Search Results (37,213)

Streptococcus uberis is a bovine mastitis pathogen with a demonstrated ability to form biofilms. However, the dynamics of this process remain poorly characterized. This study aimed to comprehensively characterize biofilm formation in four S. uberis strains that differed in their biofilm-forming capacity, from weak to strong producers, and in the presence of key virulence-associated genes, such as sua, hasA and hasC. To achieve this, we integrated structural, biochemical, physiological and transcriptional analyses using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), spectral flow cytometry and qRT-PCR. The multi-faceted analysis revealed a coordinated maturation peak at 48 h, characterized by a structured architecture with water channels, a distinct biochemical signature rich in polysaccharides and proteins, and a predominantly viable bacterial population. This peak coincided with a marked upregulation of key virulence-associated genes, with sua expression increasing 2.5-fold and hasA increasing 3-fold at 48 h. This mature biofilm conferred high tolerance to antibiotics, with eradication concentrations (>256 µg/mL) exceeding planktonic MICs, although tetracycline was notably effective. At 72 h, the biofilm entered a dispersion phase characterized by structural collapse and reduced viability. These findings establish S. uberis biofilm maturation as a highly coordinated process, providing new insights into the biofilm lifecycle of this important pathogen and identifying key temporal and molecular targets for future interventions. Full article

Background: Acacetin, a naturally occurring flavone present in various plants, is known as a promising drug candidate for cardiovascular disorders. Our previous study demonstrated that acacetin ameliorates atherosclerosis through endothelial cell protection; however, its pharmacological effects on vascular smooth muscle cells (VSMCs) remain unexplored. This study investigates the therapeutic potential of acacetin against lysophosphatidylcholine (LysoPC)-induced VSMC injury and elucidates the underlying molecular mechanisms. Methods and Results: Multiple biochemical techniques were employed in the present study. The results showed that acacetin significantly attenuated LysoPC-induced apoptosis and reactive oxygen species (ROS) generation in cultured VSMCs. Western blot analysis revealed that the cytoprotection of acacetin was associated with upregulated expression of antioxidant defense proteins, including nuclear factor erythroid 2-related factor 2 (Nrf2), catalase (CAT), NADPH quinone oxidoreductase 1 (NQO-1), and superoxide dismutase 1 (SOD1). Nrf2 silencing completely abolished these protective effects. Mechanistically, siRNA-silencing of Sirtuin 1 (Sirt1) abrogated acacetin-induced modulation of the Nrf2/Keap1/p62 signaling. In vivo validation using aortic tissues from high-fat-diet-fed ApoE^−/− mice confirmed that acacetin effectively suppressed VSMC apoptosis and ROS overproduction associated with restoring the downregulated Sirt1 expression levels. Conclusions: These findings establish a novel mechanistic paradigm wherein acacetin confers protection against LysoPC-induced VSMC apoptosis and oxidative stress through Sirt1-dependent activation of the Nrf2/p62 signaling pathway, suggesting that acacetin is a promising therapeutic drug candidate for atherosclerotic plaque stabilization. Full article

In aquaculture, feed production influences nutrition, performance, water quality, and overall profitability. This study evaluated the effects of three levels of internal fat (IF), resulting from the inclusion of 0%, 2%, or 4% fat in the preconditioner during extrusion, and their interaction with two extruder screw configurations: medium-shear (MS) and high-shear (HS), on kibble physical quality and extrusion parameters. Increasing IF resulted in a quadratic increase in amylose–lipid complexation under the HS configuration (p = 0.030; r2 = 0.9) and a linear reduction (p < 0.001) in specific mechanical energy (SME) with a strong negative Pearson correlation (r −0.9; p = 0.009) in both configurations. Fat inclusion also reduced mass temperature and die pressure (p < 0.05), leading to lower starch gelatinization degree (p < 0.05) from 87.9 ± 0.6% to 83.4 ± 0.3% in MS configuration and 95.6 ± 0.7 to 86.3 ± 0.8% in HS configuration, increased bulk and piece density (p < 0.001), and reduced radial expansion (p < 0.001). These changes decreased floatability (p < 0.05) and water stability, increasing mushiness (p < 0.01). Increased shear partially improved SME transfer, starch cooking, expansion, floatability, and mushiness; however, the negative effects of 4% IF could not be fully mitigated. Overall, higher IF compromised kibble structure, starch gelatinization, and floatability, while screw configuration resulted in only a limited compensatory effect. Full article

This study examined whether co-fermentation with Lactobacillus casei CGMCC 15956 and Lactobacillus delbrueckii CGMCC 21287 could enhance the bioactivity of peptides derived from pecan nut cake (PNC) and clarify the underlying mechanisms. The fermented hydrolysate (PNCH) was compared with an unfermented control. PNCH showed higher antioxidant and α-glucosidase inhibitory activities. Total antioxidant capacity increased from 3.17 to 4.81 mM Trolox, and DPPH radical scavenging activity increased from 62.69% to 84.12%. In addition, the IC₅₀ value for α-glucosidase inhibition decreased from 7.549 to 4.509 mg/mL. In a mouse model of acute alcohol-induced liver injury, PNCH significantly alleviated liver damage through the synergistic enhancement of antioxidant and α-glucosidase inhibitory activities. Peptidomic analysis identified two representative bioactive peptides, FAGDDAPR (from actin) and LAGNPDDEFRPQ (from cupin domain–containing protein 1), both of which exhibited antioxidant and α-glucosidase inhibitory activities. Additionally, these peptides alleviated H₂O₂-induced oxidative stress in Caco-2 cells, significantly improving GSH and MDA levels, as well as SOD activity. Molecular docking suggested potential interactions of these peptides with superoxide dismutase, Keap1, and α-glucosidase. These findings support the high-value utilization of PNC and the development of functional peptide-based ingredients. Full article

The increasing demand for sustainable and functional plant-based foods has driven interest in 3D food printing technologies, which require bioinks with tailored rheological and structural properties. This study investigated the effects of transglutaminase (TGase) on the structure–function relationships of plant protein bioinks from fava bean, mung bean, pea, and soybean. TNBS assays showed a dose-dependent increase in crosslinking (27.46–64.57%), with soybean and pea proteins exhibiting the highest reactivity (p < 0.05). ¹H-NMR confirmed protein-specific ε-(γ-glutamyl)lysine bond formation, and synchrotron FTIR revealed TGase-induced α-helix reduction and β-sheet enrichment, indicative of network formation across all proteins. SDS-PAGE analysis demonstrated TGase-mediated polymerization with high-molecular-weight aggregates, particularly pronounced in soybean, while SEM images revealed denser, more continuous protein networks compared to untreated samples. Rheological characterization showed enhanced viscoelasticity and shear-thinning behavior in all bioinks, supporting extrusion and post-printing stability. Textural analysis indicated improvements in hardness, springiness, cohesiveness, and chewiness across all proteins, with soybean and fava showing the most pronounced increases. These results demonstrate that TGase is a versatile tool for reinforcing plant protein networks, improving printability, structural integrity, and texture in 3D-printed foods, while highlighting protein-specific differences in response. Full article

The family Capitellidae performs critical roles in bioturbation and sediment remediation within global marine benthic ecosystems. However, they are a taxonomically challenging group due to their simple morphology and a ‘morphological mosaic’, where traditional classificatory traits, such as thoracic chaetiger counts, appear convergently across genera. Previous multi-locus studies (using 18S, 28S, H3, and COI) first highlighted this conflict, revealing the polyphyly of major genera like Notomastus and even Leiochrides itself (based on unidentified specimens). More recently, mitogenomic studies uncovered massive gene order rearrangements and a conflicting topology but did not include Leiochrides. Critically, with no complete mitogenome reported for a formally identified Leiochrides species, its true phylogenetic position and the validity of its polyphyly remain unresolved. To address this critical gap, we sequenced and characterized the first complete mitochondrial genome from a formally identified species, Leiochrides yokjidoensis, recently described from Korean waters. The complete mitogenome was 17,933 bp in length and included the typical 13 protein-coding genes (PCGs), 2 ribosomal RNAs (rRNAs), and 22 transfer RNAs (tRNAs). Gene order (GO) analysis revealed the occurrence of gene rearrangements in Capitellidae and in its sister clade, Opheliidae. A phylogenomic analysis using the amino acid sequences of 13 PCGs from 30 species established the first robust systematic position for the genus Leiochrides (based on this formally identified species). Phylogenetic results recovered Leiochrides as a sister group to the clade comprising Mediomastus, Barantolla, Heteromastus, and Notomastus hemipodus (BS 99%). This distinct placement confirms that Leiochrides represents an independent evolutionary lineage, phylogenetically separate from the polyphyletic Notomastus complex, despite their morphological similarities. Furthermore, our analysis confirmed the polyphyly of Notomastus, with N. hemipodus clustering distinctly from other Notomastus species. Additionally, signatures of positive selection were detected in ND4, and ND5 genes, suggesting potential adaptive evolution to the subtidal environment. This placement provides a critical, high-confidence anchor point for the genus Leiochrides. It provides a reliable reference to investigate the unresolved polyphyly suggested by previous multi-locus studies and provides compelling evidence for the hypothesis that thoracic chaetiger counts are of limited value for inferring phylogenetic relationships. This study provides the foundational genomic cornerstone for Leiochrides, representing an essential first step toward resolving the systematics of this taxonomically challenging family. Full article

Background: The zinc finger-homeodomain (ZF-HD) transcription factor family exerts pivotal regulatory functions in plant development and stress responses, yet a systematic genome-wide survey is lacking for Rosa chinensis. Methods: In this study, we performed a comprehensive genome-wide identification and analysis of RcZF-HD genes in R. chinensis using bioinformatics approaches. Nine RcZF-HD loci were mined from the rose genome and comprehensively profiled for physicochemical parameters, phylogenetic affiliations, chromosomal positions, exon–intron architectures, conserved motifs, and spatiotemporal expression landscapes. Results: The results showed that RcZF-HD genes were unevenly distributed across four chromosomes (Chr2, Chr4, Chr6, and Chr7), with tandem duplication events detected on chromosomes 2 and 7, suggesting their contribution to gene family expansion. Maximum-likelihood phylogeny placed RcZF-HD proteins within nine well-supported sub-clades alongside Arabidopsis orthologs, implying both evolutionary conservation and lineage-specific divergence. All members retain canonical zinc-finger domains, yet acquire unique motif signatures predictive of functional specialization. Gene structure analysis revealed considerable diversity in exon–intron organization. Expression profiling across six different tissues (root, stem, leaf, bud, flower, and fruit) demonstrated remarkable tissue-specific expression patterns. Notably, RchiOBHm_Chr2g0168531 exhibited extremely high expression in stem tissue, while RchiOBHm_Chr7g0181371 showed preferential expression in flower tissue, suggesting specialized roles in stem development and floral organ formation, respectively. The cold-stress challenge of ‘Old Blush’ petals further disclosed pronounced up-regulation of seven RcZF-HD genes, attesting to their critical contribution to low-temperature tolerance. Conclusions: Integrative analyses furnish a multidimensional blueprint of the rose RcZF-HD repertoire, providing molecular landmarks for future functional dissection and ornamental trait engineering. Full article

Overwinter Syndrome (OWS) affects grass carp (Ctenopharyngodon idellus) aquaculture in China, causing high mortality and economic losses under low temperatures. Failure of antibiotic therapies shows limits of the ‘low–temperature–pathogen’ model and shifts focus to mucosal barrier dysfunction and host–microbiome interactions in OWS. We compared healthy and diseased grass carp collected from the same pond using histopathology, transcriptomics, proteomics, and metagenomics. This integrated approach was used to characterize intestinal structure, microbial composition, and host molecular responses at both taxonomic and functional levels. Results revealed a three-layer barrier failure in OWS fish: the physical barrier was compromised, with structural damage and reduced mucosal index; microbial dysbiosis featured increased richness without changes in diversity or evenness, and expansion of the virobiota, notably uncultured Caudovirales phage; and mucosal immune dysregulation indicated loss of local immune balance. Multi-omics integration identified downregulation of lysosome-related and glycosphingolipid biosynthesis pathways at transcript and protein levels, with disrupted nucleotide metabolism. Overall gut microbial richness, rather than individual taxa abundance, correlated most strongly with host gene changes linked to immunity, metabolism, and epithelial integrity. Although biological replicates were limited by natural outbreak sampling, matched high-depth multi-omics datasets provide exploratory insights into OWS-associated intestinal dysfunction. In summary, OWS entails a cold-triggered breakdown of intestinal barrier integrity and immune homeostasis. This breakdown is driven by a global restructuring of the gut microbiome, which is marked by increased richness, viral expansion, and functional shifts, ultimately resulting in altered host–microbe crosstalk. This ecological perspective informs future mechanistic and applied studies for disease prevention. Full article

Dilated cardiomyopathy is characterized by progressive left ventricular dilation and impaired systolic function, with inflammation recognized as a key contributor to disease onset and adverse outcomes. C-reactive protein reflects systemic biochemical inflammation, whereas Delta Neutrophil Index represents the circulating immature neutrophil fraction and provides a cellular dimension of inflammatory burden. The combined diagnostic and prognostic value of these two biomarkers in dilated cardiomyopathy has not been adequately explored. This retrospective study included one hundred and fifty patients with dilated cardiomyopathy and one hundred and fifty age-, diabetes-, and hypertension-matched controls. Demographic, laboratory, and echocardiographic measurements were analyzed. The diagnostic and prognostic performances of C-reactive protein, Delta Neutrophil Index, and their combined model were assessed using logistic regression, receiver operating characteristic curve analysis, reclassification metrics, calibration testing, and decision curve analysis. Additional analyses were performed for patients with left ventricular ejection fraction below twenty percent, and mortality predictors were examined within the dilated cardiomyopathy cohort. Both C-reactive protein and Delta Neutrophil Index levels were significantly higher in patients with dilated cardiomyopathy than in controls and were further elevated in those with severely reduced ejection fraction. Delta Neutrophil Index remained independently associated with severe left ventricular dysfunction (ejection fraction ≤ 20%) in multivariable analysis (odds ratio 2.51). Each biomarker showed an independent association with the presence of dilated cardiomyopathy, and their combined model achieved the highest diagnostic accuracy. In receiver operating characteristic analysis, the area under the curve was 0.895 for Delta Neutrophil Index, 0.691 for C-reactive protein, and increased to 0.920 for the combined model, with a sensitivity of 81.3% and specificity of 92.0%. Delta Neutrophil Index was independently associated with severe left ventricular dysfunction and mortality, while C-reactive protein, age, ejection fraction, urea, and sodium also contributed to mortality risk. Delta Neutrophil Index was independently associated with mortality (odds ratio 2.51), while C-reactive protein, age, ejection fraction, urea, and sodium also contributed to mortality risk. The combined model provided significant improvement in risk reclassification and demonstrated superior calibration and greater net clinical benefit across a wide range of decision thresholds. C-reactive protein and Delta Neutrophil Index offer complementary diagnostic and prognostic information in dilated cardiomyopathy. Their combined use enhances diagnostic discrimination, strengthens risk stratification, and improves identification of patients at high risk for severe ventricular dysfunction and mortality. Incorporation of these accessible biomarkers into clinical evaluation may support earlier recognition and more tailored management of high-risk individuals. Full article

Over the past two decades, pharmaceutical peptides have emerged as a powerful alternative to traditional small molecules, offering high potency, specificity, and low toxicity. However, most computational drug discovery tools remain optimized for small molecules and need to be entirely adapted to peptide-based compounds. Molecular docking algorithms, commonly employed to rank drug candidates in early-stage drug discovery, often fail to accurately predict peptide binding poses due to their high conformational flexibility and scoring functions not being tailored to peptides. To address these limitations, we present PepScorer::RMSD, a novel machine learning-based scoring function specifically designed for pose selection and enhancement of docking power (DP) in virtual screening campaigns targeting peptide libraries. The model predicts the root-mean-squared deviation (RMSD) of a peptide pose relative to its native conformation using a curated dataset of protein–peptide complexes (3–10 amino acids). PepScorer::RMSD outperformed conventional, ML-based, and peptide-specific scoring functions, achieving a Pearson correlation of 0.70, a mean absolute error of 1.77 Å, and top-1 DP values of 92% on the evaluation set and 81% on an external test set. Our PLANTS-based workflow was benchmarked against AlphaFold-Multimer predictions, confirming its robustness for virtual screening. PepScorer::RMSD and the curated dataset are freely available in Zenodo Full article

Plastic pollution is a major global challenge, especially nanoplastics (NPs) emerging as harmful pollutants due to their small size, reactivity, and persistence in ecosystems. Among them, cigarette butts composed of cellulose acetate (CA) are one of the most widespread and hazardous sources of terrestrial NPs. In this study, the immunotoxic, biochemical, and histopathological effects of cellulose acetate nanoplastics (CA-NPs) derived from smoked cigarette butts (SCB-NPs), unsmoked cigarette butts (USCB-NPs), and commercial cellulose acetate (CCA-NPs) were evaluated on the earthworm Allolobophora caliginosa. Adult worms were exposed for 30 days to 100 mg/kg CA-NPs in artificial soil under controlled laboratory conditions. Results revealed that SCB-NPs induced the most pronounced alterations, including increased lysozyme and metallothionein levels, reduced phagocytic and peroxidase activities, and depletion of protein and carbohydrate reserves. Histological examination showed vacuoles in epithelial layer vacuolization, space between muscle fiber disruption, and degeneration in gut and body wall, especially under SCB-NP exposure. USCB-NPs and CCA-NPs caused milder but still significant effects. Taken together, these findings highlight that the high toxicity of SCB-NPs is due to the presence of combustion-derived toxicants (nicotine, polycyclic aromatic hydrocarbons, and heavy metals), which exacerbate oxidative stress, immune suppression, and tissue damage in soil invertebrates. This study underscores the ecological risk of cigarette butt-derived NPs and calls for urgent policy measures to mitigate their terrestrial impacts. Full article

This study evaluated the effects of replacing soybean hulls with dried grape pomace (DGP) on feed intake, milk production and composition, and rumen N degradability in dairy cows. A 3 × 3 Latin square design was used with three rumen-fistulated Holstein cows, three corn silage-based diets containing 0%, 3%, or 6% of DGP, and three 23-day periods. Measurements were taken from days 15 to 21, with in situ incubations of two protein sources during the last 2 days. At trial end, cows continued on experimental diets to determine the degradability of soybean hulls and DGP. Compared to soybean hulls, DGP presented higher fiber content and lower in situ rumen dry matter and N degradability. Condensed tannin disappearance from DGP increased over time, reaching almost 50% after 48 h. Diet did not affect feed intake, milk production, and composition, except for milk urea N and ruminal total short-chain fatty acids concentration, which showed a quadratic response, suggesting both imbalances between fermentable energy and rumen degradable protein and potential effects of DGP bioactive compounds on rumen function. The N degradation rate of rapeseed meal was unaffected, but soybean meal degradability tended to increase with higher DGP inclusion. This was also detected when combining protein sources across diets. Although DGP influenced rumen function, further research integrating omics and detailed microbiota profiling is needed. Overall, despite its low energy content and rumen degradability, DGP comprises a sustainable feed resource for high-producing animals, supporting circular economy approaches and mitigating the environmental impacts of grape pomace disposal. Full article

Background: Vedolizumab and ustekinumab are increasingly used off-label in pediatric inflammatory bowel disease (IBD) unresponsive or refractory to anti–TNFα therapy. Despite their increasing use in clinical practice, evidence in the pediatric population remains limited, especially regarding therapeutic exposure thresholds and the clinical utility of therapeutic drug monitoring (TDM). Methods: We report a series of five pediatric cases with Crohn’s disease or ulcerative colitis treated with ustekinumab or vedolizumab after anti-TNFα failure. Trough drug concentrations, anti-drug antibodies (ADAs), clinical scores (PCDAI/PUCAI), biomarkers (fecal calprotectin, C-reactive protein), and endoscopic findings were assessed longitudinally. Results: In all cases, we observed recurrent discordance between clinical indices (PCDAI/PUCAI), biochemical markers, and endoscopic activity. Clinical improvement frequently correlated with trough concentrations above commonly cited adult-derived reference ranges (>15 µg/mL for vedolizumab; >3 µg/mL for ustekinumab), although this alignment was not uniform across patients. Notably, one patient developed high-titre ADAs with undetectable ustekinumab levels, yet remained clinically stable, suggesting substantial interindividual variability in pharmacokinetics, immunogenicity, and disease control. Conclusions: Ustekinumab and vedolizumab are promising off-label options for pediatric refractory IBD. In this case series, TDM contributed to the interpretation of pharmacokinetic variability and immunogenicity, offering contextual insights that may support dose adjustments and therapeutic decision-making. Integrating TDM with clinical, biochemical, and endoscopic monitoring may improve optimize individualized treatment in this complex and vulnerable patient group. Full article

Mtx1 is a mosquitocidal protein that exhibits high toxicity toward Culex species. It is produced during the vegetative phase of Lysinibacillus sphaericus but at very low levels and is rapidly degraded. The low expression appears to result from a weak promoter and a potential regulatory stem-loop structure in the 5′ untranslated region. To investigate this regulation, promoter variants of mtx1 were constructed to disrupt stem-loop formation, and promoter activity was assessed using green fluorescent protein (GFP) as a reporter. Disruption of the inverted repeat resulted in approximately twofold higher fluorescence compared with the wild-type promoter in L. sphaericus 2297, indicating partial derepression of translation. To improve protein stability, Bacillus subtilis WB800N, a protease-deficient host, was employed for heterologous expression. Truncated Mtx1 (tMtx1) was secreted into the culture medium, and no obvious degradation products were detected by Western blot analysis under the conditions tested. Although the overall yield was low and not quantitatively determined, the secreted protein retained biological activity. Larvicidal assays showed elevated mortality in tMtx1-containing culture supernatants, with an estimated LC₅₀ at approximately a 1:83 dilution and detectable activity up to a 1:512 dilution relative to control cultures. These results demonstrate that the upstream inverted repeat contributes to partial repression of mtx1 expression in L. sphaericus and that protease-deficient B. subtilis can be used as a host for producing biologically active tMtx1, although further optimization will be required to improve yield. Full article

Breast cancer (BC) is the most frequently diagnosed malignancy in women and remains a leading cause of cancer-related mortality worldwide. Among the oncogenic microRNAs, hsa-miR-21 has been consistently implicated in tumorigenesis, yet a comprehensive network-level understanding of its regulatory landscape in BC is lacking. In this study, we performed an integrative bioinformatics analysis to characterize the molecular pathways and prognostic impact of hsa-miR-21. Experimentally validated mRNA targets were retrieved from miRTarBase and used to construct a high-confidence protein–protein interaction network via STRING, followed by hub-gene prioritization in Cytoscape. Functional enrichment analyses were conducted with DAVID to assess Gene Ontology (GO) categories and KEGG pathways. Survival analyses were performed in large BC cohorts from METABRIC and TCGA using the Kaplan–Meier Plotter. We identified 12 hub genes that are central regulators of apoptosis, proliferation, immune signaling, and transcriptional control. GO and KEGG analyses revealed enrichment in cancer-related, immune, and metabolic pathways, underscoring the pleiotropic role of miR-21. While miR-21 expression alone was not significantly associated with overall survival, a composite hub-gene signature demonstrated strong prognostic value. These findings highlight the importance of network-level biomarkers in BC and provide a reproducible framework for dissecting the clinical relevance of disease-associated miRNAs. Full article