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Data-driven modeling in wastewater treatment is increasingly constrained by the reality of small, high-dimensional data, where the abundant monitoring parameters in small-sized data sets obscure fundamental mechanistic understandings. This study proposes a knowledge-driven feature selection framework that integrates mechanistic insights with statistical correlations to identify the most informative predictive features. Using nitrous oxide (N2O) emission prediction at a full-scale plant as a case study, we compared classic deep-learning feature selection algorithms using attention mechanisms against two new knowledge-based approaches: (i) expert-guided feature selection and (ii) large language model (LLM)-augmented feature selection. Expert-knowledge-guided feature selection substantially enhances predictive accuracy, achieving a mean R2 of 0.723 and an MAE of 0.033, compared to R2 = 0.712 and MAE = 0.033 for the best-performing attention-based architecture. More importantly, the proposed framework markedly improves model generalizability: under out-of-distribution high-flow conditions where the attention-based model fails to capture N2O emission patterns, the expert-selected model continues to reproduce the dominant temporal dynamics of N2O emissions. The LLM-assisted approach also delivers competitive accuracy (mean R2 = 0.596, MAE = 0.041) and similarly preserves generalizability under an input distributional shift. By introducing mechanistic understanding into the feature selection process, this framework offers a generalizable pathway for addressing complex wastewater treatment challenges while maintaining a computational efficiency.

Olfactory dysfunction is a debilitating yet under-investigated sequela of ischemic stroke, for which no approved pharmacological intervention currently exists. Despite the documented ethnomedicinal use of Lantana camara L. in Cameroon for post-stroke impairments, its neuroprotective potential against ischemia-induced olfacto-mnemonic axis damage remains unexplored. To establish a reproducible rat model of post-stroke olfactory dysfunction and evaluate the neuroprotective effects of an aqueous extract of L. camara (AELC) against global cerebral ischemia-reperfusion-induced olfacto-cognitive deficits. A modified transient global cerebral ischemia-reperfusion model was developed in male Wistar rats via bilateral common and internal carotid artery occlusion. Animals were treated with AELC (140, 280, and 560&#x202f;mg/kg), minocycline (100&#x202f;mg/kg), or piracetam (250&#x202f;mg/kg) as reference compounds. Evaluations encompassed neurological recovery, thermoregulatory profiling, olfacto-cognitive behavioral testing, and biochemical and histological analyses of the olfactory bulb, piriform cortex, prefrontal cortex, and hippocampus. Mechanistic insights were sought through LC-MS phytochemical profiling, in silico ADMET analysis, and curvature-based blind molecular docking against the &#x3b1;7 nicotinic acetylcholine receptor (nAChR; PDB: 7KOX). Ischemia-reperfusion induced severe thermoregulatory failure, locomotor deficits, and significant olfactory dysfunction (Hedges' g > 2.00), correlating with acetylcholine depletion, oxidative stress (elevated MDA and nitrites; depleted GSH), and neuroinflammation (elevated IL-1&#x3b2; and TNF-&#x3b1;). Histological examination confirmed significant neuronal pyknosis, ghost cell formation, and architectural disorganization across the olfacto-mnemonic axis. AELC at 280 and 560&#x202f;mg/kg effectively reversed these deficits, restoring cholinergic tone (p&#x202f;<&#x202f;0.001) and preserving neuronal microarchitecture comparably to reference compounds. LC-MS and in silico analyses identified Salvigenin and Kigelinone as promising bioactive leads with favorable drug-likeness and high predicted membrane permeance. Kigelinone exhibited predicted binding interactions at a site topographically consistent with known allosteric modulators of &#x3b1;7 nAChR, forming a hydrogen bond with Ala262 of the M2 transmembrane helix, a mechanistic hypothesis warranting functional validation. These findings establish a reliable rat model of post-stroke olfactory dysfunction and demonstrate that L. camara confers significant neuroprotection by modulating the cholinergic-antioxidant-neuroinflammatory axis, validating its ethnomedicinal use and offering a promising natural scaffold for the development of targeted therapeutics against ischemic brain injury.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major multidrug-resistant human pathogen. Vancomycin has long been the cornerstone therapy for invasive MRSA infections. Although nearly all MRSA isolates are susceptible to vancomycin in standard antimicrobial susceptibility testing, vancomycin treatment failure in MRSA bloodstream infections approaches 30%, and the underlying mechanisms remain poorly defined. Here, we show that subinhibitory concentrations of vancomycin promote MRSA survival within macrophages by hijacking host macroautophagy/autophagy. Using in vitro and in vivo models, we demonstrate that vancomycin exacerbates S. aureus-induced autophagic flux blockade, facilitating bacterial persistence. Mechanistically, this effect is not caused directly by vancomycin but by extracellular vesicles (EVs) secreted by S. aureus under conditions of vancomycin stress. These EVs exhibit a potent capacity to disrupt autophagosome-lysosome fusion, leading to the accumulation of autophagosomes. Proteomic profiling revealed a significant enrichment of the toxin &#x3b1;-hemolysin (Hla) in EVs derived from vancomycin-stressed bacteria. Deletion of the hla gene in S. aureus substantially attenuates the autophagy-disrupting capacity of vancomycin-induced EVs. Furthermore, recombinant &#x3b1;-hemolysin directly impaired autophagic flux in macrophages. RNA-sequencing and mechanistic analyses identified the PI3K-AKT pathway as a key signaling axis downstream of Hla, and this mechanism was further confirmed to mediate the autophagic degradation blockade induced by EVs from vancomycin-stressed S. aureus. Collectively, vancomycin triggers S. aureus to secrete EVs enriched with &#x3b1;-hemolysin, which potently inhibit autophagosome-lysosome fusion and promote intracellular MRSA survival. These findings provide critical insights into vancomycin treatment failure and identify bacterial EVs and &#x3b1;-hemolysin as potential therapeutic targets.

Actin cytoskeleton dysregulation contributes to vascular anomalies such as cerebral cavernous malformation (CCM). Talin rod domain containing-1 (TLNRD1) has been reported to interact with cerebral cavernous malformations 2 protein (CCM2), yet the downstream signaling remains debated, as previous studies have described opposite directions of Krueppel-like factor 2/4 (KLF2/4) changes after TLNRD1 depletion. Here, we combined biochemical analyses, structural modeling, transcriptomics, and single-cell network perturbation to examine the TLNRD1-CCM2 axis in endothelial cells. Coimmunoprecipitation and mass spectrometry confirmed the association between TLNRD1 and the CCM complex. Furthermore, protein docking predicted a stable TLNRD1-CCM2 interface (&#x394;G &#x2248; -50.36 kcal/mol) supported by prominent hydrogen bonds. Bulk RNA sequencing following TLNRD1 knockdown identified 677 differentially expressed genes, which were heavily enriched for actin cytoskeleton organization, with limited support for activation of the canonical MEKK3-KLF2/4 program. To assess KLF2/4 more directly, we analyzed human CCM single-cell RNA sequencing using scTenifoldKnk alongside complementary in vitro perturbations. Across these orthogonal analyses, KLF2 and KLF4 showed little to no consistent transcriptional alterations. Instead, TLNRD1 perturbation prominently altered endothelial F-actin stress fiber formation. Together, these data support a model in which TLNRD1 preferentially modulates endothelial cytoskeletal remodeling largely independent of overt KLF2/4 transcriptional shifts, helping to contextualize previous discrepancies and refining our understanding of its role in vascular biology. Current management of cerebral cavernous malformations (CCMs) remains limited by an incomplete understanding of the molecular basis of endothelial instability. This study identifies TLNRD1 as a CCM2-associated regulator of endothelial actin organization and cytoskeletal remodeling, while showing limited support for consistent KLF2/4 transcriptional changes. For clinicians, these findings add mechanistic context to a disease with few medical options and may help explain earlier conflicting experimental observations. Although direct clinical application is not immediate, the TLNRD1-CCM2 axis warrants further study as a biologically relevant pathway that may inform future translational research on cerebrovascular lesion progression.

Typified by the Lochmann-Schlosser superbase, the use of Group 1 alkali-metal alkoxides as additives to enhance the reactivity of s-block organometallic reagents is a well-established concept in organic synthesis. However, for decades the origin of this activating effect remained unclear, and the constitution of the organometallic species involved was poorly understood. More recent studies have revealed that mixed-metal, mixed-aggregate complexes constitute the active species in these transformations. By enabling bimetallic cooperation, such combinations can display enhanced reactivity and distinct regioselectivities compared to their monometallic counterparts. This Tutorial Review highlights recent advances in this field, including the use of alkali-metal alkoxides in combination with organozinc and organomagnesium reagents to unlock new applications in deprotonative metalation and metal-halogen exchange reactions. Particular emphasis is placed on developing a mechanistic understanding of how these heterobimetallic mixtures operate. Focusing on recent synthetic studies on arene functionalisation, this Review also showcases the key role of coordination effects in finely tuning the regioselectivity of these transformations.

Converting CO2 into higher alcohols is a promising strategy for producing valuable chemicals and fuels while improving carbon utilization. However, achieving selective formation of higher alcohols remains challenging because CO2 hydrogenation is thermodynamically limited and competes with methanation, the reverse water-gas shift (RWGS), and methanol synthesis. Recent studies suggest that higher-alcohol synthesis is governed not by isolated active sites, but by cooperative interfacial ensembles with complementary catalytic functions. This review examines recent progress in identifying the active sites responsible for higher-alcohol formation in Cu&#x2500;, Co&#x2500;, and Mo-based catalysts. Particular attention is given to how interfacial structures, promoter-induced electronic effects, and metal-support interactions influence key steps, including CO2/CO activation, CHx formation, CO insertion, and C&#x2500;C coupling. Cu-based catalysts often rely on Cu&#x2500;ZnO&#x2500;FexC&#x1d67; interfaces, while Co-based catalysts are generally linked to Co0/Co&#x3b4;+ or Co2C/Co0 interfacial motifs, and Mo-based catalysts benefit from adjustable oxidation states and coordination environments that stabilize oxygenate intermediates and promote chain growth. Finally, this review highlights common mechanistic principles, unresolved challenges, and interfacial active-site engineering as a useful strategy for designing more selective catalysts.

Intrinsically disordered proteins (IDPs) and regions (IDRs) challenge the classical structure-function paradigm by fulfilling essential biological roles in the absence of a stable three-dimensional fold. Rather than occupying fixed conformations, IDPs exist as dynamic ensembles that enable high-specificity, low-affinity interactions, multivalent regulatory functions, and context-dependent binding across diverse cellular environments. This conformational plasticity underlies their central roles in signaling, transcriptional regulation, chromatin organization, and the assembly of membrane-less organelles through liquid-liquid phase separation (LLPS). The present review offers several conceptual contributions. First, we develop a cross-kingdom synthesis of disorder-based chromatin regulation, demonstrating that bacterial nucleoid-associated proteins, plant transcription factors, and mammalian chromatin regulators share a conserved charge-regulatory logic, mediated by PTM-dependent mechanisms that dynamically couple environmental signals with genome organization. Second, we integrate mechanistically related but frequently siloed disease pathways, including mitophagy dysfunction, oxidative stress signaling, neuroinflammation, and aberrant phase separation, into a unified framework linking IDP conformational dysregulation to neurodegeneration and cancer. Third, we highlight underexplored regulatory dimensions of IDP biology, including proline isomerization and ubiquitylation-driven condensate formation, that influence conformational ensembles and signaling outputs in ways not captured by conventional structural approaches. Finally, we critically evaluate recent advances in AI-assisted disorder prediction and hybrid experimental-computational ensemble characterization, emphasizing both their transformative potential and current limitations. Dysregulation of IDPs underlies a broad spectrum of human pathologies, and we discuss the emerging opportunities and persistent challenges in targeting these conformationally dynamic proteins therapeutically, including through PROTAC-based degraders, condensate modulators, and ensemble-based drug screening strategies.

Glioblastoma multiforme (GBM) remains a lethal and aggressive malignancy with limited response to standard therapies. This study hypothesized that artesunate, a ferroptosis inducer, enhances the cytotoxic and immunomodulatory efficacy of standard anticancer agents through mechanisms involving oxidative stress-mediated ferroptotic cell death and lipid peroxidation, influencing hypoxia-inducible factor-1&#x3b1;, insulin-like growth factor-1 (IGF-1), and leptin pathways. These pathways collectively regulate tumor proliferation, angiogenesis, and immune evasion; hence, their modulation may sensitize GBM to therapy. The study evaluated the therapeutic potential of artesunate in combination with anticancer drugs using in vitro GL261 glioblastoma cells and in vivo subcutaneous GL261 models. Clinically relevant agents representing distinct mechanisms, temozolomide, cyclophosphamide, paclitaxel, imatinib, and thymoquinone, were tested. Tumor volume, body weight, and serum biochemical parameters (IL-6, IGF-1, HIF-1&#x3b1;, and leptin) were analyzed, along with CD3&#x2009;+&#x2009;T cell infiltration by immunohistochemistry. Combination treatments, particularly artesunate with temozolomide or paclitaxel, produced marked tumor regression compared with controls. Serum biomarker modulation and enhanced infiltration of CD3&#x2009;+&#x2009;T cells indicated activation of ferroptotic and immune-mediated anti-tumor mechanisms, with minimal systemic toxicity. Artesunate potentiates the antitumor efficacy of standard chemotherapeutic agents through mechanisms involving ferroptosis and immune modulation, promoting a tumor-suppressive microenvironment. These findings support further mechanistic and translational studies toward developing ferroptosis-driven multimodal therapies for resistant glioblastomas.

In recent years, the incidence of allergic asthma has increased dramatically. Allergen immunotherapy (AIT) is an effective approach to achieve long-term remission of allergic asthma and has gained widespread attention. However, current AIT suffers from low efficacy and long treatment duration. In this study, we found that IL-37a transgenic mice with allergic asthma exhibited reduced pulmonary inflammation, decreased Th2 cytokines, and lower allergen-specific IgE following AIT. Mechanistic studies revealed increased IL-37+ Bregs, suggesting that IL-37a may promote AIT through Bregs. Our study demonstrates the promoting effect of IL-37 in allergen immunotherapy and suggests that Bregs may underlie this mechanism, providing insights for overcoming the limitations of AIT in the future.

The pronounced heterogeneity of bladder cancer (BLCA) drives divergent patient outcomes and therapy responses. Mitophagy, a pivotal cellular quality-control and metabolic mechanism, modulates the dynamics of the tumor microenvironment (TME). Its prognostic significance and potential for guiding precision oncology, however, remain incompletely defined. This investigation aimed to evaluate the translational utility of mitophagy-associated signatures for the risk stratification of BLCA, profiling of TME, and prediction of therapeutic response. Using The Cancer Genome Atlas (TCGA)-BLCA cohort, a prognostic signature was established via least absolute shrinkage and selection operator (LASSO) and multivariable Cox regression analyses. The robustness of the signature was externally validated using two independent cohorts from the Gene Expression Omnibus (GSE32894, n=224; GSE31684, n=93). A nomogram was established to estimate survival probability. A multi-faceted analysis of TME features was conducted to compare the subgroups, integrating three algorithms: Cell-Type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT), Tumor Immune Dysfunction and Exclusion (TIDE), and Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE). Drug sensitivity and molecular alteration profiles were interrogated to identify potential personalized therapeutic avenues. A five-core-gene signature (NTN4, AKR1B1, FBXW7, MYH10, and IGF2BP3) was established. Across all cohorts, high-risk individuals exhibited notably shorter overall survival (OS) (TCGA-BLCA: P<0.001; GSE32894: P=0.003; GSE31684: P=0.03). The areas under the curves (AUCs) for 1-, 3-, and 5-year survival were 0.645-0.654 (TCGA-BLCA), 0.625-0.785 (GSE32894), and 0.567-0.651 (GSE31684). The TME analysis suggested that high-risk tumors were associated with elevated stromal scores, enrichment of M2 macrophages, and a predicted T-cell exclusion phenotype. Conversely, low-risk individuals exhibited an 'immune-inflamed' profile, marked by elevated microsatellite instability (MSI) and infiltration of CD8+ T cells. The profiling of drug sensitivity indicated that the low-risk subgroup might be more susceptible to cisplatin, whereas high-risk cases showed potential responsiveness to epidermal growth factor receptor (EGFR) and mechanistic target of rapamycin (mTOR) inhibitors. This research developed a mitophagy-derived signature that predicts survival and is associated with a potential 'stromal barrier and immune-excluded' phenotype underlying therapy resistance in high-risk BLCA. The model offers a stratification framework for therapeutic decisions. Standard chemotherapy or immune checkpoint blockade (ICB) may lead to benefits in the low-risk subgroup, whereas high-risk individuals likely require novel combinatorial strategies targeting microenvironmental remodeling. However, as these results represent predictive associations derived from public datasets, further experimental validation is warranted to confirm the underlying biological mechanisms.

Cultured fish meat requires efficient in vitro expansion of piscine satellite cells (PSCs). However, prolonged culture impairs proliferation and differentiation. Here, astaxanthin (Ast) and naringin (Nar) were evaluated as food-grade alternatives to synthetic modulators such as MHY1485. The combination of Ast and Nar promoted proliferation by accelerating cell-cycle progression and reducing apoptosis. Synergy analysis (Bliss, HAS, and ZIP) indicated a synergistic effect between Ast and Nar. Ast combined with Nar also improved mitochondrial function and redox homeostasis, as evidenced by increased mitochondrial membrane potential, decreased mitochondrial superoxide, and decreased intracellular ROS levels. Furthermore, co-treatment upregulated the expression of myogenic markers, including MyoD1, MyoG, and MyHC, and promoted myotube formation. Mechanistically, these effects of Ast and Nar were associated with mTOR activation, as indicated by phosphorylation levels, while inhibition of mTOR with rapamycin (Rap) attenuated these effects. Collectively, these results highlighted that Ast combined with Nar was an excellent supplement for large-scale expansion and differentiation of PSCs.

Vasculogenic mimicry (VM), a process of vascular-like channel formation for nutrient acquisition, drives melanoma metastasis and poor prognosis. We demonstrate that Chlorotoxin (CTX) effectively targets VM in melanoma. In A375 and A2058 cells, CTX dose-dependently inhibits proliferation with IC50 values of 1.0 and 0.8 &#x3bc;M, respectively. Furthermore, sub-lethal CTX (0.4 &#x3bc;M) impairs cell migration and VM tube formation while downregulating VM regulators VEGFR2 and NRP1. Mechanistically, CTX suppresses the long non-coding RNA LINC01235, a competing endogenous RNA that sponges miR-128-3p. CTX-induced downregulation of LINC01235 frees miR-128-3p to target and suppress the transcription factor YY1. This reduction in YY1 diminishes its binding to the VEGFR2 promoter, ultimately decreasing VEGFR2 and NRP1 transcription. Rescue assays confirm that LINC01235 overexpression restores VM capacity. By defining the LINC01235/miR-128-3p/YY1/VEGFR2/NRP1 signaling axis disrupted by CTX, these findings establish the mechanistic basis for CTX as a promising therapeutic agent against aggressive, VM-competent melanoma.

Chronic spontaneous urticaria (CSU) involves mast cells and multiple immune pathways, yet mechanisms sustaining disease chronicity remain incompletely defined. We sought to delineate immune, stromal, and senescence-associated transcriptional programs in CSU lesional skin. Lesional biopsy samples from 28 adults with CSU and 22 matched healthy controls were analyzed by bulk RNA sequencing, guided topic model-based deconvolution, and p16INK4a immunohistochemistry. Differential expression, gene set enrichment, and immune module scoring were performed. CSU skin showed 449 differentially expressed genes (309 up, 140 down), including IL4R, LAG3, CXCL8, CASP5, and NLRP12 upregulation, with downregulation of vascular/mesenchymal (CAV1, CD34) and circadian regulators (PER1, NR1D1). Deconvolution revealed expansion of helper, cytotoxic, and regulatory T cells, macrophages, and granulocyte modules, with TH2/TH17 polarization. Fibroblasts, keratinocytes, and melanocytes displayed proinflammatory, remodeling-competent states. Senescence-associated transcripts were enriched, and p16INK4a staining confirmed higher senescent cell density in CSU dermis and hypodermis (median, 92.1 vs 36.0 cells/mm2; P = .014). Adult CSU lesions comprise a chronically inflamed immune-stromal niche, integrating T-cell activation, structural cell reprogramming, and cellular senescence. These data extend current models of CSU and identify senescence as a potential amplifier of inflammation and a testable target for future mechanistic and interventional studies.

Breast cancer remains a significant clinical challenge, with treatment failure and patient mortality primarily attributable to tumor metastasis. Daucosterol linoleate (DL) was previously isolated from sweet potato (Ipomoea batatas). Using HPLC analysis of ten commercial dried sweet potato products from different regions of China, DL was detected in all samples, with concentrations ranging from 1.24&#xa0;mg to 6.05&#xa0;mg per 100&#xa0;g, confirming its widespread presence in commercially processed sweet potato products. Pharmacokinetic analysis showed that DL exhibited a peak plasma concentration (Cmax) of 1452.28&#xa0;ng/mL, an elimination half-life (t1/2) of 11.14&#xa0;h, and a mean residence time (MRT) of 9.62&#xa0;h, supporting its potential for oral administration. DL significantly suppressed proliferation and migration of MCF-7, 4T1, and MDA-MB-231 cells in vitro and reduced lung metastasis in vivo. Proteomic analysis identified SCD1 as a key molecule mediating the effects of DL. Mechanistically, DL downregulated SCD1 to inhibit epithelial-mesenchymal transition (EMT), as evidenced by decreased expression of N-Cadherin, MMP2, Vimentin, and Snail, alongside increased E-Cadherin expression. Collectively, DL inhibits breast cancer metastasis by downregulating SCD1 and suppressing EMT, supporting its dual potential as a functional food ingredient and adjuvant therapeutic.

Intervertebral disc degeneration (IDD) is a major cause of low back pain and is closely associated with inflammatory activation, extracellular matrix (ECM) degradation, cellular senescence, and ferroptosis in nucleus pulposus cells (NPCs). In this study, we aimed to clarify the therapeutic effects and underlying mechanisms of SSK1 and to develop an injectable SSK1-loaded hydrogel for localized and sustained treatment of IDD. An interleukin-1&#x3b2; (IL-1&#x3b2;)-induced NPC degeneration model was established to evaluate ECM metabolism, ferroptosis, senescence, and inflammatory signaling. Ferroptosis-related rescue experiments, pharmacological modulation of NF-&#x3ba;B and PPAR&#x3b3;, and PPAR&#x3b3; knockdown were performed to investigate the underlying mechanism. SSK1 was incorporated into a poly (ethylene glycol) diacrylate/decellularized annulus fibrosus matrix hydrogel (SSK1@PD). The physicochemical properties, cytocompatibility, sustained-release behavior, and therapeutic efficacy of SSK1@PD were evaluated in vitro and in a puncture-induced rat model of IDD using radiographic, histological, immunohistochemical, and biochemical analyses. SSK1 markedly attenuated IL-1&#x3b2;-induced degenerative changes in NPCs, including ECM degradation, iron accumulation, lipid peroxidation, GPX4 loss, and cellular senescence. Mechanistically, SSK1 restored PPAR&#x3b3; expression and suppressed NF-&#x3ba;B activation. PPAR&#x3b3; silencing largely abolished the protective effects of SSK1, whereas pharmacological activation of PPAR&#x3b3; partially reproduced them. The SSK1@PD hydrogel exhibited favorable physicochemical properties, good cytocompatibility, and sustained drug release. In vivo, local injection of SSK1@PD preserved disc height and histological architecture, improved ECM homeostasis, and reduced ferroptosis-, senescence-, and NF-&#x3ba;B-related changes in degenerated discs. SSK1@PD alleviates IDD by enabling sustained local delivery of SSK1 and modulating degenerative cellular responses, at least in part through the PPAR&#x3b3;/NF-&#x3ba;B/ferroptosis axis. This injectable biomaterial-based platform represents a promising minimally invasive therapeutic strategy for IDD. In this study, we propose an injectable hydrogel system for localized delivery of SSK1 to treat intervertebral disc degeneration. By targeting ferroptosis and inflammation while enabling sustained drug release, this strategy effectively alleviates disc degeneration in vivo. The combination of mechanistic targeting and biomaterial-based delivery highlights its potential as a minimally invasive and clinically translatable therapy for IDD.