Search Results (347)

The type VI secretion system (T6SS) is a contact-dependent, contractile multiprotein complex widely distributed among Gram-negative bacteria. It mediates the translocation of effector proteins into bacterial competitors and eukaryotic host cells, promoting environmental fitness and contributing to virulence. In Salmonella, five pathogenicity islands encoding T6SSs (SPI-6, SPI-19, SPI-20, SPI-21, and SPI-22) have been described, along with an expanding repertoire of associated effector proteins. However, their global diversity and distribution remain incompletely resolved due to limited genomic sampling. To address this, we analyzed a curated dataset of 490 Salmonella genomes representing 45 serotypes. T6SS regions were identified using SecreT6, revealing that SPI-6 is widely distributed, whereas SPI-19, SPI-20, and SPI-21 are restricted to a subset of serotypes. SPI-20 and SPI-21 were exclusively found in S. enterica subsp. arizonae and diarizonae, while SPI-22 was absent from all analyzed genomes. All open reading frames within T6SS clusters were then analyzed for effector prediction and functional annotation. This approach recovered 32 out of 45 previously described T6SS effectors and identified several novel candidates. These included a cytidine deaminase with predicted DNase activity in SPI-6: two candidate nuclease effectors in SPI-19 with DNase and RNase activities, and four putative effectors in SPI-21, including enzymes with predicted peptidoglycan hydrolase activity, a potential inhibitor of eukaryotic ATPases, and a membrane pore-forming toxin. Additionally, a putative phospholipase effector was identified within a VgrG-associated genomic island in a subset of S. enterica subsp. diarizonae isolates. Collectively, these findings expand the known repertoire of Salmonella T6SS effector proteins and highlight their functional diversity. Full article

Background/Objectives: Prostate cancer (PCa) remains a prevalent malignancy among men, often complicated by recurrence and unfavorable clinical outcomes. Consequently, precise risk stratification and timely clinical intervention are paramount. Initially, we delineated distinct expression profiles of histone modification regulators via unsupervised clustering, identifying PCa subtypes with divergent survival probabilities and biological phenotypes. Subsequently, we sought to develop a prognostic gene signature, derived from the transcriptomic variations among these regulator-defined subtypes, to predict outcomes in PCa patients following radical prostatectomy (RP). Methods: Clinical and transcriptomic data from PCa cohorts were retrieved from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) repositories for comprehensive analysis. Subtypes driven by histone modification regulators were established using unsupervised consensus clustering, followed by in-depth characterization of their molecular features and associated pathways. A risk-scoring model was then developed to evaluate its prognostic efficacy in this patient population. Results: Stratification based on histone modification regulators yielded four distinct PCa subtypes exhibiting heterogeneous survival outcomes, functional pathways, and genomic mutational landscapes. Following rigorous feature selection, a 21-gene risk signature (HIS_score)—comprising MXD3, CCDC28B, COL11A2, SLC39A5, GPT, DNASE1L2, PIF1, KRTAP5-9, TTLL10, KRTAP5-1, KRTAP5-10, HAGHL, MSLNL, AMH, NKAIN4, CCDC114, SLC9A3, SULT1E1, ALB, SLC6A14, and RPE65—was constructed. Survival analyses demonstrated that patients assigned to the high HIS_score cohort experienced significantly worse clinical outcomes compared to their low-score counterparts. Furthermore, we integrated this signature into a novel clinical nomogram to facilitate individualized prognostic assessments. Conclusions: Derived from transcriptomic disparities between extreme epigenetic subtypes, the HIS_score and its associated nomogram serve as robust prognostic instruments. These tools effectively encapsulate the downstream transcriptional sequelae of histone modification dysregulation, offering clinicians a valuable framework to accurately predict post-RP outcomes and expedite the formulation of personalized therapeutic strategies. Full article

The hypothesis that Group A beta-haemolytic Streptococcus (GAS) triggers an autoimmune cascade targeting basal ganglia dopaminergic circuits—producing obsessive–compulsive disorder (OCD), tic disorders, or chorea depending on the receptor subtype involved—is biologically compelling and supported by emerging molecular evidence. Yet PANDAS has remained a diagnostic conundrum since its original description in 1998, with ongoing uncertainty surrounding diagnostic criteria, the interpretation of streptococcal serology, and the distinction from primary neurodevelopmental disorders. This study aimed to review the diagnostic challenges of PANDAS, with focus on streptococcal serology interpretation, advances in dopamine receptor autoantibody biology, the genetic epidemiology of primary tic disorders, and the differential diagnosis of acute neuropsychiatric presentations in children. A structured narrative review was conducted using PubMed, MEDLINE, EMBASE, and the Cochrane Library for publications from 1998 to early 2025 addressing PANDAS, PANS, streptococcal antibodies, childhood movement disorders, autoimmune encephalitis, and the genetics of tic disorders. No currently available biomarker—including ASO, anti-DNase B, anti-basal-ganglia antibodies, or the Cunningham Panel—has demonstrated adequate individual-level diagnostic accuracy for PANDAS. Emerging molecular evidence identifies anti-D1R autoantibodies, acting via G protein-and beta-arrestin-mediated signalling, as candidate biomarkers for PANDAS/PANS neuropsychiatric phenotypes, and anti-D2R autoantibodies for Sydenham chorea movement phenotypes; independent replication in unselected populations is required. Primary tic disorders carry heritability estimates of 50–80% and first-degree familial risk ratios of approximately 18-fold in large population-based cohorts. Prospective blinded studies have not demonstrated a consistent population-level association between GAS infections and tic or OCD exacerbations: PANDAS and PANS remain diagnoses of exclusion. The high background prevalence of both GAS exposure and primary neurodevelopmental disorders in overlapping paediatric age ranges creates conditions for incidental temporal co-occurrence. In the absence of validated molecular biomarkers, diagnostic imprecision carries direct clinical consequences: children may be exposed to treatments with significant risk profiles—including IVIG, plasma exchange, and prolonged antibiotic prophylaxis—while evidence-based therapies are delayed. A stepwise diagnostic approach incorporating the full differential diagnosis is both an epistemological and a patient safety imperative. Full article

Postoperative surgical draining fluid (SDF) in oral squamous cell carcinoma (OSCC) was reported to contain circulating free DNA and extracellular vesicles (EVs). The potential role of SDF DNA as a cancer biomarker in OSCC patients is investigated. To determine whether EVs contribute to the DNA content of SDF, small EVs (sEVs) were isolated by size exclusion chromatography and compared with those in paired plasma. sEV obtained from a cohort of HPV(+) and HPV(−) patients were evaluated for the endocytic origin, size, vesicular morphology, and the DNA content. The sEV from HPV(+) and HPV(−) SDF were similar. The sEVs from SDF were larger and contained more DNA than the sEV from plasma. Treatments of sEV with the DNase/RNase cocktail established the presence of DNA on the surface and in the sEV lumen. Furthermore, 20 to 30% of the total SDF DNA was associated with sEV. Fragmentomic analyses identified the largest DNA (>10,000 bp) on the vesicle surface. The DNA in the vesicle lumen consisted of smaller (~5000 bp) DNA fragments. Electron microscopy of the enzyme-treated vesicles indicated that surface DNA/RNA may be involved in maintaining vesicular integrity and modulating the vesicle entry into cells. The sEV-carrying surface and luminal DNA emerge as significant contributors to the total SDF DNA and, following validation, might serve as useful biomarkers of disease presence/progression in OSCC. Full article

Background: Eosinophilic chronic rhinosinusitis (ECRS) is characterized by refractory nasal polyps and severely impaired mucociliary clearance (MCC). The molecular mechanisms underlying the modulation of mucociliogenesis following IL-4/13 blockade with dupilumab remain poorly understood, notwithstanding its proven clinical efficacy. Methods: Bulk RNA Barcoding and sequencing (BRB-seq) was performed on nasal polyp tissues collected from healthy controls (n = 6), patients with non-ECRS (n = 8), and patients with ECRS both before and four weeks after dupilumab treatment (n = 9) to identify the early molecular drivers underlying ciliary regeneration. Comprehensive gene-set scoring systems were developed to evaluate multiciliogenesis master regulators, master regulators of core/ciliary planar cell polarity (PCP) and PCP components. Interaction scores for epithelial-derived cytokines—thymic stromal lymphopoietin (TSLP), IL-25, and IL-33—were calculated based on ligand and cognate receptor subunit expression. Results: The ciliary master regulatory hierarchy (e.g., FOXJ1, RFX2/3), PCP components (CELSR1 and the ciliogenesis and planar polarity effector (CPLANE) module: FUZ, INTU, WDPCP), and structural ciliogenesis pathways were robustly restored following IL-4/13 blockade. The TSLP interaction score correlated with global mucosal damage, serving as a trigger for compensatory multiciliogenesis. The pre-treatment IL-33 interaction score emerged as a significant predictor of transcriptomic ciliary recovery (p < 0.05). DNASE1L3—the primary endonuclease for degrading eosinophilic extracellular traps (EETs)—remained persistently downregulated post-treatment. Conclusions: IL-4/13 blockade successfully restores the structural and directional “hardware” of the respiratory epithelium but fails to rectify the enzymatic “software” required for mucus degradation. This “residual molecular scar” may explain the persistent mucus hyperviscosity observed in some ECRS patients even after clinical polyp resolution. Full article

Canine urolithiasis is a common and highly recurrent urinary tract disease, with struvite and calcium oxalate being the predominant stone types. Stone formation has traditionally been attributed to urinary physicochemical factors, including urine pH, mineral supersaturation, and urinary tract infection. However, these factors alone cannot fully explain the persistent growth and rapid progression of stones in affected dogs. In this study, we provide evidence that innate immune mechanisms, particularly neutrophil extracellular traps (NETs), are associated with canine urolith formation. We found that neutrophils, key cells of the innate immune system, release neutrophil extracellular traps composed of DNA and antimicrobial proteins, which are consistently present within urinary stones and their surrounding microenvironments. Common canine urinary pathogens were shown to trigger this response, and these immune-derived structures promoted crystal nucleation, aggregation, and stone growth in experimental systems. Importantly, enzymatic degradation of NETs by DNase I attenuated NET-associated stone growth under in vitro conditions. These findings suggest that canine urinary stones develop not only as a consequence of physicochemical factors, but also in association with inflammation-driven biomineralization processes involving NETs. Recognizing the contribution of innate immunity provides new insight into stone recurrence and may inform future preventive and therapeutic strategies. Full article

Background: Tumor-derived small extracellular vesicles (sEV), which we call TEX, carry a cargo of molecules that resembles the producer tumor cells. Circulating freely in body fluids, TEX potentially serve as a liquid tumor biopsy. TEX horizontally transfer their cargo to various recipient cells, imparting to them pro-tumor activity. Mechanisms of TEX-driven reprogramming might involve nucleic acids, especially double-stranded (ds)DNA. Methods: TEX isolated from supernatants of human tumor cells were identified as sEV, based on their size, endocytic origin and morphology. TEX treated with DNase/RNase cocktail were examined by transmission and cryo-electron microscopy and tested for biologic activity. DNA was extracted from enzyme-treated TEX, quantified by Qubit and analyzed for fragment sizes. The presence of genomic DNA in TEX was confirmed by PCR, and sequencing of the TP53 gene fragment for a mutational signature was performed. Results: Enzymatic and microscopic studies of TEX showed that nucleic acids are present in the biocorona on the outer surface. Their removal interfered with the biocorona integrity. A short TEX exposure to DNase/RNase altered their morphology without impairing vesicle functions; longer treatments induced TEX re-organization into smaller membrane-bound vesicles. The TEX lumen contained long fragments of protected genomic DNA with a mutational signature reflecting that of the tumor. Conclusions: Nucleic acids present on the TEX surface support the vesicular integrity. The TEX lumen contains membrane-protected large (ds)DNA fragments with the mutational signature of the parent tumor. The presence of surface and luminal nucleic acids in TEX, and especially their mutational signature, suggests that TEX may serve as highly promising cancer-specific biomarkers. Full article

Introduction: Recombinant human deoxyribonuclease I (rhDNase) cleaves DNA in mucus, facilitating increased mucociliary clearance of purulent sputum. In cystic fibrosis (CF), rhDNase improves pulmonary function and decreases exacerbations. Conversely, rhDNase use in non-CF bronchiectasis (NCFB) patients has not yielded similarly effective results. We explored the safety and feasibility of rhDNase in patients with bronchiectasis due to primary ciliary dyskinesia (PCD). Methods: In this real-life pilot study, patients with PCD received rhDNase to treat viscous mucus. We compared pulmonary function tests and pulmonary exacerbations for these patients over six months of use of rhDNase. Results: Eight PCD patients with symptomatic bronchiectasis commenced use of rhDNase at variable dosing (ranging from at least twice weekly to a full 2.5 mg dose daily). Over a six-month period, pulmonary function tests, as measured by mean FVC and FEV₁, remained relatively stable compared to prior to commencing rhDNase. Mean pulmonary exacerbations decreased from 3.1 to 2.3 in the six-month period after commencing rhDNase, as compared to the six-month period prior to rhDNase. Conclusions: Use of rhDNase in PCD patients was safe and did not adversely impact lung function or increase pulmonary exacerbations, in contrast to earlier trial results in NCFB patients with heterogeneous etiologies. Further clinical data is required to identify the population of PCD patients who can benefit from rhDNase, as well as the appropriate dosing and timing. Full article

Semi-aquatic turtles are popular companion animals and represent an important One Health interface in Portugal due to their potential to harbour zoonotic pathogens. This study aimed to characterize the virulence and antimicrobial resistance (AMR) profiles of Aeromonas spp. isolated from captive semi-aquatic turtles in Portugal. Cloacal swabs (n = 31) were collected from turtles under human care, and Aeromonas spp. were isolated using selective media and identified by multiplex PCR. Antimicrobial susceptibility was assessed by disk diffusion using eleven antibiotics, while the phenotypic virulence profile was evaluated by determining isolates’ ability to express five hydrolytic enzymes and to form biofilm. A total of 86 Aeromonas isolates were recovered, with A. hydrophila being the most prevalent (77.9%). Most isolates displayed high pathogenic potential: over 87% produced DNase, haemolysin, and lecithinase, while nearly all produced protease and gelatinase. Also, 44.2% of the isolates were resistant to at least one antibiotic, and 12 (14.0%) were multidrug resistant. Higher Multiple Antibiotic Resistance (MAR) indices were significantly associated with turtles housed in indoor aquariums (p < 0.05). These findings indicate that captive semi-aquatic turtles may act as reservoirs of virulent and antimicrobial-resistant Aeromonas spp., highlighting potential zoonotic risks and supporting the need for a One Health approach in their management. Full article

This study evaluated the probiotic, technological, and safety properties of 124 lactic acid bacteria isolated from traditional Beyaz cheese, and the in situ survival of selected strains in fermented milk. Eighteen isolates showing over 80% tolerance in simulated gastric juice (pH 3.0) were subjected to further characterization. On the basis of 16S rRNA gene sequencing, most isolates belong to Lacticaseibacillus paracasei and Lactiplantibacillus plantarum, while Lactobacillus helveticus, Lentilactobacillus kefiri, and Limosilactobacillus fermentum were also identified. L. plantarum EH140 showed the greatest resistance to the simulated gastric environment (pH 2.0), whereas L. paracasei EH131 exhibited the highest bile salt tolerance. L. plantarum EH106 demonstrated strong auto-aggregation, and L. fermentum EH132 displayed notable hydrophobicity. Nine isolates exhibited bile salt hydrolase activity, but none showed γ-hemolysis, gelatinase, or DNase activity. All the isolates were susceptible to ampicillin, erythromycin, clindamycin, and chloramphenicol. Organic acid analysis revealed lactic acid as the major metabolite, followed by acetic acid. Virulence gene screening identified the efaAfs gene only in L. paracasei strains, and no biogenic amine genes were detected. The selected isolates maintained viability above 6 log CFU/mL in milk during storage. Overall, L. plantarum EH106, EH109, EH140, and EH141 were identified as the most promising candidates because of their safety and superior probiotic potential. Full article

Methicillin-resistant Staphylococcus aureus (MRSA) produces virulence factors and causes hard-to-treat infections. This study aimed to evaluate the effect of trans-cinnamaldehyde (TC) on the selected virulence factors of MRSA: adhesion to host plasma and extracellular matrix proteins, protease, DNase and esterase production, and hemolytic activity. Our results showed that TC at ½ MBIC (Minimum Biofilm Inhibition Concentration) of 240 µg/mL or 60 µg/mL, depending on the isolate, significantly reduced MRSA adhesion. Inhibition varied between isolates, ranging from 26.1% to 41.3% (fibrinogen), 18.2% to 34.9% (elastin), 26.5% to 32.4% (laminin), and 17.1% to 30.5% (collagen). TC at ½ MIC (Minimum Inhibitory Concentration) of 30 µg/mL also significantly inhibited MRSA enzyme production, and reduced hemolytic activity (by 80.0–83.1%, depending on the isolate). TC may be an alternative to antibiotics for combating infections caused by S. aureus, as it not only reduces bacterial survival in the host but also reduces S. aureus virulence at subinhibitory concentrations. TC at higher concentrations exhibits cytotoxicity in human fibroblasts, limiting its topical use. Therefore, to exploit TC’s antibacterial potential, it is necessary to identify substances that act synergistically with it, enabling reduced effective doses. Full article

Neutrophil extracellular traps (NETs) are web-like DNA–protein structures released by activated neutrophils. Initially recognized for their antimicrobial roles, NETs are now known to drive sterile inflammation, thrombosis, and tissue remodeling. This review highlights their involvement in key pancreatic diseases, including acute pancreatitis (AP) and pancreatic ductal adenocarcinoma (PDAC). In AP, early NET formation correlates with disease severity and septic complications, contributing to acinar injury, microvascular thrombosis, ductal obstruction, and organ dysfunction. In PDAC, NETs shape a fibrotic and immune-resistant tumor microenvironment by promoting stromal activation, immune exclusion, metastasis, and hypercoagulability. Tumor- and stroma-derived signals sustain NET formation within this niche. We also discuss NET-related biomarkers for risk assessment and therapy monitoring, and explore therapeutic strategies that target NETs—ranging from their degradation with DNase to their inhibition of upstream pathways such as PAD4, autophagy, and oxidative signaling. Targeting NETs may also reduce their downstream effects on thrombosis and immune suppression. Overall, NETs emerge as critical drivers of pancreatic disease progression and represent promising therapeutic targets. Full article

Bioresorbable scaffolds (BRS; also referred to as bioresorbable vascular scaffolds, BVS) represent a promising approach in interventional cardiology, offering theoretical advantages such as temporary mechanical support followed by complete resorption. However, clinical experience has revealed challenges, including late-stage scaffold thrombosis and heterogeneous scaffold discontinuity during degradation, prompting investigation into host immune responses. Neutrophil extracellular traps (NETs), which are network-like structures composed of decondensed chromatin decorated with antimicrobial proteins, have emerged as critical mediators of vascular inflammation and thrombosis. This review explores the intersection between NET biology and BRS performance, investigating how NETosis affects biocompatibility, degradation kinetics, and device-related complications. We discuss the molecular mechanisms that trigger neutrophil activation and NET formation in scaffold materials, the effect of NET components on polymeric and metallic scaffold degradation, and emerging biomarkers to monitor NET-mediated complications. We also evaluate therapeutic strategies targeting NET pathways, including DNase-based therapies, peptidylarginine deiminase 4 (PAD4) inhibitors, and anti-inflammatory coatings that can optimize next-generation BRS outcomes. Understanding the immunological environment surrounding bioresorbable vascular devices is crucial for developing scaffolds that deliver predictable degradation while minimizing adverse inflammatory responses. Full article

Ménière’s disease (MD) is thought to involve dysfunction of the blood–labyrinth barrier, but circulating mechanisms of endothelial injury remain poorly understood. The present study investigated whether cell-free DNA (cfDNA) and inflammatory mediators in plasma contribute to vascular stress and barrier disruption in MD. cfDNA levels were significantly elevated in plasma from patients compared with plasma from healthy controls. Exposure of primary human stria vascularis endothelial cell monolayers to plasma from MD patients led to decreased transepithelial electrical resistance and a significant increase in FITC-dextran permeability, indicating impaired barrier function. MD plasma also induced higher lactate dehydrogenase release and pronounced F-actin disorganization with reduced syndecan-1 expression, consistent with endothelial cytotoxicity and glycocalyx degradation. DNase I partially reversed these effects, implicating extracellular DNA as a key driver. Furthermore, IL-1β, CCL3 (MIP-1α), and CCL27 were elevated in MD plasma. Collectively, our data support a model in which cfDNA and inflammatory mediators cooperatively induce endothelial injury, cytoskeletal remodeling, and glycocalyx shedding, leading to blood–labyrinth barrier weakening. Targeting extracellular DNA or glycocalyx preservation may represent a novel strategy to protect inner ear vascular integrity and modify disease progression in MD, and cfDNA-related readouts may be promising biomarkers of endothelial damage. Full article

Hyperuricemia (HUA) is a metabolic disorder that can easily lead to gout or kidney disease, and it is believed that it can be treated effectively using probiotics. This study evaluated the safety, probiotic, and functional properties of Wickerhamomyces anomalus YFJ252, isolated from pickled vegetables, including its in vitro inhibitory activity against xanthine oxidase (XO) and in vivo uric acid-lowering activity in mice, using virulence factor screening, plate counting, and colorimetric assays. Meanwhile, the potential anti-HUA mechanism was also investigated using untargeted metabolomics and whole-genome analysis. The results show that YFJ252 is non-hemolytic and does not produce DNase, gelatinase, or biogenic amine. It has potential probiotic properties: 85.83% DPPH radical scavenging, 39.94% α-amylase inhibition, 35.32% α-glucosidase inhibition, 20.73% anti-inflammatory ability, and 84.15% XO inhibition capacity. Animal experiments indicated that early intake of YFJ252 could maintain serum uric acid levels at 165.08 μmol/L (p < 0.05), lower than the HUA group (212.19 μmol/L), and significantly decrease creatinine and urea nitrogen levels (p < 0.05). The hypothetical anti-HUA potential of YFJ252 might be due to the production of antioxidant, hypoglycemic, and XO-inhibitory metabolites during growth, as well as a purine-degrading pathway that the strain inherited. This study provides a theoretical basis for using W. anomalus YFJ252 as a food ingredient with preventive effects against HUA. Full article