Search Results (654)

This study presents the first documented case of a disease syndrome in cultured largefin longbarbel catfish (Hemibagrus macropterus). The condition is characterized by massive abdominal pseudotumor formation, severe cachexia, and functional gonadal arrest. Comprehensive pathological investigation revealed that the pseudotumor was encapsulated by fibroblasts and primarily composed of host-derived, poorly differentiated hyperplastic cells, interspersed with invasive, basophilic Type III cells. These cells and associated inflammatory–fibrotic lesions were also disseminated in the gill, kidney and spleen. Systematic diagnostic approaches, including microbiology and transmission electron microscopy, found no evidence of conventional bacterial or viral pathogens. Metagenomic analysis further supported these findings and suggested a link to infection by an as-yet-unidentified eukaryotic parasite, with Microsporidia or Ichthyosporea being the primary candidates. Functional (KEGG) profiling of the pseudotumor tissue further revealed a molecular signature consistent with active cellular proliferation and metabolism. We propose that the pseudotumor acts as a metabolically active “nutrient sink,” driving the systemic catabolism that underlies the severe cachexia and reproductive arrest. This work provides the first case of a eukaryotic parasite-induced pseudotumorous syndrome in fish, which represents an emerging threat to conservation aquaculture and offering novel insights into parasite-mediated host metabolic hijacking and tumor-mimicry. Full article

Background/Objectives: Streptococcus pyogenes (Group A beta-hemolytic streptococcus, GAS) is the most common cause of bacterial pharyngitis and a major contributor to morbidity and mortality worldwide. There has been an increase in invasive GAS infections and related deaths in several European countries post-COVID-19 pandemic. We aimed to assess parental knowledge, attitudes, and practices regarding GAS pharyngotonsillitis, with a focus on antibiotic use and misuse. Methods: A cross-sectional questionnaire-based study was conducted on a convenience sample of parents of children admitted to the Pediatric Ward or visiting the Pediatric Emergency Department of the University General Hospital of Patras, Greece (September 2024–February 2025). For knowledge assessment, the questionnaire consisted of 10 True/False questions, based on which a total knowledge score was calculated. For attitude and practice assessment, the questionnaire consisted of 10 Likert scale questions. A Negative Practice Score was calculated as a sum of the answers in five practices with a negative perspective, with higher scores indicating worse practices and lower scores indicating better practices. Results: The study enrolled 378 parents, 79% of them were aware that not all children with a sore throat need antibiotics, and 61% believed asymptomatic children with a positive strep antigen test should receive antibiotics. Concerns about GAS transmissibility were high (76%), while attitudes about severity were mixed. A median GAS total knowledge score of 6 (IQR: 4–7) indicated moderate knowledge. Multivariable analysis revealed that male parents, non-immigrants, those previously hospitalized for GAS infection, and those informed by pediatricians or reliable websites had significantly higher knowledge scores. Regarding practices, most parents (72%) disagreed with requesting antibiotics from pediatricians, and 93.9% did not administer leftover antibiotics. Additionally, 58% expressed more concerns in recent years due to the increase in invasive infections. The median GAS Negative Practice Score was 10.5 (IQR: 7.0–13.0), indicating generally good practices, as lower scores correspond to fewer negative practices. Older parents and those with higher knowledge scores were also linked to fewer negative practices in multivariable analysis. Conclusions: These findings highlight the importance of targeted education on GAS pharyngotonsillitis and the need to focus on specific population groups to reduce antibiotic misuse. Full article

Severe COVID-19 and bacterial sepsis share clinical manifestations of systemic inflammation and organ dysfunction. Yet, early differentiation between these conditions and timely identification of patients at risk of deterioration remain major clinical challenges. Extracellular vesicle (EV)-associated microRNAs (miRNAs) have emerged as promising biomarkers of host immune dysregulation. In our study, we have characterized circulating EV-miRNAs in patients with COVID-19, bacterial sepsis, localized bacterial infections, and healthy subjects to assess their diagnostic and prognostic utility. After EV isolation from plasma and characterization by nanoparticle tracking analysis and flow cytometry, a panel of 12 inflammation-related miRNAs were individually quantified by qRT-PCR. Four EV-miRNAs—miR-28-5p, miR-199a-5p, miR-200a-3p, and miR-369-3p—were significantly elevated in COVID-19 patients, with higher levels in those with poor prognosis. miR-199a-5p and miR-200a-3p were increased in bacterial sepsis compared with COVID-19, enabling discrimination between viral and bacterial sepsis. Three EV-miRNAs—miR-28-5p, miR-199a-5p, and miR-200a-3p—were markedly higher in bacterial sepsis than localized infections, and ROC analysis showed a strong diagnostic performance, particularly for miR-199a-5p, alone or in combination with other EV-miRNAs. The increased expression of selected EV-miRNAs was associated with higher SOFA scores and in-hospital mortality. These findings indicate that EV-miRNAs reflect pathogen-specific and severity-related immune responses, supporting their potential as minimally invasive biomarkers for early diagnosis and risk stratification in severe infections. Full article

Alcohol-associated hepatitis (AH) is the most severe clinical manifestation of alcohol-associated liver disease. Corticosteroids are the only disease-specific therapy shown to improve short-term survival. Currently, no non-invasive markers are available to predict patient response to corticosteroids or long-term survival in AH. This study investigates whether surface antigens on plasma extracellular vesicles (EVs), key mediators of intercellular communication, can reflect the underlying immune dysregulation in AH and serve as prognostic markers. Patients with AH were prospectively enrolled between 2020 and 2024. Blood samples were collected before corticosteroid initiation during the first 24 h of hospitalization. EVs were characterized using nanoparticle tracking analysis, cryo-electron microscopy, and flow cytometry. Interleukin-6 (IL-6), soluble (s)CD62p, Circulating Vascular Cell Adhesion Molecule-1 (sVCAM), tumor necrosis factor receptor superfamily member 1 (TNRFS1a), and Intercellular Adhesion Molecule 1 (ICAM-1) were quantified by ELISA. Key outcome variables included response to corticosteroids and mortality. A total of 46 patients with AH and 28 healthy donors (HD) were included. EV concentration was significantly higher in AH patients than in HD (9.3 × 10¹¹ [IQR 4–24] versus 2.4 × 10¹¹ [IQR 2–4], p = 0.03). Specific EV antigens were associated with key clinical outcomes: CD20 and CD2 levels differed between patients with or without infections (bacterial, viral, and fungal) developed during hospitalization; CD40 and CD146 were elevated in patients who developed acute kidney injury. EVs enriched in monocyte (CD14) and T-reg (CD25) markers were associated with plasma IL-6 levels, while endothelial markers CD105 and CD146 correlated with sVCAM and sCD62p. EVs enriched in platelet (CD49e) and endothelial (CD31) markers were associated with corticosteroid response, whereas EVs enriched with endothelial (CD105 and CD146) and B lymphocyte (CD19) markers were associated with mortality. Overall, EVs enriched in endothelial and monocyte markers may represent a candidate non-invasive tool for predicting corticosteroid response and mortality in AH, aiding risk stratification and early identification of non-responders for timely transplant evaluation. Full article

Background: Airway bacterial infection and inflammation are often present in children with bronchiectasis. Systemic inflammation has also been reported. Currently, there are no data on the association between systemic inflammatory markers with airway pathogens or neutrophilia in children with bronchiectasis. We aimed to define the bronchoalveolar lavage (BAL) pathogens (bacteria and viruses), and cytology in children with bronchiectasis and to explore any association between peripheral inflammatory markers and airway neutrophilia. Methods: Participants numbering 402, aged <18 years, with peripheral blood and BAL results within 3 months of diagnosis of bronchiectasis were included. Blood and BAL results were reviewed and analysed for possible associations. Results: Of 355 children (88.31%), cultured bacteria from BAL and Haemophilus influenzae (n = 185) were the most frequent. A virus was identified in 131 (32.59%). Adenovirus (n = 69) was most common. Children numbering 279 (69.40%) had airway neutrophilia (neutrophils > 15%) which was associated with the presence of H. influenzae (OR 2.03 95% CI 1.31–3.15, p = 0.002), S. pneumonia 2.41 (95% CI 1.36–4.29, p = 0.003), and Adenovirus (OR 2.06 95% CI 1.06–4.04, p = 0.033). Airway neutrophilia was associated with raised CRP (OR 2.26 95% CI 1.14–4.49, p = 0.019), but there were no other systemic inflammatory markers including monocyte/lymphocyte ratio, neutrophil/lymphocyte ratio, platelet/lymphocyte ratio, and platelet/mean platelet volume ratio. Conclusions: In children, there is an association between airway neutrophilia and raised CRP in bronchiectasis, but not with other peripheral inflammatory markers. There is a need to identify non-invasive inflammatory markers in children with bronchiectasis. Full article

Between 2015 and 2022, we evaluated a novel broad-range (BR) 16S PCR rDNA PCR/Sanger sequencing assay to improve diagnosis of invasive infections in culture-negative specimens. Using dual-priming oligonucleotides (DPO), this assay analyzed ribosomal DNA from sterile fluids or tissues. A total of 762 specimens were analyzed from 661 patients: 61% had negative cultures and BR 16S PCR tests; 35% had negative cultures but positive BR 16S PCR tests; and only 4% had negative cultures with indeterminate BR 16S PCR results. After resolution of indeterminate BR 16S PCR results (i.e., 29 negative, 1 false-positive, and 1 positive) the assay showed a sensitivity of 98.26% (95% CI = 96.00–99.43%), specificity of 99.79% (95% CI: 99.82–99.99%), positive predictive value of 99.65% (95% CI: 97.56–99.95%), negative predictive value of 98.94% (95% CI: 97.51–99.55%), and accuracy of 99.21% (95% CI: 98.28–99.71%) for a disease prevalence of 38.10% (95% CI: 34.62–41.66%). Gram stain purulence predicted the BR 16S PCR result better (69.4%) than organisms (24.6%), but the latter had a higher PPV (78.5%). Increased peripheral WBC (86.1%) or CRP (71.8%) predicted positive BR 16S PCR results. Our DPO BR 16S PCR assay improved pathogen detection over culture and minimized contamination. Broad range 16S rDNA PCR/sequencing (BR 16S PCR) is an important diagnostic technique in cases with invasive infection due to fastidious or uncultivatable pathogens. However, appropriate case selection, the quality of clinical specimen, and the specific assay primers affect its performance. Our novel BR 16S PCR assay uses unique dual-priming oligonucleotides (DPO) primers and fast protocols for rapid, optimal detection of bacterial pathogens, while minimizing contamination. Fast BR 16S PCR assay reports occurred within 24–48 h. BR 16S PCR and culture analyzed a diverse range of clinical specimens from patients with invasive infections. BR 16S PCR demonstrated a high performance for accurately detecting pathogens, ruling out infections, and minimizing contamination. BR 16S PCR detection of a pathogen allowed the appropriate clinical management of one-third of patients in this cohort. BR 16S PCR is an essential tool for the clinical management of patients with invasive infection when primary cultures are negative or contaminated. Full article

Background: Fungal infection of pancreatic fluid collections (PFCs) in severe acute pancreatitis (SAP) is under-recognized and associated with poor outcomes. Overlap with bacterial infections and the need for invasive sampling often delay diagnosis, leading to prolonged antibiotic use without the use of antifungal agents. Methods: We report three cases of SAP complicated by fungal infection of PFCs. Two patients, one with alcohol-related pancreatitis and the other with biliary pancreatitis, developed symptomatic encapsulated necrosis. Both were successfully managed with endoscopic drainage and targeted antifungal therapy against Candida albicans, achieving full resolution. The third patient, with necrotizing biliary pancreatitis, underwent multiple surgical and endoscopic interventions and developed an infection with a non-albicans Candida species. Reduced susceptibility requires individualized antifungal adjustment guided by susceptibility testing. Despite aggressive multimodal therapy, the patient progressed to multiorgan failure and died subsequently. Results: These cases emphasize the clinical impact of fungal infections in patients with SAP, particularly their association with severe disease, prolonged hospitalization, and prior antibiotic exposure. These findings highlight the prognostic value of early microbiological sampling, species-level identification, and prompt initiation of antifungal therapy. Infections caused by non-albicans species pose additional challenges due to their reduced sensitivity to standard antifungal agents. Conclusions: Fungal infection of PFCs is a clinically significant and frequently underestimated complication of SAP. Early recognition and species-directed antifungal therapy are critical for improving outcomes in high-risk patients. Full article

Antimicrobial resistant (AMR) infections are a persistent public health issue causing excess death and economic impacts globally. Because AMR in clinical settings is often acquired from nonpathogenic bacteria that surround us, environmental surveillance must be better characterized. It has been well established that metals can co-select for bacterial AMR. Furthermore, recent studies have shown that compromised microbial community diversity may lead to community invasion by antibiotic resistance genes (ARGs). Widespread legacy mining has led to acid mine drainage and metal contamination of waterways and sediments throughout the western United States, potentially compromising microbial community diversity while simultaneously selecting for AMR bacteria. Our study objectives were to survey metal contaminated sediments from the Bonita Peak Mining District (BPMD) in southwestern Colorado, USA, compared to sites downstream in Durango, CO for bacterial and ARG diversity. Sediment bacteria were characterized using 16S rRNA Ilumina and metagenomic sequencing. We found that overall, bacterial diversity was lower in metal-contaminated, acidic sites (p = 0.04). Metagenomic sequencing revealed 31 different ARGs, with those encoding for efflux pumps (mex and spe gene families) substantially more prevalent in the BPMD sites, elucidating a specific AMR marker fingerprint from the high metal concentration sediments. Raising awareness and providing antimicrobial tracking techniques to resource limited communities could help provide information needed for better antibiotic use recommendations and environmental monitoring. Full article

Adherent-invasive E. coli (AIEC) is closely related to inflammatory bowel disease (IBD). However, its pathogenic mechanism has not yet been fully elucidated. Using a BLASTP search, we discovered that the amino acid sequence of a putative protein (UFP37798.1) in the AIEC LF82 strain is highly homologous to some regulators in the SlyA family. We named it EruA. We displayed the secondary structures of EruA using bioinformatics, overexpressed the His₆-tagged EruA protein using SDS-PAGE, and dissected the genetic organization of the eruA chromosomal region using 5′RACE. We constructed an eruA deletion mutant (ΔeruA) and a complementary strain (CΔeruA) of the LF82 strain. The transcriptomes of wild-type (WT) and ΔeruA bacteria were compared using RNA sequencing and qRT-PCR, thereby identifying 32 differentially expressed genes (DEGs). Based on YASARA software and EMSA analysis, EruA directly binds to the consensus sequences (P_fimA and P_tnaB) in the promoter region of the fimA and tnaB genes from these DEGs. By using a super-resolution confocal microscope (SCM), counting CFUs of colonies on plates, indole quantification, and crystal violet staining of biofilms adhered to tubes or 96-well plates, we found that EruA activates the fimA to promote bacterial adhesion to intestinal epithelial cells and activates the tnaB to enhance bacterial indole production and biofilm formation. Moreover, EruA helps AIEC resist environmental stress and enhances bacterial survival within macrophages as well as loading in mouse tissues. Notably, EruA promotes AIEC colonization in the colons of mice and exacerbates intestinal inflammation caused by bacterial infection in mice with DSS-induced inflammatory colitis, manifested by weight loss, colon length shortening, and pathological changes in colon tissues. Therefore, EruA plays a key role in the pathogenicity of AIEC. Full article

Listeria monocytogenes (LM) is a lethal foodborne intracellular pathogen. Internalins A and B (inlA and inlB) are critical virulence factors that promote LM’s adhesion and invasion into host cells. InlA is covalently anchored to the cell wall by LM SortaseA (SrtA), while inlB is anchored to the cell wall via non-covalent bonds. Therefore, inhibiting SrtA and inlB is expected to suppress LM’s adhesion and invasion of host cells, enabling the prevention and control of infections. This study demonstrated that Luteolin inhibited the activity of purified LM SrtA protein in vitro. Interactive mechanism analysis indicated that Luteolin generates interaction with the critical active sites of SrtA, which may affect its binding to its natural substrates, thereby reducing the anchoring of inlA on the cell wall and achieving the inhibition of bacterial adhesion and invasion. In addition, Luteolin binds to the groove at the binding interface between inlB and its host receptor. The key residues in inlB that interact with the host receptor form weak interactions (Hydrogen bonds and van der Waals interactions) with Luteolin, this binding may inhibit their binding, suppressing LM’s adhesion and invasion of host cells. At the tested concentrations, Luteolin did not affect the growth of LM, but remarkably reduced the mortality and alleviated the infection symptoms of LM-infected Galleria mellonella. These results provide additional theoretical evidence for the application of Luteolin in the prevention and control of LM infections, which is expected to accelerate its application progress. Full article

Methicillin-resistant Staphylococcus aureus (MRSA), a major global public health threat due to its broad resistance, urgently requires the development of new antibiotic alternatives. (‒)‒Epigallocatechin-3-gallate (EGCG) is considered a natural bioactive compound with anti-MRSA properties. The L-Lectin module of serine-rich adhesin for platelets (SraP) is considered an important target for blocking MRSA-infected hosts. This study aims to investigate the mechanism of action of EGCG against MRSA. Surface plasmon resonance (SPR), cell adhesion and invasion, biofilm formation, checkerboard assays, RNA sequencing (RNA-seq) and quantitative real-time polymerase chain reaction (qRT-PCR) were performed. The results showed that EGCG bound to SraP L Lectin with high affinity and effectively inhibited MRSA colonization. Additionally, EGCG significantly suppressed pyrimidine metabolism and downregulated related genes, thereby potentially inhibiting bacterial growth. It also markedly reduced the expression of multiple genes associated with β-lactam resistance and inhibited biofilm formation. A strong synergistic effect was observed between EGCG and the bactericidal agent ceftriaxone (CRO). When combined with 10 μg/mL EGCG, CRO required 75% less dosage and exhibited a prolonged antimicrobial effect. In conclusion, EGCG exerts anti-MRSA effects through multiple pathways and represents a promising candidate as an alternative therapeutic agent against MRSA infections. Full article

Down Syndrome Regression Disorder (DSRD) is a rare but severe neuropsychiatric condition characterized by abrupt loss of speech, autonomy, and cognitive abilities in individuals with Down syndrome, often associated with immune dysregulation and gut–brain axis dysfunction. We report the case of an 11-year-old girl with Down syndrome who developed developmental regression at age five, in temporal proximity to a family transition (the birth of a younger sibling), with loss of continence, language, and comprehension, alongside persistent behavioral agitation and gastrointestinal symptoms. Laboratory assessment revealed Giardia duodenalis infection, elevated fecal calprotectin and secretory IgA, and microbial imbalance with overgrowth of Streptococcus anginosus and S. sobrinus. The patient received a single oral dose of tinidazole (2 g), daily folinic acid (1 mg/kg), and a 90-day course of transcranial and abdominal photobiomodulation (PBM) (1064 nm, 10 min per site). Post-treatment, stool analysis showed normalized inflammation markers and restoration of beneficial bacterial genera (Bacteroides, Bifidobacterium, Lactobacillus) with absence of Enterococcus growth. Behaviorally, she exhibited marked recovery: CARS-2-QPC decreased from 106 to 91, ABC from 63 to 31, and ATEC from 62 to 57, alongside regained continence, speech, and fine-motor coordination. These outcomes suggest that abdominal and transcranial PBM, by modulating mitochondrial metabolism, mucosal immunity, and microbiota composition, may facilitate systemic and neurobehavioral recovery in DSRD. This translational case supports further investigation of PBM as a non-invasive, multimodal therapy for neuroimmune regression in genetic and developmental disorders including validation through future randomized controlled clinical trials. Full article

The phage-resistant mutant (PRM) strains of Escherichia coli (E. coli) exhibited abundant genetic and phenotypic diversity. IS elements played a vital role in creating various genetic divergences and regulating gene functions under phage infection stress. Genetic variations of PRM strains derived from E. coli MG1655 and mutation frequencies of coevolved E. coli populations with phages were explored by high-throughput sequencing and resequencing. Infrequent-restriction-site PCR (IRS-PCR) and carbon utilization test revealed the genetic and phenotypic diversity of the PRM strains. Numerous and discrepant mutation sites (MSs) were observed in the PRM strains and the coevolved populations, and many MSs were related to the synthesis of flagella and LPS, which often serve as receptors in a phage invasion. The insertions of various IS elements in key gene locations were also frequently found in the PRM strains, which indicate for the first time that IS elements played a vital role in generating genetic divergence and regulating gene functions under phage infection stress. Resequencing revealed that the coevolved populations at three evolving stages had discrepant profiles of MSs, and nearly all detected MSs occurred in the coevolved populations, which led to coexisting phages that increased the mutation rates and expedited the occurrence of the defective MSs in E. coli populations. In summary, our results reveal that the widespread and abundant presence of phages may provide one important force driving bacterial genomic evolution and prompt bacterial genetic divergence via accelerated mutation and increased mutation rates in the E. coli genome. Full article

This article describes the microbiome of the outer ear and the earwax in the ear canal, which performs various protective functions against bacterial infections. This article is based on an analysis of literature gathered from databases including PubMed, Google Scholar, Web of Science, and Scopus, primarily from the last 15 years. The search strategy included MeSH terms: ear canal, microbiome, earwax, cerumen, antibacterial peptides, ear infections, biofilm. Only peer-reviewed articles were included. The natural ear canal microbiota provides so-called colonization resistance, which protects against invasion by pathogenic microorganisms. Earwax is composed primarily of keratin secreted by epithelial cells and substances secreted by sweat and apocrine glands. It plays a key role in the physiology of the ear canal, maintaining a low pH, limiting moisture, and exhibiting antimicrobial properties. Both an excess and a deficiency of earwax can lead to dysbiosis of the outer ear, and consequently to the development of various infections. In an era of increasing antibiotic resistance and the search for new solutions in the fight against pathogenic microorganisms, understanding the natural properties of earwax is becoming increasingly important. Full article

Background: Immune aging is a complex process involving various cellular changes, such as a myeloid bias, decreased functional activity of immune cells, accumulation of senescent cells, and alterations in serum levels of bactericidal humoral factors. As believed, these changes contribute to increased susceptibility of older adults to infectious diseases. Myeloid cells are considered the first line of defense against bacterial invasion. However, it remains unclear whether the protective functions of myeloid cells diminish in active older adults and whether potential age-related changes are evolutionarily conserved across primates. Methods: In this study, myeloid cell populations from peripheral blood and bone marrow of cynomolgus macaques and human peripheral blood were analyzed across a broad age range for phenotypic and functional characteristics, e.g., E. coli phagocytosis, secretion of proinflammatory factors, genetic instability, and signs of cellular aging. Results: Despite minor interspecies phenotypic differences in granulocyte populations, both the quantity and functions of myeloid cells were remarkably stable during aging in both species. Myeloid cells maintained genetic stability, and high SA-β-Gal activity was observed, likely reflecting metabolic traits rather than age-related changes. Importantly, a predominant and age-independent role of humoral factors, rather than cellular mechanisms, was identified in the initial control of bacterial infection. Conclusions: These findings suggest that innate immune functions remain stable for a long time during aging in both species. Full article