Search Results (4,548)

Objective: In this study, Dehydroepiandrosterone (DHEA-induced Polycystic Ovary Syndrome (PCOS) mice were used as models to evaluate the improvement effect of Vitexin (Vit) on ovarian fibrosis and explore the mechanism of action of the NR4A1/NLRP3 signaling pathway. Method: Sixty 4-week-old female ICR mice of the same batch number were selected and their systems were divided into 6 groups (n = 10): normal (Control, Ctrl) group, model (Polycystic Ovary Syndrome, PCOS) group, treatment (Vitexin, The Vit group, normal NR4A1 gene silencing group (Ctrl NR4A1^-/-), NR4A1 gene silencing model group (PCOS NR4A1^-/-), and NR4A1 gene silencing treatment group (Vit NR4A1^-/-). Silencing gene modeling was performed by tail vein injection of adeno-associated virus (serotype AAV-8), and the mouse genotypes were detected by qRT-PCR technology 14 days after injection. After the genotype was determined, the PCOS group and the PCOS NR4A1^-/- group were administered dehydroepandrosterone (6 mg/100 g/d) by gavage for 28 consecutive days for modeling, while the Vit group and the Vit NR4A1^-/- group were treated with dehydroepandrosterone + vitexin (10 mg/kg/d) by gavage for 28 consecutive days. All mice were raised with pure water and regular maintenance food. After 4 weeks of drug intervention, the mice were euthanized and samples were collected. The pathological changes in ovarian tissue were observed by H&E staining, and the degree of ovarian tissue fibrosis was observed by Masson staining. The levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in mouse serum were detected by biochemical kits. The levels of inflammatory factors (IL-1β, IL-6, IL-18, TNF-α) in mouse serum were determined by enzyme-linked immunosorbent assay. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect oxidative kinase (Gsta4, Prdx3, Mgst1, Gpx3, Gsr), inflammatory factors (Nlrp3, Caspase-1, Asc, Il-1β, Il-18, Tnf-α) and fibrotic pathway-related genes (Tgf-β1, Smad3, Collagen1, CTGF, α-SMA, Mmp-13, and β-catenin) in ovarian tissues. The levels of inflammatory factors (NLRP3, Caspase-1, ASC, IL-1β, IL-18, TNF-α, IκBα) and fibrosis in mice were determined by Western blot method, and statistical description and analysis were performed using SPSS software. Result: In the wild-type genotype group, compared with the PCOS group, Vit treatment could effectively regulate the metabolic abnormalities of PCOS mice, including inhibiting excessive weight gain, restoring normal glucose tolerance, and reducing body fat content. After Vit treatment, the levels of MDA, TC, TG, LDL, IL-1β, IL-6, IL-18 and TNF-α in the serum of PCOS mice were significantly reduced, while the levels of SOD and HDL in the serum of PCOS mice were increased. The staining results indicated that Vit treatment could significantly inhibit the process of ovarian fibrosis in PCOS mice. The results of WB and PCR demonstrated that after Vit gavage treatment in mice, inflammatory and fibrotic factors such as Nlrp3, Caspase-1, Asc, Il-1β, Il-18, Tgf-β1, Smad3, Collagen1, CTGF, and α-SMA in ovarian tissues could be significantly down-regulated, and the fibrotic level of ovarian tissues could be reduced. Among the same measurement indicators, the silenced NR4A1 group showed a certain degree of increase compared with the wild genotype group, but there was no significant difference. Conclusions: Vit intervention can restore the sex hormone levels and follicular development in ovarian tissues of PCOS mice, regulate reproductive endocrine disorders and abnormal lipid metabolism levels, and regulate the expression of Collagen I, a-SMA and CTGF in the ovaries by inhibiting the NR4A1/NLRP3 signaling pathway, thereby improving the ovarian fibrosis level of PCOS mice. It is suggested that it may play a key role in the treatment of PCOS and the prevention and delay of its long-term complications. Full article

Objectives: An infectious trigger can initiate a systemic inflammatory response, which in turn activates immune cells and causes the release of various mediators. Local mediators, such as resolvin D1 (RvD1), actively interact with immune cells to promote the resolution of inflammation. This study aimed to determine the impact of RvD1 on the inflammatory response mediated by monocytes in response to LPS. Methods: To investigate the mechanism by which RvD1 affects the monocyte-mediated inflammatory response to LPS, human THP-1 monocytic cells were treated with LPS, RvD1, or vehicle for 24 h. Inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor (TNF-α), were measured using enzyme-linked immunosorbent assay (ELISA). RNA sequencing (RNA-seq) was used to identify differentially expressed genes (DEGs). The NF-κB and MAPK p38 signaling pathways were validated using real-time quantitative PCR (RT-qPCR) and Western blotting (WB). Results: RvD1 diminished the levels of IL-1β and TNF-α in LPS-induced inflammation. RvD1 significantly enhanced the mRNA expression of CREB, NRF2, and BCL-2. In addition, RvD1 significantly decreased the mRNA expression of CASP3. RvD1 regulated the inflammatory process in human monocytic THP-1 cells via the NF-κB p65 (MyD88, p65) and p38 MAPK signaling pathways (p38, BCL-2) and further suppressed the expression of apoptotic factors (PI3K, caspase-3). Conclusions: RvD1 has been shown to exert pro-resolving effects by regulating the anti-apoptotic gene BCL-2 and activating the NF-κB p65 and MAPK p38 signaling pathways. Full article

Background: Cytomegalovirus (CMV) viral load monitoring forms the basis of preemptive treatment strategies in patients undergoing solid organ and hematopoietic stem cell transplantation. This study aimed to evaluate the analytical performance and inter-method agreement of a laboratory-developed CMV real-time PCR (qPCR) test compared to a commercial reference method using plasma samples. Methods: A total of 100 EDTA plasma samples were analyzed in parallel using a laboratory-developed CMV qPCR test and the reference method (Roche Cobas^® CMV). Analytical sensitivity was determined us-ing synthetic DNA cloned into the pUC57 plasmid backbone containing the US17 region of the CMV genome, and the limit of detection (LoD₉₅) was calculated using probit regression analysis. The relationship between the quantitative results obtained from clinical samples was evaluated using the Spearman rank correlation coefficient, while inter-method clinical agreement was assessed using the Bland–Altman method. Results: The limit of detection (LoD₉₅) of the laboratory-developed CMV qPCR test, as determined by probit regression analysis, was 63.8 copies/µL. A weak and statistically non-significant correlation was ob-served between the laboratory-developed CMV qPCR test and the reference method in Spearman rank correlation analysis of samples for which numerical quantitative results were available from both methods (ρ = 0.32; p = 0.22; n = 16). Bland–Altman analysis showed a mean difference of −0.48 log₁₀ units, with the vast majority of measurements falling within the 95% limits of agreement. Conclusions: The assay demonstrated measurable analytical performance and inter-method agreement; however, its use for quantitative viral load monitoring, particularly at low CMV DNA levels, should be interpreted with caution. Full article

Background/Objectives: As a global chromatin organizer, SATB1 is increasingly implicated in neurodevelopmental disorders (NDDs). This study aims to delineate the clinical and molecular characteristics of a novel de novo SATB1 variant in a patient presenting with epilepsy-dominant NDDs phenotypes. Methods: Triggered by the onset of seizures, trio-based whole-exome sequencing (Trio-WES) was performed to identify the genetic etiology. Subsequent sleep electroencephalogram (EEG) and magnetic resonance imaging (MRI) were then conducted to further characterize the patient’s clinical phenotypes. Pathogenicity was assessed through structural modeling and functional characterization. Nonsense-mediated mRNA decay (NMD) status, protein expression profiles, and subcellular localization were determined by reverse-transcription quantitative PCR (RT-qPCR), Western blotting, and immunofluorescence staining. The transcriptional regulatory impacts of the variant were quantified using dual-luciferase reporter system targeting known downstream regulatory elements. Clinical responses to antiepileptic intervention was also monitored. Results: We identified a novel de novo heterozygous pathogenic frameshift variant in SATB1 (NM_002971.5: c.1718_1719insCA; p.Val574Argfs*134) in a patient presenting with early-onset epilepsy, mild intellectual developmental disorder (IDD), speech delay, and dental anomalies. Functional assays demonstrated that the variant-derived transcript escaping NMD, yielding a truncated protein that forms irregular punctate aggregates within nuclei. Dual-luciferase assays revealed significantly increased transcriptional activity, indicating a loss of the protein’s innate transcriptional regulatory capacity. Clinically, treatment with sodium valproate (VPA) successfully stabilized seizures of the patient, markedly reducing both frequency and intensity. Conclusions: The study reports a novel SATB1 frameshift variant that exerts pathogenicity significant functional impairment by disrupting protein localization and transcriptional regulation. These findings expand the genetic spectrum of SATB1-related NDDs and underscore the efficacy of targeted antiepileptic management in genetic diseases. Full article

Background: We recently identified multiple alternative circRNAs generated through alternative circularization of primary transcripts of the BCL2-related ovarian killer (BOK) gene. In the present study, we evaluated the prognostic potential of a recently discovered BOK circRNA, namely circ-BOK-6, in breast cancer (BC). Methods: Tumor specimens from a cohort of 172 female BC patients were analyzed, including paired adjacent non-cancerous breast tissue from 44 cases. circ-BOK-6 expression levels were quantified using an in-house–developed quantitative real-time PCR assay, followed by comprehensive biostatistical and survival analyses. Results: circ-BOK-6 expression differed between matched tumor and normal breast tissues. However, no significant associations were observed between circ-BOK-6 expression and BC clinicopathological characteristics. Importantly, after splitting at the median value, low circ-BOK-6 expression was associated with poorer disease-free survival (DFS) and overall survival (OS). Multivariate Cox regression analysis further demonstrated that the prognostic value of low circ-BOK-6 expression was independent of established prognostic factors included in the models, including the prognostic stage or the molecular subtype and the anatomic stage of the disease. Additionally, stratified analyses based on these key prognostic parameters revealed that low circ-BOK-6 expression retains prognostic significance within specific patient subgroups. Conclusions: These findings suggest that low circ-BOK-6 expression represents a promising independent biomarker of adverse prognosis in BC. Full article

Reusing waste materials is a sustainable practice to reduce the impact of human activities on the global ecosystem. In particular, agro-industrial waste can be explored as an innovative and green source of beneficial bioactive compounds. For this purpose, seventeen different varieties of wine grape pomace were collected from the Apulia region (Italy) to perform extractions and qualitative–quantitative phenolic profile characterization. To ensure an environmentally friendly extraction process, natural deep eutectic solvents (NADESs) were used as privileged media. After an initial screening, Merlot (MEA) and Sangiovese (SGA) varieties were then selected for extraction with betaine/lactic acid 1:4 (mol/mol) +40% water (w/w) and choline chloride/ascorbic acid 2:1 (mol/mol) +40% water (w/w). They were further investigated by LC-QTOF-MS for a comprehensive metabolomic evaluation. Betaine-based extracts from both cultivars exhibited higher polyphenol contents than choline chloride-based ones: 175.0 and 161.8 mg/kg compared to 59.6 and 40.4 mg/kg. Qualitative antimicrobial assays confirmed the effectiveness of betaine/lactic acid (MEA) and choline chloride/ascorbic acid (SGA) extracts on B. allii and E. coli, respectively, as well as the NADES themselves, even though it is difficult to discriminate the individual effects. Finally, the evaluation of both antioxidant activity and total polyphenol content led to good results for betaine/lactic acid (2.8 and 3.2 mg TEAC/g DW and 0.92 and 0.93 mg GAE/g DW for SGA and MEA, respectively), while the content of ascorbic acid, used as a component of NADES, substantially influenced the relevant obtained results in choline chloride/ascorbic acid samples. These findings support the potential of combining NADES with polyphenols recovered from grape pomace as a promising approach for further optimization and application-oriented investigation. Full article

Maize (Zea mays L.), a typical thermophilic crop originating from tropical regions, exhibits an inherent sensitivity to low-temperature stress. Cold stress severely restricts maize seed germination, seedling growth, the physiological metabolism, and the final grain yield, which greatly limits its geographical cultivation range and sustainable industrial development. Elucidating the molecular regulatory mechanisms underlying maize cold tolerance and excavating cold-resistant functional genes are essential for the molecular breeding of cold-tolerant maize varieties and expanding maize planting areas in high-latitude and low-temperature-prone regions. In this study, using the strongly cold-tolerant maize inbred line B144 as the experimental material, we cloned the ZmMAPKKKA gene (NCBI accession: LOC103651289) and systematically screened and verified its cold-stress-specific interacting proteins via multiple molecular biological assays. The full-length coding sequence (CDS) of ZmMAPKKKA is 1134 bp, encoding a 377-amino-acid protein with a predicted molecular weight of 40.37 kDa. The quantitative real-time PCR (qRT-PCR) results demonstrated that the ZmMAPKKKA expression was significantly upregulated by 16.56-fold in maize roots after 12 h of low-temperature treatment, indicating a tissue-specific and robust cold response in root tissues. A total of 25 interacting proteins were identified through yeast two-hybrid screening, among which three stress-responsive proteins, including a protein kinase (LOC100286253), a protein phosphatase 2C (PP2C) (LOC542176), and a NAC transcription factor (LOC118474710), were selected for subsequent verification. The Pull-Down, Co-immunoprecipitation (Co-IP), and bimolecular fluorescence complementation (BiFC) assays consistently confirmed that ZmMAPKKKA specifically interacts with these three proteins both in vitro and in vivo under cold stress conditions. This study is the first to construct a ZmMAPKKKA-centered protein interaction module in the maize mitogen-activated protein kinase (MAPK) cascade under cold stress, establishing a novel kinase–phosphatase–transcription factor regulatory cascade that improves the current understanding of cold signal transduction mechanisms in maize. Homologous genes of ZmMAPKKKA in gramineous crops including rice (Oryza sativa) and sorghum (Sorghum bicolor) have been proven to participate in diverse abiotic stress responses, suggesting the conserved functional roles of MAPKKK family genes across gramineous species. Collectively, our findings provide comprehensive insights into the molecular mechanism of the maize MAPK signaling pathway mediating cold stress adaptation and supply valuable functional gene resources for cold-tolerant maize germplasm innovation and molecular breeding. Full article

Biophysical sensing technologies have become central to modern drug discovery because they enable direct, quantitative characterization of ligand–target interactions. In contrast to conventional biochemical and cellular assays that infer binding from downstream functional responses, biophysical methods detect interaction events through measurable physical changes such as refractive index, heat, fluorescence, mass, or protein stability. This review surveys the major classes of biophysical sensors used in drug discovery, including surface-based optical methods, calorimetry, solution-state spectroscopic techniques, mass spectrometry-based approaches, and cellular target engagement assays. For each modality, we outline the measurement principle, the key parameters obtained, and its value across hit identification, hit validation, lead optimization, and mechanism-of-action studies. We also emphasize the growing importance of combining orthogonal methods to improve confidence in binding data, resolve assay artifacts, and strengthen early decision-making. Finally, we discuss how biophysical measurements are increasingly integrated with structural biology and computational analysis to support more predictive and mechanism-driven discovery workflows. Collectively, these technologies provide a richer and more reliable understanding of molecular recognition and thereby improve the progression of drug candidates. Full article

Type-1 ryanodine receptor (RyR1) is essential for skeletal muscle contraction. This Ca²⁺ release channel is expressed in cardiac myocytes; however, its function remains elusive. Cardiac-specific RyR1 overexpression (OE) mice were generated under the cardiac-specific Myh6 promoter. Cardiac hypertrophy (CH), cardiac functions, and mechanistic changes in RyR1 OE and control (wildtype, WT) mice were assessed using hematoxylin and eosin staining, echocardiography, electrocardiogram, quantitative RT-PCR, Western blotting, [³H]-ryanodine binding assay, confocal microscope, ROS dye Amplex Red and 2′,7′-dichlorofluorescein diacetate. RyR1 OE mice had increased whole heart, left ventricular weight, and left ventricular wall thickness, but decreased cardiac output and stroke volume, thereby presenting CH and heart failure (HF). CH markers like ANF, BNF, and aSKA mRNAs were increased in RyR1 OE heart. RyR1, but not RyR2 or RyR3, expression was increased in the RyR1 OE mouse heart. Similar results were found in mice with TAC-induced CH. RyR1, but not RyR2 mRNA, was increased in cardiac muscle from dogs and humans with CH and/or HF. Maximum [³H]-ryanodine binding was increased, whereas the binding dissociation constant decreased in left ventricular cardiomyocytes from RyR1 OE mice. RyR2-dependent Ca²⁺ sparks were increased, which was blocked by riluzole, a small molecule known to inhibit RyR2. Consistently, ROS was remarkably increased in RyR1 OE cardiac cells. We first generated cardiac-specific RyR1 OE mice; these mice had CH, HF, and increased RyR1 expression with no RyR2 or RyR3 alteration. Similar changes were observed in mice, dogs, and humans with CH and HF. Increased mitochondrial ROS-dependent RyR2 Ca²⁺ release was essential for RyR1-induced CH and HF. Full article

Background/Objectives: Electrolytes used in in vitro corrosion testing critically determine the behavior inferred for metallic dental implants, yet formulations and their justifications are inconsistently reported across the literature. This review compiles and compares electrolytes employed to simulate the oral cavity and the bone–implant interface, linking their chemical composition to the corrosion mechanisms they target. Methods: This structured narrative review synthesized peer-reviewed literature on simulated electrolytes used for in vitro corrosion testing of metallic dental implants and implant-related alloys. Literature was identified using database searches and targeted reference screening, with emphasis on artificial saliva formulations, physiological simulated fluids, challenge chemistries, protein-containing media, hydrodynamic conditions, and microbiological models. Relevant formulations were standardized to grams per liter and grouped according to application domain and targeted corrosion mechanisms. Results: The analysis maps electrolyte selection to corresponding corrosion modes, including uniform dissolution, pitting, crevice, galvanic, and microbiologically influenced corrosion. Consolidated composition tables highlight how pH, halide concentration, calcium–phosphate balance, proteins, gas control, and flow conditions modify passive-film stability and metal-ion release. Dental-specific gaps are identified, notably the lack of a standardized fluoride–pH matrix and limited guidance for microbiome-integrated assays. Conclusions: Aligning electrolyte formulations with the research question enhances reproducibility and mechanistic interpretation. However, current in vitro corrosion data should be interpreted cautiously because quantitative links between simulated-fluid testing and clinical outcomes such as peri-implantitis, peri-implant bone loss, and implant failure remain insufficiently established. The adoption of shared reporting standards, dynamic programmable chemistries, and interoperable datasets may improve the translational value of future corrosion studies. Full article

Background: To investigate whether coating xenogeneic bone grafts with hyaluronic acid influences early osteogenic and fibrotic marker expression in vitro. Methods: Three xenograft materials were evaluated, including one hyaluronic acid-coated product and two uncoated deproteinized bovine bone mineral products, all commercially available. Human osteoblast-like cells (U2OS) and bone marrow stromal cells (HS5) were cultured with material extracts. Proliferation was assessed using XTT assay at 24 and 48 h. Cell adhesion was evaluated through fluorescence microscopy. Osteogenic markers (RUNX2, COL1A1) and fibrotic markers (COL3A1, TGF-β3) were quantified using quantitative real-time PCR. Statistical analysis employed one-way ANOVA with Benjamini–Krieger–Yekutieli (BKY) two-stage FDR correction for datasets that met the normality assumption, and the Kruskal–Wallis test with Dunn’s post hoc test for non-normally distributed data (HS5 XTT assay). Pairwise comparisons were restricted to each xenograft group versus the untreated control; an adjusted p-value < 0.05 was considered statistically significant. Results: At 48 h, the HA-coated xenograft (Xeno1) showed the highest mean metabolic activity in U2OS cells (0.538 ± 0.056) compared with the uncoated Xeno2 (0.450 ± 0.120) and Xeno3 (0.439 ± 0.073); however, after FDR correction no statistically significant differences were observed between groups. The coated material was associated with upregulation of early osteogenic markers, 2.61-fold RUNX2 upregulation (p = 0.01) compared to untreated cells. Both coated and uncoated xenografts demonstrated equivalent suppression of fibrotic markers in HS5 cells, reducing COL3A1 by 92.7% (p = 0.001) and TGF-β3 by 92.1% (p = 0.001). Conclusions: These exploratory in vitro findings suggest that HA coating may enhance early osteogenic marker expression. The observed effects on stromal markers warrant further investigation using primary cells, additional fibrotic endpoints (e.g., TGF-β1, ACTA2), and in vivo models before translational conclusions can be drawn. Full article

The qualitative detection of circulating tumor human papillomavirus DNA (ctHPV) has shown promise in HPV-associated cancers. We performed a systematic review and meta-analysis to evaluate the association of pretreatment ctHPV levels with survival outcomes and quantitative tumor burden metrics. Databases were searched through 5 January 2026. Studies were eligible if they included patients with HPV-associated cancers and reported quantitative pretreatment ctHPV levels in relation to survival outcomes or tumor burden. Twenty-three studies were included in the quantitative synthesis. Higher pretreatment ctHPV levels were associated with poorer progression-free survival (PFS) (8 studies, n = 883; HR = 1.86, 95% CI 1.19–2.90; p = 0.013). This association was primarily driven by studies in oropharyngeal cancer (OPC; HR = 2.47, 95% CI 1.59–3.84; p = 0.007), whereas no significant association was observed in cervical cancer. In multivariable analyses, elevated ctHPV remained associated with shorter PFS (HR = 1.87, 95% CI 1.66–2.10; p = 0.002). No significant association was observed for overall survival. Pretreatment ctHPV correlated with nodal volume in OPC (r = 0.45), nodal and tumor volume in OPC/anal cancer (r = 0.39), primary tumor volume in OPC (r = 0.22), and tumor diameter in cervical cancer (r = 0.44). Higher pretreatment ctHPV levels are associated with greater tumor burden and poorer PFS in HPV-associated OPC. CtHPV shows potential as a prognostic biomarker, although further prospective studies and assay standardization are needed. Full article

Atherosclerosis is caused by inflammatory processes that alter the permeability of arterial wall cells and leucocyte recruitment, leading to oxidation of low-density lipoproteins in the artery. The use of dietary polyphenols as antioxidants seems promising. Herein, molecular docking-based screening was initially used to predict the interactions of epicatechin and hydroxytyrosol on multiple cytokines that can trigger atherosclerosis development. Computational results show that epicatechin and hydroxytyrosol interact with the cytokines, namely, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, monocyte chemoattractant protein 1 (MCP-1), granulocyte–macrophage colony-stimulating factor, leukocyte differentiation antigen CD36, and oxidized low-density lipoprotein receptor-1. Cytotoxicity of both the bioactive compounds to human monocytic THP-1 macrophages was evaluated by lactate dehydrogenase and crystal violet assays. ROS activity evaluation was done for the phytocompounds followed by monocyte migration assay for MCP-1. The expression levels of selected biomarkers were further assessed by quantitative polymerase chain reaction. Inhibition of these atherosclerotic biomarkers may limit the atherogenic effect. Notably, these two polyphenols at a concentration of 0–125 µg/mL for 24 h showed no cytotoxicity on THP-1 macrophages and exhibited decreased ROS production and MCP-1 levels. The genes implicated in the early stages of inflammation are potential therapeutic targets to effectively reduce atherogenesis and prevent CVD. The interaction between the selected cytokines and the two natural compounds indicates their potential ability to inhibit the inflammation in vitro and exhibit anti-atherogenic effects. Hence, epicatechin and hydroxytyrosol possess significant anti-atherosclerotic effects and, in combination, could contribute positively to the treatment of atherosclerosis. Full article

The growing threat of antimicrobial resistance necessitates alternative strategies to conventional antibiotics. Insects represent a promising source of antimicrobial peptides (AMPs) due to their potent innate immune responses. In this study, we investigated the hemolymph peptide extracts from Hermetia illucens larvae as a bioresource of infection-induced AMPs. Larvae were challenged with Gram-negative (Escherichia coli) and Gram-positive (Micrococcus flavus) bacteria, and hemolymph-derived peptides were extracted and fractionated by RP-HPLC. Peptide fractions were screened in vitro by agar diffusion assay against a panel of pathogenic strains. While extracts from uninfected larvae were essentially inactive, fractions 9–13 from infected larvae showed broad-spectrum antibacterial activity, including against multidrug-resistant clinical isolates such as carbapenem-resistant Klebsiella pneumoniae, suggesting an inducible immune response. Fractions were further analyzed by SDS-PAGE and LC-MS/MS to identify candidate AMPs, while shotgun proteomics on unfractionated extracts enabled quantitative comparison among conditions. Proteomics raw data are available via ProteomeXchange Consortium, with the dataset identifier PXD071144. In silico prediction using ProP 1.0, APD3, and CAMP_R4 supported the antimicrobial potential of candidate peptides. Overall, our integrative approach demonstrates that bacterial infection induces a diverse and bioactive immunopeptidome in H. illucens, supporting its potential as a sustainable platform for the discovery of novel antimicrobial agents. Full article

Hydrodistillation of aromatic plants for essential oil production generates substantial amounts of solid and liquid residues that are commonly discarded despite their potential value as sources of bioactive compounds. In this study, the essential oil and post-distillation residues of Rhododendron tomentosum Harmaja were evaluated within a waste-to-value framework to recover phenolic compounds with antioxidant and antimicrobial properties. Dry extracts obtained from liquid (DEA) and solid (DEE) residues were characterized in terms of total phenolic and flavonoid contents, antioxidant capacity (DPPH assays), and antimicrobial activity against selected microorganisms. Quantitative HPLC–PDA analysis revealed multiple phenolic compounds. Extracts derived from solid residues exhibited significantly higher phenolic and flavonoid contents and stronger antioxidant activity than those obtained from liquid residues, indicating that solid by-products constitute a richer phenolic matrix. Antimicrobial assays revealed pronounced activity for extracts prepared from plant material harvested in October, particularly those based on propylene glycol and glycerin, which were effective against both Gram-positive bacteria and selected Gram-negative clinical isolates. The essential oil showed broad-spectrum antimicrobial activity, including inhibition of Aspergillus niger. Stability studies demonstrated that the phenolic composition and bioactivity of the dry extracts were largely preserved after one year of storage. These findings demonstrate that R. tomentosum hydrodistillation residues represent a promising source of natural antioxidants and antimicrobial agents, supporting their potential utilization as value-added ingredients in food and cosmetic applications and contributing to circular economy strategies. Full article