Search Results (4,871)

Background/Objectives: One of the ongoing clinical constraints is limiting microbial growth on prostheses, justifying the need for material surface enhancements to reduce microbial complications. This study aimed to investigate a potentially applicable and reproducible coating technique to overcome clinical microbial challenges. Methods: Silver (Ag) nanoparticles (NPs) were applied to three types of materials through spray, spin, and dip coating techniques. Scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared (FTIR) spectroscopy, energy-dispersive X-ray fluorescence (EDXRF), and inductively coupled plasma optical emission spectroscopy (ICP-OES) were performed. Subsequent optimization of spray numbers was determined. Antimicrobial performance of one- and three-layered coatings was evaluated through agar diffusion, direct contact, and adhesion (time-dependent) assays against Pseudomonas aeruginosa (P. aeruginosa). Results: Spray coating exhibited superior coating uniformity. In total, 15 sprays were determined as an effective number for a single-layer coating. EDS confirmed Ag NP presence; FTIR revealed no chemical alteration. Disk diffusion tests showed no inhibition zones. Adhesion and direct contact tests displayed antibacterial activity. The effect was superior in direct contact test. Short-term time-dependent adhesion test of one-layer coating of acrylic and silicone had a consistent decrease in bacterial amount, whilst zirconium had only a strong initial activity. In general, the three-layer coating did not reveal a higher antimicrobial activity, suggesting that the increase in layering can negatively impact surface effectiveness. Conclusions: Spray coating of Ag NPs represents a potentially feasible and relevant strategy for enhancing the antibacterial properties of dental and maxillofacial prosthetic materials without compromising their inherent physicochemical characteristics, pending further cytotoxicity and in vivo validation. Full article

Bacterial infections pose a serious threat to human health, and antibiotics remain the first-line therapeutic agents in clinical practice. However, the vast majority of antibiotics lack the ability to penetrate cell membranes, which severely limits the number of clinically available options for treating intracellular bacterial infections. Developing efficient intracellular antibiotic delivery strategies is therefore of considerable clinical significance, both for reducing antibiotic dosage and for expanding the repertoire of drugs applicable to intracellular infections. To address this challenge, we constructed a protein-based delivery platform mediated by a cell-penetrating miniprotein for efficient intracellular antibiotic delivery. In this system, bovine serum albumin (BSA), which possesses broad antibiotic-binding capability, was employed as the drug carrier, while the cell-penetrating miniprotein ZF5.3, which is capable of endosomal escape, served as the transmembrane delivery mediator. ZF5.3 was conjugated to BSA via a bioorthogonal reaction, and ceftriaxone (CRO) was selected as the model antibiotic to construct a nanoscale delivery system. The binding interaction between CRO and BSA was characterized using UV-Vis, HPLC, and molecular docking techniques. The assembly of the ZF5.3–BSA delivery platform was confirmed by UV-Vis absorption spectroscopy and gel electrophoresis. Intracellular delivery efficiency was evaluated by confocal fluorescence imaging and flow cytometry, and the results demonstrated that ZF5.3 conjugation enhanced intracellular protein delivery efficiency by over 5-fold. Fluorescence co-localization analysis revealed that ZF5.3-mediated cargo is mainly distributed in the cytoplasm and does not completely co-localize with lysosomal markers, suggesting its ability to effectively escape from lysosomes. An intracellular infection model using Staphylococcus aureus was established. Colony-forming unit (CFU) counting experiments confirmed that the delivery system significantly enhanced the intracellular antibacterial activity of ceftriaxone. CCK8 cytotoxicity assays confirmed that the system is non-toxic to cells. Full article

Drought impairs osmotic adjustment and photosynthetic performance in legumes; however, the role of micronutrients in modulating these responses across genotypes remains unclear. This study investigated the effects of selenium on the osmolytes and photosynthetic efficiency in two vegetable-soybean (Glycine max L. Merrill) cultivars differing in drought responses: UVE14 (drought-tolerant) and UVE17 (drought-susceptible). Plants were grown under well-watered (100% soil water-holding capacity, WHC) and water-limited (30% soil WHC) conditions, with or without soil-applied selenium. Free amino acids, soluble sugars, chlorophyll pigments, vegetation indices, and chlorophyll fluorescence parameters were assessed at the flowering and pod-filling stages. Under drought conditions, selenium enhanced tolerance primarily by modulating free amino acid metabolism at flowering, increasing aspartate, asparagine, glutamine, and glutamate levels, alongside improvements in chlorophyll content, canopy greenness, and PSII photochemical efficiency. These responses indicate a coordinated adjustment between nitrogen metabolism and photosynthetic function. Both cultivars benefited from selenium application, although the responses were more pronounced in the susceptible cultivar (UVE17). Selenium-induced changes in soluble sugar content were greater under well-watered conditions in both cultivars. The limited accumulation of stress-associated osmolytes, such as proline, following selenium soil drench suggests reduced cellular disruption and mitigation of drought-induced stress. These findings highlight selenium as a context-dependent modulator of drought resilience and emphasize cultivar- and developmental stage-specific effects. Full article

New copper(II) (C1) and palladium(II) (C2) complexes with S,O-tetradentate ligand (L) derived from thiosalicylic and thiopropionic acids were synthesized. In cell-based assays, (C1) exhibited the most pronounced activity within the tested compound series and was therefore advanced for mechanistic evaluation in 4T1 triple-negative breast cancer cells. (C1) significantly reduced 4T1 cell viability by inducing early and late apoptosis, accompanied by mitochondrial membrane depolarization and enhanced cytochrome C release. Consistently, (C1) increased the Bax/Bcl-2 ratio, promoting a pro-apoptotic shift. In parallel, (C1) triggered autophagy, as evidenced by decreased p62 and LC3B levels, induced G0/G1 cell-cycle arrest, and suppressed proliferative signaling by downregulating Ki67, cyclin D, and phosphorylated AKT. The DNA-binding studies showed moderate to strong affinity, favoring minor groove binding, with higher affinity for (C1) than for (C2). Tryptophan fluorescence quenching indicated a strong interaction with BSA via a predominantly static mechanism, more pronounced for (C1). Molecular docking at the DNA and BSA binding sites corroborated experimental findings and suggested favorable interactions between the complexes and apoptosis-related proteins (CASP3, BAX, and BCL2). The integrated experimental and computational data identify (C1) as a biologically active compound with multimodal biological effects in vitro, supporting further structural optimization and mechanistic investigation. Full article

Rapid detection of heavy metals is vital for monitoring surface water contamination and preventing environmental and health risks. Traditional detection methods for metals such as lead and copper often require sophisticated, costly instrumentation, limiting their use in routine analyses. To address this challenge, we developed a cost-effective fluorescence-based approach using semiconductor quantum dots (QDs) as nanosensors for metal ion detection. The QDs were synthesized directly in aqueous medium through a reflux-assisted process employing cadmium precursors, selenium, thioglycolic acid (TGA), and branched polyethyleneimine (PEI, Mw ~25,000) as stabilizing agents. Structural analysis revealed nanoparticles with diameters below 5 nm, spherical morphology, and a zinc blende (face-centered cubic) crystalline structure. Optical characterization by UV–Vis, photoluminescence (PL), and FTIR spectroscopy confirmed effective surface functionalization and strong quantum confinement. PEI-capped QDs exhibited enhanced colloidal stability and showed pronounced fluorescence quenching in the presence of Pb²⁺ ions, indicating high sensitivity and selectivity toward lead. Both TGA- and PEI-capped QDs also demonstrated moderate responses to Co²⁺ but negligible interaction with Sn²⁺, confirming ion-specific detection. Overall, this study demonstrates that surface-engineered QDs constitute a simple, accessible platform for selective detection of toxic metals, with promising applications in environmental monitoring and water quality assessment. Full article

Grapevine black-foot disease is a destructive trunk disease with a complex pathogen composition that often involves mixed and latent infections, making timely field diagnosis challenging. To improve rapid field detection, we developed a rapid, sensitive, and low instrument-dependent nucleic acid assay. The assay integrates recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)–Cas12a for the detection of Ilyonectria and Dactylonectria, two genera associated with grapevine black-foot disease. Conserved regions of the histone H3 and β-tubulin genes were selected for the design of specific RPA primers and corresponding CRISPR RNAs (crRNAs) for Ilyonectria and Dactylonectria, respectively. A workflow integrating RPA, Cas12a-mediated recognition, and lateral flow assay (LFA)-based visualization was established. The reaction conditions were optimized to enhance amplification efficiency and Cas12a recognition stability. Specificity was evaluated using DNA from target and non-target fungi, and sensitivity was determined using serially diluted templates. Under optimized conditions, the assay detected Ilyonectria DNA at concentrations as low as 3.6 ng/μL within 1 h at 39 °C. For Dactylonectria, the detection limit reached 80 fg/μL within 50 min at 41 °C. No cross-reactivity was observed. The LFA strips exhibited positive and negative bands within minutes, enabling rapid visual interpretation. This RPA-CRISPR/Cas12a-LFA system provides a rapid, visually interpretable approach for detecting selected grapevine black-foot disease-associated species in China. The workflow reduces the requirement for specialized thermocycling and fluorescence detection equipment during amplification and readout, following DNA extraction. Full article

Copper ions (Cu²⁺) and intracellular biothiols are tightly coupled in cellular redox regulation, where copper–thiol coordination governs oxidative stress and metal homeostasis. However, analytical platforms capable of sequentially monitoring Cu²⁺ and biothiols within a single molecular system remain scarce. Herein, we report a coumarin-based fluorescent probe XDP that enables sequential ON–OFF–ON sensing of Cu²⁺ and biothiols through a coordination–competition mechanism. The imine (C=N) site of XDP selectively coordinates Cu²⁺, leading to fluorescence quenching arising from coordination-induced electronic perturbation and enhanced nonradiative decay. The probe exhibits a linear response toward Cu²⁺ over 1–80 μM with a detection limit of 0.108 μM. Subsequent competitive binding of biothiols (GSH, Cys, and Hcy) releases Cu²⁺ from the complex, thereby restoring fluorescence and enabling detection within 1–30 μM with submicromolar sensitivity. XDP also displays a large Stokes shift (135 nm), which minimizes spectral overlap and improves signal reliability. Notably, Cu²⁺ binding triggers a distinct color change that supports naked-eye detection and smartphone-based RGB quantification. The probe further enables visualization of Cu²⁺ and thiol-triggered signal recovery in living cells and zebrafish. This work establishes a versatile analytical platform for probing copper–thiol interactions in environmental and biological systems. Full article

Cooking liquid from Manila clam (Ruditapes philippinarum) is an underutilized byproduct rich in water-soluble taste compounds, representing a potential source of natural umami peptides. In this study, peptide fractions were separated from the cooking liquid. A total of 764 peptide sequences were identified from the most potent fraction, F3 (<3 kDa), by UPLC-ESI-Q-TOF-MS/MS. Machine learning prediction and molecular docking were further used for screening. Five candidate peptides were selected: TQDTVVALDA, KEY, YKD, RND, and GEAF. Sensory evaluation (on a 0–5 scale) and electronic tongue measurements independently confirmed that peptide YKD possessed the strongest taste profile, with an electronic tongue relative umami score of 8.81 ± 0.22. Furthermore, cell-based assays demonstrated that YKD effectively up-regulated the transcriptional expression of taste-related receptors, including GPRC6A, in STC-1 cells, revealing a multi-receptor synergetic mechanism for umami perception. In STC-1 cells, all peptides induced intracellular Ca²⁺ responses and showed no obvious cytotoxicity at 0.5–8.0 mmol/L. YKD produced the highest fluorescence response (0.59) at 1.0 mmol/L. Quantitative RT-PCR analysis suggested that YKD was associated with T1R1/T1R3-related expression, whereas TQDTVVALDA induced stronger CaSR expression. These findings elucidate the specific peptide sequence that engages multiple receptors to create complex tastes, providing a theoretical basis for converting seafood processing byproducts into natural flavor enhancers. Full article

In vitro maturation (IVM) is a critical step affecting the efficiency of bovine in vitro embryo production; however, oxidative stress during in vitro culture can impair oocyte quality and subsequent developmental competence. This study investigated the effects of cysteine supplementation on bovine oocyte IVM, redox homeostasis, mitochondrial status, and transcriptomic changes. Bovine cumulus-oocyte complexes were cultured in IVM medium supplemented with 0, 25, 50, 75, 100, or 125 μM cysteine, and 75 μM was identified as the optimal concentration. Compared with the control group, 75 μM cysteine increased the first polar body extrusion rate from approximately 78% to 81% and improved the fertilization/cleavage rate from approximately 74% to 82%. It also significantly increased the proportions of 2-cell, 4-cell, and 8-cell embryos, whereas morula and blastocyst rates were not significantly affected. At the cellular level, 75 μM cysteine significantly reduced ROS levels and increased GSH content, as indicated by changes in relative fluorescence intensity. JC-1 staining showed that the JC-1 monomer signal decreased from approximately 16.0 to 13.5, whereas the JC-1 aggregate signal increased from approximately 13.2 to 14.8, indicating improved mitochondrial membrane potential status. In addition, lipid droplet fluorescence intensity increased from approximately 11.8 to 13.4, mitochondrial fluorescence intensity increased from approximately 6.0 to 7.0, and cytoskeletal fluorescence intensity showed no significant difference between groups. Smart-seq2 transcriptomic analysis identified 1935 differentially expressed genes, including 1778 upregulated and 157 downregulated genes, which were mainly enriched in translation, ribosomal structural components, RNA binding, oxidative phosphorylation, and metabolism-related pathways. qRT-PCR further confirmed the upregulation of key genes, including NDUFS2, VDAC3, ANXA2, MTHFD1L, and SCD. Overall, 75 μM cysteine improves bovine oocyte IVM quality by enhancing antioxidant capacity, improving mitochondrial membrane potential, increasing lipid-derived energy substrate storage, and regulating genes related to energy metabolism and developmental competence. Full article

Introduction: Establishing baseline descriptions of inorganic elements in the early life stages of sharks and in their respective nursery areas is essential for assessing anthropogenic impacts and supporting conservation strategies. Objectives: This study presents the first baseline of plasma trace element concentrations (Al, Zn, As, Cu, Cr, Cd, Co, Mn, Ti, Ni, Hg, Pb) for four juvenile shark species (Carcharhinus limbatus, Paragaleus pectoralis, Rhizoprionodon acutus, and Sphyrna lewini) from Sal Rei Bay, Boa Vista Island, Cabo Verde—the first multi-species shark nursery area described in Atlantic Africa. Methodology: Seawater and sediment samples were collected from eight sites and analyzed along with plasma samples using total reflection X-ray fluorescence spectroscopy (TXRF). Sediment granulometry and pollution indices, including the enrichment factor (EF), ecological risk index (RI), and metal pollution index (MPI), were used to characterize habitat contamination. Data were analyzed using statistical models to explore spatial and element-specific patterns. Results: Overall, environmental contamination was low, with slight increases in Cd, Co, and Hg at sites 1 and 2, near the fishing port, and at site 5, likely reflecting natural transport, sediment redistribution, and enhanced nearshore deposition. Juvenile sharks exhibited generally low plasma trace element concentrations, although species-specific elemental signatures were evident: elevated levels of Al and Cu in C. limbatus, Zn in S. lewini, and As in R. acutus and P. pectoralis. Conclusions: These findings establish critical baseline reference values for trace elements in juvenile sharks from a key Atlantic nursery area. The results provide an essential framework for future biomonitoring efforts and contribute to the management and conservation of Cabo Verdean shark nursery habitats. Full article

(1) Background: Provisional restorations play a crucial role in maintaining oral function and must exhibit adequate mechanical properties, particularly fracture resistance, to ensure structural integrity throughout the provisional phase. The aim of this study was to compare the flexural strength and modulus of materials used for provisional dental prostheses, with and without fiber or metal reinforcement. (2) Methods: Standardized specimens (2 × 2 × 25 mm) were fabricated from an acrylic resin (Unifast LC), a 3D-printed resin (NextDent C&B), and a bis-acryl resin (Luxatemp Fluorescence). For each material, four experimental subgroups were established: no reinforcement, two types of glass fiber reinforcement (EverStick C&B and EverStick Post NET), and metal reinforcement. Specimens were subjected to a three-point bending test. Flexural strength and flexural modulus were analyzed using a two-way, non-parametric ANOVA with the aligned rank transform. The significance level was set at 0.05. (3) Results: Material type and reinforcement strategy significantly affected flexural strength and flexural modulus. Fiber reinforcement with EverStick C&B yielded the highest values across all materials, particularly in the acrylic resin. Metal reinforcement showed moderate improvements, whereas EverStick NET had limited or no effect and reduced strength in the bis-acryl resin. Reinforced specimens exhibited altered fracture behavior, preventing complete separation after failure. (4) Conclusions: Fiber reinforcement, particularly with EverStick C&B, significantly enhances the flexural strength and modulus of provisional materials. The reinforcement performance is dependent on its type and material interaction, modifying fracture behavior by preventing complete separation. Full article

Biomass-derived carbon dots (CDs) have attracted increasing attention in agriculture due to their simple synthesis and low environmental impact. In this study, CDs were synthesized from guava (Psidium guajava) leaves using a hydrothermal method (200 °C, 15 h). The particles had an average size of 6.17 nm and a quantum yield of 2.46%, confirming the successful synthesis of fluorescent carbon nanomaterials from the natural precursor. The effects of CDs on rice (Oryza sativa L., variety HT1) were evaluated through both seed treatment and field application. Soaking seeds in a 200 ppm CD solution for 24 h significantly enhanced shoot and root lengths (28.87 mm and 34.00 mm, respectively) among the tested treatments. In field trials, applying CDs at the same concentration also promoted plant growth, as evidenced by improvements in plant height, leaf development, tillering, and flag leaf characteristics. These changes were reflected in yield, with the highest grain yield of 6.13 t ha⁻¹ at 200 ppm, exceeding that of the control treatment. The observed positive effects may be due to enhanced photosynthetic activity and better control of oxidative processes in plants. Nevertheless, the effect was less pronounced at higher concentrations. This trend suggests a dose-dependent response. Full article

Ubiquitin-specific protease 22 (USP22) regulates epigenetic gene expression by deubiquitinating histone H2B (H2Bub1) and upregulating oncogenic proteins and pathways, while antagonizing p53-mediated tumor suppression. USP22 is frequently overexpressed in cancers and associated with therapy resistance and poor prognosis yet remains largely untargeted pharmacologically. Here, using a fluorescence-based USP22 deubiquitinase assay to screen the LOPAC^®1280 library, we identified β-Lapachone, a natural ortho-naphthoquinone with strong anticancer activities, as a potent USP22 inhibitor. β-Lapachone potently inhibited USP22 enzymatic activity, with a half-maximal inhibitory concentration (IC₅₀) of ~0.75 μM, and molecular docking revealed its occupation of the catalytic pocket adjacent to the USP22 active-site triad, supporting a potential binding mode. Functionally, β-Lapachone suppressed proliferation and induced apoptosis in A549 and H1299 RAS-mutant lung adenocarcinoma (LUAD) cells, while USP22 knockout conferred marked resistance, indicating partial USP22 dependence. In patient-derived LUAD models, β-Lapachone inhibited sphere formation and reduced CD133⁺ cancer stem cell populations. Notably, it synergized with cisplatin to enhance DNA damage and apoptosis. In vivo, β-Lapachone significantly suppressed tumor growth in a syngeneic KRAS-mutant/p53-Null mouse lung cancer model and further potentiated cisplatin-induced antitumor effects. Collectively, these findings identify β-Lapachone as a potent inhibitor of USP22 and validate USP22 inhibition as a key mechanism underlying its anticancer activity in LUAD cells, both in vitro and in vivo. Full article

Since its introduction in 2006, DNA origami has enabled the fabrication of a wide variety of two- and three-dimensional DNA nanostructures. From the very beginning, researchers have explored these nanostructures as programmable nanobreadboards with hundreds of uniquely addressable positions, allowing precise spatial arrangement of biomolecules, fluorophores, and nanoparticles. This capability has been leveraged to create functional DNA nanomachines capable of single-molecule detection. Here, DNA origami is utilized to precisely engineer various nanoarchitectures, such as conformational switches and plasmonic hotspots. Through coupling of these concepts with tailored readout strategies, true single-molecule detection can be achieved. This literature review systematically examines the development of DNA origami-based single-molecule detection concepts. We first explore general design principles to produce functional DNA nanostructures, followed by an overview of non-fluorescence-based approaches employing atomic force microscopy, nanopores, and optical nanoantennas with surface-enhanced Raman spectroscopy readout, as well as fluorescence-based approaches relying on dynamic DNA nanostructures and optical nanoantennas with fluorescent readout. We highlight key trends as well as the remaining technology gaps that should be bridged to further advance DNA origami towards next-generation single-molecule detection. Full article

The valorization of culinary by-products into functional bioactive resources represents a significant advancement in sustainable biotechnology. This study characterizes an extract derived from “battuto toscano” by-products, a traditional blend of garlic, onion, carrot, and celery trimmings, recovered through circular economy principles. Comprehensive antioxidant profiling was performed alongside biological evaluations on human cell lines and anti-glycation assays. Results from Folin–Ciocalteu, FRAP, and TEAC assays confirmed a high concentration of secondary metabolites with significant scavenging capacity. In vitro testing on primary human fibroblasts and HaCaT keratinocytes revealed a concentration- and time-dependent biological response, with lower concentrations showing better compatibility and transiently enhancing HaCaT metabolic activity. Furthermore, BTE reduced AGE-associated fluorescence in the BSA–glucose model, particularly at 5 mg/mL, supporting its potential anti-glycation activity. These findings establish “battuto toscano” by-products as a reservoir of sustainable biomolecules. This study offers a transformative resource for the pharma/nutraceutical sectors by bridging culinary tradition with biomedical innovation. Full article