Search Results (5,897)

Pulmonary arterial hypertension (PAH) is marked by vascular remodeling, yet the role of adventitial fibrosis—and its modulation by sex and hormonal status—remains unclear. We examined stage-specific adventitial remodeling and pulmonary artery adventitial fibroblast (PAAF) mechanosensitivity in male, ovary-intact female, and ovariectomized (OVX) female Sprague–Dawley rats with SuHx-induced PAH. Hemodynamics, pulmonary artery histology, and adventitia-specific transcriptional profiling were integrated with in vitro assays of PAAFs exposed to defined substrate stiffness and stretch. All groups developed comparable increases in mean pulmonary arterial pressure, but vascular resistance shift and adventitial fibrosis diverged by sex: intact females showed attenuated increase in pulmonary vascular resistance and transient collagen accumulation, whereas OVX females mirrored the sustained, male-like progression. Extracellular matrix (ECM) gene activation occurred without smooth muscle actin induction, suggesting noncanonical fibrotic pathways. In vitro, intact female PAAFs required higher substrate stiffness to induce profibrotic gene expression, indicating a hormone-modulated stiffness threshold. OVX PAAFs showed persistent transcriptional reprogramming, while stretch-induced ECM upregulation occurred predominantly in male-derived PAAFs. These findings demonstrate that adventitial fibrosis in PAH is shaped by both hormonal and chromosomal sex, independent of hemodynamic severity, and highlight fibroblast mechanosensitivity as a potential target for stage- and sex-specific interventions. Full article

Antimicrobial resistance (AMR) was associated with 4.95 million deaths in 2019 and may cause 10 million deaths annually by 2050. We synthesize evidence on how the black soldier fly (Hermetia illucens) has evolved an expanded antimicrobial peptide (AMP) repertoire, which structural features drive family-specific activity, what mechanisms are directly demonstrated in H. illucens, and how AI contributes. PubMed, Web of Science, and Scopus (plus targeted Google Scholar) were searched from inception to 1 February 2026; studies were included when they reported BSF peptide identities, expression/proteomics, evolutionary analyses, quantitative activity, mechanistic assays, or BSF-focused computation, and claims were tiered as predicted, expression-supported, or experimentally supported. The literature supports 50–80 BSF AMP genes, plausibly shaped by gene duplication and balancing/diversifying selection in microbe-rich substrates, with marked induction plasticity across tissues, development, diet, and challenge. SAR is family-dependent: defensin-like peptides rely on disulfide-stabilized CSαβ folds and cationic surface topology; cecropin-like peptides on amphipathic α-helices with selectivity trade-offs; attacin-like peptides on β-architecture where charge-based heuristics are weak; and diptericin/proline-rich peptides remain largely inference-driven in BSF. Mechanistic evidence is strongest for membrane/envelope-centered killing by DLP4 and pore-associated envelope disruption by a recombinant attacin-like peptide, whereas pore geometry, oligomerization, intracellular targets, and broad “resistance-proof” claims remain unresolved. Key gaps include assay heterogeneity, salt/serum stability, selectivity/toxicity, resistance-risk testing, and limited in vivo validation, which must be addressed for credible AMR-relevant translation. Full article

Inonotus hispidus is an important medicinal and edible fungus within the “Sanghuang” category, featuring a broad host range and rapid fruiting body growth. However, its wild resources are currently threatened by overharvesting. Simultaneously, large-scale jujube (Ziziphus jujuba) cultivation generates substantial pruning waste, often burned. This study explored the feasibility of using jujube wood as a cultivation substrate for I. hispidus. Three I. hispidus strains, Z1, Z2, and ZL, were cultivated on substrates with varying proportions of jujube wood replacing cottonseed hulls. The biological efficiency, nutritional components, active compounds, and free amino acid profiles of the resulting fruiting bodies were analyzed. Non-targeted metabolomics was used to investigate global metabolic changes. Results indicated that all strains successfully colonized the jujube-based substrates and produced fruiting bodies. Strain ZL exhibited the highest biological efficiency and the shortest growth period on the 48% jujube wood substrate, while others showed significantly increased triterpenoids and flavonoids content. Metabolomic analysis revealed substrate-dependent and strain-specific alterations in metabolic pathways, particularly in amino acid biosynthesis, the TCA cycle, and secondary metabolism. This study confirms jujube wood as a viable alternative substrate for the edible (ZL) and medicinal (Z1, Z2) cultivation of I. hispidus, providing a sustainable production method while establishing a valuable utilization pathway for jujube wood waste. Full article

Thyroid hormones profoundly modulate hepatic fatty acid and cholesterol synthesis and turnover. Although nonalcoholic fatty liver disease (NAFLD) shows epidemiological links to hypothyroidism, the genetic substrates of this relationship remain unresolved. Integrating large-scale genome-wide association studies with single-cell transcriptomics, spatial transcriptomics, and single-cell chromatin accessibility via state-of-the-art computational approaches, we interrogated the association between NAFLD and hypothyroidism across organ systems, cellular expression landscapes, and molecular–genetic strata. We uncovered pronounced spatial specificity in genetic risk within the liver, prioritized hepatocytes as the principal shared cell type affected, and, leveraging spatial transcriptomics, advanced a dynamic spatiotemporal two-hit model. We further nominated MAGI3, RRNAD1, and PRCC as high-confidence candidate genes and pinpointed a key risk locus, rs926103. These findings deliver a dynamic, testable framework for the full pathophysiological continuum linking NAFLD and hypothyroidism and yield new targets and leads for precision intervention. Full article

General-purpose large language models excel at open-domain question answering, but in railway operation and maintenance (O&M) scenarios they still suffer from hallucinated knowledge and poor domain adaptation. In practice, railway O&M knowledge mainly arises from two heterogeneous sources: spatio-temporal data such as train trajectories, which are organized along the spatial layout of railway lines, and domain documents such as operating rules, which exhibit varying degrees of structural regularity. Traditional retrieval-augmented generation (RAG) systems usually flatten these multi-source data into a single unstructured text space and perform global retrieval in one embedding space, which easily introduces noisy context and makes it difficult to precisely target knowledge for specific lines, sections, or equipment states. To overcome these limitations, we propose CARAG, a context-aware RAG framework tailored to railway O&M data. CARAG treats domain documents and spatial data as a unified knowledge substrate and builds a spatial knowledge graph with concept and instance levels. On top of this knowledge graph, a GraphReAct-based multi-turn interaction mechanism guides the LLM to reason and act over the concept knowledge graph, dynamically navigating to spatially and semantically relevant candidate regions, within which vector retrieval and instance-level graph retrieval are performed. Experiments show that CARAG significantly outperforms baseline RAG methods on RAGAS metrics, confirming the effectiveness of structure-guided multi-step reasoning for question answering over multi-source heterogeneous railway O&M data. Full article

Amid accelerating global urbanization, the Qinghai–Tibet Plateau, as a repository of multi-ethnic architectural heritage, plays a crucial role in preserving plateau cultural diversity and sustaining harmonious human–environment relationships. A critical research gap persists, however, in the systematic, comparable, and quantitative assessment of urban architectural character across plateau towns, particularly in high-altitude, ecologically sensitive, and multi-ethnic regions such as Haixi Mongol and Tibetan Autonomous Prefecture. This study takes the Haixi Mongol and Tibetan Autonomous Prefecture as a case to address the specific paradox between the homogenization of urban architectural styles and the erosion of cultural authenticity in plateau towns. We develop and apply an innovative three-dimensional evaluation model—encompassing natural substrate, built environment, and cultural context—to 22 towns. For the first time in research on this region, a chained methodological approach integrating descriptive statistics, principal component analysis (PCA), and cluster analysis is employed to systematically examine the spatial differentiation of architectural character. The analysis reveals three key findings. First, it delineates a regional composite landscape characterized by mountain-basin enclosures, seasonal arid rivers and lakes, small-scale towns with expansive layouts, and multi-ethnic cultural fusion. Second, it identifies a clear ternary differentiation in urban style dominance: nine towns are nature-dominated, nine are human-made (built environment) dominated, and only four are culture-dominated, quantitatively highlighting a significant weakness in the cultural dimension. Third, cluster analysis objectively classifies the towns into eight distinct character groups—for instance, Category I towns exhibit strong architectural regionalism and traditional continuity, whereas Category V towns integrate modern relics with adjacent mountain-water features. Methodologically, this study contributes by providing a replicable, chained quantitative framework that addresses a critical gap in comparative urban studies of high-altitude, underdeveloped regions. Empirically, it reveals the specific “nature > human-made > culture” dominance pattern in Haixi and offers a scientific foundation for formulating differentiated conservation and development strategies tailored to distinct town types in the ecologically fragile areas of western China. Full article

Heroin is a highly addictive drug that exerts its primary effects through activation of μ-opioid receptors. Its principal active metabolite, 6-monoacetylmorphine (6-MAM), significantly contributes to heroin’s neurological effects and acute toxicity. Current pharmacotherapies for heroin use disorder, employing opioid receptor agonist or antagonist, are often limited by risks of dependence, tolerance, and/or adverse side effects. In this context, enzyme-based therapy emerges as a promising alternative by rapidly converting drugs into inactive or less harmful metabolites in the blood. As a macromolecule, the enzyme does not cross the blood–brain barrier, thereby avoiding side effects in CNS. Through structure-based computational screening, Xoo-PepA (PDB ID: 3JRU), a leucine aminopeptidase from Xanthomonas oryzae pv. oryzae, was identified as a potential enzyme capable of hydrolyzing heroin and 6-MAM. Computational and experimental analyses confirm that Xoo-PepA hydrolyzes heroin sequentially to 6-MAM and subsequently to morphine. Enzymatic properties including dependence on metal ions, optimal pH, thermal stability, and substrate specificity were characterized accordingly. Notably, supplementation with Ni²⁺ or Zn²⁺ and TCEP extended Xoo-PepA’s half-life at 37 °C from 1 h to over 24 h, highlighting the essential role of metal ions in maintaining structural stability. Moreover, Ni²⁺ enhanced Xoo-PepA’s hydrolysis toward peptidase substrate L-leucine-p-nitroaniline by 770-fold, yet conferred no significant activation toward heroin. Mutations in metal ion-coordination residues (e.g., K262A, D267A/E346L) exhibited different activity profiles toward these two types of substrates, suggesting a distinct regulatory mechanism of metal ions may be involved in these activities. This study provides the first demonstration that Xoo-PepA, a non-mammalian, metal-dependent aminopeptidase, can hydrolyze heroin and 6-MAM, shedding light on its functional versatility and biochemical characteristics. Full article

Endoplasmic reticulum-mediated protein quality control (ERQC) safeguards secretory pathway proteostasis by recognizing, retaining, repairing, and removing misfolded proteins, and is therefore essential for plant growth, development, and stress tolerance. This system relies on ER-associated degradation (ERAD), in which irreparably misfolded proteins are first recognized in the ER, then exported across the ER membrane to the cytosol, where they are ubiquitinated by ER membrane-anchored ubiquitin ligases, and subsequently degraded by the cytosolic proteasome. Studies in yeast and mammals have defined several conserved ERAD branches, including a multiprotein ERAD complex centered on the polytopic ER membrane E3 ligase HMG-CoA reductase degradation protein 1 (Hrd1), which integrates substrate recognition, membrane retrotranslocation, ubiquitin conjugation, and cytosolic extraction. Recent advances in Arabidopsis show that plants retain the core Hrd1 ERAD architecture while incorporating additional regulatory elements that adapt this machinery to plant-specific physiological demands. Genetic and biochemical analyses of misfolded receptor kinases and engineered substrates have uncovered conserved and plant-specific components of the plant Hrd1 complex, revealing how the plant ERAD pathway integrates ERQC with hormone signaling, stress adaptation, immune responses, and growth regulation. This review synthesizes recent advances in plant ERAD research and highlights key conceptual and mechanistic questions that remain to be resolved. Full article

India’s diverse culinary heritage includes a wide spectrum of traditional fermented foods that harbour complex microbial communities essential for flavour development, preservation, and nutritional enhancement. These microorganisms—primarily lactic acid bacteria, yeasts, and molds—contribute functional properties that extend beyond food transformation to confer health benefits, including probiotic potential and metabolic regulation. This review integrates classical microbiological studies with modern molecular approaches such as metagenomics, metatranscriptomics, and metabolomics to elucidate the microbial diversity of Indian fermented foods. It highlights how geography, substrates, and ethnic traditions shape region-specific microbial consortia sustained through long-standing ethno-microbiological practices. Special focus is given to the glycemic modulation achieved through microbial fermentation, wherein organic acid production and resistant starch formation lower glycemic index and improve glucose metabolism. These processes, along with enhanced nutrient bioavailability, vitamin synthesis, and immunomodulation, illustrate the broader functional potential of fermentation. The review also examines interactions between food-borne microbes and the human gut microbiota, underscoring implications for personalized nutrition. Finally, it discusses modernization and commercialization strategies and outlines future directions involving multi-omics integration, indigenous starter cultures, and microbiome-based innovations to harness India’s microbial heritage for improved health and sustainable food development. Full article

Background: Pathogenic ST3GAL3 variants cause neurological and cognitive impairment, defining a distinct congenital disorder of glycosylation (ST3GAL3-CDG). Nonetheless, limited enzyme characterization exists due to the lack of a non-radiochemical assay. Methods: Here, we developed an LC-MS/MS-based method using the artificial substrate para-nitrophenyl-lacto-N-biose (LNB-pNP; Galβ1,3GlcNAcβ1-O-C6H4NO2) to measure ST3GAL3 activity in vitro. Results: A peak corresponding to sialyl-LNB-pNP was detected in reactions with homogenate from HEK-293T cells transfected with pCDNA3 ST3GAL3 plasmid, but was virtually absent in mock-transfected cells. A substrate dependence curve provided an apparent Km value for the substrate (0.40 mM) and closely matched values from prior radiochemical methods. No activity was detected with homogenates from cells expressing pathogenic ST3GAL3 variants, except p.A13D, which is known to retain about 10% of residual activity. Compared to ST3GAL4 and ST3GAL6, ST3GAL3 showed markedly higher specificity toward LNB-pNP, lactotetraosylceramide (Lc4) and asialo-GM1, which are rather specific substrates. Instead, neo-lactotetraosylceramide (neoLc4) was processed by all three ST3GALs. Conclusions: These findings suggest that ST3GAL4 or ST3GAL6 cannot compensate for ST3GAL3 loss in the biosynthesis of gangliosides sialyl-Lc4 and GM1b, but may do so for sialyl-neoLc4. This non-radiochemical assay enables screening and diagnostic evaluation of novel ST3GAL3 variants potentially associated with ST3GAL3-CDG. Full article

The growing demand for clean-label plant-based cheese alternatives underscores the need for products with desirable properties. A key technological challenge is replicating the firmness of traditional cheese without synthetic additives. This study explores lactic acid bacteria (LAB) fermentation as a natural texturizing method for clean-label plant-based cheeses. We investigated the link between LAB metabolic traits and the firmness of three substrates: cashew, soybean, and sunflower seed. A strong correlation (r ≈ −0.88) was found between final pH and firmness in cashew paste, where strains achieving a pH of ~4.0–4.5 (near the protein isoelectric point) produced the firmest gels (up to 3.35 N). Lactiplantibacillus plantarum 729/23 and Lacticaseibacillus helveticus NK-1 were most effective. Soybean paste firmness increased moderately (to 1.93 N), while sunflower seed paste showed no significant improvement (≤1.14 N) despite active acidification, indicating substrate-specific limitations. This study, supported by a comprehensive analysis of the literature on firmness measurement in plant-based cheeses, demonstrates the potential of targeted selection of natural starter cultures to create clean-label products with minimal ingredients. Full article

The aza-Michael reaction is a fundamental transformation for carbon–nitrogen bond formation, providing efficient access to β-amino carbonyl compounds, nitriles, and related nitrogen-containing building blocks of broad importance in medicinal chemistry and organic synthesis. Over the past two decades, ionic liquids (ILs) have attracted considerable attention as alternative reaction media, promoters, and catalysts for aza-Michael reactions, owing to their distinctive physicochemical properties and tunable structures. This review presents a comprehensive and critical overview of ionic-liquid-mediated aza-Michael reactions, emphasizing the evolution of IL design from early imidazolium-based systems to modern task-specific, supported, and bio-derived ionic liquids. Conventional room-temperature ionic liquids are discussed as non-innocent solvents capable of stabilizing charged intermediates and enhancing electrophilicity, thereby enabling catalyst-free or metal-assisted aza-Michael additions. Subsequent sections focus on task-specific ionic liquids incorporating Brønsted acidic, basic, hydrogen-bond-donating, or bifunctional motifs, highlighting how rational structural design translates into improved activity, selectivity, and substrate scope. Particular attention is devoted to guanidine-, DABCO-, and DBU-based ionic liquids, where mechanistic studies reveal cooperative activation modes rather than simple acid–base catalysis. Recent advances in supported and polymeric ionic liquids are also reviewed, demonstrating effective strategies to combine IL-like reactivity with enhanced recyclability and operational simplicity. Overall, this review clarifies the diverse roles of ionic liquids in aza-Michael chemistry and outlines current challenges and future perspectives toward more sustainable and efficient C–N bond-forming methodologies. Full article

Global food security is an increasing challenge due to population growth and the limited availability of natural resources, driving the search for sustainable protein sources. In this context, edible insects such as yellow mealworms (Tenebrio molitor) have emerged as a promising alternative, while probiotics have been widely applied in animal production to enhance growth performance and nutritional quality. This study aimed to evaluate the effects of probiotic supplementation on the growth performance, biomass yield, and nutritional composition of yellow mealworm larvae at laboratory and pilot scales. Three probiotic strains—Bacillus velezensis, Bacillus coagulans, and Pediococcus pentosaceus—were tested at four different dosage levels, using wheat bran and brewer’s spent grain as feed substrates. Larval growth was monitored weekly, and total harvested biomass, proximate composition (dry matter, protein, fat, and ash), amino acid profile, and mineral composition were determined using standardized analytical methods. At the laboratory scale, probiotic supplementation did not result in significant differences in mean larval weight or total biomass (p > 0.05). In contrast, at the pilot scale, significant improvements in larval growth and biomass were observed for specific probiotic treatments, with mean larval weights reaching approximately 140–150 mg and total harvest biomass increases of up to ~15% compared to the control (p < 0.05). Growth curves at both scales followed a sigmoidal pattern with a high correlation between laboratory and pilot experiments (R² = 0.98). Probiotic supplementation did not significantly affect crude protein content, but alterations in fat content, specific amino acid concentrations, and mineral composition were observed at the pilot scale, depending on strain and dosage. Overall, the results demonstrate that probiotic supplementation can enhance yellow mealworm production under pilot-scale conditions, while laboratory-scale trials may not fully capture these effects. These findings highlight the importance of scale when evaluating probiotic strategies and support the potential application of Bacilli-based probiotics to improve the efficiency and nutritional quality of industrial insect production systems. Full article

Enzyme kinetic parameters $(k_{\mathit{cat}}, K_m, \mathrm{and} k_{\mathit{cat}}/K_m)$ are fundamental for quantifying catalytic efficiency and substrate specificity in biochemistry and drug discovery. However, experimental determination is resource intensive, and accurate prediction remains a persistent challenge due to the complex spatial nature of catalysis. In this paper, we present UniKineG, a novel deep learning framework that redefines kinetic prediction by modeling the explicit spatial state of enzyme–substrate complexes. Unlike conventional methods that treat proteins and ligands as isolated modalities, UniKineG employs molecular docking to embed both entities into a unified 3D coordinate system. Within this shared geometric context, we utilize a heterogeneous graph neural network integrated with geometric vector perceptrons (GVPs) to capture intricate vector-based interactions, such as directional hydrogen bonds, hydrophobic contacts, and electrostatic complementarity. This structure-based approach confers exceptional robustness: UniKineG effectively overcomes the dependency on high-sequence homology, demonstrating superior generalization on out-of-distribution (OOD) datasets encompassing both unseen enzyme sequences and diverse substrate scaffolds. Consistently outperforming state-of-the-art predictors, UniKineG achieves high-precision predictions. This work establishes a solid foundation for understanding enzyme–small molecule interactions in 3D space and offers a transformative tool for computational enzymology. Full article

Nuclear receptor HR97 is considered as a non-insect arthropod–specific receptor, but its roles in decapod reproduction remain poorly understood. Here, we identified and characterized an HR97 ortholog from the swimming crab Portunus trituberculatus (PtHR97) and verified its placement within the NR1L nuclear receptor family by phylogenetic analysis. PtHR97 encodes a canonical nuclear receptor with a conserved DNA-binding domain (DBD) and ligand-binding domain (LBD). Quantitative PCR revealed predominant PtHR97 expression in the ovary and stage-dependent changes during ovarian development. Using an ovarian explant culture system, we found that arachidonic acid (AA) consistently suppressed PtHR97 transcript levels, while methyl farnesoate (MF) and pyriproxyfen (P) had no significant effect, indicating a potential inhibitory role for AA in PtHR97 expression. RNA interference of HR97 caused significant changes in ovarian development, including reduced GSI, smaller oocytes, and uneven eosinophilic granule distribution. Transcriptomic profiling of HR97-silenced ovaries indicated that the major responses involved genes associated with substrate transport/exchange, cell boundary–related signaling and transduction, and disturbed nuclear transcriptional regulation. Short-term in vivo perturbations (HR97 RNAi and AA treatment) further supported these expression changes and revealed that AA- and HR97 RNAi–elicited transcriptional responses only partially overlapped. Taken together, these results suggest that HR97 contributes to ovarian development, potentially through broad transcriptional responses related to transport, signaling, and gene regulation. Although AA may suppress HR97 expression, HR97 does not fully explain AA-mediated regulation of ovarian development. Full article