Search Results (5,482)

The earliest gut and nasal bacterial community profiles after birth remain incompletely characterized, particularly when both sites are examined in parallel. We aimed to characterize these profiles according to delivery mode within the first 48 h of life in a Mexican cohort. Meconium and nasal wash samples from 60 neonates (30 vaginal and 30 cesarean deliveries) were analyzed by V3–V4 16S rRNA gene amplicon sequencing on an Illumina MiSeq platform, followed by QIIME 2 processing. All 60 meconium samples yielded analyzable 16S rRNA profiles. Nasal washes yielded analyzable profiles in 10/30 vaginally delivered and 12/30 cesarean-delivered neonates (p = 0.79). No statistically significant differences in alpha diversity were detected by delivery mode in gut samples (observed ASVs, p = 0.842; Shannon index, p = 0.183; Pielou’s evenness, p = 0.121; Faith’s phylogenetic diversity, p = 0.574) or nasal samples (p = 0.356, 0.947, 0.644, and 0.114, respectively). In unadjusted PERMANOVA comparisons, gut community structure differed by delivery mode for Bray–Curtis (p = 0.001), weighted UniFrac (p = 0.021), and unweighted UniFrac (p = 0.013) distances, whereas nasal comparisons were not statistically significant. Traditional ANCOM identified Staphylococcus as the feature with the highest W statistic in cesarean-delivered neonates (W = 65). Descriptive phylum-level profiles were examined across maternal urinary tract infection, premature rupture of membranes, maternal comorbidities, maternal BMI, hours since birth, and recorded antibiotic exposure; these comparisons were not multivariable and cannot exclude residual confounding. Delivery mode was associated with differences in early gut bacterial community structure in this cohort. Nasal findings were inconclusive because only 22 washes yielded analyzable 16S rRNA profiles and the workflow was not optimized to quantify ultra-low-biomass bacterial DNA. These cross-sectional data do not establish persistence, causality, or the timing at which nasal communities become stable. Full article

The ongoing Bundibugyo virus disease outbreak in the Democratic Republic of the Congo and Uganda has become the largest documented outbreak caused by this orthoebolavirus and already ranks as the second-largest Ebola disease outbreak recorded in history. This review highlights its major public health relevance, given its cross-border spread, substantial mortality burden, high clinical severity, infections among healthcare workers, and the lack of any approved vaccine or specific treatment for Bundibugyo virus disease. We discuss the epidemiological and virological significance of this exceptional event, the limitations of current diagnostic and therapeutic preparedness beyond Ebola virus, and the urgent need for broader filovirus-focused strategies, including improved diagnostics, genomic surveillance, candidate vaccines, antiviral platforms, and resilient local health systems. The outbreak illustrates how preparedness efforts centered mainly on the Ebola virus have left substantial gaps for other human-pathogenic orthoebolaviruses. Full article

Background and Objectives: Inborn errors of immunity (IEI), also referred to as primary immunodeficiencies, are chronic conditions characterized by recurrent infections and long-term treatment requirements that may adversely affect psychological well-being. Although mental health outcomes in patients with IEI have been increasingly recognized, data regarding the psychological impact on family members remain limited. This study aimed to evaluate anxiety and depressive symptoms in patients with IEI and their family members compared with healthy controls and to examine the association of sociodemographic and clinical factors with anxiety and depressive symptom scores. Methods: A total of 375 participants were included, comprising 34 patients with IEI, 102 family members, and 239 healthy controls. Anxiety and depressive symptoms were assessed using the Beck Anxiety Inventory (BAI) and the Beck Depression Inventory (BDI). Group comparisons were performed using non-parametric statistical tests. Multivariable linear regression analyses were conducted to identify independent predictors of anxiety and depressive symptoms. The internal consistency of the scales was evaluated using Cronbach’s alpha, McDonald’s omega, and Guttman’s lambda-2 coefficients. Results: Both patients with IEI and their family members demonstrated significantly higher BAI and BDI scores than healthy volunteers in the unadjusted comparisons (all p < 0.001), with no statistically significant differences between patients and relatives. After multivariable adjustment using HC3 robust standard errors, healthy volunteers had significantly lower BAI and BDI scores than patients with IEI. Patient relatives had significantly lower BDI scores, but their adjusted BAI scores did not differ significantly from those of patients. Female sex remained independently associated only with higher BAI scores. The BAI and BDI showed excellent internal consistency, with all reliability coefficients exceeding 0.90. Conclusions: Patients with IEI and their family members had higher anxiety and depressive symptom scores than healthy volunteers. Although patients and relatives had comparable BAI and BDI scores in the unadjusted comparisons, adjusted analyses showed significantly lower BDI scores among relatives, whereas the adjusted BAI difference was not statistically significant. Healthy volunteers had significantly lower adjusted BAI and BDI scores than patients, and female sex was independently associated with higher BAI scores. These findings support consideration of psychological well-being at both the patient and family levels in IEI, although the cross-sectional design precludes causal inference. Full article

Although flaviviruses, including DENV, ZIKV and JEV, remain important causes of febrile, congenital, and neurological diseases, treatment options remain largely supportive, with limited availability of virus-specific antiviral therapies. Marine organisms and marine-derived microorganisms produce chemically distinct antiviral materials, including sulfated polysaccharides, terpenoids, alkaloids, peptides, cyclodepsipeptides, and polyketides. However, their activities range from preliminary extract-level inhibition to direct biochemical target validation, making mechanistic comparison difficult. This review critically evaluates marine-derived anti-flaviviral agents using two complementary dimensions, the infection stage implicated by experimental assays and the strength of evidence supporting that assignment. DENV evidence is dominated by sulfated algal macromolecules that interfere with adsorption or internalization, whereas ZIKV studies encompass lipophilic algal metabolites, fungal alkaloids, cyclodepsipeptides, and a few target-oriented candidates. Across the field, most reports remain stage-associated rather than target-validated. Cross-study potency comparisons are constrained by differences in virus strains, cell models, assay formats, and treatment schedules. JEV-specific evidence is particularly sparse. Based on the DENV and ZIKV evidence map, we propose concise priorities for JEV-oriented discovery: early compound-level dereplication, parallel cytotoxicity testing, orthogonal confirmation of productive infection, stage-resolved assays, and biochemical or genetic validation of conserved flaviviral targets. This evidence-based framework can help distinguish promising chemical candidate scaffolds from preliminary antiviral signals and guide mechanism-informed development of marine-derived natural products against flaviviruses. Full article

Background/Objectives: Acute lower respiratory infections (ALRIs) are an important cause of morbidity among children under five years of age. This study aimed to identify factors associated with ALRI among children under five treated at a Peruvian emergency hospital. Methods: We conducted an observational, analytical, retrospective cross-sectional study using medical records of children treated in the pediatric emergency department of Hospital de Emergencias José Casimiro Ulloa, Lima, Peru, between January and December 2024. ALRI was defined as a physician-recorded diagnosis of pneumonia or bronchiolitis; the comparison group comprised children with acute bronchitis or upper respiratory tract infection. Crude and adjusted prevalence ratios (PRs) with 95% confidence intervals (CIs) were estimated using Poisson regression with robust variance. Results: The final sample included 322 children, of whom 154 (47.8%) had ALRI. In the adjusted model, age 0 to <2 years (aPR = 1.27; 95% CI: 1.03–1.56), absence of exclusive breastfeeding (aPR = 1.24; 95% CI: 1.02–1.51), birth weight < 1000 g (aPR = 1.43; 95% CI: 1.09–1.87), incomplete immunizations (aPR = 1.28; 95% CI: 1.07–1.53), and passive smoking exposure (aPR = 1.38; 95% CI: 1.15–1.65) were independently associated with higher ALRI prevalence. Conclusions: ALRI accounted for nearly half of the ARI diagnoses in this pediatric emergency analytical sample. Younger age, absence of exclusive breastfeeding, extremely low birth weight, incomplete immunizations, and passive smoking exposure were independently associated with ALRI and may help identify children requiring closer clinical assessment and preventive interventions. Full article

Background/Objectives: Post-COVID syndrome (PCS) remains a clinically heterogeneous condition. In patients with chronic diseases, real-world evidence is needed to describe its frequency and to identify clinical factors associated with its current presence. This study assessed the prevalence of PCS and factors associated with current PCS status among patients with chronic diseases. Methods: This observational cross-sectional study included a consecutively enrolled source clinical sample of 850 adults receiving inpatient or outpatient care in Almaty and the Almaty Region between June and December 2025. The primary analytic cohort comprised participants with documented prior COVID-19 (n = 250). PCS was assessed at study enrollment according to the national clinical protocol as current symptoms persisting for more than 12 weeks and not explained by an alternative diagnosis. Multivariable logistic regression was used to estimate adjusted odds ratios (ORs) and 95% confidence intervals (CIs). COVID-19 vaccination was excluded from the primary model and evaluated only in a sensitivity analysis because vaccination timing relative to infection was unavailable. The exploratory association-based probability model was evaluated using the AUC with 95% CI, Brier score, calibration assessment, bootstrap internal validation with 1000 resamples, assessment of age nonlinearity, and decision curve analysis. Results: PCS was identified in 147 of 250 participants with prior COVID-19 (58.8%). In the revised primary multivariable model, age (OR = 1.24 per 10-year increase; 95% CI: 1.01–1.54; p = 0.040) and endocrine diseases (OR = 3.36; 95% CI: 1.69–6.98; p < 0.001) remained associated with current PCS status. The association with endocrine diseases persisted after adjustment for body mass index. The apparent AUC was 0.687 (95% CI: 0.620–0.753), and the Brier score was 0.218. Bootstrap internal validation yielded an optimism-corrected AUC of 0.655, a calibration intercept of 0.061, and a calibration slope of 0.816. Conclusions: PCS was frequent among patients with chronic diseases who had prior COVID-19. Older age and endocrine pathology were the most consistent factors associated with current PCS status. All model-performance estimates were obtained within the development sample; the model should not be used for individual prediction or follow-up planning without prospective external validation. Full article

Raccoon dog and fox amdoparvovirus (RFAV) has been reported in raccoon dogs and foxes, but natural infection in domestic dogs has not previously been documented. During March–April 2026, samples from four affected Dobermann dogs from a kennel near Novi Sad, Serbia, were submitted for laboratory investigation. After negative testing for canine adenovirus, canine coronavirus, herpesvirus, parvovirus, distemper virus, influenza A virus, and leptospirosis, metagenomic sequencing was performed on selected tissues, followed by bioinformatic analysis and targeted RFAV PCR screening of additional outbreak-associated samples. Affected dogs had prolonged illness characterized by conjunctivitis with ocular and nasal discharge, occasional blue eye appearance, progressive weight loss, poor coat quality, jaundice and biochemical evidence of hepatic injury, and neurologic signs including paraplegia in advanced cases. Sequencing generated 434,220 reads and identified multiple RFAV hits; pooled assembly produced a 4799 bp consensus genome with approximately 97% similarity to known RFAV strains and genome organization consistent with the genus Amdoparvovirus. RFAV DNA was subsequently detected by virus-specific PCR in an epidemiologically linked dog and across diverse specimen types including blood, urine, kidney, spleen, brain, lung, testicle, ileocecal lymph node, and throat swabs, whereas clinically healthy unrelated dogs were PCR-negative. Full article

Background/Objectives: Celiac disease (CeD) is a complex autoimmune enteropathy triggered by gluten intake in genetically predisposed individuals. While the bacterial microbiome has been extensively studied in CeD, the role of the gut virome and mycobiome is less well defined. Emerging evidence suggests that viruses and fungi may influence mucosal immunity, intestinal permeability and gluten tolerance, acting as potential environmental cofactors in the onset of this disease. Methods: A narrative review of the literature was conducted to summarize current knowledge of the composition, development and function of the gut virome and mycobiome and to explore their potential involvement in CeD pathogenesis. Results: The gut virome shows high interindividual variability and dynamic maturation early in life. Multiple studies have associated early enterovirus or parechovirus infections with an increased risk of celiac autoimmunity, supporting a virus–gluten “double hit” model. Viral dysbiosis in CeD has been reported to include an enrichment of human polyomavirus 2 (JC polyomavirus) and of specific enterobacterial phages. The mycobiome, although less abundant, also displays disease-associated alterations: cross-sectional mycobiome studies have reported higher relative abundance of Saccharomyces, Candida, and Tricholomataceae and lower relative abundance of Pichia and Pneumocystis taxa in children with CeD. Conclusions: Current evidence, although preliminary, supports associations between selected viral exposures and CeD-related immune alterations, whereas current phageome and mycobiome findings mainly describe cross-sectional compositional differences. These data do not establish temporal direction or causality, and further longitudinal and mechanistic investigations are required before preventive or therapeutic implications can be considered. Full article

Japanese encephalitis virus (JEV) is a zoonotic pathogen transmitted primarily by Culex mosquitoes and causes severe neurological diseases in humans and animals. The main endemic areas are the Western Pacific and Southeast Asia, and its geographical distribution has expanded in recent years. The development of a diagnosis for orthoflavivirus infections is hampered by two main problems: the short duration of viremia, resulting in a narrow detection window, and severe cross-reactivity. NS1, a secreted nonstructural protein of orthoflavivirus, holds promise as a new target for overcoming these limitations. In this study, we established a highly specific and sensitive capture ELISA for the JEV NS1 protein. First, the JEV NS1 protein was successfully expressed in mammalian cells and purified using affinity chromatography. Seven mAbs recognizing JEV NS1 were generated, and the mAb 20B6 exhibited the strongest binding affinity. Based on 20B6, a capture ELISA was developed with an optimal coating concentration of 3 μg/mL and an optimal detection antibody working concentration of 0.432 μg/mL. No cross-reactivity was observed with other orthoflaviviruses (including WNV, KUNV, USUV, MVEV, SLEV, and ZIKV) or common porcine viruses. The method could effectively detect NS1 protein in cell culture medium, cell lysates, mouse tissues, and porcine serum samples from JEV-infected subjects. This study provides a solid foundation that may be further developed into an efficient and specific tool for epidemiological surveillance of Japanese encephalitis. Full article

CD163 not only acts as an essential receptor for porcine reproductive and respiratory syndrome virus (PRRSV) infection but also serves as a barrier to cross-species cellular infection by arteriviruses. Among its domains, the scavenger receptor cysteine-rich 5 (SRCR5) domain of CD163 is functionally indispensable. In this study, two monoclonal antibodies (mAbs), designated 5A and 11D, were developed against the SRCR4-6 region of porcine CD163. The linear epitopes recognized by mAbs 5A and 11D were identified as ⁵⁴⁶CEGHESHLSLCPVAP⁵⁶⁰ in SRCR5 and ⁴⁴⁹WDCKNW⁴⁵⁴ in SRCR4, respectively. Crystal structural analysis and sequence alignment showed that the ⁵⁴⁶CEGHESHLSLCPVAP⁵⁶⁰ epitope resides in the long loop 5-6 of SRCR5, which is a key structural region responsible for ligand binding. Meanwhile, the ⁴⁴⁹WDCKNW⁴⁵⁴ epitope is located within a highly conserved region of SRCR4, which confers broad cross-species reactivity. Notably, mAb 5A reduced PRRSV infection, while mAb 11D exhibited no anti-PRRSV activity in target cells. Collectively, this study provides essential molecular tools for exploring CD163-domain-dependent cross-species reactivity and developing anti-PRRSV strategies. Full article

Cucurbit downy mildew, caused by the oomycete Pseudoperonospora cubensis, is a devastating disease of cucurbit crops requiring rapid detection for effective management. Current molecular diagnostics often rely on complex laboratory infrastructure, limiting their on-site applicability. To address this limitation, we developed a highly specific, field-deployable loop-mediated isothermal amplification (LAMP) assay targeting the P. cubensis-specific c2555.2e1 marker gene. The assay’s optimal reaction conditions were determined, and its specificity was evaluated using closely related oomycetes, including P. humuli and P. belbahrii, alongside other fungal species. Assay sensitivity was compared to conventional PCR using a pathogen DNA dilution series, and diagnostic efficacy was validated using field-collected winter squash samples suspected of infection and environmental spore-trap samples. The optimal LAMP reaction conditions were 71 °C for 60 min. The assay accurately amplified P. cubensis with no observed cross-reactivity. Sensitivity analysis demonstrated a detection limit of 100 pg/µL, and was at least 10-fold more sensitive than conventional PCR. Furthermore, the LAMP assay exhibited greater detection sensitivity than conventional PCR when analyzing environmental and field samples. This rapid and sensitive LAMP assay provides a rapid and highly sensitive tool for on-site diagnostics, offering a crucial advancement for the early detection and management of cucurbit downy mildew. Full article

Background/Objectives: Strongyloides stercoralis may persist through autoinfection and is clinically important before corticosteroid exposure, but its relationship with airway obstruction in children with sickle cell disease (SCD) is uncertain. We examined whether routine-detected S. stercoralis infection was associated with spirometry-defined airway obstruction in children with SCD in French Guiana. Methods: This multicentre cross-sectional secondary analysis used an availability-based sample from a routine respiratory screening database. Eligible children aged 5–17 years had SCD, spirometry, and stool parasitology documented during routine care. The primary outcome was FEV1/FVC < 80%. Logistic regression adjusted for age, sex, genotype, hydroxyurea treatment, previous acute chest syndrome, and atopy. Results: Stool parasitology was available for 160 children; 26 (16.2%) had S. stercoralis detected. Among 138 children with complete spirometry and covariate data, 45 (32.6%) had obstruction. S. stercoralis was not associated with obstruction in univariable analysis (OR 0.75, 95% CI 0.30–1.91) or after adjustment (adjusted OR 1.02, 95% CI 0.37–2.86; p = 0.964). Male sex was independently associated with obstruction (adjusted OR 2.32, 95% CI 1.11–4.83). Conclusions: Routine-detected S. stercoralis infection was not an independent correlate of airway obstruction. The wide confidence interval, limited exposure ascertainment, and small infected subgroup mean that clinically important protective or harmful associations cannot be excluded. Full article

Adult plant resistance (APR) is widely used in wheat breeding, but its behaviour across genetic backgrounds remains poorly understood. In this study, we analysed the expression of three APR loci (Lr34, Lr46, Lr67) following their introgression into elite winter wheat cultivars. BC₂F₁ populations derived from crosses between donor lines and commercial cultivars were evaluated under controlled infection with Puccinia triticina (Pt). Gene expression was assessed using RT-qPCR, and miRNA abundance was quantified by ddPCR at five time points (0–48 hours post-inoculation, hpi). Expression patterns differed markedly between genetic backgrounds, affecting both the magnitude and timing of gene activity. Lr34 and Lr67 showed the highest expression prior to inoculation, with no consistent or sustained induction following infection. In contrast, the candidate gene Lr46-Glu2 displayed a clear tendency towards early induction, with peak expression typically observed at 6–12 hours post-inoculation, although the amplitude of this response varied among genotypes. Levels of miRNA varied across genotypes and time points and did not consistently reflect mRNA expression. The results suggest that the expression of APR-associated loci is influenced by genetic background. Lr34 and Lr67 showed predominantly baseline-associated expression, whereas the Lr46-associated candidate gene Lr46-Glu2 displayed a more infection-responsive transcriptional profile following inoculation. These patterns are consistent with different transcriptional behaviours of the analysed genes but do not establish their functional contribution to resistance. Further phenotypic and functional validation will be required to determine the biological significance of these expression patterns. Full article

Background/Objectives: Aneurysmal subarachnoid hemorrhage (aSAH) carries high morbidity and mortality, with delayed cerebral ischemia (DCI) as a major secondary complication. Although infection has increasingly been implicated in DCI pathogenesis, the role of central nervous system (CNS) infection remains unclear. We evaluated CNS infection-related parameters for predicting DCI and functional outcome after aSAH using machine learning. Methods: This retrospective single-center study included 191 patients with confirmed aSAH treated in the intensive care unit between 2019 and 2024. Demographic, clinical, radiographic, treatment-related, microbiological, cerebrospinal fluid (CSF), and longitudinal laboratory data were extracted from electronic records. DCI was analyzed as a binary outcome, and discharge functional outcome was dichotomized using the modified Rankin Scale (0–2 vs. 3–6). Twelve feature selection methods and twelve classifiers were evaluated using five-fold cross-validation. Results: For DCI prediction, the models achieved a mean accuracy of 0.96, F1-score of 0.96, and AUC of 0.98. Key predictors included antibiotic therapy, aneurysm treatment modality, history of thrombosis, and EVD revision, along with mean albumin, red cell distribution width, INR, minimum aspartate aminotransferase, and CSF glucose, lactate, nucleated cell count, and cellular debris. For functional outcome prediction, accuracy was 0.948, F1-score 0.95, and AUC was 0.97. Predictors comprised cerebral vasospasm, number of spasmolysis procedures, pre-admission anticoagulation, CSF pathogen detection and type, aPTT, fibrinogen, aspartate aminotransferase, lactate dehydrogenase, and CSF mononuclear cell proportion, leukocyte count, and glucose concentration. Conclusions: Machine learning models integrating clinical, systemic, and CSF-derived parameters demonstrated excellent performance in predicting both DCI and functional outcome after aSAH. The identified predictors highlight the importance of the neurocritical care course, including infection-related and thromboinflammatory processes, and extend beyond traditional vasospasm-centered paradigms. These findings may inform risk stratification, although the temporal relationship between several predictors and outcome onset means the models should be regarded as hypothesis-generating rather than tools for early clinical prediction. Full article

Background: Nosocomial infections, or healthcare-associated infections (HAIs), represent a significant global health challenge frequently associated with invasive medical procedures and extended hospitalizations. This study was designed to evaluate the current level of knowledge and the perceptions of inpatients regarding HAIs within a high-risk tertiary hospital in the Qassim region of Saudi Arabia. Method: A descriptive cross-sectional study was conducted from August 2024 to February 2025 at a tertiary facility in the Qassim region. The study population included 500 inpatients aged 18 years and older who consented to participate. Data were collected via individual interviews using a closed-ended questionnaire, which assessed demographics, knowledge and perceptions of HAIs on a 4-point Likert scale. Results: Among 500 study participants, good knowledge, fair knowledge and poor knowledge regarding HAIs were demonstrated by 57%, 38.4% and 4.6%, respectively. More than 98% of participants achieved a perception score of above 70%, indicating a profoundly positive perception level. Knowledge was significantly associated with age (p = 0.018), educational status (p < 0.01), and nationality (p < 0.01). Participants most commonly recognized contaminated hospital beds (95.6%), airborne transmission (91.2%), and unsterilized equipment (90%) as the potential modes of infection transmission. Conclusions: Sustaining ongoing infection prevention efforts and fostering improved awareness among inpatients regarding HAIs are of paramount importance in implementing effective public health prevention measures. Full article