Search Results (459)

Lytic polysaccharide monooxygenases (LPMOs) are promising enzymes for lignocellulose degradation; however, wild-type LPMOs often exhibit limited catalytic activity and stability. In this study, computer-aided virtual saturation mutagenesis was applied to AnLPMO15g from Aspergillus niger, and eight potentially beneficial mutants (S197H, S197F, E185V, E185L, E185M, E185I, Q108M, and A249P) were identified based on predicted changes in unfolding free energy (∆∆G). Six mutants demonstrated enhanced activity in a 2,6-dimethoxyphenol (2,6-DMP) oxidation assay, which serves as a proxy for peroxidase-like activity. The E185V mutant exhibited a 45% increase over the wild type. The triple mutant E185V/Q108M/A249P further increased the catalytic efficiency by 56%. Notably, when combined with cellulase, E185V/Q108M/A249P enabled a 202.5% increase in reducing sugars from wheat straw, achieving a synergy degree of 1.83, highlighting its potential to improve agricultural residue conversion. Molecular dynamics simulation suggested that the E185V/Q108M/A249P triple mutant induced flexible conformational changes in six residues, which may improve substrate binding affinity. This study presents an effective strategy for engineering AA9 family LPMOs to enhance catalytic performance, facilitating efficient and cost-effective degradation of lignocellulosic biomass with implications for sustainable agricultural waste management and circular bioeconomy. Full article

Phthalic acid esters (PAEs), ubiquitous plasticizers and recognized endocrine-disrupting chemicals, pose a protracted threat to aquatic ecosystems and biodiversity. However, current ecotoxicological assessments often focus on isolated chemicals at exceedingly high laboratory doses, failing to reflect true environmental risks. This review systematically evaluates and compares the multisystemic toxicological effects of six priority PAEs (DEHP, DBP, BBP, DNOP, DEP, and DMP) using the zebrafish biological model. The synthesized evidence reveals a distinct structure–activity relationship, where long-chain and highly hydrophobic congeners exhibit substantially higher toxicity than their short-chain counterparts. Exposure to these PAEs induces severe developmental, cardiovascular, neurobehavioral, and reproductive anomalies. Specifically, DBP and BBP display the most potent cardiotoxic and neurotoxic effects, while DEHP and DBP drive profound reproductive decline and endocrine disruption at concentrations as low as 0.5–20 μg/L. Crucially, comparative environmental relevance assessments indicate that real-world PAE concentrations in industrial hotspots frequently meet or exceed these laboratory-derived lowest observed effect concentrations. These findings underscore the severe ecological risks posed by PAE contamination and position the zebrafish as a vital biological sentinel. Future ecotoxicological evaluations must prioritize chronic, low-dose mixture exposures and transgenerational toxicity to fully characterize the protracted legacy of these pollutants on zebrafish populations. Full article

Background: Plasticizers, including phthalate esters and phthalate-free alternatives, are widely detected environmental chemicals. Although increasing evidence suggests that plasticizers may disrupt gastrointestinal homeostasis, their potential molecular links with inflammatory gastrointestinal disorders (IGDs) remain unclear. Methods: This study aimed to systematically identify potential molecular targets and pathways linking representative plasticizers with IGDs. An integrative network toxicology framework was applied to investigate four plasticizers, including dimethyl phthalate (DMP), diethyl phthalate (DEP), dioctyl phthalate/di(2-ethylhexyl) phthalate (DOP/DEHP), and acetyl tributyl citrate (ATBC), in relation to Crohn’s disease (CD), ulcerative colitis (UC), esophagitis, and gastritis. Plasticizer- and disease-related targets were collected from public databases, followed by overlapping target screening, protein–protein interaction network analysis, functional enrichment analysis, GEO-based transcriptomic validation, molecular docking, molecular dynamics simulation, and single-cell RNA-seq analysis. Results: Disease-specific candidate targets were identified, including CXCL8 and FN1 for CD, IL1B for UC, MAPK3, FASN, FN1, PPARG, CXCL8, FOS, and HIF1A for esophagitis, and MMP9, TNF, TLR4, IL6, CCR2, IFNG, and PTGS2 for gastritis. Cross-disease analysis further identified plasticizer-associated signature targets, including MMP7 for DMP, HMOX1 and NOS2 for DEP, and LTF and CCL11 for ATBC. Enrichment analysis indicated that these targets were mainly involved in inflammatory, chemokine, MAPK-related, and xenobiotic response pathways. Molecular docking and dynamics simulations suggested stable interactions between selected plasticizers and candidate targets, while single-cell analysis revealed their cell-type-specific expression patterns in epithelial, immune, and stromal compartments. Conclusions: This study provides an exploratory network toxicology framework for identifying potential molecular associations between plasticizer exposure and IGDs. The findings highlight disease-specific and plasticizer-associated candidate targets that may guide future experimental validation and environmental risk assessment. Full article

Real-time deployment of modern object-detection networks on edge devices is challenging because of limited compute resources, external-memory bandwidth, and strict power constraints. To address these issues, this paper presents a host–FPGA collaborative accelerator for quantized YOLOv5n on a Xilinx Zynq-7100 platform. The proposed design includes a modular multi-operator neural processing unit supporting seven atomic operators, a Dual-Multiply Packing (DMP) scheme to improve DSP48E1-based INT8 convolution density, a cache–compute–cache dataflow with global ping-pong buffering to overlap DMA transfers and computation, and a Multi-Quantization Domain Alignment (MQDA) pipeline to preserve accuracy at Add and Cat fusion nodes. Implemented at 200 MHz, the prototype achieves 24.617 ms FPGA-side forward-inference latency, 36.686 ms end-to-end single-frame latency, 27.2 FPS system-level performance, 182.8 GOPS equivalent throughput, and 8.536 W on-chip power consumption, corresponding to 21.42 GOPS/W. Experimental results also show that INT8 quantization causes only limited accuracy degradation, while MQDA improves quantized detection accuracy by reducing cross-domain fusion error. These results demonstrate that the proposed architecture provides an effective balance among throughput, energy efficiency, hardware cost, and quantized accuracy for real-time edge object detection. Full article

Periodontal ligament mesenchymal stem cells (PL-MSCs) are vital for both periodontal regeneration and alveolar bone maintenance, including their turnover during orthodontic therapy. Chronic periodontal inflammation, mainly caused by Gram-negative bacterial lipopolysaccharides (LPS), interferes with osteogenic differentiation and leads to bone loss. Increasing evidence indicates that long non-coding RNAs (lncRNAs) link inflammatory signaling to osteogenic regulation, but their specific role in LPS-driven modulation of PL-MSC osteogenesis is not well understood. The aim of this study was to assess the effects of LPS from two bacterial strains on PL-MSCs differentiation. Human PL-MSCs were cultured under standard stem cell or osteogenic conditions and treated with LPS from Escherichia coli or Porphyromonas gingivalis. Mineralization was assessed using Alizarin Red staining. Osteogenic differentiation was evaluated through immunocytochemical analysis of osteopontin, collagen type 1, osteocalcin, osteonectin, and dentin matrix protein-1 (DMP-1). Expression levels of lncRNAs growth arrest-specific transcript 5 (GAS5), Metastasis-Associated Lung Adenocarcinoma Transcript 1 (MALAT1), maternally expressed gene 3 (MEG3) and Nuclear Enriched Abundant Transcript 1 (NEAT1) were measured by real-time PCR at 6, 24 and 48 h of LPS exposure. Exposure to E. coli LPS significantly inhibited extracellular matrix mineralization and decreased the expression of key osteogenic markers, indicating impaired osteoblast maturation. In contrast, P. gingivalis LPS caused a partial, dysregulated osteogenic response, marked by increased expression of osteopontin, osteonectin, and dentin matrix protein-1 (DMP-1), but without complete differentiation. LPS types altered lncRNA expression profiles, suggesting that non-coding regulatory networks are involved in inflammation-induced osteogenic dysregulation. Multivariate analyses showed decreased expression of GAS5, MEG3, and MALAT1 in the LPS vs. CTR comparison, decreased COL1A1 in LPS-PG vs. CTR, and increased OSTEOPONTIN in LPS vs. CTR. Differentiation was significantly associated with reduced expression of XIST and NEAT1. Time exerted significant effects on GAS5, MEG3, XIST, and MALAT1, with lower expression at 48 h compared with 6 h, and on COL1A1, which was significantly reduced at both 24 h and 48 h relative to 6 h. Bacterial LPS disrupt osteogenic differentiation of PL-MSCs depending on the species, affecting matrix formation, mineralization, and lncRNA expression. These findings highlight lncRNA-mediated communication between inflammatory signals and osteogenic pathways, providing new insights into the molecular mechanisms of inflammation-related bone remodeling in periodontal disease and orthodontic movements. Targeting lncRNA-regulated pathways could be a promising strategy to enhance periodontal regeneration during inflammation and also ensure optimum outcomes in orthodontic therapy. Full article

Glatiramer acetate (GA) is a first-line disease-modifying therapy for multiple sclerosis (MS) with well-established moderate efficacy and high safety, yet its mechanisms of action remain incompletely understood. DNA methylation plays a significant role in MS development and is modulated by various environmental factors, including therapeutic drugs. In this pilot study, we report the first prospective analysis of genome-wide DNA methylation changes in peripheral blood mononuclear cells (PBMCs) from four female relapsing-remitting MS patients before GA initiation and after approximately four and eight months of therapy. We identified 365 loci that are characterized by differential methylation, distinguishing post-treatment time points from baseline, with significant enrichment in CpG islands, shores, and promoter regions. Two distinct temporal patterns emerged: (1) non-monotonic DNA methylation changes peaking at four months and associated with response to foreign antigenic stimuli, and monotonic changes progressively increasing by eight months and related to mTOR-associated pathways relevant to chronic inflammation and neurodegeneration. Integration of DNA methylation and transcriptomic data revealed significant methylation-expression correlations for eight genes, including HLA-DMA, PDE4A, and SMOX—genes with established roles in MS-associated antigen presentation, immunoregulation, and neuroinflammation. Cell composition of PBMCs remained stable throughout treatment. In general, GA therapy for MS appears to induce dynamic, locus-specific DNA methylation changes in PBMCs, with distinct temporal patterns suggesting a biphasic response of the immune system. However, given that none of the individual DMPs reached genome-wide significance, the results presented in this pilot study strongly require validation in larger independent cohorts. Nevertheless, we believe that our findings provide insights into the immunomodulatory effects of GA and lay the foundation for future hypothesis-driven studies to develop epigenetic biomarkers for therapeutic monitoring and generic GA product assessment. Full article

High-temperature sintered conductive silver paste serves as a critical material in the fabrication of electronic components, with its performance directly influencing device reliability and integration density. In this work, conductive silver paste was prepared via a ball milling method by dispersing silver powder (conductive filler), glass powder (binder), and ethyl cellulose (EC, thickener) in an organic carrier composed of α-terpineol, diethylene glycol butyl ether acetate (DBA), and dimethyl phthalate (DMP) at specific ratios. The effects of the formulation composition and preparation process on the rheological properties of the paste as well as the electrical and mechanical properties of the resulting films were systematically investigated. The results indicated that sintering time and temperature exerted regular effects on the resistance of the silver paste; ball milling speed and duration influenced the particle size distribution, thereby affecting the resistance behavior; thixotropy significantly impacted the resistance characteristics. Under optimal conditions, where the organic carrier consisted of α-terpineol, DBA, and DMP at a ratio of 6:3:1, with 30 wt.% silver powder, 18 wt.% glass powder, and 4 wt.% EC, combined with a sintering temperature of 500 °C for 50–60 min, a ball milling speed of 500–600 r/min, and a ball milling time of approximately 1.5 h, the obtained silver paste exhibited pronounced shear-thinning behavior and excellent thixotropy, indicating favorable processability. The corresponding silver paste film demonstrated the lowest resistivity, superior bending resistance, and good adhesion to both PET and glass substrates. This study provides valuable insights for the design and preparation of high-performance, high-temperature sintered conductive silver pastes. Full article

The mealworm beetle, Tenebrio molitor, is a coleopteran of importance for both immunological and biotechnological research, and it has even been considered as a potential nutraceutical. In recent years, the study of T. molitor has undergone significant development, including immune response, host–parasite interactions, and physiological approaches. However, to perform this type of study, one of the main obstacles is obtaining sufficient hemolymph and viable hemocytes. Thus, we developed a protocol for adult specimens that enables the collection of up to 300 μL of the hemolymph–anticoagulant buffer mixture per specimen, containing approximately 1.5 × 10⁵ hemocytes, with viability ranging from 85% to 90%. The technique involves a double mesothoracic puncture (DMP) and the use of a modified anticoagulant buffer that prevents hemolymph clotting, enabling continuous extravasation and ensuring high yields. Additionally, the hemocytes recovered with this protocol are intact and can be used for subsequent analysis. The hemolymph obtained using this protocol and its applications will help to better understand the processes involving hemolymph and its components in T. molitor, paving the way for further applications. Full article

Dynamic Movement Primitives (DMPs) are widely used for learning and generalizing robot skills. However, standard quaternion DMPs, when modeling orientation trajectories, constrain only the final orientation and cannot freely specify the final angular velocity. This limitation restricts its application to dynamic tasks requiring precise boundary conditions, such as hitting or throwing. Although existing improved methods achieve velocity generalization to some extent, they often struggle to balance trajectory shape preservation with dynamic smoothness, frequently causing significant deviation from demonstrations or abrupt acceleration discontinuities. In this paper, we propose a novel robot skill generalization method that enables controllable final angular velocity for quaternion DMPs. Specifically, we construct a dynamic goal system driven by a quintic polynomial in Lie algebra space, analytically planning the target orientation’s evolution based on given multi-order boundary constraints. This mechanism not only achieves precise control over the final angular velocity but also inherently guarantees global C² continuous dynamics across primitive segments. Comparative simulations and real-world robot hitting experiments demonstrate that, compared to existing approaches, our proposed method effectively satisfies dynamic boundary constraints while exhibiting superior shape preservation, minimal trajectory deviation, and higher smoothness, thereby significantly improving skill generalization performance in complex dynamic tasks. Full article

Phthalate acid esters (PAEs) are persistent organic pollutants (POPs) widely prevalent in industrial wastewater, posing significant threats to both ecological environments and human health. Although Advanced Oxidation Processes (AOPs) are recognized as efficient technologies for PAE degradation, conventional synergistic systems typically employ a simultaneous dosing mode. This approach often leads to the instantaneous quenching of excess radicals, low oxidant utilization, and imbalanced degradation kinetics. Despite its critical role in determining efficiency and costs, the dosing strategy remains an overlooked factor in current research. In this study, dimethyl phthalate (DMP) was selected as the target pollutant to evaluate a synergistic FeSO₄/H₂O₂/K₂S₂O₈ system. An innovative continuous dosing strategy was implemented to optimize radical utilization. A laboratory-scale continuous flow apparatus was developed to simulate industrial onsite conditions, enabling a systematic comparison of degradation kinetics, mineralization characteristics, and radical evolution between the two dosing modes. Results indicated that the degradation rate constant for the continuous dosing system reached 0.659 h⁻¹, representing a 21.1% increase over the simultaneous dosing system (0.544 h⁻¹). Electron Paramagnetic Resonance (EPR) analysis confirmed that the continuous dosing mode maintains a sustained and stable radical flux (^•OH and SO₄^•−) during the critical mid-stage of the degradation, effectively mitigating radical–radical quenching. When applied to real industrial wastewater (salinity: 2083 mg/L), the continuous dosing system achieved a Total Organic Carbon (TOC) removal efficiency of 86.0% at ambient temperature and initial raw water pH, outperforming the simultaneous dosing system (82.0%). GC-MS analysis further confirmed the thorough mineralization of complex organic compounds, especially those containing ester groups and aromatic rings. This research addresses a critical gap in dosing strategy studies, providing an efficient, cost-effective, and industrially viable solution for recalcitrant wastewater treatment while establishing a theoretical foundation for large-scale continuous dosing applications. Full article

Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder caused by the interaction between genetic and environmental influences, potentially mediated by epigenetic mechanisms such as DNA methylation. Genome-wide DNA methylation profiling was performed using the Infinium MethylationEPIC v2.0 array on peripheral blood mononuclear cells (PBMCs) from 100 children with ASD and 50 typically developing controls. Differential methylation analyses were conducted by adjusting for age, sex, and estimated blood-cell-type composition as covariates. Functional enrichment, SFARI gene-overlap analysis, and cross-cohort validation were performed. We identified 3507 differentially methylated positions (DMPs) in the ASD cohort. Functional enrichment revealed pathways involved in neuronal signaling, synaptic activity, and immune regulation, suggesting coordinated neurodevelopmental and immune processes in ASD. Stratification by clinical severity demonstrated common and unique biological characteristics between the moderate and severe ASD groups. Furthermore, DMP-associated genes significantly overlapped with high-confidence ASD risk genes from the SFARI database and established transcriptomic signatures of neurodevelopmental disorders. Comparisons with independent post mortem brain tissue and peripheral blood datasets revealed partial overlap and directional concordance. However, the strength of concordance varied across datasets and was limited in the most directly comparable peripheral blood cohort. Our findings suggested that DNA methylation profiling of PBMCs provided peripheral epigenetic signatures and candidate loci for further validation in larger independent cohorts. Full article

Background/Objectives: Opioid use disorder (OUD) is caused by a complex interplay between genetic and non-genetic factors. DNA methylation is an epigenetic mechanism that modulates gene expression. Data on DNA methylation and opioid addiction and treatment are limited. This association study was designed to assess the difference in genome-wide methylation patterns between individuals with OUD in methadone maintenance treatment (MMT) (n = 114) and those with OUD who achieved long-term abstinence (>10 years) without mu opioid receptor agonist treatment (n = 136). Methods: Differential DNA methylation analysis was performed in whole blood using the Illumina EPIC array. Results: A total of 135 differentially methylated probes (DMPs) reached epigenome-wide significance (p < 1 × 10⁻⁷), controlling for sex, age, estimates of blood cell proportions, and the first two principal components based on genome-wide SNP genotypes. The methylation sites were annotated to 157 genes, including 32% long non-coding RNAs. These genes are related to several systems, including cell adhesion (e.g., SAXO4), immune system and inflammation (e.g., UBTF, USP39, C10orf90, PRKCA), stress response (e.g., CRHR1, GPR19), and spermatogenesis (e.g., SPATA16, COX7B2). DMP cg11641410 is located in lncRNA ENSG00000254687, an antisense to OPRK1. Six of the DMPs were also identified in a related longitudinal study of MMT. Conclusions: At this point, it is not possible to determine whether the minor methylation differences observed in this study cause clinically relevant changes in gene expression. However, these findings have the potential to identify biomarkers and to provide new targets for treatment optimization. Full article

In the context of the increasing digitalization of urban cultural heritage communication, public acceptance, identification, and dissemination of symbolic cultural heritage content exhibit pronounced structural differences across sustainability levels. Taking Xuzhou—a national historical and cultural city in China—as the empirical context, this study conceptualizes cultural heritage as symbolic carriers of cultural meaning and constructs a sustainability-stratified analytical framework. By integrating the Theory of Planned Behavior (TPB) and Cultural Identity (CI) theory, and incorporating Perceived Sustainability of Cultural Heritage (PSC) and Digital Interactive Media Participation (DMP), the study develops a comprehensive model of public communication acceptance mechanisms. Based on 931 valid questionnaires collected from local residents and visitors, exploratory and confirmatory factor analyses, structural equation modeling (SEM), and permutation-based multi-group analysis (MGA) are employed to examine both overall behavioral pathways and cross-group structural heterogeneity across symbolic heritage contexts with different sustainability tiers. The results indicate that: (1) PSC significantly influences communication intention through attitude, subjective norms, and perceived behavioral control, with cultural identity playing a central mediating role; (2) digital interactive media participation primarily functions as a contextual enabler, significantly moderating the relationship between perceived behavioral control and communication intention; and (3) substantial structural differences exist across sustainability tiers, with medium-sustainability symbolic contexts demonstrating the strongest psychological activation effects in attitude formation, identity internalization, and intention conversion. Theoretically, this study extends the integrative application of TPB and cultural identity theory by embedding sustainability perception as an upstream cognitive trigger and repositioning cultural identity as a mediating mechanism within symbolic heritage communication processes. Methodologically, it establishes a systematic “sustainability evaluation–stratified modeling–multi-group comparison” analytical framework. Practically, the findings provide empirical guidance for differentiated communication strategies and digital media interventions tailored to symbolic cultural heritage systems. Full article

The aim of this study was to evaluate the biological effects of bioactive glass-containing self-adhesive resin cements (SARCs) on human dental pulp stem cells (DPSCs), focusing on cytocompatibility, odontogenic differentiation, and mineralization. Experimental SARCs containing 0–5 wt% BAG (BG0–BG5) were compared with two commercially available SARCs, RelyX U200 and TheraCem. Eluates were prepared and applied to DPSCs for the methylthiazol tetrazolium (MTT) assay, quantitative real-time polymerase chain reaction (qRT-PCR), immunofluorescence (IF) staining, and Alizarin Red S (ARS) staining. The result showed there were no significant differences in cell viability across all groups (p > 0.05), indicating that the addition of BAG did not affect cell viability, while the early odontogenic differentiation markers, such as RUNX2, ALP, and COL1A1, showed no clear trend among the groups. However, late-stage markers (DMP-1 and DSPP) were significantly higher in the BG2–BG5 groups relative to the OM group (p < 0.05). IF staining revealed intense signals in the BG2–BG5 groups (p < 0.05) and also ARS staining showed a time-dependent increase in mineral deposition. Within the limitations of this study, BAG-containing SARCs do not negatively impact cytocompatibility and promote late-stage odontogenic differentiation and mineral deposition. Full article

This study evaluated whether three-dimensional alginate–gelatin hydrogels (AGHs) crosslinked with calcium chloride (CaCl₂) enhance the osteo-odontogenic differentiation of odontoblast-like cells in vitro. Two seeding configurations were compared: inter-hydrogel (INT) surface seeding and intra-hydrogel (INTR) encapsulation. Here, the MDPC-23 cells were cultured in AGHs crosslinked with 70 or 100 mM CaCl₂ and assessed for proliferation, cytoskeletal morphology, alkaline phosphatase (ALPase) activity, osteo-odontogenic gene expression, and mineralized nodule formation. After 7 days, cell proliferation was significantly greater in the alginate–gelatin hydrogel (AGH) groups than in the control group. Cells in the intra alginate–gelatin hydrogel 100 (INTR-AGH100) remained predominantly rounded, whereas those in the inter alginate–gelatin hydrogel 100 (INT-AGH100) formed irregular clusters on the hydrogel surface. ALPase activity was highest in INTR-AGH100 at the early stage of culture. Both INT-AGH100 and INTR-AGH100 showed significantly increased expression of DSPP, DMP-1, BSP, OCN, OPN, and Runx-2, together with enhanced mineralized nodule formation. Although no significant differences were detected between the two seeding strategies in all assays, distinct morphological patterns were observed, and the INTR configuration showed relatively greater early differentiation-related activity. These findings suggest that 100 mM CaCl₂-crosslinked AGHs provide a favorable three-dimensional microenvironment under the present experimental conditions and represent a promising in vitro scaffold platform to support future studies of scaffold-guided dentin regeneration. Full article