Search Results (548)

Background: In an earlier murine model of myocardial infarction (MI), we showed that CD8 cells and myeloid dendritic cells (mDCs) infiltrate the infarcted myocardium within the first week. However, in humans, the spatial interplay between CD8⁺ T cells and dendritic cells in the spatial context of human myocardial infarction remains underexplored. Objective: In the present study, we applied spatial transcriptomics and functional assays to characterize immune–stromal dynamics in infarcted myocardium and peripheral blood. Methods & Results: Spatial transcriptomics analysis of infarcted human myocardium at days 2 and 6 post-MI, combined with peripheral blood flow cytometry and EPC colony-forming assays, was performed. Cell composition, pathway enrichment, and cell-to-cell communication analyses were conducted to map immune–stromal cells’ dynamics across time points. Spatial mapping identified dynamic shifts in immune, fibroblast, and endothelial populations, with fibroblasts and endothelial cells remaining abundant throughout. CD8⁺ T cells accumulated in ischemic regions while their circulating levels declined. Gene Ontology and pathway analyses of CD8A⁺ transcripts revealed enrichment of proinflammatory and NF-κB survival programs. ITGAX/CD33/THBD⁺ APCs progressively increased within infarct zones, activating antigen-presentation and leukocyte chemotaxis pathways. Early (day 2) APC–endothelial crosstalk showed the strongest predicted recruitment signals for CD8⁺ T cells, which diminished by day 6. Finally, EPC colony-forming capacity showed a tendency for reduction in MI patients and inversely correlated with coronary lesion burden, indicating impaired vascular repair potential. Conclusions: This integrative spatial and functional study demonstrates that APC-driven CD8⁺ recruitment and EPC dysfunction are key features of human MI. Immune–endothelial niches facilitate early cytotoxic T-cell infiltration, while progenitor depletion limits vascular regeneration. These findings provide mechanistic insight into immune–vascular imbalance during infarct healing and highlight potential therapeutic targets to modulate inflammation and restore vascular repair. Full article

Fermentation is a type of biological process conducted domestically or commercially to preserve foods and beverages, produce alcohol, add nutritional value and improve aroma and flavor. The natural fermentation of flour in water to obtain a leaven for baking, lately scrutinized in the laboratory with the application of metagenomic methods, has been ubiquitous since the dawn of civilization. Commercially, single culture or defined mixtures of microorganisms are used for their predictability, but regularly fed two-domain microorganism cultures are favored in less industrialized and domestic operations. Fungi principally produce the carbon dioxide responsible for leavening. The bacteria produce acid in the bread commonly known as sourdough for its aroma and flavor. A leaven made by fermentation using flour and water can be stored while it is dormant. We studied a mature culture that is fed twenty-fold with water and flour by incubating it for 24 h, sampling it regularly for pH measurements, and plating it. The colonies were suspended for micrography and DNA extraction for PCR and Sanger sequencing. The metagenomic DNAs were analyzed for bacterial and fungal composition. The proportions of the plant and microbial DNA endogenous to the flour decline rapidly, and the predominant bacteria and fungi in mature leaven propagate, without overlap between the respective microbiomes. Full article

Cancer remains a significant challenge to global public health. Preliminary studies indicate that the protein Golgi-associated, Gamma-adaptin Ear Containing, ARF Binding Protein 2 (GGA2) may influence various cancers. However, the potential role of GGA2 in oncogenesis remains unknown. We utilized data from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) projects to analyze GGA2 expression levels. Genetic variations and protein expression of GGA2 in human tissues were assessed using the cBioPortal. Gene Set Enrichment Analysis (GSEA) provided deeper insights into GGA2’s oncogenic functions. Comprehensive analysis of TCGA datasets combined with ESTIMATE and TIMER tools demonstrated significant correlations between GGA2 expression levels and clinical outcomes, survival metrics, genomic instability markers (microsatellite instability (MSI)/tumor mutational burden (TMB)), and immune microenvironment composition. Functional validation in prostate cancer models employed qRT-PCR quantification, immunoblotting verification, and cellular behavior assessments through colony formation, Transwell migration, and wound closure assays. Our findings suggest GGA2 could serve as a prognostic biomarker in various cancers. Abnormal levels of GGA2 promoter methylation and genetic alterations may contribute to its dysregulated expression in some cancers. Distinctly, GGA2 expression correlates with MSI and TMB across different cancers and is linked to the expression of immune checkpoint genes. Functionally, GGA2 is instrumental in inhibiting oncogenic mechanisms by diminishing the proliferation, colony formation, invasion, and migratory capabilities of prostate cancer cells. Our study shows that the oncogenic role of GGA2 in various cancers and GGA2 could be served as a biomarker of PARD. Full article

Mobile robot path planning using Ant Colony Optimization (ACO) has the disadvantages of slow convergence, local optima, and unsmooth paths because of fixed heuristics and constant pheromone updating. In this paper, Self-Adaptive Risk-Aware Bidirectional updating ACO (SAR-BACO) is proposed with three improvements: (1) composite heuristic incorporating target attraction, obstacle repulsion and direction consistency to minimize early blind searching; (2) dynamic pheromone updating based on solution quality and number of iterations to balance exploration and exploitation; (3) triangular pruning to remove redundant turning points and become smoother. Theoretical analysis verifies convergence. Our experimental results on grids up to 50 × 50 demonstrate that SAR-BACO performs much better than classical and heuristic-improved algorithms with respect to the length of a path, convergence rate, smoothness and efficiency. Using SAR-BACO on a 50 × 50 map, the path lengths, convergence iterations and turning points decreased by 60.68%, 48.96%, and 96.00% respectively compared to Basic ACO (after triangular pruning, values averaged over 50 runs). The framework provides a generalizable solution to autonomous navigation with the need to consider both search efficiency and path executability. Full article

This study evaluated Salmonella behavior during Sardinian fermented sausage (SFS) production through a challenge test on experimentally inoculated raw meat. The objectives were to (i) determine the survival and reduction kinetics of Salmonella during fermentation and ripening and (ii) evaluate the relationship between pathogen behavior and the evolution of key chemical-physical parameters (pH, water activity). Three batches of SFS were produced, and the meat mixture was inoculated with a three-strain Salmonella cocktail (reference and field strains) to 10² CFU/g. After 20 days of ripening, sausages were vacuum-packed and stored under refrigerated conditions (+4 ± 2 °C). For each batch, triplicate samples were collected and analyzed at different production stages (mixing, after overnight rest, and 24 h after stuffing) and during shelf life (days 6, 21, 30, and 40). Analyses included Salmonella detection and enumeration by direct plating, aerobic colony count, Enterobacteriaceae, staphylococci, lactic acid bacteria, molds and yeasts, as well as pH, water activity, and gross composition. Salmonella counts increased by approximately one log unit after stuffing, before the onset of acidification. During fermentation and ripening, pathogen levels declined but remained detectable, even after prolonged refrigerated storage. These findings indicate that although ripening, and particularly fermentation, significantly (p < 0.05) reduce Salmonella levels, complete inactivation is not achieved. The study highlights the importance of controlling initial contamination levels, validating fermentation and ripening conditions, and the application of additional post-process hurdles to ensure product safety. Full article

The use of microalgae as a food source is limited by consumers’ dislike of their organoleptic traits, primarily the intense green color and bitter taste associated with high chlorophyll content. The eukaryotic microalgae Chlorella vulgaris can grow under heterotrophic conditions, providing the opportunity to cultivate chlorophyll-less strains. In this work we applied random mutagenesis for breeding chlorophyll-deficient C. vulgaris strains. Wild-type strain was UVC-radiated, and 12 colonies with changed pigmentation were selected. Based on phenotypic stability two mutants, M6 and M11, were selected for characterization of growth, pigment and biomass accumulation. Cultivation under photo-, mixo- and heterotrophic conditions revealed distinct phenotypes for the two mutants. M6 remained chlorophyll-deficient in all cultivation conditions tested, while chlorophyll was observed in M11 when grown under light. Under heterotrophic and mixotrophic growth conditions, both mutants were chlorophyll-deficient while biomass productivity, protein content, and amino acid composition remained similar to wild type. Characterization of the cellular ultrastructure of the wild type and mutants using cryo Focused Ion-Beam Scanning Electron Microscopy revealed that functional chloroplasts and thylakoid membranes were absent in the mutants. Our work demonstrates how a simple approach using UV mutagenesis and visual screening can provide novel strains of C. vulgaris with traits for improved consumer acceptance, without compromising the use of the algae biomass as a protein-rich food source. Full article

The Esox lucius is a high-quality fish species endemic to northern Xinjiang, having developed into a regional specialty industry with significant market value. However, during storage, it is prone to microbial growth that elevates biogenic amine levels, posing potential food safety risks. Therefore, this study systematically evaluated the effects of protamine—extracted from Esox lucius byproducts and used as a natural preservative—on the succession of microbial communities and biogenic amine accumulation in fish muscle under storage conditions of 4 °C, −3 °C, and −18 °C. A detection method for biogenic amines was also established. Results revealed characteristic changes in fish muscle microbial community α-diversity over storage time. Protamine treatment significantly delayed increases in total colony counts and microbial succession processes without altering the final dominant microbial community composition. By optimizing ultrasonic-assisted extraction and derivatization steps, an analytical method suitable for detecting eight biogenic amines in fish muscle matrices was established. Results indicate that protamine effectively inhibits the accumulation of all eight biogenic amines, with the 1% treatment group showing the most significant effect (p < 0.05). This study not only provides basis for the precise application of protamine in seafood preservation but also offers guidance for the resource utilization of aquatic by-products. Full article

To address the challenges of dense structural layouts, limited path feasibility, and stringent assembly constraints in cable routing within complex compartments of aerospace equipment, this paper proposes a cable path planning method that integrates Bidirectional Crossing Line Pruning (BCLP) with an improved ant colony optimization (IACO) algorithm. First, a hierarchical activation strategy for key obstacles is realized by constructing primary and extended crossing lines. On this basis, the BCLP algorithm is introduced, combining global perspective with local reduction capability to significantly reduce the complexity of the search space. Second, in line with cable assembly process requirements, a composite heuristic function is formulated by integrating obstacle-crossing cost and bending penalty. Additionally, a multi-objective-driven pheromone update model is developed to enhance the routing process’ feasibility and convergence performance. Experimental results across various aerospace cabling simulation scenarios demonstrate that the proposed method achieves an average reduction of 19.6% in multi-objective process cost and a 68.5% improvement in convergence efficiency compared to traditional visual graph methods combined with standard ACO. The approach provides effective support for the automation and intelligent planning of cable layouts in complex environments, offering strong potential for engineering applications. Full article

The incorporation of spore-forming probiotics into thermally processed foods represents a promising strategy to expand functional food availability. In this study, probiotic snacks were formulated from oat and rice-bean matrices as delivery vehicles for Heyndrickxia coagulans (formerly Bacillus coagulans) BC4 spores. The effects of baking and microwave processing, packaging, and storage conditions on spore viability and functionality were evaluated. While oven baking (180 °C) preserved viability in both matrices under mild conditions (survival > 90%), dielectric heating induced significant viability loss depending on the matrix. The starch-based rice-bean matrix, characterized by higher post-processing water activity (Aw), suffered a thermal runaway effect, resulting in significant spore inactivation (viability decreased to 6.08 log colony forming units/g (CFU/g); 1.5 min). Conversely, the oat matrix acted as a thermo-physical stabilizer, maintaining high viability (9.41 log CFU/g; 1.5 min) by limiting dielectric energy absorption via its fiber-lipid composition. Oxidative stress and premature germination likely contributed to the viability loss observed in atmospheric packaging during the 30-day storage. The oat matrix mitigated this effect through a dual-protective mode: active radical scavenging (validated by superior ferric reducing ability of plasma (FRAP) values) and passive water binding. Simulated digestion data align with the functional preservation observed, resulting in increased survival for oat-based formulations. Overall, the results demonstrate the feasibility of developing oat and rice-bean snacks enriched with H. coagulans spores and highlight the critical role of matrix and processing conditions in preserving probiotic viability. Full article

The rapid growth of e-commerce has intensified operational and sustainability challenges in urban last-mile delivery, necessitating routing methods that perform reliably under realistic traffic and spatial conditions. This study evaluates three routing algorithms, Nearest Neighbor (NN), Clarke–WrightSavings (CWS), and Ant Colony Optimization (ACO), using 1764 real-world Amazon delivery stops grouped into ten operational clusters in the Nashville metropolitan area. Travel distances and times were obtained through the Google Maps Distance Matrix API in driving mode to reflect actual road network structure and typical traffic conditions. Substantial performance differences were observed across algorithms and cluster configurations. NN achieved a strong performance in compact clusters (18.43 miles and 58.48 min in Cluster 4) but performed poorly in dispersed clusters (82.44 miles and 196.48 min in Cluster 9), reflecting high sensitivity to spatial dispersion. In contrast, CWS consistently reduced travel distance and time across clusters, achieving the shortest observed route (18.50 miles and 47.82 min in Cluster 10). Relative to ACO, CWS reduced travel distance by up to 42% (Cluster 9) and reduced travel time by over 45% in high-dispersion clusters. ACO exhibited the highest variability, with distances reaching 98.77 miles and travel times exceeding 218 min. Multi-criteria evaluation using efficiency ratios, distributional analysis, performance quadrant visualization, and a Composite Performance Index (CPI) confirmed the dominance of CWS. CPI scores of 1.00 (CWS), 0.78 (NN), and 0.00 (ACO) reflected balanced spatial and temporal efficiency under identical traffic-informed inputs. The results demonstrate that deterministic savings-based routing provides superior stability, efficiency, and scalability in semi-static urban delivery systems. However, the present study did not benchmark the evaluated algorithms against state-of-the-art exact TSP solvers (e.g., Concorde, LKH) or more recent metaheuristics such as Genetic Algorithms or Variable Neighborhood Search. The objective was to provide a controlled empirical comparison under consistent traffic-informed cost matrices rather than to establish global optimality bounds. Consequently, while the findings strongly support the relative superiority of the Clarke–Wright Savings approach within the evaluated framework, future research incorporating advanced exact and hybrid optimization methods would further contextualize algorithmic performance. Full article

Background/Objectives: Chlorhexidine (CHX) and alcoholic (A+) mouthwashes are associated with adverse oral effects. Therefore, this study compared the efficacies of non-alcoholic mouthwashes, including fluoride (A−) and herbal (Hr) rinses, for preventing bacterial accumulation and enamel demineralization around metal brackets (MBs), ceramic brackets (CBs), and resin composite attachments (RCAs). Methods: Following the exposure to CHX, A+, A−, and Hr rinses for 1 min, the growth of Streptococcus mutans on MB, CB, and RCA was assessed using colony-forming units and scanning electron microscopy (SEM). Controls included attachments without intervention. In another setting, enamel with bonded attachments was exposed to mouthwashes for 1 min and subjected to cariogenic demineralization for 24 h. Enamel’s Vickers microhardness was measured before and after the demineralization challenge. Data were analyzed using paired t-tests and one-/two-way ANOVA with Tukey’s tests. Results: CHX mouthwash demonstrated superior antimicrobial efficacy against S. mutans biofilms across all orthodontic attachments (p < 0.05). On metallic brackets, CHX (0 ± 0 log₁₀) and A− (1.7 ± 0.4 log₁₀) significantly (p < 0.001) outperformed controls (6.9 ± 0.1 log₁₀), Hr (6.08 ± 0.2 log₁₀), and A+ (6.2 ± 0.6 log₁₀). Similar patterns emerged for ceramic brackets, with CHX (0 ± 0 log10) and A− (1.4 ± 0 log10) superior to controls (6.6 ± 0.4 log₁₀). On resin composite attachments, CHX (2.9 ± 0.05 log₁₀) and Hr (3.4 ± 0.08 log₁₀) exceeded controls (5.4 ± 0.09 log₁₀) in inhibiting the biofilm growth (p < 0.05). Enamel microhardness reduction was significantly influenced by attachment type (p < 0.0001) and mouthwash type (p = 0.0063), with significant interaction between variables (p = 0.0052). Conclusions: CHX and A− mouthwashes effectively inhibited S. mutans biofilms on orthodontic attachments, while attachment type and mouthwash significantly influenced enamel microhardness reduction. Full article

Barley grown in temperate regions is often challenged by powdery mildew disease. An effective solution is mildew resistance locus o (mlo)-based resistance, which is monogenic, durable, and broad-spectrum. While the pleiotropic effects of mlo mutations on above-ground tissues are well documented, their impact on the root-associated microbiome remains underexplored. We utilized CRISPR/Cas9 to generate novel mlo mutant lines and evaluated their resistance to causal fungus Blumeria hordei. We further examined if mlo knockout has any impact on the overall root microbiome diversity and composition under field-like conditions and applied DESeq2 to compare the abundance of microbial taxa between mutants and wild type. We created five novel resistant mlo lines, including the first mutants with amino acid alterations in the protein’s extracellular region. Mutant lines showed significantly reduced B. hordei colony formation (0.5–5%). While microbial alpha and beta diversity were not significantly altered, a few microbial taxa displayed time-dependent shifts in abundance. Overall, our study demonstrates the effectiveness of CRISPR/Cas9 in generating mlo-based resistance. Moreover, the study revealed functionally important residues in the protein’s extracellular region. Finally, we present the first evidence of limited mlo-associated effects on root microbiome diversity and relative abundance of microbial taxa. Full article

This study evaluated the viability, microbiological composition, functional traits, and safety of probiotic bacteria isolated from commercial products marketed as containing Limosilactobacillus reuteri. Viable cell counts, biochemical characterization, strain-level identification, functional properties, gastrointestinal tolerance, and safety attributes were assessed. Among the evaluated products, only four presented colony-forming units (CFU) counts consistent with label claims (products E, F, G, and H), while two showed no detectable viable microorganisms (products B and L). All isolates were Gram-positive, catalase-negative, and predominantly rod-shaped. rep-PCR analysis revealed strain homogeneity in most products, whereas others (products A and K) exhibited heterogeneous microbial compositions. Molecular identification based on 16S rRNA sequencing showed a predominance of Lmb. reuteri and Lacticaseibacillus rhamnosus, with some products containing additional species such as Lactiplantibacillus plantarum and Lactobacillus acidophilus. Functional assays demonstrated strain-dependent proteolytic and diacetyl-producing capacities, as well as variable tolerance to simulated gastrointestinal conditions. Most strains preferentially produced L-lactate, although some generated substantial amounts of D-lactate. All isolates were susceptible to antibiotics recommended by EFSA, except for intrinsic vancomycin resistance, and no transferable virulence markers, biogenic amine production, or Salmonella contamination were detected. Furthermore, virulence-related genes such as hdc, tdc, odc, hyl, cylA, and ace were not identified. Overall, the results highlight pronounced discrepancies between label claims and microbiological quality among commercial probiotic products and reinforce the importance of strain-level characterization to ensure safety, functional performance, and regulatory compliance. Full article

While harmful cyanobacterial blooms (HCBs) are extensively characterized in eutrophic lakes, the ecological dynamics of connected river networks remain oversimplified, obscuring the mechanisms of community assembly and toxin distribution across the lake–river interface. This study investigated the spatial heterogeneity of HCBs and microcystins (MCs) in the Lake Taihu watershed, revealing a complex functional divergence between lotic and lentic ecosystems. The rivers functioned as primary nutrient sources, with Total Nitrogen (3.35 ± 1.52 mg·L⁻¹) and Total Phosphorus (0.21 ± 0.22 mg·L⁻¹) concentrations being 1.7-fold and 1.8-fold higher, respectively, than those in the lake during peak periods. Conversely, the lake acted as a biological sink, supporting a peak cyanobacterial density (3.32 × 10⁷ cells·L⁻¹) nearly 1.5 times that of the river network. Phytoplankton community analysis revealed distinct ecological niches: while the lentic lake environment was almost exclusively dominated by colonial Microcystis (>90% relative abundance), the lotic river networks harbored a diverse assemblage with significant contributions from filamentous Oscillatoria and Dolichospermum. Correspondingly, intracellular MC (IMC) in the lake (up to 14.5 μg·L⁻¹) significantly exceeded riverine levels (generally <1.0 μg·L⁻¹). Despite these compositional differences, toxin dynamics exhibited strong bidirectional coupling (r > 0.75, p < 0.01), suggesting a spillover effect where the lake determines the watershed’s toxin burden during rivers outflow period. Redundancy Analysis (RDA) further elucidated that limnetic blooms were primarily regulated by water temperature and pH, whereas riverine communities were strictly constrained by hydrodynamic flow. Consequently, the health risk assessment revealed a highly heterogeneous landscape where, beyond the northern lake bays, specific semi-lentic river segments emerged as cryptic hotspots. These findings demonstrate that while nutrients fuel the system, hydrodynamic conditions act as the ultimate ecological filter determining the spatiotemporal distribution of cyanobacterial risks, necessitating an integrated approach to monitoring the entire lake–river continuum. Full article

Spent coffee grounds are an abundant agro-industrial by-product with considerable potential as a functional food ingredient. This study investigated the effects of lactic acid fermentation on the antioxidant and anti-inflammatory activities of spent coffee grounds, as evaluated using their extracts, with a focus on fermentation-induced remodeling of phenolic compounds and the functional implications. Fermentation was conducted using Lactobacillus plantarum, and changes in microbial growth, pH, reducing sugar content, phenolic composition, antioxidant capacity, and anti-inflammatory activity were evaluated. During fermentation, viable cell counts increased from 6.73 log colony-forming units (CFU)/mL at 0 h to 9.27 log CFU/mL at 48 h, accompanied by a decrease in pH and an increase in reducing sugar content, indicating active microbial metabolism. Total polyphenol content increased markedly, reaching 97.44 mg gallic acid equivalents (GAE)/100 g in water extracts fermented for 48 h compared with 62.96 mg GAE/100 g in non-fermented controls. High-performance liquid chromatography analysis revealed significant enrichment of phenolic acids, including caffeic, ferulic, and protocatechuic acids. Correspondingly, fermented extracts exhibited enhanced antioxidant activities, as determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), superoxide dismutase (SOD-like, and catalase assays. In addition, fermented extracts showed improved cellular compatibility and significantly inhibited nitric oxide production (approximately 50–60% at 200–300 μg/mL) and pro-inflammatory cytokine production, with interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) inhibition rates exceeding 60% at 200–300 μg/mL in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. These biological effects were closely linked to fermentation-induced qualitative and quantitative changes in phenolic composition, providing mechanistic insight beyond simple activity enhancement. Overall, lactic acid fermentation enhances the functional properties of spent coffee grounds, highlighting their potential as upcycled, value-added ingredients for functional food and nutraceutical applications. Full article