Search Results (7,632)

Urinary epidermal growth factor (uEGF) and clusterin (uCLU) are emerging biomarkers in chronic kidney disease (CKD), but rigorous analytical validation is required before clinical implementation. We evaluated intra-individual variability and long-term storage stability of uEGF and uCLU in CKD. In the prospective, multicenter UVALID study, 60 adults with CKD stages 2–4 underwent urine sampling at three visits over 8 weeks. First-morning and 24-h urine samples were collected to assess intra-individual variability over 24 h, 3 days and 8 weeks. Biomarkers were measured in duplicate by ELISA and normalized to urinary creatinine (/Cr). Inter-laboratory performance was assessed using quality control samples. Stability after 12 and 15 months of storage at −20 °C and −80 °C and the influence of pH were evaluated. Over 24 h, 3 days, and 8 weeks, uEGF/Cr demonstrated low variability and remained stable after long-term storage at both temperatures. In contrast, uCLU/Cr showed greater variability and pronounced instability at −20 °C, whereas stability was preserved at −80 °C. Samples with pH > 6 partially preserved uCLU stability at −20 °C. Inter-laboratory reproducibility was acceptable for uEGF but suboptimal for uCLU at low concentrations. Thus, uEGF showed robust analytical performance, supporting its potential clinical applicability in CKD, whereas uCLU exhibited important analytical and pre-analytical limitations, warranting further assay optimization. These findings underscore the need for rigorous validation to facilitate biomarker implementation in clinical practice. Full article

Purpose: To investigate the association between COVID-19 infection and the 1-year risk of acute urinary retention (AUR) and related urological complications in patients with benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTs). Materials and Methods: Using the TriNetX global network, patients with BPH and LUTs between January 2020 and January 2024 were identified. Participants were classified into a COVID-19 cohort (N = 32,948) and a non-COVID control cohort (N = 434,123). Propensity score matching (1:1) balanced demographics, comorbidities, medications, and laboratory parameters. The primary outcome was AUR within one year. Secondary outcomes included Foley catheterization, urinary tract infection (UTI), gross hematuria, bladder stones, and prostate-related surgery. Results: After matching, 32,918 patients remained in each cohort. The COVID-19 group showed a significantly higher 1-year incidence of AUR compared with controls (2.18% vs. 0.32%; aHR 6.89, 95% CI 5.62–8.45; p < 0.0001). Increased risks were also observed for Foley catheterization (aHR 4.10), UTI (aHR 3.52), and prostate-related surgery (aHR 6.02). Kaplan–Meier analysis demonstrated persistent divergence in AUR-free survival. Conclusion: COVID-19 infection is independently associated with a markedly increased risk of AUR and urological complications in patients with BPH, highlighting the need for closer post-infection monitoring. Full article

Chronic kidney disease (CKD) is a prevalent condition in cats and recognized as a leading cause of mortality among cats generally, yet it is difficult to detect by cat caregivers, especially in its early stages. This retrospective study aimed to enhance detection of CKD by cat caregivers through a smart litter box monitor technology that tracks feline elimination behaviors. We established cohorts of cats with CKD and cats with no known conditions and analyzed behavioral features captured by the monitor. A mixed-effects model identified significant differences, which were then integrated into a machine learning framework for predictive modeling. The model achieved a weighted F1-score of 92.7% in training using cross-validation and 89.9% in validation, demonstrating high precision for CKD predictions. Key findings included a behavioral profile unique to cats with CKD characterized by increased urination frequency, longer elimination durations, and reduced post-elimination covering behavior. These results suggest that the smart litter box monitor can provide valuable, continuous, and non-invasive data for CKD detection and management. 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

Per- and polyfluoroalkyl substances (PFASs) are a large class of thousands of synthetic organofluorine chemical compounds used for many industrial applications. Humans are exposed to PFASs mainly through diet and contaminated drinking water. Studies show that PFASs induce several adverse effects on humans. A great number of human biomonitoring studies have been widely conducted with the aim of estimating exposure to PFASs. The matrices mainly investigated are blood, serum and breast milk. However, in many cases, the need for non-invasive sampling methods with a minimal impact on donors has become paramount to comply with modern ethical standards and regulations. For this reason, we developed a streamlined and efficient method for the analysis of eight perfluorocarboxylic and perfluorosulfonic acids (PFHpA; PFHxS; PFOA; PFHpS; PFNA; PFOS; PFDA; and PFUdA) in human urine samples by UPLC chromatography tandem mass spectrometry. Chromatographic and MS parameters were optimized; the method was validated for: repeatability (<20%), within-lab reproducibility (<20%), trueness (within the set 20% variation limit of agreement between the mean of the data set and the true value), efficiency (51–97%), linearity (R² > 0.99), limits of detection (0.0003 ng/mL), and limits of quantification (0.001 ng/mL). To our knowledge, this is the first published method in Italy for the detection of PFASs in human urine. 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

Tuberculosis (TB) remains a major global cause of morbidity and mortality. Current tools for monitoring treatment response rely on sputum-based microscopy and culture, which are often insensitive, time-consuming, and impractical in extrapulmonary or pediatric TB and in individuals unable to produce sputum. Metabolomics has emerged as a promising approach for identifying host-derived biomarkers that reflect treatment-associated immunometabolic changes; however, the available evidence remains heterogeneous and has not been comprehensively synthesized. We conducted a comprehensive literature review of human studies evaluating metabolomic biomarkers in relation to TB treatment response or outcomes. PubMed, Scopus, and EMBASE were searched for human studies evaluating targeted or untargeted metabolomics (NMR, LC-MS, GC-MS, CE-MS) in relation to treatment response or outcomes. Two reviewers independently screened studies, extracted data, and assessed risk of bias using QUIPS and PROBAST. Findings were synthesized using a structured framework organized across treatment stages and outcomes. Of 218 records identified, 139 titles and abstracts were screened and 42 full texts assessed; 15 studies met the inclusion criteria. Recurrent treatment-associated signals involved amino acid metabolism, particularly the tryptophan–kynurenine pathway, as well as vitamin and cofactor metabolites (pyridoxate, nicotinamide, trigonelline). Plasma studies frequently reported lipid remodeling and bile acid perturbations, whereas urine studies highlighted polyamine metabolism (e.g., N¹,N¹²-diacetylspermine) and fatty acid β-oxidation markers. Common limitations included inadequate adjustment for confounders and, in prediction models, small sample sizes and limited external validation. Metabolomics reveals reproducible but heterogeneous immunometabolic changes during TB therapy. Key pathways include tryptophan–kynurenine metabolism, vitamin and cofactor metabolism, lipid remodeling, and urine polyamine pathways. Standardization and prospective multicenter validation are needed for clinical translation. Full article

Background: Urinary tract infections (UTIs) are prevalent bacterial infections associated with significant morbidity and healthcare burden. Traditional diagnosis relies on urine culture, which is limited by long turnaround times and potential contamination. Automated urine flow cytometry, particularly the UF-5000 (Sysmex Corporation, Kobe, Japan), offers rapid and precise screening through bacterial and white blood cell (WBC) quantification. This systematic review and meta-analysis evaluates the diagnostic accuracy of the UF-5000 for UTI screening. Methods: We searched PubMed, Embase, Science Direct, and Web of Science for studies assessing the UF-5000’s performance, including sensitivity and specificity, with a minimum sample size of 40 and at least 10 UTI cases. Quality assessment was performed using QUADAS-2. Pooled estimates for sensitivity, specificity, and agreement (kappa) were calculated using random-effects models. Results: Eighteen studies, encompassing 25,337 samples, were included in the analysis. Pooled sensitivity and specificity for bacterial count (nine studies) were 0.927 (95% CI, 0.872–0.959) and 0.751 (95% CI, 0.558–0.878), respectively. For WBC count (four studies), sensitivity was 0.897 (95% CI, 0.755–0.961) and specificity was 0.600 (95% CI, 0.293–0.844). The UF-5000 also demonstrated moderate agreement (pooled kappa 0.52, 95% CI, 0.08–0.79) in distinguishing Gram-negative bacteria. Conclusions: Despite heterogeneity across studies, the UF-5000 demonstrates high diagnostic accuracy, particularly high sensitivity, supporting its role as a useful UTI screening tool to rule out infection in clinical settings. The device further provides clinical value through its ability to assist in the differentiation of Gram-negative bacteria. Full article

Background/Objectives: Synthetic stimulants represent the most prevalent subclass on the new psychoactive substances (NPSs) market. However, the toxicokinetic properties of M-ALPHA, a regioisomer of MDMA and N-methyl-cyclazodone a pemoline derivative, are not yet characterized. Methods: Therefore, this study investigated the metabolism of both NPSs in pooled liver S9 fraction and rat urine, characterized cytochrome P450 (CYP) kinetics and plasma protein binding (PPB), and assessed the CYP inhibition potential of M-ALPHA, using high-performance liquid chromatography coupled to high resolution tandem mass spectrometry (HPLC-HRMS/MS). Results: Four metabolites of M-ALPHA were detected including one phase I and three phase II metabolites, resulting from demethylenation followed by subsequent methylation or glucuronidation. For N-methyl-cyclazodone, one phase I metabolite formed via N-demethylation was identified. The primary enzymes involved in M-ALPHA metabolism were CYP2B6 and CYP2D6. Notably, M-ALPHA inhibited these enzymes to a strong or moderate extent, respectively. In contrast, the metabolism of N-methyl-cyclazodone was primarily mediated by CYP2A6. PPB studies indicated low-to-moderate binding for both compounds, suggesting that significant protein-binding interactions are unlikely. Conclusions: As M-ALPHA only formed metabolites that overlapped with those of MDMA, differing only by minor retention time shifts, reliable HPLC-HRMS/MS-based identification may be challenging in clinical and forensic toxicology settings as well as doping analysis. Furthermore, drug–drug interactions following polydrug use cannot be excluded for either NPS, particularly when co-ingested with other CYP substrates metabolized by the same isoforms. 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

Sepsis-associated acute kidney injury (SA-AKI) remains a major diagnostic challenge in critically ill patients, as conventional functional criteria—serum creatinine and urine output—often detect AKI after clinically relevant pathophysiological derangement has already evolved. Increasing evidence suggests that SA-AKI reflects a heterogeneous process characterized by early cellular stress, microcirculatory dysfunction, inflammation-associated injury, and maladaptive repair preceding overt functional decline. In this context, biomarker-based approaches have been investigated to improve early risk stratification, phenotypic characterization, and prognostic assessment in septic patients. This narrative review synthesizes current evidence on established and emerging biomarkers relevant to SA-AKI, encompassing stress markers ([TIMP-2]•[IGFBP7]), tubular injury markers (e.g., NGAL, KIM-1, IL-18), functional markers (e.g., proenkephalin/penKid, cystatin C), and exploratory molecular signatures such as circulating microRNAs (miRNAs). We examine their temporal dynamics, performance estimates, and context-dependent applicability in sepsis, and discuss limitations related to heterogeneity, assay variability, and threshold standardization. Particular attention is given to multimodal and longitudinal strategies integrating biomarkers with KDIGO criteria and clinical phenotyping. Finally, we outline a stratified framework for biomarker interpretation in SA-AKI anchored to pathophysiological windows and clinical decision points. While available evidence supports the potential of selected biomarkers for short-term risk stratification and trajectory assessment, implementation requires prospective validation demonstrating incremental value beyond established models and measurable impact on patient-centered outcomes. Full article

The conventional window for ultrasonic pregnancy diagnosis in sows is 22–25 days post-insemination, which often results in missed opportunities for the optimal re-insemination of non-pregnant sows and elevated production costs. This present study aimed to establish an early pregnancy detection method for sows at 12–18 days post-insemination, thereby providing a reference for efficient reproductive management. Saliva, urine and vaginal secretions were collected from sows during this period, and seven metabolites were quantified. Seven machine learning models were employed for data analysis, after which the optimal combination was determined, and the detection protocol was refined using recursive feature elimination. The results revealed that the majority of metabolites in saliva and urine differed significantly between pregnant and non-pregnant groups (p < 0.05). Among the models evaluated, the random forest algorithm exhibited the best predictive performance, with accuracy ranging from 0.59 to 1.00. Saliva sampled at 17 days post-insemination was identified as the optimal diagnostic medium, and 100% prediction accuracy was achieved by measuring only three metabolites: Glc, Ste, and Xan. The diagnostic approach established in this study allows pregnancy detection 5–8 days earlier than conventional methods, with the additional benefits of non-invasive sampling and minimal stress to sows. Accordingly, it provides a novel reference for enhancing the efficiency of swine production. Full article

Porcine reproductive and respiratory syndrome virus (PRRSV)-positive pigs can be exposed to high concentration of gases, such as ammonia and hydrogen sulfide when manure and urine stored in the pits beneath them is agitated and pumped. Such acute exposure can lead to adverse health effects such as respiratory system irritation. This study aimed to explore whether PRRS-positive growing pigs experience changes in viremia detection after manure pit agitation and pumping has been performed. To address this objective, two PRRS-positive growing pig farms were conveniently selected and visited twice during the week before and after manure agitation and pumping. Blood samples were collected to assess detection of viremia, using reverse transcription-polymerase chain reaction (RT-PCR). A logistic regression model was used to evaluate serum detection of PRRSV before and after the manure management event, accounting for pig age. Although PRRSV was detected in the serum of some pigs, under the conditions of the study, there were no statistically significant changes that would indicate that viremia detections change after the pigs had been exposed to barn manure pit agitation and pumping. Full article

Background and Objectives: Acute kidney injury (AKI) is a frequent and life-threatening complication in patients with liver cirrhosis (LC). Nephrotic-range proteinuria may reflect underlying structural renal vulnerability; however, its association with AKI severity in cirrhosis remains unclear. Materials and Methods: We conducted a retrospective cohort study of 408 adults with LC admitted to a tertiary referral hospital between January 2016 and December 2025. Nephrotic-range proteinuria was defined as a urine protein-to-creatinine ratio (UPCR) ≥3.5 g/g measured within 7 days before or at admission. AKI was staged using serum creatinine-based Kidney Disease: Improving Global Outcomes criteria. Baseline creatinine was defined as the lowest value within 7 days before admission; if unavailable, the lowest stable value within the preceding 3 months was used. Severe AKI was defined as KDIGO stage 2–3. Multivariable logistic regression was performed to evaluate the association between nephrotic-range proteinuria and severe AKI after adjustment for age, sex, diabetes mellitus, hypertension, chronic kidney disease (CKD), sepsis, ICU admission, and Child–Pugh class. Results: Of the 408 patients, 56 (13.7%) had nephrotic-range proteinuria. Severe AKI occurred more frequently in patients with nephrotic-range proteinuria than in those without (39.3% vs. 21.9%), corresponding to an absolute risk difference of 17.4 percentage points (p = 0.008). In the adjusted model, nephrotic-range proteinuria was associated with a higher likelihood of severe AKI (adjusted odds ratio [OR], 2.27; 95% confidence interval [CI], 1.17–4.41; p = 0.015). CKD (adjusted OR, 2.26; 95% CI, 1.33–3.81; p = 0.002), ICU admission (adjusted OR, 2.03; 95% CI, 1.22–3.39; p = 0.007), and Child–Pugh class C versus A (adjusted OR, 3.50; 95% CI, 1.37–8.93; p = 0.009) were also significantly associated with severe AKI. Conclusions: Among hospitalized patients with LC, nephrotic-range proteinuria was associated with a higher likelihood of severe AKI. Quantitative proteinuria assessment may help identify patients at increased risk of advanced renal dysfunction, although causal inference is limited by the retrospective observational design. Full article