Search Results (299)

Backround: The agr (accessory gene regulator) quorum sensing (QS) system of Staphylococcus aureus participates significantly in its virulence and biofilm formation—either through its activation or suppression. The aim of this study was to investigate the impact of photoactive nanomaterials that have been functionalized with erythrosine B (EryB) on the modulation of this agr QS system on three methicillin-resistant S. aureus (MRSA). Methods: The functionality of the agr system was determined by the CAMP test and by quantitative PCR (qPCR) to analyze the expression of the hld gene, which is located within the RNAIII and encodes δ-hemolysin. The biofilm was evaluated by crystal violet assay and fluorescence microscopy. The anti-biofilm activity was determined by calculating the colony-forming units. The relative expression of the hld gene, determined by qPCR. Results: Using the CAMP test, S66 and S68 strains were found to be agr-positive, and strain S73 was agr-negative. The relative expression of the hld gene increased only in the agr-positive strains (600- and 1000-fold). In these strains, the biofilm was less compact compared to the dense biofilm formed by the agr-negative strain. The anti-biofilm effectiveness on the nanocomposite with EryB after irradiation reduced the growth of biofilm cells by 100- to 1000-fold compared to the biofilm on polyurethane alone. The qPCR results showed a significant decrease in the relative expression of the hld gene in the agr-positive strains after irradiation compared to the non-irradiated samples. Conclusions: These results suggest that photoactive nanocomposites with EryB can significantly reduce biofilm formed by MRSA strains, regardless of the functionality of the agr QS system. Full article

This study investigates the rheological, thermal, and microstructural performance of three novel high-elasticity polymer modifiers (HEMs) incorporated into asphalt binders. The modifiers were evaluated at their recommended dosages using a multi-scale framework combining rotational viscosity, dynamic shear rheometry (frequency sweeps, Cole-Cole plots, Black diagrams, and master curves), bending beam rheometry, differential scanning calorimetry (DSC), fluorescence microscopy (FM), atomic force microscopy (AFM), and Fourier transform infrared spectroscopy (FTIR). Results show that HEM-B achieved the highest values of the superpave rutting parameter (G*/sinδ = 5.07 kPa unaged, 6.73 kPa aged), reflecting increased high-temperature stiffness but also higher viscosity, which may affect workability. HEM-C exhibited the lowest total enthalpy (1.18 W·g⁻¹) and a glass transition temperature of −7.7 °C, indicating improved thermal stability relative to other binders. HEM-A showed the greatest increase in fluorescent area (+85%) and the largest reduction in fluorescent number (−60%) compared with base asphalt, demonstrating more homogeneous phase dispersion despite higher enthalpy. Comparison with SBS confirmed that the novel HEMs not only meet but exceed conventional performance thresholds while revealing distinct modification mechanisms, dense cross-linking (HEM-B), functionalized thermoplastic compatibility (HEM-C), and epoxy-tackified network formation (HEM-A). These findings establish quantitative correlations between rheology, thermal stability, and microstructure, underscoring the importance of dosage, compatibility, and polymer network architecture. The study provides a mechanistic foundation for optimizing high-elasticity modifiers in asphalt binders and highlights future needs for dosage normalization and long-term aging evaluation. Full article

The role of mesenchymal-to-endothelial transition in the angiogenic response remains controversial. In this study, we investigated whether human fetal lung fibroblasts (MRC-5 cells) exhibit morphological plasticity in a biomimetic extracellular matrix environment. To this end, MRC-5 cells were first cultured on and within Matrigel hydrogel and then studied with tube formation assays, confocal/fluorescence microscopy, invasion assays, and transcriptomic profiling. In addition, quantitative assessment for cord formation and gene expression was conducted via qPCR and RNA sequencing. In this study, MRC-5 cells quickly self-organized into cord-like networks, resembling early stages of vascular patterning, and at higher densities, invaded the hydrogel and formed spheroid-like aggregates. Transcriptomic analysis revealed upregulation of genes related to nervous system development and synaptic signaling in Matrigel-grown MRC-5 cultures. Collectively, these findings suggest that MRC-5 fibroblasts display structural and transcriptional plasticity in 3D Matrigel cultures, forming vascular-like cords that are more likely to resemble early developmental morphologies or neuroectodermal-like transcriptional signatures than definitive endothelial structures. This work underscores the potential of fibroblasts as an alternative cell source for vascular tissue engineering and highlights a strategy to overcome current limitations in autologous endothelial cell availability for regenerative applications. Full article

Background: It is crucial to evaluate liver fibrosis in metabolic dysfunction-associated steatotic liver disease (MASLD). Digital pathology, an automated method for quantitative fibrosis measurement, provides valuable support to pathologists by providing refined continuous metrics and addressing inter-observer variability. Although non-invasive tests (NITs) have been validated as consistent with manual pathology, the relationship between digital pathology and NITs remains unexplored. Methods: This study included 99 biopsy-proven MASLD patients. Quantitative-fibrosis (Q-Fibrosis) used second-harmonic generation/two-photon excitation fluorescence microscopy (SHG/TPEF) to quantify fibrosis parameters (q-FPs). Correlations between eight NITs and q-FPs were analyzed. Results: Using manual pathology as standard, Q-Fibrosis exhibited excellent diagnostic performance in fibrosis stages assessment with area under the receiver operating characteristic curves (AUCs) ranging from 0.924 to 0.967. In addition, magnetic resonance elastography (MRE) achieved the highest diagnostic accuracy (AUC: 0.781–0.977) among the eight NITs. Furthermore, MRE-assessed liver stiffness measurement (MRE-LSM) showed the strongest correlation with q-FPs, particularly adjusted by string length, string width, and the number of short and thick strings within the portal region. Conclusions: Both MRE and digital pathology demonstrated excellent diagnostic accuracy. MRE-LSM was primarily determined by collagen extent, location and pattern, which provide a new perspective for understanding the relationship between the change in MRE and histological fibrosis reverse. Full article

Studies assessing the long-term effects of brushing with cleaning and disinfecting agents on surface roughness and biofilm accumulation on the three-dimensionally (3D) printed resins remain scarce. This study aimed to evaluate the effects of different solutions and simulated brushing times on the surface roughness and biofilm formation on heat-cured and 3D-printed denture base resins. Discs samples (10 mm × 1.2 mm) were prepared and randomly assigned to brushing treatments using the following solutions: distilled water, disinfectant liquid soap and dentifrice. The specimens (n = 9) were subjected to 10,000, 20,000 and 50,000 cycles to simulate 1, 2 and 5 years of brushing, respectively. The control group consisted of unbrushed samples. Surface roughness (Ra) was measured. Candida albicans biofilm formation was evaluated by counting colony forming units, cellular metabolic activity, and qualitative and quantitative analysis through confocal fluorescence microscopy. A significant increase in roughness was observed in both resins after two years of simulated brushing, mainly with dentifrice. After two years of brushing, an increase in the number of cells and metabolism of C. albicans was observed, in agreement with the fluorescence and biofilm thickness results. Brushing with dentifrice increased the roughness of heat-cured and 3D-printed resins and potentially increased C. albicans biofilm formation. Full article

Cellular organization relies on both membrane-bound organelles and membraneless biomolecular condensates formed through liquid–liquid phase separation. Recent discoveries reveal intricate coupling between lipid membrane organization and condensate assembly, reshaping our understanding of cellular compartmentalization. This review synthesizes multidisciplinary research using advanced techniques including super-resolution microscopy, fluorescence recovery after photobleaching, and in vitro reconstitution to examine lipid-condensate interactions. Lipid membranes serve as nucleation platforms that reduce critical concentrations for condensate formation by orders of magnitude through membrane anchoring and thermodynamic coupling, creating specialized microenvironments that substantially enhance enzymatic activities. Key regulatory mechanisms include phosphorylation-driven assembly and disassembly, membrane composition effects from cholesterol content and fatty acid saturation, and environmental factors such as calcium and pH. These interactions drive signal transduction through receptor clustering, membrane trafficking via organized domains, and stress responses through protective condensate formation. Dysregulation of lipid-condensate coupling, including aberrant phase transitions and membrane dysfunction, underlies metabolic disorders and neurodegenerative diseases. This coupling represents a fundamental organizing principle with significant therapeutic potential. Current challenges include developing quantitative methods for characterizing condensate dynamics in complex cellular environments and translating molecular mechanisms into clinical applications. Future progress requires interdisciplinary approaches combining advanced experimental techniques, computational modeling, and standardized protocols to advance both fundamental understanding and therapeutic innovations. Full article

Alzheimer’s disease (AD) is characterized by amyloid-beta (Aβ) plaque accumulation and neurodegeneration. This study identified gyrophoric acid, a lichen-derived phenolic metabolite, as a dual-action Aβ42 inhibitor preventing aggregation and disassembling of mature Aβ42 fibrils. Integrated in silico studies revealed that gyrophoric acid was a strong thermodynamic stabilizer of Aβ42 (MM–GBSA: −27.3 kcal/mol) via entropically driven hydrophobic interactions and disruption of aggregation-prone conformations (100 ns MD simulations). Through biochemical analysis of the fluorescent dye thioflavin T (ThT), gyrophoric acid induced rapid Aβ42 fibril disassembly within 5 h, with time-lapse confocal microscopy quantitatively confirming the near-complete dissolution of large aggregates by 24 h. ADMET profiling revealed favorable pharmacokinetics (moderate oral absorption: 48.5–57.3%; low toxicity) and Lipinski’s rule compliance. These results establish gyrophoric acid as a promising natural bioactive compound for anti-AD therapeutics with a unique hydrophobic-stabilization mechanism. Full article

This study presents a novel method for high-precision detection and quantitative evaluation of the spatial relationship between cells and amyloid-$\beta$ (A$\beta$) in time-lapse brightfield microscopy images. Achieving accurate detection of non-fluorescent cells and A$\beta$ deposits requires high-quality video images free from noise, distortion, and frame-to-frame luminance flicker. To this end, we employ a robust preprocessing pipeline that combines multi-frame integration with vignetting correction to enhance image quality and reduce luminance variability across frames. Key preprocessing steps include background correction via two-dimensional polynomial fitting, temporal smoothing of luminance fluctuations, histogram matching for luminance normalization, and dust artifact removal based on intensity thresholds. This enhanced imaging approach enables accurate identification of A$\beta$ aggregates, which typically appear as jelly-like structures and are difficult to detect under standard brightfield conditions. Furthermore, we introduce a quantitative index to assess the spatial relationship between cells and A$\beta$ concentrations, facilitating detailed analysis under varying A$\beta$ levels. Full article

During large-scale pavement construction, the preparation of SBS-modified asphalt typically produces large amounts of harmful fumes. The emergence of deodorants can effectively alleviate the problem of smoke emissions during the asphalt manufacturing process. On the basis of ensuring the original road performance, exploring more suitable dosages and types of deodorant is urgently needed. Five commercial deodorants were evaluated using an asphalt smoke collection system, and UV-visible spectrophotometry (UV) was employed to screen the deodorants based on smoke concentration. Gas chromatography–mass spectrometry (GC-MS) was used to quantitatively analyze changes in harmful smoke components before and after adding two deodorants. Subsequently, the mechanisms of action of the two different types of deodorants were analyzed microscopically using fluorescence microscopy. Finally, the performance of bitumen and asphalt mixtures after adding deodorants was evaluated. The results showed that deodorant A (reactive type) and D (adsorption type) exhibited the best smoke suppression effects, with optimal addition rates of 0.6% and 0.5%, respectively. Deodorant A reduced benzene homologues by nearly 50% and esters by approximately 40%, while deodorant D reduced benzene homologues by approximately 70% and esters by approximately 60%, without producing new toxic gases. Both deodorants had a minimal impact on the basic properties of bitumen and the road performance of asphalt mixtures, with all indicators meeting technical specifications. This research provides a theoretical basis for the effective application of deodorants in the future, truly enabling a transition from laboratory research to large-scale engineering applications in the construction of environmentally friendly roads. Full article

Porcine reproductive and respiratory syndrome virus (PRRSV), an enveloped single-stranded positive-sense RNA virus, poses a significant threat to global swine production. Despite the availability of modified live virus and inactivated vaccines, their limited efficacy and safety concerns highlight the urgent need for novel antiviral therapeutics. This study aimed to investigate the molecular mechanisms by which lycopene inhibits PRRSV replication. Initial assessments confirmed that lycopene did not adversely affect cellular viability, cell cycle progression, or apoptosis. Using fluorescence microscopy, flow cytometry, immunoblotting, quantitative real-time PCR (qRT-PCR), and viral titration assays, lycopene was shown to exhibit potent antiviral activity against PRRSV. Mechanistic studies revealed that lycopene suppresses reactive oxygen species (ROS) production, which is critical for PRRSV proliferation. Additionally, lycopene attenuated PRRSV-induced inflammatory responses, as demonstrated by immunoblotting, ELISA, and qRT-PCR assays. These findings suggest that lycopene inhibits PRRSV replication by modulating ROS levels and mitigating inflammation, offering a promising avenue for the development of antiviral therapeutics. This study provides new insights and strategies for combating PRRSV infections, emphasizing the potential of lycopene as a safe and effective antiviral agent. Full article

Asphalt plant reclaimed powder is a common solid waste in road engineering. Reusing reclaimed powder as filler holds significant importance for environmental protection and resource conservation. The key factors affecting the feasibility of reclaimed powder reuse are its acidity/alkalinity and cleanliness. Traditional evaluation methods, such as the methylene blue test and plasticity index, can assess reclaimed powder properties to guide its recycling. However, these methods suffer from inefficiency, strong empirical dependence, and high variability. To address these limitations, this study proposes a rapid and precise evaluation method for reclaimed powder properties based on Fourier transform infrared spectroscopy (FTIR). To do so, five field-collected reclaimed powder samples and four artificial samples were evaluated. Scanning electron microscopy (SEM), X-ray fluorescence spectroscopy (XRF), and X-ray diffraction (XRD) were employed to characterize their microphase morphology, chemical composition, and crystal structure, respectively. Subsequently, FTIR was used to establish correlations between key acidity/alkalinity, cleanliness, and multiple characteristic peak intensities. Representative infrared characteristic peaks were selected, and a quantitative functional group index (Is) was proposed to simultaneously evaluate acidity/alkalinity and cleanliness. The results indicate that reclaimed powder primarily consists of tiny, crushed stone particles and dust, with significant variations in crystal structure and chemical composition, including calcium carbonate, silicon oxide, iron oxide, and aluminum oxide. Some samples also contained clay, which critically influenced the reclaimed powder properties. Since both filler acidity/alkalinity and cleanliness are affected by clay (silicon/carbon ratio determining acidity/alkalinity and aluminosilicate content affecting cleanliness), this study calculated four functional group indices based on FTIR absorption peaks, namely the Si-O-Si stretching vibration (1000 cm⁻¹) and the CO₃²⁻ asymmetric stretching vibration (1400 cm⁻¹). These indices were correlated with conventional testing results (XRF for acidity/alkalinity, methylene blue value, and pull-off strength for cleanliness). The results show that the Is index exhibited strong correlations (R² = 0.89 with XRF, R² = 0.80 with methylene blue value, and R² = 0.96 with pull-off strength), demonstrating its effectiveness in predicting both acidity/alkalinity and cleanliness. The developed method enhances reclaimed powder detection efficiency and facilitates high-value recycling in road engineering applications. Full article

Low-level laser therapy (LLLT) is gaining attention as an effective adjunct to non-surgical periodontal treatment. This study evaluates the potential of LLLT to reduce bacterial load in a clinically relevant in vitro subgingival biofilm model and its impact on the inflammatory response. A subgingival biofilm model consisting of seven bacterial species was established. Primary human gingival fibroblasts (GFs) and periodontal ligament cells (PDLs) were cultured. Both biofilms and host cells were treated with the DenLase Diode Laser (980 nm) under various clinically relevant settings. The composition and structure of the seven-species biofilms were evaluated using quantitative PCR and fluorescence microscopy, respectively. The inflammatory response in host cells was analyzed by measuring the gene and protein expression levels of various inflammatory mediators. Laser treatment at power outputs ranging from 0.3 to 2 W had no significant effect on biofilm composition or architecture. LLLT, particularly at higher power settings, reduced the viability in both GFs and PDLs up to 70%. Gene expression levels of inflammatory mediators were only minimally influenced by laser treatment. However, LLLT significantly decreased the secretion of all examined cytokines. These findings suggest that LLLT with a 980 nm diode laser, under clinically relevant conditions, exerts anti-inflammatory rather than antimicrobial effects. Full article

Orthohantaviruses are tri-segmented negative-sense RNA viruses that can cause severe pathologies in humans. Currently, limited information exists on the molecular interactions driving orthohantavirus assembly in infected cells. Specifically, it is not clear how its glycoproteins (i.e., Gn and Gc) interact with other viral or host molecules. In this study, we use one- and two-color Number and Brightness fluorescence microscopy approaches to quantitatively characterize the interactions between orthohantavirus glycoproteins and the nucleoprotein in transfected cells. Our results indicate that orthohantavirus nucleoprotein homo-interactions are strongly affected by the host environment. Furthermore, we report evidence of Gc–nucleoprotein interactions, based on (i) the high fluorescence cross-correlation between these two proteins and (ii) the increased Gc-Gc interactions observed in the presence of nucleoprotein. Finally, experiments on a Gc deletion mutant suggest that the observed protein–protein interactions are mediated by the cytoplasmic tail of Gc. In conclusion, this study provides new insights into the role of the interactions between orthohantavirus glycoproteins and nucleoprotein in the context of viral assembly. Full article

Total internal reflection fluorescence–structured illumination microscopy (TIRF-SIM) can enhance the lateral resolution of fluorescence microscopy to twice the diffraction limit, enabling subtler observations of activity in subcellular life. However, the lack of an axial resolution makes it difficult to resolve three-dimensional (3D) subcellular structures. In this paper, we present an alternative TIRF-SIM axial resolution enhancement method by exploiting quantitative information regarding the distance between fluorophores and the surface within the evanescent field. Combining the lateral super-resolution information of TIRF-SIM with reconstructed axial information, a 3D super-resolution image with a 25 nm axial resolution is achieved without attaching special optical components or high-power lasers. The reconstruction results of cell samples demonstrate that the axial resolution enhancement method for TIRF-SIM can effectively resolve the axial depth of densely structured regions. Full article

This study characterises low-grade copper ore tailings from a conventional flotation circuit to evaluate their feasibility for further processing. A suite of advanced analytical techniques, such as X-ray fluorescence (XRF), inductively coupled plasma (ICP), X-ray diffraction (XRD), and the quantitative evaluation of minerals by scanning electron microscopy (QEMSCAN), was employed to assess the elemental, chemical, and mineralogical composition of the tailings. Chalcopyrite was identified as the dominant copper-bearing mineral phase, predominantly locked within iron oxides and silicate gangue minerals. The QEMSCAN results showed that chalcopyrite was only partially liberated, which highlights the complex mineral intergrowths that hinder efficient recovery. Based on the mineralogical characteristics, the applicability of various processing techniques, including conventional froth flotation, advanced flotation methods [including Hydrofloat^TM, Jameson, and the Reflux Flotation Cell (RFC)], magnetic separation, and gravity separation, was evaluated. Overall, this study indicates that incorporating HydroFloat™, the Jameson Cell, and the RFC into the flotation circuit could greatly improve copper recovery from tailings. This study also identified rare earth elements (REEs) as potential by-products of copper recovery, so it is an additional opportunity for resource recovery. This paper contributes to sustainable mining practices and resource optimization by highlighting the characteristics and recovery of valuable minerals from tailings. Full article