Search Results (11,158)

The broad substrate specificity of enzymes, while advantageous for catalytic diversity, often leads to undesired side reactions and reduced product yields in engineered metabolic pathways. To address this challenge, we developed a programmable protein scaffold based on a self-assembled γPFD-SpyCatcher hydrogel for the in vivo co-immobilization of SpyTag-cyclized cascade enzymes, enabling the co-immobilization of cascade enzymes in a spatially organized manner. Enzymes with broad substrate specificities were linearly fused with SpyTags, facilitating their spatial organization on the nanoscaffold within engineered E. coli to ensure directed catalytic flux. Using this strategy, the yields of pinene and caffeoyl-CoA were enhanced by 5.8-fold (reaching 94.5 mg/L) and 2.4-fold (reaching 78.6 mg/L), respectively, compared to free enzyme systems. This work establishes an effective approach to mitigate the limitations posed by broad enzyme specificity and demonstrates its potential for applications in synthetic biology and industrial biotechnology. Full article

This study investigates the influence of design factors and key process parameters—including explosive welding (EXW), rolling, and laser processing—on the formation, microstructure, and tribological properties of Ti–Cu–Ni intermetallic coatings. A combined manufacturing approach was employed, starting with the EXW of an MN19 cupronickel alloy to a VT1-0 titanium substrate, followed by multi-pass rolling to achieve a cladding thickness of approximately 0.3 mm. Subsequently, laser surface remelting was performed to facilitate controlled mass transfer and homogenization within the reaction zone. Numerical simulation using COMSOL Multiphysics v. 5.4 was utilized to optimize the thermal cycles and determine the ideal energy density (42 J/mm²) for phase formation. The results demonstrate that the primary structural components of the coatings produced under optimal conditions are solid solutions based on the ternary-modified titanium cuprides Ti₂Cu(Ni) and TiCu(Ni). The transition from a layered bimetal to a finely dispersed intermetallic structure significantly enhances the surface characteristics. This specific phase composition provides a sustained microhardness of ~5 GPa across the coating cross-section. Comparative wear tests against fixed abrasive revealed that the wear resistance of the Ti–Cu–Ni coatings is 2.5 times higher at room temperature and 1.5 times higher at 600 °C than that of the base VT1-0 titanium. Full article

Microbial contamination in aquatic environments poses severe threats to aquaculture sustainability, ecological balance and public health. Traditional culture-based detection methods, while standardized, are time-consuming and labor-intensive, often failing to meet the urgent need for rapid on-site monitoring required to prevent disease outbreaks and manage water quality effectively. By integrating latest research advances (2020–2025), this study reviews advances in rapid detection technologies for aquatic microorganisms, including the evolution of nucleic acid amplification strategies, with a focused comparison of the analytical sensitivity and field deployability of quantitative polymerase chain reaction (qPCR) and mainstream isothermal amplification techniques (loop-mediated isothermal amplification, LAMP; recombinase polymerase amplification, RPA). Furthermore, this study reports on the emergence of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated protein (Cas) systems as next-generation diagnostic tools, highlighting their integration with microfluidic Lab-on-a-Chip (LOC) platforms to achieve attomolar sensitivity. We also consider the application of portable nanopore sequencing for real-time pathogen identification and the growing role of Artificial Intelligence (AI) in analyzing complex diagnostic datasets. Advanced molecular methods have achieved significant reductions in time consumption—from days to less than one hour—while challenges regarding sample preparation and environmental matrix inhibition remain. The future of aquatic monitoring lies in integrated, automated systems that combine the specificity of CRISPR-Cas diagnostics with the connectivity of IoT-enabled biosensors. Comparative analysis indicates that isothermal amplification methods (LAMP, RPA) coupled with CRISPR-Cas systems offer the optimal balance of sensitivity, speed, and field deployability for point-of-care aquaculture diagnostics, while qPCR/dPCR remain indispensable for quantitative regulatory applications. We propose a structured technology selection framework to guide researchers and practitioners in choosing appropriate detection modalities based on specific sensitivity, cost, throughput, and deployment requirements. Full article

Background: Sensor-based systems and virtual reality (VR) technologies provide new opportunities for the objective, technology-driven assessment and training of visuomotor performance in applied contexts such as sport. Methods: This study examined the effects of an integrated visual training program combining stroboscopic stimulation, VR-based vergence exercises, and instrumented reaction-light tasks in adolescent handball players. Twenty-eight adolescent handball players (under-18 competitive level) completed two baseline assessments separated by six weeks, followed by a six-session training program (approximately 15 min per session) integrated into regular team practice. The intervention targeted visuomotor reaction speed, accommodative dynamics, and peripheral visual responsiveness using sensor-based and virtual reality–assisted stimuli. Results: Compared with both baseline measurements, the intervention produced selective improvements in accommodative facility (cycles per minute, cpm)—particularly near–far focusing speed—and in multiple reaction-time conditions (milliseconds, ms) involving manual and decision-based responses. Specific peripheral-field locations showed increased response scores, whereas binocular alignment, AC/A ratio, near phoria, and stereoscopic acuity remained unchanged. Conclusions: These findings indicate that technology-supported visual training protocols incorporating sensor-based reaction systems and VR stimuli were associated with measurable adaptations in dynamic visuomotor processing while preserving fundamental binocular vision parameters. Full article

Extracellular vesicles (EVs) are promising biomarkers for liquid biopsy, but their clinical application is limited by intrinsic heterogeneity and the lack of methods capable of resolving functionally distinct EV subpopulations at the single-vesicle level. Conventional bulk analyses obscure rare but clinically relevant EV subsets, while most single-EV approaches focus on physical properties or surface markers, with limited access to intravesicular functional information. Here, we report a fusion-enabled EV detection strategy at the single-particle level for functional profiling of macrophage-derived EVs. Liposomal probes encapsulating L-arginine, NADPH, and a nitric oxide (NO)-responsive fluorescent dye are engineered to fuse with EV membranes, delivering substrates into the vesicle lumen. In macrophage-derived EVs, inducible nitric oxide synthase (iNOS) catalyzes NO production, activating the fluorescent probe and generating a localized signal within individual vesicles. Signal generation is confined to vesicle-restricted reactions, ensuring specificity and minimizing background. The formation of hybrid vesicles further facilitates optical detection using conventional fluorescence microscopy. Full article

This study demonstrates the feasibility of encapsulating Paracoccus yeei VKM B-3302 cells, which contain palladium nanoparticles, within an organosilicon matrix synthesized using the sol–gel method. The resulting organosilicon material is characterized by a well-developed porous structure and a high specific surface area, ensuring the formation of a catalytic system with accessible active sites. Kinetic studies of the Mizoroki–Heck reaction showed that, although encapsulating the Pd/P. yeei catalyst in an organosilicon matrix slightly decreases its initial reaction rate, it increases the selectivity of the process and reduces the leaching of the active metal during repeated use. These results suggest the potential of encapsulating microorganisms containing metal nanoparticles in organosilicon materials to create stable hybrid catalytic systems. Full article

The European Regulation (UE) 432/2012 includes a specific health claim for olive-oil-associated with its phenolic content, which is based on its protective role against lipid oxidation in the blood. To make use of the health claim, olive oil must have a minimum concentration in phenolic compounds of 250 mg/kg. Reviewing the health claim, the phenolic compounds referred to are the secoiridoid derivatives of hydroxytyrosol and tyrosol. A method based on absolute quantification of phenolic compounds in olive oil is proposed for the recognition of the health claim. The method involves liquid–liquid extraction with a 1:8 (v/v) oil:extract ratio to avoid saturation of the extract in oils with a higher phenolic content and its subsequent determination through LC–MS/MS in multiple reaction monitoring (MRM) mode, the gold standard technique in many application fields because of its analytical features. The optimized method was applied to a set of 100 extra virgin olive oils (EVOOs), and the results obtained were compared with the classic Folin–Ciocalteu method. The comparison between the two methods showed that the classic method is a non-selective method that can be affected by many interferences and that the Folin method underestimates the real phenolic content. Full article

Copper is a crucial cofactor that sustains multiple cellular electron-transfer reactions, making it an essential element for life. However, cytotoxic levels of copper can cause structural damage and cell death through the production of reactive oxygen species (ROS) and nonspecific attacks on proteins. Moreover, immune cells, including neutrophils and macrophages, accumulate copper to induce oxidative bursts that kill engulfed pathogens. Therefore, a well-regulated copper homeostasis system is required for the human commensal fungus Candida albicans to thrive in extreme host environments. Remarkably, C. albicans exhibits higher copper tolerance than the nonpathogenic model yeast Saccharomyces cerevisiae, suggesting the presence of a specific copper tolerance mechanism that supports its adaptability to copper stress. Ndt80 is a versatile transcription factor that regulates several biological processes in C. albicans, ranging from morphological control to drug resistance. This study further reveals that Ndt80 may contribute to copper tolerance by regulating copper transporters and copper-dependent superoxide dismutases (Sods). Additionally, RNA sequencing and complementary approaches uncovered the involvement of Ndt80 in plasma membrane integrity and mitochondrial respiration under copper stress, further linking Ndt80 to copper tolerance. Together, these results broaden our understanding of Ndt80 functions and provide new insights into copper tolerance in C. albicans. Full article

Common bunt of wheat (Tilletia spp.) remains a significant threat to wheat production in low-input and organic farming systems, where chemical seed treatments are restricted or avoided. Host resistance represents a key component of sustainable disease control, but its effective deployment requires detailed knowledge of race-specific virulence and the genetic basis of resistance. In this study, we analysed the reaction of a large and diverse wheat germplasm collection to current European populations of common bunt and mapped the underlying resistance genes using SNP-based approaches. A total of 2731 wheat accessions were phenotyped from 2012 to 2025 using up to 42 purified bunt races with well-defined virulence profiles. Based on phenotypic responses to race-specific resistance patterns, accessions were grouped and compared with established differential lines. A total of 1504 selected accessions were genotyped using Illumina 26k SNP arrays, and resistance loci were identified by genome-wide association studies followed by fine mapping using recombination analysis. All classical Bt resistance genes from Bt1 to Bt10 and Bt13 and BtZ were mapped to defined physical intervals, and the genomic positions of 18 additional race-specific resistance genes were identified in a panel of germplasm. Our results confirm that several historically defined Bt genes, including Bt11 and Bt12, represent multi-gene resistance complexes rather than single loci. Also, genes established as separate genes may possibly be identical, including Bt4 being identical to Bt6, Bt10 being identical to BtZ, and Bt9 possibly being identical to one of the genes in the Bt11 complex. These findings highlight the need for a revised nomenclature of genes and a differential set of varieties. The identified resistance haplotypes provide an improved tool for marker-assisted selection and support the development of wheat cultivars with durable resistance to common bunt. Full article

Coumarins represent a distinctive class of non-classical carbonic anhydrase inhibitors that interact with the entrance region of the catalytic pocket rather than directly coordinating the catalytic Zn²⁺ ion. In this study, a series of glycosylated coumarins was synthesized through a copper-catalyzed multicomponent reaction involving propargyl glycosides, salicylaldehyde, and tosyl azide, providing efficient access to iminocoumarin-based glycosides derived from natural carbohydrates. The inhibitory activity of the synthesized compounds was evaluated against human carbonic anhydrase isoforms I, II, IX, and XII using a stopped-flow CO₂ hydrase assay. The compounds showed negligible inhibition of the cytosolic isoforms hCA I and hCA II, while displaying moderate activity toward the tumor-associated isoforms hCA IX and hCA XII, with Ki values ranging from 12.9 to 41.8 μM. Among the series, 6-O-(2H-chromene-2-one-3-yl-methyl)-D-galactopyranose (10a) emerged as the most potent inhibitor of hCA IX and XII. Structure–activity relationship analysis indicated that deprotected glycosyl derivatives exhibit improved inhibitory activity compared to protected analogues. To rationalize these observations, molecular docking followed by molecular dynamics simulations and MM-GBSA binding free energy calculations were performed for both anomeric forms of compound 10a. The computational results revealed a clear preference for the β-anomer, particularly in hCA IX and XII, where favorable interactions with catalytic threonine residues and isoform-specific aromatic residues stabilize the ligand within the active-site entrance. These findings provide a molecular explanation for the experimentally observed selectivity and highlight glycosyl coumarins as potential starting points for further optimization toward selective inhibitors of tumor-associated carbonic anhydrases. Full article

Metal ion-based chemo-immunotherapy is often limited by rigid intracellular metal homeostasis, insufficient reactive oxygen species (ROS) accumulation, and an immunosuppressive tumor microenvironment (TME). To overcome these limitations, we engineered an ATP-responsive, core–shell bimetallic nanoreactor (Cu/ZIF@PDA, termed CZP) featuring a precisely controlled ~25 nm biomimetic polydopamine (PDA) coating. Triggered by elevated tumoral ATP levels, CZP undergoes coordination-induced disassembly and promotes oxidative stress amplification. Specifically, the PDA shell acts as a superoxide dismutase (SOD) mimetic to continuously supply H₂O₂, fueling Cu²⁺-mediated Fenton-like reactions to unleash highly toxic hydroxyl radicals (•OH) while aggressively depleting the intracellular glutathione (GSH) pool. This irreversible oxidative damage, coupled with Zn²⁺-induced mitochondrial dysfunction, triggers profound mitochondrial DNA (mtDNA) leakage. Crucially, this cytosolic DNA robustly activates the cGAS-STING signaling axis, driving a massive surge in immunogenic cell death (ICD) and significantly promoting dendritic cell (DC) maturation. Furthermore, CZP markedly inhibited primary tumor growth in vivo and showed protection in a tumor re-challenge model, accompanied by enhanced dendritic cell maturation. These findings support the potential of this ATP-responsive bimetallic nanoplatform to promote antitumor immune activation. Full article

Background/Objective: In recent years, epidemiological and clinical evidence has suggested an association between the use of second-generation antipsychotics (SGAs) and hyperglycemic complications: notably, diabetic ketoacidosis (DKA) and hyperglycemic hyperosmolar state (HHS). However, the role of first-generation antipsychotics (FGAs) remains less well understood. To conduct a systematic review of evidence established in case reports (CRs) on adverse drug reactions, specifically DKA and HHS, associated with the use of both FGAs and SGAs in order to identify patterns that may inform clinical awareness and future research. Methods: Pertinent bibliographic databases (MEDLINE, EMBASE, PsycINFO and the Cochrane Central Register of Controlled Trials (CENTRAL)) were searched using index phrases and keywords up until 17 October 2025. Eligible CRs discussed exposure to at least one US FDA-approved antipsychotic drug (APD) and assessed either DKA or HHS. Results: A total of 151 CRs were included in the systematic review (DKA, n = 121; HHS, n = 28; both conditions, n = 2). Patients aged 30 to 39 years accounted for the highest number of emergencies (n = 49, 32.5%), which occurred mostly in males (n = 108, 71.5%). The most common mental health diagnosis was schizophrenia (n = 77, 51%), followed by bipolar disorder (n = 26, 17.2%). Olanzapine was associated with the highest number of DKA cases (n = 53, 43.1%), followed by clozapine (n = 24, 19.5%). The average blood glucose at presentation was 842.8 mg/dL for DKA patients and 1252.8 mg/dL for HHS patients. The average hemoglobin A1c levels (HbA1c) were 11.5% and 12%, respectively, for these two conditions. Of the 12 reported fatalities, treatment with olanzapine was noted in four DKA cases and in one HHS case. Conclusion: This analysis provides additional evidence of an association between the use of atypical APDs and DKA or HHS. Clinicians should continue to monitor metabolic risk factors for these conditions, as well as educating patients about the prevention of acute diabetic complications. Full article

Anti-A and anti-B antibodies are essential for monitoring adverse reactions in organ transplants and transfusions. However, their importance is also growing due to their involvement in the pathophysiology of various human diseases, such as infections, although this is currently the subject of heated debate. A characteristic heterogeneity in the titers and classes of anti-A and anti-B antibodies is observed among individuals. Several factors appear to be responsible, such as everyone’s specific immune profile, age, sex, microbiota composition, lifestyle, and health status. The immune profile, the result of a specific genetic predisposition and mediated and controlled by cytokines, shows a bidirectional relationship with ABO antigen expression, the gut microbiota, and the levels and class switching of anti-ABO antibodies. Associations between ABO groups and circulating levels of cytokines and chemokines further highlight this complex interaction. To better understand the role of the immune profile in this context, we evaluated, for the first time, the possible association between polymorphic variants in the regulatory regions of the genes encoding the cytokines IL-8, IL-1, IL-4, IL-6, IFN-γ, and IL-10 and anti-A and anti-B antibody titers and classes by group and in total. We also assessed the levels of these cytokines in each group and their correlations with anti-A and anti-B antibodies, as well as with age and associations with gender. Significant data were obtained that may contribute to a better understanding of the other roles of ABO antibody titers. Full article

Bacillus and Paenibacillus species are common and widely distributed microorganisms in food systems, often implicated in food spoilage and quality issues. Bacillus subtilis, in particular, has been associated with gas production and package bulging in seasoned foods. In this study, we developed a rapid and visual detection method for Bacillus subtilis by integrating (Recombinase Polymerase Amplification) RPA with (Clustered Regularly Interspaced Short Palindromic Repeats) CRISPR/Cas12a technology (designated as RPA-CRISPR/Cas12a). Specific RPA primers and probes were designed based on the conserved gyrB gene of Bacillus subtilis. Two sets of crRNA were designed according to the number of T-rich PAM sites on the RPA-amplified target sequence, and the reaction conditions were optimized in combination with the CRISPR/Cas12a trans-cleavage detection technology. Under optimized conditions, the crRNA3 guide (with a TT-rich PAM site) demonstrated superior cleavage efficiency compared to crRNA2 (TTT-rich PAM), while crRNA1 (TTTT-rich PAM) showed no activity. The assay achieved a detection limit of 150 pg/μL for genomic DNA and 5.5 CFU/mL for bacterial suspensions within 10 min at 37 °C. The method exhibited high specificity and sensitivity, providing a robust tool for early and on-site detection of Bacillus subtilis in food products. Full article

Diagenetic anomalies within the Upper Paleozoic coal-bearing strata of the Longdong area, Ordos Basin, represent a complex interplay between thermal maturation and fluid evolution, yet their governing mechanisms remain poorly understood. This study integrates petrographic analysis, X-ray diffraction, vitrinite reflectance (Ro) measurements, and fluid inclusion microthermometry to evaluate the discrepancy between organic thermal maturity and mineralogical diagenetic records. The results indicate that the mudstones achieved high thermal maturity, with mean Ro and Tmax values of 2.3% and 555.1 °C, respectively. However, the associated sandstones exhibit anomalous mineral assemblages, characterized by persistent high levels of illite/smectite (I/S) mixed-layer minerals and authigenic kaolinite, which are inconsistent with the anticipated advanced diagenetic stage. Furthermore, homogenization temperatures (Th) of fluid inclusions are significantly lower than expected, implying a localized suppression of illitization. We propose that this atypical diagenetic trajectory is governed by sluggish fluid–rock interactions in a confined diagenetic environment. Specifically, the dissolution of feldspars during acidic diagenesis provided a localized Al³⁺ supply, favoring kaolinite precipitation, while the limited availability of reactive feldspar precursors and pore-fluid retention effectively stalled the progression of illitization. These findings demonstrate that reactant availability and reaction kinetics can decouple mineralogical evolution from organic thermal maturation in coal-bearing sequences. This study provides a novel mechanistic framework for interpreting anomalous diagenetic signatures in heterogeneous sedimentary basins, offering significant implications for reservoir quality prediction in deep-seated, thermally mature strata. Full article