Search Results (1,846)

Trans-epithelial permeability is a critical functional parameter for reconstructed tissues, particularly in genitourinary tissue engineering, where urine leakage must be avoided. Although Franz diffusion cells are considered the gold standard for permeability measurements, their cost and limited accessibility restrict their widespread use. In parallel, the reliable quantification of urea in culture media remains challenging due to protein interference and assay cost. The Inexpensive Trans-Epithelial Permeability (I-TEP) test is a simple and a low-cost Franz-like permeability system which can be combined with an optimized diacetyl monoxime–thiosemicarbazide (DAMO–TSC) colorimetric urea assay. I-TEP system relies on readily available laboratory components to create physically separate donor and receiver compartments, with the tissue acting as the sole diffusion interface. The DAMO–TSC assay was optimized through systematic evaluation of deproteinization, incubation time, storage conditions, and serum interference. The I-TEP test showed a strong correlation with conventional Franz diffusion cells when testing similar tissue samples. Deproteinization was identified as a mandatory step for accurate urea quantification in serum-containing media. The combined approach was successfully applied on engineered genitourinary tissues, demonstrating sensitivity to tissue maturation and cellular composition. This protocol provides a proof of concept for an affordable, robust, and autonomous method for routine permeability assessment, bridging the gap between costly commercial systems and high-throughput experimental needs. Full article

Background: Acinetobacter baumannii is a non-fermenting Gram-negative bacillus responsible for severe nosocomial infections, particularly in intensive care units (ICUs). The increasing prevalence of multidrug-resistant (MDR) and carbapenem-resistant A. baumannii (CRAB) strains has become a significant challenge for infection control and antimicrobial therapy worldwide. Objectives: This study aimed to analyze the antimicrobial susceptibility patterns of clinical A. baumannii isolates recovered from a multi-profile hospital in years 2020–2024 in Lodz, Poland. Methods: Clinical isolates from various specimen types (blood, urine, wound swabs, biopsies, sputum, and bronchoalveolar lavage fluid) were obtained during routine microbiological diagnostics. Identification was performed using MALDI-TOF MS. Antimicrobial susceptibility testing (AST) was conducted using the automated VITEK^®2 system with EUCAST/CLSI interpretive criteria. Minimum inhibitory concentrations (MICs) for colistin were determined by broth microdilution. Carbapenemase production was assessed using the Carbapenem Inactivation Method (CIM) and immunochromatographic assays for OXA-23, OXA-40/58, and NDM detection. Results: A total of 244 A. baumannii isolates were recovered over the study period. Susceptibility to carbapenems (meropenem, imipenem) declined markedly, with resistance exceeding 90% by 2023–2024. Aminoglycosides exhibited variable activity, with gentamicin demonstrating the highest susceptibility rates (up to 88% in 2022). Resistance to ceftazidime and cefepime remained consistently high (>90% in 2023–2024). No fully susceptible isolates were identified for ciprofloxacin. Conclusions: The high prevalence of CRAB strains highlights the urgent need for effective infection control measures, optimized antimicrobial stewardship, and consideration of novel treatment options in the clinical setting. Full article

Urinary Tract Infections (UTIs) represent one of the most prevalent bacterial infections globally, posing significant health burdens, especially in low- and middle-income countries (LMICs), due to delayed diagnoses, limited access to laboratory services, and rising antimicrobial resistance. This study presents a machine learning (ML)-based diagnostic support framework for early UTI detection, leveraging structured clinical data and explainable artificial intelligence (XAI) techniques to enhance interpretability and trust among healthcare providers. A patient dataset containing 4865 records was used in the study to train and test Extreme Gradient Boosting (XGBoost), Decision Tree (DT) and Random Forest (RF) classifiers, while class imbalance was addressed using Synthetic Minority Over-sampling Technique (SMOTE). The performance of the models was evaluated through accuracy, precision, recall, F1-score, Log Loss, and AUC-ROC, and random forest showed the best results (accuracy: 86.43%, F1-score: 86.71%, AUC-ROC: 0.8695). To ensure that such models can be adopted by stakeholders in the health sector, Local Interpret-able Model-agnostic Explanations (LIME) were integrated, which identified painful urination, urinary frequency, and suprapubic pain as primary predictors in the model. This study shows that interpretable ML models can be helpful in resource-limited regions in predicting UTIs, thereby rendering a solution to improve the management of infections in these regions. Full article

Early-onset infection caused by Streptococcus agalactiae (Group B Streptococcus, GBS) may occur during gestation or delivery and can lead to severe neonatal sepsis, meningitis, or pneumonia. Discordant GBS infections in twin gestations are rare. We report a fatal case of early-onset GBS infection in dichorionic–diamniotic twins conceived via IVF and delivered by caesarean section at 32 weeks’ gestation due to discordant fetal growth and abnormal Doppler indices in Twin A (Umbilical Artery PI = 1.4; Middle Cerebral Artery PI = 1.5). Twin A had Apgar scores of 3, 5, and 5 and rapidly developed tachycardia, respiratory distress, and systemic infection, while Twin B, with Apgar scores of 7, 8, and 9, remained clinically stable. Both infants were admitted to the NICU and underwent routine blood, urine, and cerebrospinal fluid testing. Despite the prompt initiation of parenteral ceftriaxone and respiratory support, Twin A deteriorated rapidly and died within 28 h. GBS was isolated from Twin A’s blood culture, and maternal placental tissue and high vaginal samples collected before antibiotic administration also grew GBS, with all isolates demonstrating identical antimicrobial resistance profiles. Molecular analysis revealed matching rib1 and alp2/3 gene patterns in isolates from the mother and Twin A. Maternal anovaginal immunochromatography at delivery was positive, whereas screening cultures obtained at 29 weeks’ gestation were negative. This case highlights the limitations of culture-based GBS screening in high-risk pregnancies and preterm deliveries and underscores the potential value of molecular assays and point-of-care testing to improve detection of S. agalactiae throughout pregnancy and the peripartum period. Emerging preventive strategies, including modulation of the genital microbiome and maternal vaccination aligned with WHO recommendations, may further reduce the burden of neonatal GBS disease. Full article

Background and Objectives: Urinary Tract Infections (UTIs) are a major concern worldwide due to increasing antimicrobial resistance. Even though sex-based differences in antimicrobial resistance are recognized worldwide, there is a lack of data in the literature. This study aims to evaluate differences in the distribution of uropathogens and antibiotic resistance across large groups of Romanian males and females. Materials and Methods: This retrospective descriptive analysis included 2567 positive urine cultures collected over a 1-year period from hospitalized and outpatient patients at two representative urology centers in Bucharest. Only urine tests with ≥10⁵ CFU/mL and monomicrobial growth were included. Results: The sex distribution showed a predominance of male patients (62.2%). Also, their age tended to be higher than that of females. Escherichia coli remained the most common pathogen, with a higher prevalence in females (54.08% vs. 32.54%), while Klebsiella and Pseudomonas were more frequently noted in males. The Gram-negative analysis revealed higher resistance rates in male patients, particularly for common antibiotics such as amoxicillin-clavulanic acid (50.37% vs. 35.77%), trimethoprim-sulfamethoxazole (46.69% vs. 34.64%), and levofloxacin (45.55% vs. 34.71%). Notably, carbapenem resistance in Klebsiella exceeded 30% in males, indicating major concerns about multidrug resistance in this case. In contrast, Gram-positive bacteria showed more stable resistance patterns among the studied groups. Preserved sensitivity was found to linezolid, vancomycin, fosfomycin, and nitrofurantoin. Conclusion: These findings demonstrate clinically relevant sex-based differences in both pathogen distribution and antimicrobial resistance, particularly among Gram-negative uropathogens. Also, it highlights the importance of developing sex-adapted antibiotic strategies in conjunction with the local epidemiological data. Full article

In Cambodia, farmers construct artificial household bat roosts to collect and sell guano as fertilizer. We investigated farming practices and attendant spillover risks using (1) surveys on guano production; (2) an estimation of bat population size and species present using carcasses, visual identification, and audio recordings; (3) surveys of guano-producing and neighboring households on water, sanitation, and hygiene practices; and (4) the testing of guano and household food, water, and surfaces for coronaviruses using RT-qPCR. Bat roosts are constructed using dried palm leaves with coconut tree and/or steel/concrete supports. Roosting areas ranged from 42 to 327 m², bat abundance varied from 0 to 11,187, guano production was between 5 and 120 kg/week, guano yields were from 0.15 to 0.4 kg/m²/week, and farmers earned USD ~100–200/household/month. Higher guano production in the peak (normally wet) season was associated with greater bat abundance (p = 0.016). The lesser Asiatic yellow house bat (Scotophilus kuhlii) was the only bat species identified. Roosts were <20 m from guano-producing households. Neighbors and households’ hygiene risks included not having handwashing stations and not covering food in storage/while drying. Coronaviruses (Alphacoronaviruses or Infectious Bronchitis Virus) were detected in 14.6%, 17.3%, 2.9%, 1.4%, and 0.0% of guano, urine, household surface, food, and water samples, respectively. While guano farming offers economic benefits, spillover risks exist. Safe guano collection and storage, handwashing, and food covering in guano-producing communities are necessary to mitigate spillover risks. Full article

West Nile virus (WNV) diagnosis relies on nucleic acid amplifications, but these techniques do not discriminate between infectious and non-infectious viral particles. This limitation can be bypassed by using a genome-binding dye (PMAxx) that is unable to cross membranes and can only bind to the genomes of non-intact (i.e., non-infectious) viral particles. This study evaluated a workflow combining PMAxx treatment with digital PCR to improve the molecular discrimination of intact WNV particles. Fifty-five samples (35 plasma, 20 urine) from 41 patients with WNV fever (WNF) or WNV neuroinvasive disease (WNND) were analyzed. Samples were tested with/without PMAxx treatment. Overall, PMAxx treatment resulted in a significant reduction in detectable viral RNA (median reduction: 1.0 Log copies/mL; p < 0.0001), indicating that a substantial fraction of RNA detected by standard methods originated from non-infectious particles. This reduction was more visible in urine (1.8 Log copies/mL) than in plasma (0.4 Log copies/mL), suggesting a higher proportion of degraded viral particles or free RNA in urine. Stratification by clinical presentation showed significant reductions in both WNF and WNND patients, with no significant differences between groups. This approach may represent a valuable adjunct for improving diagnostic interpretation and epidemiological assessment of WNV infection, particularly in matrices characterized by prolonged RNA persistence. Full article

Autism spectrum disorder (ASD) has been associated with alterations in the gut microbiota and its metabolites, particularly short-chain fatty acids (SCFAs) and microbiota-derived tryptophan catabolites, which may influence neurodevelopment through immune and epigenetic mechanisms. We investigated whether stool SCFAs and tryptophan-pathway metabolites differ between children with ASD and typically developing controls, and whether these metabolites associate with ASD severity and systemic biochemical signatures. In this cross-sectional study, we analyzed stool samples from 229 children (160 with ASD, 69 controls) with complete SCFA and tryptophan-metabolite data, while urine metabolomics data were available for a subset and were used for exploratory stool–urine integration analyses. Children with ASD and controls were similar in age, but the ASD group had a higher proportion of males. Absolute concentrations of individual SCFAs, total SCFAs, and derived indices were broadly comparable between groups; nominal differences in propionate/acetate ratio and caproate did not remain significant after false discovery rate correction. Similarly, stool tryptophan-pathway metabolites reported as ng/a.u. based on the NanoDrop-derived proxy (tryptophan, kynurenine, indole-3-acetic, indole-3-lactic, indole-3-propionic, indole-3-aldehyde, N-acetyl-tryptophan, serotonin, melatonin, tryptamine) and functional ratios (kynurenine/tryptophan, indole-derived/tryptophan, serotonin/tryptophan) showed no robust ASD–control differences; N-acetyl-tryptophan was nominally higher in ASD but did not survive multiple-testing correction. In the ASD subgroup with available Childhood Autism Rating Scale (CARS) data (n = 34), SCFA and tryptophan indices showed only weak, non-significant correlations with global ASD severity. In contrast, correlation analyses revealed two coherent metabolic modules, i.e., an SCFA block with very strong internal correlations among individual SCFAs and total SCFAs and a tryptophan block with strong correlations between metabolites and their normalized ratios, while cross-module correlations were modest. These results indicate that stool SCFA and microbiota-derived tryptophan profiles do not robustly distinguish ASD from controls in this cohort, but they form stable metabolic modules compatible with microbiome–epigenome frameworks. Full article

Antimicrobial resistance (AMR) is an increasing concern in veterinary medicine and may compromise empirical treatment of feline urinary tract infections (UTIs). This retrospective observational study evaluated antimicrobial resistance patterns and their association with multidrug resistance (MDR) in bacterial isolates obtained from urine cultures of cats presented to a feline referral practice in Lisbon, Portugal, between January 2023 and December 2025. A total of 174 cats with positive urine cultures were included, yielding 178 bacterial isolates. Escherichia coli was the most frequent pathogen (57.30%), followed by Enterococcus spp. (16.98%) and Staphylococcus spp. (14.61%). Antimicrobial resistance was detected in 107/178 isolates (60.11%), and 76/178 (42.70%) were classified as multidrug-resistant. Extended-spectrum beta-lactamase-producing Escherichia coli and methicillin-resistant Staphylococcus strains were also identified. Resistance to penicillins and fluoroquinolones was significantly associated with MDR (p < 0.001). These findings highlight the high burden of antimicrobial resistance in feline urinary isolates in this clinical setting and support routine urine culture and susceptibility testing to guide therapeutic decision-making, particularly in recurrent, complicated, or high-risk cases. Full article

It is well known that gas sensor responses are affected by the presence of humidity in the analyzed gas. This is particularly true when dealing with biological fluid samples, whose high moisture content interferes with the adsorption of the trace volatile organic compounds (VOCs) on the sensors’ active layer. To address this challenge, this study focuses on designing and testing a novel sampling system for the dehumidification of biological fluid headspace to be characterized by an electronic nose (e-Nose). Such a system, based on the use of disposable polymeric sampling bags purged with dry air, exploits the polymers’ permeability to water vapor to reduce sample humidity. Tested materials included Nalophan^TM (20 μm), high-density polyethylene (HDPE, 8, 9, 10 and 11 μm), low-density polyethylene (LDPE, 12 and 50 μm), and biodegradable polyester (Bio-PS, 15 μm). First, dehumidification performance was characterized as a function of dry air flow rate and film type. A purge of 1 L/min accelerated the sample humidity removal compared to passive storage of bags from >2 h to <1 h (from 80% to 20% RH). Second, a mass-balance model was applied to dedicated experiments to decouple water losses due to diffusion and adsorption, showing that diffusion through the polymer wall dominates, while adsorption occurs in the early stages of conditioning. Third, because these materials are not selectively permeable to water, potential loss of water-soluble VOCs during dehumidification was investigated. Pooled urine headspace samples—both raw and spiked with a metabolite mix of VOCs—were dried using each material and analyzed using a photo-ionization detector (PID) and an e-Nose. Results were compared against a Nafion^TM dryer. Comparison was based on the e-Nose’s ability to discriminate between pooled vs. spiked samples and reveal real-life metabolomic changes. Nalophan^TM bags and Nafion^TM dryer provided the highest VOC fingerprint to support discrimination by the e-Nose, while Bio-PS provided the fastest sample dehumidification. The proposed bag-based system offers a cost-effective, disposable, and contamination-free solution to humidity interference in e-Noses. Full article

Background: Effective sick-day management, including ketosis home management aimed at preventing diabetic ketoacidosis (DKA), is essential for families living with a child/adolescent with type 1 diabetes (T1D). Methods: Adolescents living with T1D and caregivers of younger children living with T1D were invited to participate in an interview consisting of five parts: (I) demographic data, (II) subjective self-ratings on competence in ketosis home management, (III) objective assessment of competence in ketosis home management using a standardized clinical case scenario consisting of 10 management steps, in which participants were asked to describe the actions they would take to prevent DKA, and (IV) practical demonstrations to objectively assess skills in (IVa) urine dipstick self-testing and (IVb) insulin administration, (V) household availability of (Va) urine dipsticks and (Vb) insulin cartridges. Results: (I) We enrolled 61 adolescents and 79 caregivers. (II) Competence in ketosis home management was subjectively self-rated as good to very good. (III) Adolescents reported 4 (median; Q25/Q75 3/5) and caregivers 5 (4/5) of 10 management steps. Never self-testing ketone levels was reported by 33% of adolescents and 11% of caregivers. (IVa) At least one handling error occurred in 100% of adolescents’ and in 98% of caregivers’ practical demonstrations of urine dipstick self-testing and in (IVb) 98% of adolescents’ and 98% of caregivers’ insulin administrations. (Va) Altogether urine dipsticks were available in 43% of households, whereas (Vb) insulin cartridges were available in 78% of households. Conclusions: Our results demonstrate a mismatch between challenges in ketosis home management and high subjective self-ratings. Full article

Paper-based microfluidic devices (μPADs) have attracted significant attention for point-of-care testing (POCT), environmental monitoring, and food safety due to their low cost, ease of use, and minimal instrument dependence. However, fabricating high-resolution and reproducible microchannels on paper remains challenging. Conventional methods such as wax printing, photolithography, and inkjet printing are limited by resolution or equipment cost. Here, we present a low-cost, high-resolution fabrication method for μPADs, termed wax soft lithography, which combines wax printing with soft lithography. Through this method, microchannels with a minimum width of 234 ± 62 μm were consistently produced, and complex patterns were successfully fabricated, demonstrating high precision and reproducibility. As a proof-of-concept demonstration of device functionality, the fabricated μPADs were used to detect glucose in spiked urine samples, showing a concentration-dependent colorimetric response. This method provides an effective route for rapid production of high-resolution μPADs in resource-limited settings. With further validation before practical applications, this method shows promise for future development in POCT. Full article

The genus Siccibacter consists primarily of environmental bacteria, with strains of Siccibacter colletis previously isolated only from plant materials and related environments. This study aims to characterize the first clinical isolate of S. colletis and explore its genomic evolution and clinical relevance. Strain S25242 was isolated from the urine of a 64-year-old male with a severe urinary tract infection. The genome of S25242 is 4.19 Mb, containing 4012 coding sequences, 73 tRNAs, 10 rRNAs, and 38 snRNAs. Phylogenetic and phylogenomic analyses indicated that strain S25242 is closely related to S. colletis type strain 1383^T. The strain shared >70% of digital DNA-DNA hybridization (dDDH) values and >96% of average nucleotide identity (ANI) values with the type strain of S. colletis 1383^T, thereby confirming its taxonomic status. The isolate was susceptible to all 11 tested antimicrobials. Comparative genomics identified 1942 S. colletis-specific genes (including multidrug efflux systems) and 13 unique genes in S25242 related to transposition and DNA integration. This study reports the first clinical isolate of S. colletis, providing evidence that genomic plasticity facilitates its transition from an environmental inhabitant to an opportunistic pathogen. The findings highlight the need for enhanced clinical surveillance of the Siccibacter genus and offer insights into its genomic evolution and clinical adaptation. Full article

Prostate cancer (PCa) remains one of the most prevalent urological malignancies worldwide, with early and accurate diagnosis being critical for improving patient outcomes. Traditional screening approaches, such as digital rectal examination and prostate-specific antigen (PSA) testing, have long served as frontline tools; however, their limited specificity and sensitivity contribute to high rates of false positives, unnecessary biopsies, and overtreatment. Recent UK guidelines and international consensus increasingly question the role of PSA-based population screening, advocating for risk-stratified pathways and multiparametric MRI as first-line investigations. In parallel, advances in molecular biology have identified promising cancer-specific biomarkers, such as prostate cancer antigen 3 (PCA3) and transmembrane protease serine 2 (TMPRSS2:ERG), that outperform PSAs in terms of specificity and prognostic value. These developments have catalysed innovation in biosensor technologies, enabling rapid, cost-effective, and non-invasive detection of single and multiplex biomarkers in urine and serum. Electrochemical and optical affinity-based biosensors offer transformative potential for the development of personalised point-of-care platforms and diagnostics, reducing the reliance on invasive procedures and improving clinical decision-making. The latter can be augmented with artificial intelligence (AI) tools. This review critically examines the limitations of PSAs, synthesises evidence on novel biomarkers and imaging-led strategies, and evaluates the design, performance, and translational challenges of biosensor-based assays. Furthermore, it outlines future directions, including standardisation, large-scale clinical validation, and integration of multiplex biosensors with AI for precision diagnostics. By bridging molecular insights with engineering innovations, these approaches promise to redefine PCa screening and enable accurate, patient-centred care. Full article

Dopamine is a key neurotransmitter and neuromodulator that regulates many critical brain functions. Accurate monitoring of its level is essential for neuroscience as well as the diagnosis and treatment of many brain diseases. In this work, we developed a new electrochemical sensor, comprising phosphorus-doped graphitic carbon nitride (P-g-C₃N₄) and zeolitic imidazolate framework 67 (ZIF-67), for dopamine detection. In this composite electrode material, ZIF-67 provides numerous adsorption and sensing sites, while P-g-C₃N₄ enhances overall electrical conductivity and stability. Cyclic voltammetry tests reveal the redox behavior of dopamine at the surface of the composite electrode across various pH values and scan rates. Using differential pulse voltammetry, the sensitivity and selectivity of this dopamine sensor were assessed, identifying a limit of detection of 0.39 nM. Further successful quantification of dopamine in urine samples suggests the potential practical use of this new composite electrochemical sensor for detecting dopamine and/or other neurotransmitters. Full article