Search Results (96)

Music festivals have emerged as venues for the consumption of recreational drugs and novel psychoactive substances. This systematic review provides the first critical evaluation and synthesis of published wastewater analyses for detecting recreational drug use at music festivals worldwide. Following Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines, we systematically searched PubMed, Embase, MEDLINE, and SCOPUS databases using terms combining music festivals, drugs, and wastewater analysis. Twenty-three studies were included, spanning festivals with 6200 to 600,000 attendees. Two primary sampling approaches emerged: wastewater sampling (16 studies) and pooled urine sampling (7 studies), using Liquid Chromatography–Tandem Mass Spectrometry or Liquid Chromatography–High-Resolution Mass Spectrometry for chemical analysis. 3,4-Methylenedioxymethamphetamine (MDMA; ecstasy) emerged as the most consistently detected substance. Regional variations included a dominance of methamphetamine in Eastern Europe, MDMA use in Western Europe, and a high prevalence of cocaine use in South America. Regarding the music genre, electronic dance music events showed markedly higher MDMA rates. Limitations include geographic underrepresentation of African and Asian countries and a gender bias in pooled urine sampling. Future research should work on enhancing sampling infrastructure, analytical capabilities, public health surveillance, and harm reduction strategies. Full article

West Nile virus (WNV) is a mosquito-borne pathogen of the Flaviviridae family that poses recurring public health threats in Tunisia, where Culex pipiens is recognized as the primary vector. Identification of circulating strains in different mosquito species is essential for targeted prevention and control. Between November 2021 and October 2022, mosquitoes were collected at four high-risk sites, and human samples were obtained through the national meningitis surveillance program. Human serum, cerebrospinal fluid (CSF), and urine samples were tested for WNV-specific IgM and IgG antibodies using ELISA, and molecular diagnosis was performed using Real-time RT-PCR (RRT-PCR). Positive samples underwent sequencing for phylogenetic characterization. Serological investigation on human serum revealed the presence of IgM and/or IgG antibodies reactive to WNV antigens, which may indicate exposure to WNV or related flaviviruses. RNA of WNV was detected in 21 mosquito pools (10.19%) belonging to Culex pipiens, Cx. perexiguus, Aedes caspius, and Ae. detritus, as well as in three human cases. Phylogenetic analysis of positive human and mosquito samples showed that all detected WNV strains belonged to sublineage 1a. The concurrent detection of WNV in vectors and humans confirms active circulation in Tunisia and underscores the role of Culex spp. Mosquitoes in transmission. Sustained multidisciplinary surveillance integrating entomological and clinical data is critical for early detection, guiding control measures, and preventing future outbreaks in humans and animals. Full article

Background/Objectives: The limited specificity of prostate-specific antigen (PSA) drives unnecessary biopsies in prostate cancer (PCa). Urinary extracellular vesicles (uEVs) provide a non-invasive reservoir of tumor-derived nucleic acids and proteins. Aptamers selected by SELEX enable highly specific capture, and artificial intelligence (AI) can accelerate their optimization. This systematic review evaluated AI-assisted SELEX for urine-derived and exosome-enriched aptamer panels in PCa detection. Methods: Systematic searches of PubMed, Scopus, and Web of Science (1 January 2010–24 August 2025; no language restrictions) followed PRISMA 2020 and PRISMA-S. The protocol is registered on OSF (osf.io/b2y7u). After deduplication, 1348 records were screened; 129 studies met the eligibility criteria, including 34 (26.4%) integrating AI within SELEX or downstream refinement. Inclusion required at least one quantitative metric (dissociation constant K_d, SELEX cycles, limit of detection [LoD], sensitivity, specificity, or AUC). Risk of bias was appraised with QUADAS-2 (diagnostic accuracy studies) and PROBAST (prediction/machine learning models). Results: AI-assisted SELEX workflows reduced laboratory enrichment cycles from conventional 12–15 to 5–7 (≈40–55% relative reduction) and reported K_d values spanning low picomolar to upper nanomolar ranges; heterogeneity and inconsistent comparators precluded pooled estimates. Multiplex urinary panels (e.g., PCA3, TMPRSS2:ERG, miR-21, miR-375, EN2) yielded single-study AUCs between 0.70 and 0.92 with sensitivities up to 95% and specificities up to 88%; incomplete 2 × 2 contingency reporting prevented bivariate meta-analysis. LoD reporting was sparse and non-standardized despite several ultralow claims (attomolar to low femtomolar) on nanomaterial-enhanced platforms. Pre-analytical variability and absent threshold prespecification contributed to high or unclear risk (QUADAS-2). PROBAST frequently indicated high risk in participants and analysis domains. Across the included studies, lower K_d and reduced LoD improved analytical detectability; however, clinical specificity and AUC were predominantly shaped by pre-analytical control (matrix; post-DRE vs. spontaneous urine) and prespecified thresholds, so engineering gains did not consistently translate into higher diagnostic accuracy. Conclusions: AI-assisted SELEX is a promising strategy for accelerating high-affinity aptamer discovery and assembling multiplex urinary panels for PCa, but current evidence is early phase, heterogeneous, and largely single-center. Priorities include standardized uEV processing, complete 2 × 2 diagnostic reporting, multicenter external validation, calibration and decision impact analyses, and harmonized LoD and K_d reporting frameworks. Full article

Urine analysis is a straightforward, non-invasive testing method that, when integrated with metabolomics, shows great potential for detecting small-molecule metabolites as biomarkers of abnormal metabolic activity in the urinary tract, including drug interactions, toxicity, and diseases. However, integrated and comparative analyses of multiple urinary tract pathologies are currently limited. In this study, ¹²C2/¹³C2-chemical dansylation labeling was used to explore the urinary amine/phenol-metabolome profiles of eight urological conditions compared with normal profiles. We obtained ten samples for each condition (disease and normal) from a total of 90 participants, pooling them as representative samples, and constructed metabolite panels to differentiate various urological conditions. We discovered nine metabolites that were dysregulated between urine samples from patients with and without cancer. Another seven metabolites were differentially expressed between the benign prostatic hyperplasia group and the prostate cancer group. Among 1854 peak pairs of metabolites in an amine/phenol submetabolome analyzed by dansyl chloride derivatization coupled with LC–MS/MS, 1747 (94.2%) were detectable in urine specimens from all nine groups. Notably, 18 identified metabolites showed substantial stability across all urological conditions. Given the considerable variability in urine metabolite composition, these metabolites could potentially be used for normalization in urine metabolome analysis, addressing the need for stably expressed molecules as internal standards in the development of urinary biomarkers. Our findings provide the preliminary insights into the stability of urinary metabolomics and the metabolic perturbations associated with different urinary tract-related pathologies. Full article

Background: Quantitative urine culture (CFU/mL) remains the reference standard for diagnosing urinary tract infections (UTIs) but is limited by delayed turnaround times and sensitivity to pre-analytic factors. Multiplex PCR panels offer rapid detection; however, standardized mappings between molecular signals and viable bacterial burdens are insufficiently defined. We used the multicenter NCT06996301 paired dataset to evaluate the analytical validity (AV), clinical validity (CV), and pre-analytic robustness of ΔCt (Ct_target − IC_Ct) as a semi-quantitative indicator of bacterial load. Methods: We analyzed 1027 paired PCR and quantitative urine culture specimens from six sites. The primary molecular predictor was ΔCt (Ct_target − IC_Ct). Species-level Spearman and Pearson correlations, species-specific linear mixed-effects calibration models (log₁₀CFU ~ ΔCt + (1|site)), and ROC analyses were performed for the taxa meeting pre-specified sample thresholds. A pooled multilevel model assessed the collection method and time-to-processing (hours) effects (log₁₀CFU ~ ΔCt × collection_method + ΔCt × time_to_processing_h + (1|site) + (1|run)). AV was assessed via reproducibility, internal control normalization, and site run variance. CV was assessed by ΔCt calibration and discrimination. Clinical utility (CU) was contextualized using outcomes from the parent randomized trial. Results: PCR positivity exceeded culture positivity across all sites (PCR ~82–88% vs. culture ~66–70%); this excess likely reflects a combination of molecular detection of non-viable DNA, detection of fastidious taxa less readily recovered by culture, and pre-analytic effects. For six common uropathogens (n = 90 pairs/species), ΔCt correlated strongly with log₁₀CFU (Spearman ρ = −0.64 to −0.75; Pearson r = −0.75 to −0.83). Species-specific mixed models yielded slopes of −0.746 to −0.922 log₁₀CFU per ΔCt unit (all p < 0.001), indicating that each 1 unit ΔCt change corresponds to a ~5.6–8.4-fold CFU difference. ROC AUCs for ΔCt discrimination were 0.78–0.84 (interpreted as good discrimination, i.e., ΔCt meaningfully improves the clinician’s probability estimate of a high CFU but does not perfectly classify every specimen). The collection method (catheter vs. clean-catch) did not materially modify the ΔCt→CFU relationship, whereas the processing delay was associated with reduced recovered CFU (~0.048 log₁₀CFU lost per hour) and a significant ΔCt × time interaction, consistent with time-dependent viability loss driving the PCR⁺/culture⁻ discordance. Conclusions: ΔCt from the DOC Lab UTM 2.0 panel shows a reproducible, analytically valid semi-quantitative measure of urinary bacterial load. Laboratories can derive assay- and workflow-specific ΔCt cut points for semi-quantitative reporting, but thresholds must be validated prospectively and paired with operational controls for specimen handling. Full article

Background: Here, we validate a unique and rapid susceptibility assay, Pooled Antibiotic Susceptibility Testing (P-AST), used for complicated, persistent, and recurrent urinary tract infections (UTIs), following Clinical and Laboratory Standards Institute (CLSI) protocols and performance metrics. Methods: P-AST™ was validated against the standard disk diffusion method with discrepancy resolution by the broth microdilution reference method. Performance was evaluated for five groups of non-fastidious uropathogenic organisms (Enterobacterales, Enterococci, Staphylococci, Pseudomonas aeruginosa, and Acinetobacter species) for up to 20 antibiotics, as clinically relevant per group. Fresh (144 monomicrobial and 49 polymicrobial) and frozen (78 monomicrobial and 7 polymicrobial) clinical urine specimens, as well as contrived specimens from pre-characterized frozen “challenge” isolates (52 monomicrobial and 37 polymicrobial), were included. Results: P-AST met CLSI target performance criteria of ≥90.0% categorical agreement, <3.0% very major error, <3.0% major error, minor error ≤ 10.0%, or within laboratory standards, and precision > 95.0% across all analysis groups. Across all monomicrobial analyses, there were no very major errors (VMEs), and two major errors (MEs). Across all polymicrobial analyses, there were three VMEs and two MEs. No organism–antibiotic pair analysis had more than a single VME or ME. Conclusions: P-AST, a component of the Guidance^® UTI assay, demonstrates acceptable performance within the thresholds established by CLSI when compared against standard and reference methods for antibiotic susceptibility testing. Appropriate performance was established in both monomicrobial and polymicrobial specimens for five CLSI-defined groups of uropathogenic bacteria, against up to 20 antibiotics as clinically relevant to each organism group. Full article

Vicadrostat, a selective aldosterone synthase inhibitor, reduced albuminuria with concurrent renin–angiotensin system inhibition and empagliflozin, suggesting additive efficacy for chronic kidney disease (CKD) treatment. Specific urinary extracellular vesicle microRNAs (uEV miRNAs) may reflect key mechanisms of kidney injury. We investigated how vicadrostat alone or with empagliflozin affected uEV miRNA expression in study participants. Small RNA sequencing was conducted on uEV miRNAs from 435 participants with CKD who completed 14 weeks treatment in the phase II trial of vicadrostat given with or without empagliflozin. Differentially expressed uEV miRNAs in participants with ≥30% UACR (urine albumin–creatinine ratio) reduction treated with 10 or 20 mg vicadrostat were pooled and evaluated with or without empagliflozin. Changes in miRNA-142-5p correlated significantly with changes in UACR in participants treated with vicadrostat alone, whereas changes in expression of eight additional uEV miRNAs (miR-192-5p, miR-194-5p, miR-6882-5p, miR-27a-5p, miR-381-3p, miR-192-3p, miR-513a-5p, and miR-199b-3p) correlated with ≥30% UACR improvements in patients treated with vicadrostat plus empagliflozin. Cellular deconvolution revealed that these miRNAs were expressed in various kidney cell types. Vicadrostat plus empagliflozin altered uEV miRNAs involved in immunomodulatory and fibrotic pathways irrespective of participant diabetes status. Regulation of miRNAs may provide insights into synergistic mechanisms of vicadrostat and empagliflozin in CKD treatment. Full article

Background/Objectives: The association between plasma metabolites derived from dietary substrates and inflammatory processes remains underexplored, despite its potential relevance in the prevention of non-communicable diseases. This systematic review aimed to examine the relationship between blood metabolites and the modulation of inflammatory biomarkers. Methods: A total of 25 randomized controlled trials, published between 2019 and 2024, were included from an initial pool of 111 records. These studies investigated the effects of dietary patterns, specific food groups, or nutritional supplements on the human metabolome and their potential links to inflammation. Results: Metabolomic analyses were predominantly performed using mass spectrometry (MS)-based platforms (17 out of 25), with liquid chromatography–mass spectrometry as the most frequently employed method. Both targeted (n = 14) and untargeted (n = 11) approaches were represented, and samples were drawn from plasma, urine, and feces. Across the interventions, 64 metabolites were modulated, including fatty acyls, glycerolipids, benzenoids, and organic acids, reflecting potential changes in pathways related to oxidative stress, lipid and carbohydrate metabolism, and inflammatory signaling. Several studies also assessed classical inflammatory biomarkers such as C-reactive protein (CRP), tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1). Interventions involving healthy traditional dietary patterns, improvements in dietary fat quality, or the use of specific probiotic strains were often associated with favorable immunometabolic outcomes. In contrast, some interventions, such as Mohana Choorna, elicited upregulation of immune-related gene expression in adipose tissue without improvements in glucose or lipid metabolism. Conclusions: While metabolomic responses varied across studies, the evidence highlights the value of dietary interventions in modulating systemic metabolism and inflammation. These findings support the integration of metabolomics into clinical nutrition to define more personalized and effective dietary strategies for inflammation-related chronic disease prevention. Full article

Coeliac disease (CD) is a chronic immune-mediated enteropathy triggered by gluten ingestion in genetically predisposed individuals carrying HLA-DQ2 or HLA-DQ8 haplotypes, characterized by small intestinal mucosal damage and systemic manifestations. This systematic review and meta-analysis aimed to compare the effectiveness of urine versus stool GIPS testing for monitoring gluten intake in coeliac patients, providing evidence-based recommendations for clinical practice. A comprehensive literature search was conducted in databases like PubMed and Embase. Studies evaluating urine or stool GIPS testing in coeliac patients were included, focusing on sensitivity, specificity, and patient adherence. The meta-analysis included six studies with a total of 572 participants. The stool GIPS testing demonstrated a pooled sensitivity of 85.1% (95% CI: 79.0–89.9%) and a specificity of 92.5% (95% CI: 88.3–95.6%), making it highly reliable for detecting gluten exposure and ruling out false positives. It also achieved an AUC of 0.9853, indicating excellent diagnostic performance. In contrast, the urine GIPS testing showed a pooled sensitivity of 55.4% (95% CI: 49.6–61.2%) and a specificity of 73.0% (95% CI: 67.4–78.1%), with an AUC of 0.7898. The heterogeneity across the studies was significant (I² > 80%), driven by variations in the population characteristics, sample handling, and testing protocols. These findings emphasize the need for standardized methodologies to enhance the reliability and comparability of results. Full article

Background: Dehydration is a prevalent and potentially serious condition, particularly affecting vulnerable populations such as children and older adults. Prompt recognition and intervention are critical for preventing associated complications. Methods: A systematic review and meta-analysis were conducted, registered in PROSPERO (CRD42024594780), to identify key clinical and demographic risk factors associated with dehydration. A comprehensive search of PubMed, Scopus, and the Cochrane Library was performed for studies published between 2000 and 2024. The risk of bias in included studies was assessed using the Newcastle–Ottawa Scale and the Cochrane Risk-of-Bias (RoB) tool. Ten studies met the inclusion criteria for quantitative synthesis. Based on pooled diagnostic metrics, a preliminary scoring tool was developed for dehydration risk stratification. Results: The pooled sensitivity and specificity of common clinical signs, such as thirst, dry mouth, and dark urine, were 85% (95% CI: 80–90%) and 70% (95% CI: 65–75%), respectively. The positive predictive value (PPV) was 75%, and the negative predictive value (NPV) was 80%. Pediatric subgroup analysis yielded the most robust data, while data for adult and elderly populations were limited. A conceptual risk scoring system was proposed based on relative diagnostic utility, though it has not yet been externally validated. Conclusions: Simple clinical signs demonstrate reasonable diagnostic accuracy for identifying individuals at risk of dehydration. The proposed scoring system offers a promising, evidence-informed framework for early risk assessment but requires further validation in prospective studies before integration into clinical practice. Full article

Background/Objectives: The emergence of cathinone-based psychostimulants necessitates ongoing research and analysis of the characteristics and properties of novel derivatives. The metabolic fate of five novel cathinone-derived substances (ASProp, MASProp, MASPent, PySProp, and PySPent) containing a selenophene moiety was investigated in vitro and in vivo. Methods: All compounds were incubated individually with pooled human liver S9 fraction. A monooxygenase activity screening investigating the metabolic contribution of eleven recombinant phase I isoenzymes was conducted. Rat urine after oral administration was prepared by urine precipitation. Liquid chromatography–high-resolution tandem mass spectrometry was used for the analysis of all samples. Reversed-phase liquid chromatography (RPLC) and zwitterionic hydrophilic interaction liquid chromatography (HILIC) were used to evaluate and compare the metabolites’ chromatographic resolution. Results: Phase I reactions of ASProp, MASProp, MASPent, PySProp, and PySPent included N-dealkylation, hydroxylation, reduction, and combinations thereof. The monooxygenase activity screening revealed the contribution of various isozymes. Phase II reactions detected in vivo included N-acetylation and glucuronidation. Both chromatographic columns complemented each other. Conclusions: All substances revealed metabolic reactions comparable to those observed for other synthetic cathinones. Contributions from isozymes to their metabolism minimized the risk of drug–drug interactions. The identified metabolites should be considered as targets in human biosamples, especially in urine screening procedures. RPLC and HILIC can both be recommended for this purpose. Full article

This study evaluated the performance of urine reagent strips (URSs) in detecting Schistosoma haematobium infection in individual and pooled urine samples. Between June 2022 and April 2023, 2634 urine samples (10 mL each) from school-age children (5–15 years) in 15 villages across Ethiopia’s Afar, Benishangul-Gumuz, and Gambella regions were tested using urine filtration microscopy (UFM) and URSs for blood, a marker of S. haematobium eggs. Pooled samples from 5, 10, 20, and 40 individuals (one positive, others negative) were examined with both methods. UFM results were used to calculate URSs’ sensitivity, specificity, and predictive values for detecting infection. A total of 2634 children were screened for S. haematobium infection. UFM detected S. haematobium eggs in 370 samples, while URSs identified infection in 414 children. URSs showed 64% sensitivity and 92% specificity for individual samples. The positive and negative predictive values for individual samples were 57% and 94%, respectively. Sensitivity for pooled samples ranged from 47% (pools of 40) to 53% (pools of 20). In pools with one positive sample, URSs misclassified 220 (50%), 109 (49.5%), 52 (47.0%), and 28 (50.9%) pools as negative for S. haematobium eggs for pool sizes 5, 10, 20, and 40, respectively. Sensitivity for individual samples was higher in children with heavy infection (92.5%) compared to light infection (55.9%), and sensitivity in pooled samples increased with infection intensity (p < 0.001). In conclusion, URSs may misclassify S. haematobium infection in children when samples are examined individually or in pools, potentially leading to unnecessary treatment or missed cases. However, URSs shows promise as a screening tool for detecting S. haematobium infection in areas with high infection intensity. Full article

Background/Objectives: Urinary tract infections (UTIs) pose an increasing risk of antimicrobial resistance, and novel diagnostic tests have been developed to address the limitations of standard urine culture in these cases. It is important that these novel tests be validated for agreement and error rates against the standard antibiotic susceptibility testing (AST) methods. Methods: Polymicrobial (≥two non-fastidious microorganisms) consecutive clinical urine specimens submitted for UTI diagnostic testing were included in this analysis. Specimens were tested with Pooled Antibiotic Susceptibility Testing (P-AST) and with broth microdilution/disk diffusion (BMD/DD) in parallel. Performance characteristics, such as essential agreement (EA%), very major errors (VMEs), and major errors (MEs), were assessed using Clinical and Laboratory Standards Institute (CLSI) standards. Specimens with P-AST-resistant and BMD/DD consensus-sensitive results were assessed for heteroresistance. Real-world clinical sample data were used to assess associations between increasing organism counts and average “sensitive” antibiotic count per sample. Results: The essential agreement between P-AST and standard isolate AST was ≥90%, VMEs were <2.0%, and MEs were <3.0%, meeting the CLSI guidelines for AST verification and validation studies. When heteroresistance was accounted for, overall VMEs and MEs were both <1.5%. The presence of additional non-fastidious organisms dropped the number of average “sensitive” antibiotics from 9.8 with one organism to 2.5 with five or more organisms. The presence of fastidious organisms did not have any meaningful impact. Conclusions: P-AST, a component of the Guidance^® UTI assay (Pathnostics, Irvine, CA, USA), performed within CLSI standards for AST in polymicrobial UTI diagnostic urine specimens. Full article

Background/Objectives: While new methods for measuring antimicrobial susceptibility have been associated with improved patient outcomes, they should also be validated using standard protocols for error rates and other test metrics. The objective of this study was to validate a novel susceptibility assay for complicated and recurrent urinary tract infections (UTIs): pooled antibiotic susceptibility testing (P-AST). This assay was compared to broth microdilution (BMD) and disk diffusion (DD), following Clinical and Laboratory Standards Institute (CLSI) guidelines for assessment of error rates and agreement. Methods: This study analyzed consecutive fresh clinical urine specimens submitted for UTI diagnostic testing. Upon receipt, the urine samples were subjected in parallel to standard urine culture and multiplex polymerase chain reaction (M-PCR) for microbial identification and quantification. Specimens with the same monomicrobial non-fastidious bacteria detected by both M-PCR and standard urine culture (SUC) underwent standard antibiotic susceptibility testing (AST) and P-AST antibiotic susceptibility testing. Analysis was also undertaken to assess the presence of heteroresistance for specimens with P-AST-resistant and BMD/DD consensus-susceptible results. Results: The performance measures without correction for heteroresistance showed essential agreement (EA%) of ≥90%, very major errors (VMEs) of <1.5%, and major errors (MEs) of <3.0% for P-AST, all meeting the threshold guidelines established by CLSI for AST. The categorical agreement (CA%) also met acceptable criteria (>88%), as the majority of the errors were minor (mEs) with essential agreement. The very major and major error rates for P-AST decreased to <1.0% when heteroresistance was accounted for. Conclusions: The P-AST assay methodology is validated within acceptable parameters when compared to broth microdilution and disk diffusion using CLSI criteria. Full article

Background/Objectives: We aimed to compare the prescribing behavior and clinical experience of urology providers when using the combined multiplex polymerase chain reaction (M-PCR)/Pooled Antibiotic Susceptibility Testing (P-AST) diagnostic test versus the standard urine culture (SUC) in the same set of patients previously reported to have improved clinical outcomes with M-PCR/P-AST. Methods: We conducted a multi-centered, prospective, observational study (clinical trial registration: NCT05091931) with Western Institutional Review Board (IRB) approval (20214705). Adult subjects were split between the M-PCR/P-AST (n = 250) and SUC arms (n = 135). Treatment details were determined by clinician and subject surveys. Differences in prescribed antibiotics were compared using the Chi-square or Fisher’s exact test. Results: There was no significant difference in the overall use of “access” antibiotics (p = 1.0) or first-line drugs (p = 0.4483) between M-PCR/P-AST and SUC. Nitrofurantoin (p = 0.0172) and metronidazole (p = 0.0309) were more frequently used with M-PCR/P-AST, while amoxicillin/clavulanate (p = 0.0008), cefuroxime (p = 0.0378), and ertapenem (p = 0.0378) were more frequently used with SUC. Conclusions: The use of M-PCR/P-AST to guide complicated UTI management was not associated with the increased use of non-first-line antibiotics, such as carbapenems, compared to SUC. Combined with the prior reported evidence of improved patient outcomes in this same set of patients, this test should be considered for utilization when managing complicated UTI cases. Full article