression, Random Forest, AICc model comparison, and partial least-squares regression converged on carbon metabolism, transport, energy metabolism, and acid- or bile-stress functions as the principal exploratory signals; however, sparse LASSO regression for variable selection was unstable, and prediction of unobserved species was limited. None of the five targeted pathways: acetaldehyde, citrate, diacetyl, exopolysaccharide, and lactose metabolism, remained significant after multiple-testing correction. These findings provide a genomic and phenotypic basis for selecting native starter-culture candidates while emphasizing the need for independent functional and safety validation. Full article

Background and Objectives: Impaired autophagy has been implicated in neurodegeneration, but circulating autophagy-related proteins have shown inconsistent associations with cognitive impairment. We compared serum autophagy-related protein 5 (ATG5) among older adults with normal cognition, mild cognitive impairment (MCI), and clinically diagnosed Alzheimer’s disease (AD), and examined its relationship with global cognitive performance. Materials and Methods: This single-centre cross-sectional study enrolled 164 older adults (55 with normal cognition, 55 with MCI, and 54 with clinically diagnosed AD). Cognitive status was determined through integrated clinical assessment rather than a single test cut-off. An enzyme-linked immunosorbent assay was employed to quantify serum ATG5, yielding quantifiable values for 156 participants, right-censored observations above the highest calibrator for six, and unmeasurable results for two. Since the two 96-well plates, drawn from one ELISA kit lot and processed on the same day, exhibited a substantial discrepancy in measurement scale across runs, they were analysed as distinct strata, with the principal group contrast estimated via a right-censored Tobit regression of log-transformed ATG5 within each plate, controlling for cognitive group, age and sex; a pooled plate-adjusted model also provided support. Supportive analyses used within-plate z-standardised ATG5. Associations with clinical variables were assessed using age- and sex-adjusted partial Spearman correlations with false discovery rate correction. Results: Across both analytical strata (plate 1 omnibus p = 0.293; plate 2 p = 0.264) and within the supportive pooled model (p = 0.143; AD vs. normal geometric mean ratio 0.89, 95% CI 0.73–1.09), serum ATG5 concentrations did not differ significantly between cognitive groups. Concentrations on plate 2 were 47% lower (GMR 0.53, 95% CI 0.45–0.62), a gap exceeding the inter-assay imprecision declared by the manufacturer and unexplained by the calibrator values from plate 2; since relative dispersion matched across both plates (likelihood ratio p = 0.730), the pattern suggests a multiplicative scale shift of unknown origin. Serum ATG5 levels were weakly associated with global cognitive performance assessed via the S-MMSE (partial ρ = 0.283, p < 0.001, FDR p = 0.010), an association that persisted after adjusting for education, albumin and folate and retained a similar magnitude within a linear model (β = 0.044, p = 0.014). Incorporating ATG5 into a model that included age, sex and education failed to enhance discrimination (ΔAUC +0.012 and +0.000). Conclusions: Serum ATG5 did not discriminate normal cognition, MCI, and clinically diagnosed AD. A weak rank-based association with S-MMSE was observed, but the effect was small. These findings do not support serum ATG5 as a diagnostic discriminator for clinically defined cognitive status. 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

Background/Objectives: Osteoarthritis (OA) is biologically heterogeneous, and the relationship between MRI-detected inflammation and circulating innate immune activity remains uncertain. This study aimed to determine whether serum soluble Toll-like receptor 4 (sTLR4) and soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) distinguish between the study-specific MRI-defined INF-OA and non-INF-OA categories, and whether these biomarkers are associated with OA disease burden. Methods: This prospective, single-center, cross-sectional study included 30 patients with knee OA and 30 asymptomatic healthy controls. Patients with active OA flare-up, defined by a Knee Osteoarthritis Flare-Ups Score (KOFUS) ≥ 7, were excluded before MRI assessment and category assignment. Patients with OA were assigned to study-specific MRI-defined categories using MRI Osteoarthritis Knee Score (MOAKS) features: INF-OA (n = 16) required effusion-synovitis and/or Hoffa-synovitis-related signal alteration graded ≥ 1, whereas non-INF-OA (n = 14) required grade 0 for both features. Biomarkers were measured by enzyme-linked immunosorbent assay. Primary within-OA biomarker comparisons were Holm-corrected, and adjusted models included age, sex, and body mass index (BMI). Results: Serum sTLR4 and sTREM-1 did not differ between INF-OA and non-INF-OA after Holm correction (adjusted p = 0.710 for both). In age-, sex-, and BMI-adjusted models, the INF-OA versus non-INF-OA geometric mean ratios were 1.15 (95% CI 0.92–1.44; p = 0.219) for sTLR4 and 1.09 (95% CI 0.89–1.35; p = 0.398) for sTREM-1. Given the modest within-OA subgroup sizes, smaller or moderate between-category differences cannot be excluded. After covariate adjustment, the overall OA cohort had higher geometric mean concentrations than controls for sTLR4 (ratio 1.85, 95% confidence interval 1.56–2.20; p < 0.001) and sTREM-1 (ratio 1.67, 95% confidence interval 1.43–1.97; p < 0.001). None of the systemic inflammatory measurements significantly distinguished INF-OA from non-INF-OA after correction for multiple comparisons. Within the OA cohort, each 1 ng/mL increase in sTLR4 was associated with a 4.99-point higher Western Ontario and McMaster Universities Osteoarthritis Index total score after adjustment for age and body mass index (95% confidence interval 2.28–7.70; p < 0.001). Conclusions: Circulating sTLR4 and sTREM-1 concentrations were higher in patients with OA than in asymptomatic healthy controls but did not distinguish the study-specific INF-OA and non-INF-OA categories in this sample. These findings should not be interpreted as evidence of biological equivalence between the MRI-defined categories. sTLR4 warrants further evaluation as a circulating correlate of current OA burden in larger longitudinal and multimodal studies. Full article

Antibiotic resistance represents a pressing global health challenge, with polymyxins serving as one of the last lines of defense against multidrug-resistant Gram-negative pathogens. Among them, polymyxin B (PMB) has gained renewed attention; however, its clinical use is limited by systemic toxicity. This study developed porous maize starch microparticles with tunable structure and surface chemistry to modulate PMB loading and release. Porous normal maize starch (NMS) and high-amylose maize starch (HAMS) were prepared by enzymatic treatment and modified by 2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO)-mediated oxidation to introduce anionic carboxyl groups for PMB binding. Structural characterization revealed distinct pore morphology and lamellar organization in NMS- and HAMS-derived microparticles, while oxidation increased surface charge without disrupting granular integrity. TEMPO oxidation substantially enhanced PMB loading, suggesting that electrostatic interactions between PMB and oxidized starch contributed to adsorption. In an enzyme-assisted in vitro release model, oxidized porous starches exhibited biphasic profiles, with an initial rapid release followed by a plateau, indicating heterogeneous PMB binding. Oxidized porous NMS (NMS-P-T) and oxidized porous HAMS (HAMS-P-T) showed more sustained PMB release than non-oxidized particles. PMB-loaded HAMS-P-T maintained prolonged antibacterial activity against Escherichia coli in vitro relative to NMS-based matrices. These findings demonstrate that structural modification and surface-charge influence PMB loading and release behavior in starch microparticles, enabling the design of starch-based antimicrobial peptide delivery systems. Full article