Search Results (236)

Exosomes are nanosized vesicles that carry intracellular mediators and their abundance in urine opens new and intriguing possibilities in nephrology since they provide a non-invasive insight into kidney diseases. The aim of this review is to examine the main applications of urinary exosomes in nephropathies. Urinary exosomes are isolated through ultrafiltration, ultracentrifugation, precipitation, and immunoaffinity chromatography. After isolation they are characterized through Western blotting, flow cytometry, and, more recently, with mass spectrometry. Through the analysis of urinary exosomes, it has been possible to distinguish patients with IgA nephropathy from healthy controls. Different profiles of expression have been identified between patients with MCD and FSGS. A distinct exosomal composition has been discovered in patients with lupus nephropathy when compared to those without renal involvement. Significant findings have been reported also in patients with monoclonal gammopathy of renal significance, allowing a differential diagnosis between LCDD and amyloidosis. Among kidney transplant recipients, the analysis of urinary exosomes highlighted differences between antibody-mediated rejection and cell-mediated rejection. Urinary exosomes are new non-invasive, promising biomarkers and potential therapeutic options that have already shown interesting results in the nephrological field. Further studies are needed to harness their potential and diffusion. Full article

Today’s linear nutrient flows are rooted in a long history of agronomic and wastewater engineering strategies that have created cascading environmental, social, and economic side effects, signaling the need for more holistic and circular approaches. Our examination of the regulatory pathways that enable and constrain urine recycling—an underutilized approach to repurposing human waste as fertilizer—addresses a persistent research gap related to the mainstreaming of transformative technologies. Framed around policy process theories—Street Level Bureaucracy and Multiple Streams Theory—our methods include a review and mapping of 54 regulatory documents; action research where we reflect on our own efforts to expand urine recycling; and interviews with 16 practitioners and regulators in four states which, to our knowledge, are the only places in the US with efforts to scale up urine recycling in community settings. Given its circular nature, a key challenge we find is a lack of clarity around which sectors, or what scales of government, “own” the decision to allow the collection and use of urine as a fertilizer. Working around these challenges, we show how practitioners use many practical strategies to simplify the approval process and reduce the risk aversion regulators face when confronted with ambiguous rulemaking. Full article

Objectives: Silodosin, a selective α1A-adrenoceptor antagonist, is used to treat lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH). Genetic polymorphisms in drug-metabolizing enzymes and transporters may contribute to interindividual variability in its efficacy and safety. This study aimed to investigate the influence of CYP3A4, CYP3A5, UGT2B7, and ABCB1 polymorphisms on silodosin pharmacokinetics, efficacy, and safety in Russian patients with BPH. Methods: A prospective observational study included 103 Russian male patients with moderate-to-severe LUTS (IPSS > 8) due to BPH, treated with silodosin (8 mg daily) for 8 weeks. Genotyping for CYP3A4*1B, CYP3A4*22, CYP3A5*3, UGT2B7 (rs73823859, rs7439366, and rs7668282), and ABCB1 (rs4148738, rs1045642, rs2032582, and rs1128503) was performed using real-time PCR. The silodosin minimum steady-state plasma concentration (Css min) was measured via HPLC-MS. Efficacy was evaluated by the International Prostate Symptom Score (IPSS), quality of life scale, maximum urinary flow rate (Qmax), residual urine volume (RUV), and prostate volume at the baseline and week 8. Adverse drug reactions (ADRs) were recorded. Results: CYP3A4*22 CT carriers (n = 6) exhibited higher Css min (17.59 ± 2.98 vs. 9.0 ± 10.47 ng/mL, p = 0.049) but less absolute IPSS improvement (p < 0.05), likely due to higher baseline symptom severity. However, the change in IPSS (ΔIPSS_1–4) from the baseline to week 8 did not differ significantly (−5.78 ± 5.29 vs. −6.0 ± 4.54, p = 0.939). CYP3A5*3 GG homozygotes (n = 96) showed greater ΔIPSS_1–4 improvement (−6.25 ± 4.60 vs. 0.0 ± 9.53, p = 0.042) and a lower IPSS at day 28 (7.64 ± 4.50 vs. 20.0 ± 6.55, p < 0.001). UGT2B7 rs7439366 TT carriers (n = 34) had an improved Qmax (ΔQmax_1–4 5.4 vs. 3.3 and 2.0 mL/s for CC and CT, p = 0.041). ABCB1 1236C>T TT homozygotes (n = 25) showed a trend toward reduced RUV (p = 0.053). No polymorphisms were associated with adverse drug reactions (15 events in 42 patients, 35.7%). Conclusions: Genetic polymorphisms CYP3A4*22, CYP3A5*3, and UGT2B7 rs7439366 may modulate silodosin pharmacokinetics and efficacy parameters in BPH patients but not safety. Larger-scale studies are warranted to validate these initial findings. Full article

Introduction: Endometriosis is a chronic inflammatory disease affecting women of reproductive age, often accompanied by chronic pelvic pain and infertility. Despite numerous studies, its pathogenesis remains incompletely understood. Increasing evidence indicates the important role of immunological disorders, especially in the mechanisms of innate immunity and Toll-like receptors (TLRs). Study objective: This study aimed to assess the expression of selected TLRs (TLR2, TLR3, TLR4, TLR7, TLR8, and TLR9) on peripheral blood lymphocyte subpopulations (CD4+, CD8+, and CD19+ cells) in patients diagnosed with endometriosis and to quantify the levels of their soluble forms in serum and urine. This study was conducted on patients who were not undergoing hormonal bridging therapy and were not using any form of hormonal contraception to eliminate potential confounding effects on immune parameters. Methods: Flow cytometric analysis of TLR expression on peripheral blood lymphocytes was performed, and the levels of their soluble forms in serum and urine samples were determined. Additionally, ROC curve analysis was used to evaluate the diagnostic potential of the studied parameters. Results: We found increased expression of TLRs in lymphocyte populations in patients with endometriosis compared to the control group, suggesting their involvement in both local and systemic immune responses. In addition, ROC analysis showed the diagnostic potential of TLR expression in differentiating patients with endometriosis from healthy women, and it may also identify disease subtypes. Conclusions: The findings support the role of TLRs in the immunopathogenesis of endometriosis and highlight their promise as diagnostic biomarkers and therapeutic targets. Further studies on larger patient cohorts and functional signaling analyses are warranted. Full article

Collagen type I (COL(I)) is a key component of the extracellular matrix (ECM) and is involved in cell signaling and migration through cell receptors. Collagen degradation produces bioactive peptides (matrikines), which influence cellular processes. In this study, we investigated the biological effects of nine most abundant, naturally occurring urinary COL(I)-derived peptides on human endothelial cells at physiological concentrations, using cell migration assays, mass spectrometry-based proteomics, flow cytometry, and AlphaFold 3. While none of the peptides significantly altered endothelial migration by themselves at physiological concentrations, full-length COL(I) increased cell migration, which was inhibited by Peptide 1 (²²⁹NGDDGEAGKPGRPGERGPpGp²⁴⁹). This peptide uniquely contains the DGEA and GRPGER motifs, interacting with integrin α2β1. Flow cytometry confirmed the presence of integrin α2β1 on human endothelial cells, and AlphaFold 3 modeling predicted an interaction between Peptide 1 and integrin α2. Mass spectrometry-based proteomics investigating signaling pathways revealed that COL(I) triggered phosphorylation events linked to integrin α2β1 activation and cell migration, which were absent in COL(I) plus peptide 1-treated cells. These findings identify Peptide 1 as a biologically active COL(I)-derived peptide at a physiological concentration capable of modulating collagen-induced cell migration, and provide a foundation for further investigation into its mechanisms of action and role in urine excretion. Full article

Background: Urine microbial analysis is a frequently requested test that is often associated with contamination during specimen collection or storage, which leads to false-positive diagnoses and delayed reporting. In the era of digitalization, machine learning (ML) can serve as a valuable tool to support clinical decision-making. Methods: This study investigates the application of a simple artificial neural network (ANN) to pre-identify negative and contaminated (false-positive) specimens. An ML model was developed using 8181 urine samples, including cytology, dipstick tests, and culture results. The dataset was randomly split 2:1 for training and testing a multilayer perceptron (MLP). Input variables with a normalized importance below 0.2 were excluded. Results: The final model used only microbial and either urine color or urobilinogen pigment analysis as inputs; other physical, chemical, and cellular parameters were omitted. The frequency of positive and negative specimens for bacteria was 6.9% and 89.6%, respectively. Contaminated specimens represented 3.5% of cases and were predominantly misclassified as negative by the MLP. Thus, the negative predictive value (NPV) was 96.5% and the positive predictive value (PPV) was 87.2%, leading to 0.82% of the cultures being unnecessary microbial cultures (UMC). Conclusions: These results suggest that the MLP is reliable for screening out negative specimens but less effective at identifying positive ones. In conclusion, ANN models can effectively support the screening of negative urine samples, detect clinically significant bacteriuria, and potentially reduce unnecessary cultures. Incorporating morphological information data could further improve the accuracy of our model and minimize false negatives. Full article

Prostate cancer diagnostics are rapidly advancing through innovations in nanotechnology, biosensing strategies, and molecular recognition. This review analyzes studies focusing on quantum dot (QD)-based biosensors for detecting prostate cancer biomarkers with high sensitivity and specificity. It covers diverse sensing platforms and signal transduction mechanisms, emphasizing the influence of the QD composition, surface functionalization, and bio interface engineering on analytical performance. Key metrics such as detection limits, dynamic range, and compatibility with biological samples, including serum, urine, and tissue, are critically assessed. Recent advances in green-synthesized QDs and smartphone-integrated diagnostic platforms are highlighted, including lateral flow assays, paper-based devices, and pH-responsive hydrogels for real-time, low-cost, and decentralized cancer screening. These innovations enable multiplexed biomarker detection and tumor microenvironment monitoring in point-of-care settings. This review concludes by addressing the current limitations, scalability challenges, and future research directions for translating QD-enabled biosensors into clinically viable diagnostic tools. Full article

This paper describes a novel wearable personal uroflowmeter and its use to log urine leakage episodes in women. Consisting of a miniature flow rate sensor attached under the urethral meatus, it recorded both urine flow rate and volume during activities of daily living. The sensor communicated with a determining unit incorporating a microcontroller and an inertial measurement unit worn at the waist, facilitating the post-hoc determination of which activities and changes in pose caused leakage. Six women participated in a feasibility study performed in a clinical setting. The results indicate that the uroflowmeter was 97.5% accurate in assessing micturition flow compared to gold standard uroflowmetry and leakage measurements. The system also provides subject-specific information on the relationship between physical activity and urine leakage, thereby eliminating errors due to missing data and recall bias in bladder leakage diaries and circumventing the limitations of office-based uroflowmeters. Full article

Background/Objectives: The clinically validated multiplex Oncuria bladder cancer (BC) assay quickly and noninvasively identifies disease risk and tracks treatment success by simultaneously profiling 10 protein biomarkers in voided urine samples. Oncuria uses paramagnetic bead-based fluorescence multiplex technology (xMAP^®; Luminex, Austin, TX, USA) to simultaneously measure 10 protein analytes in urine [angiogenin, apolipoprotein E, carbonic anhydrase IX (CA9), interleukin-8, matrix metalloproteinase-9 and -10, alpha-1 anti-trypsin, plasminogen activator inhibitor-1, syndecan-1, and vascular endothelial growth factor]. Methods: In a pilot study (N = 36 subjects; 18 with BC), Oncuria performed essentially identically across three different common analyzers (the laser/flow-based FlexMap 3D and 200 systems, and the LED/image-based MagPix system; Luminex). The current study compared Oncuria performance across instrumentation platforms using a larger study population (N = 181 subjects; 51 with BC). Results: All three analyzers assessed all 10 analytes in identical samples with excellent concordance. The percent coefficient of variation (%CV) in protein concentrations across systems was ≤2.3% for 9/10 analytes, with only CA9 having %CVs > 2.3%. In pairwise correlation plot comparisons between instruments for all 10 biomarkers, R² values were 0.999 for 15/30 comparisons and R² ≥ 0.995 for 27/30 comparisons; CA9 showed the greatest variability (R² = 0.948–0.970). Standard curve slopes were statistically indistinguishable for all 10 biomarkers across analyzers. Conclusions: The Oncuria BC assay generates comprehensive urinary protein signatures useful for assisting BC diagnosis, predicting treatment response, and tracking disease progression and recurrence. The equivalent performance of the multiplex BC assay using three popular analyzers rationalizes test adoption by CLIA (Clinical Laboratory Improvement Amendments) clinical and research laboratories. Full article

Oxidative stress and hypoxia-induced inflammation contribute to benign prostatic hyperplasia (BPH) progression. This study investigated the roles of urinary inflammatory and oxidative stress biomarkers in BPH patients. This prospective study enrolled 62 clinical BPH patients (33 treated medically, 29 surgically) and 20 controls. Symptom scores, uroflowmetry, and urinary biomarker levels were assessed at baseline and three months post-treatment. Before treatment, BPH patients exhibited elevated urinary levels of total antioxidant capacity (TAC), PGE2, IL-1β, and IL-6. Post-treatment, successful outcomes were reported in 63.6% of the medical treatment group and 86.2% of the surgical treatment group, with improvements in symptom scores and urinary flow rate, along with reductions in urinary 8-isoprostane, TAC, and IL-1β. Prior to treatment, voiding efficiency (VE) was negatively correlated with urinary IL-1β, IL-6, and IL-8 levels, while bladder wall thickness was positively correlated with TAC. After treatment, changes in VE were negatively correlated with changes in IL-1β, and changes in post-void residual urine were positively correlated with changes in IL-1β, IL-6, IL-8, and TNF-α. Urinary inflammatory and oxidative stress biomarkers may serve as non-invasive indicators of disease severity and treatment response in clinical BPH. Their significant correlations with clinical improvements underscore their potential utility in monitoring treatment efficacy. Full article

Background/Objectives: Challenges in normothermic machine perfusion (NMP) remain, particularly concerning the duration for which individual organs can be safely preserved. We hypothesize that optimal preservation can be achieved by perfusing organs together in a multivisceral block. Therefore, our aim was to establish a platform for ex situ multivisceral organ perfusion. Methods: Multivisceral grafts containing the liver, kidneys, pancreas, spleen, and intestine were obtained from Yorkshire pigs. Three generation (gen) set-ups were tested during the iterative design process, and minor changes were made throughout. Gen 1 (n = 4) used a custom-designed single perfusion circuit. Gen 2 (n = 3) employed a dual perfusion circuit. Gen 3 (n = 4) featured a single perfusion circuit with an optimized basin and reservoir. Grafts underwent NMP using an autologous blood-based perfusate, while hemostatic parameters and function were assessed. Results: Comparing Gen 1 versus Gen 3, the mean aortic flow improved (1.018 vs. 2.089 L), resistance decreased (0.224 vs. 0.038), urine output increased (51.90 vs. 271.3 mL), oxygen consumption rose (43.56 vs. 49.52 mL O₂/min), perfusate lactate levels dropped (10.44 vs. 3.10 mmol/L), and the pH became more physiological (7.27 vs. 7.30). Cellular injury trended lower in Gen 3. Histological evaluation demonstrated minimal differences in Gens 2 and 3. Conclusions: We demonstrate the feasibility of abdominal multivisceral NMP for up to 8 h. Adequate arterial flow, stable perfusate pH, and high oxygen consumption in setup 3 indicated organ viability. Multivisceral perfusion may serve as a plat-form for long-term NMP. Full article

Preeclampsia, one of the leading causes of maternal and fetal morbidity and mortality, affects approximately 3–5% of pregnancies worldwide. However, its etiology remains poorly understood. The aim of this study was to identify molecular markers of preeclampsia. Protein concentrations in blood and urine were determined using the Bio-Plex Kidney Toxicity 1 assay Bio-Rad, Hercules, CA, USA followed by magnetic separation and flow cytometry. This study included 51 patients with preeclampsia and 25 healthy pregnant women. The results revealed that five out of the six serum biomarkers of kidney injury were elevated in the preeclampsia group compared to the control group (calbindin 1, clusterin, glutathione transferase pi (GSTP1), monocyte chemotactic protein 1 (MCP-1), and kidney injury molecule type 1 (KIM-1)). Additionally, the serum concentrations of calbindin 1, clusterin, GSTP1, and KIM-1 were significantly higher in both early-onset and late-onset preeclampsia compared to the control group. The analysis of urinary proteins showed that only the KIM-1 concentration was elevated in late-onset preeclampsia compared to the control group. These findings suggest that the calbindin 1, clusterin, GSTP1, KIM-1, and MCP-1 concentrations in maternal plasma could serve as potential biomarkers for monitoring kidney injury in preeclamptic women. This study provides a foundation for future research to explore novel biomarkers of preeclampsia and renal injury in pregnant women. Full article

Acute coronary syndrome (ACS) refers to a spectrum of conditions characterized by a sudden decrease in blood flow to the heart. This includes unstable angina, the mildest form, as well as non-ST- and ST-segment elevation myocardial infarction. The primary cause of ACS is typically the rupture or erosion of an atherosclerotic plaque in a coronary artery, resulting in the formation of a blood clot that can, partially or completely, block the blood flow to the heart muscle. The ongoing discovery and comprehension of emerging biomarkers for atherosclerosis could enhance our capacity to predict future events, particularly when integrated alongside traditional risk factors in assessing overall risk profiles. With advancements in proteomic technologies, large-scale approaches have been increasingly instrumental in unraveling pathways implicated in atherosclerotic degeneration and identifying novel circulating markers, which may serve as early diagnostic indicators or targets for innovative therapies. Over recent decades, numerous matrices including plasma, urine, microparticles, lipoproteins, atherosclerotic plaque extracts and secretomes, as well as thrombi, have been examined to address these questions. Furthermore, proteomics has been applied to various experimental models of atherosclerosis to deepen our understanding of the mechanisms underlying atherogenesis. This review offers a critical overview of the past two decades of untargeted omics research focused on identifying circulating and tissue biomarkers relevant to ACS. Full article

Tritrichomonas foetus is a sexually transmitted protozoan that causes early embryonic death in cattle. A challenge in trichomonosis research is that in vivo studies of treatments, diagnostic strategies, and vaccines are severely hampered by the logistical challenge and cost of maintaining adult bulls. Since natural infections are diagnosed in postpubescent animals, the paradigm is that only mature breeding bulls can be infected. In this study, we hypothesized that prepubescent bull calves could be artificially infected with T. foetus trophozoites for the purpose of conducting research trials. Initial attempts to directly infect bull calves with two different parasite isolates resulted in the sporadic and transient detection of parasite DNA but not culturable trophozoites. In vitro and in vivo studies suggested that urine directly inhibited trophozoites, likely by osmotic damage and mechanical flushing action. Studies utilizing a perineal urethrostomy to remove urine flow from the prepuce resulted in the ability to colonize the prepuce, with live organisms being cultured for as long as 15 days post-inoculation. Future studies optimizing this technique have the potential to accelerate the pace of bovine trichomonosis research and may have applications in the study of human trichomoniasis. Full article

Background: Vesicoureteral reflux (VUR) is a common childhood condition where urine flows backward from the bladder to the kidney, affecting 2–5 per 1000 live births. There is still no consensus on VUR diagnostic procedures, treatment options, or the most effective timing of treatment. This study aimed to identify factors influencing VUR management timing, with a focus on the role of urodynamic assessment. Methods: We retrospectively analyzed 100 children with VUR, splitting them into two groups: 50 who received urodynamic assessment and 50 who did not. The median age at diagnosis for both groups was 2 years (ranging from 1 month to 14 years). The children’s medical records were analyzed to determine gender, age at VUR diagnosis and its resolution, grade, and laterality. Additionally, we recorded the clinical history of bladder dysfunction, recurrent UTIs, and renal function parameters. For those who had urodynamic assessments, we also evaluated the age at initial testing and its findings. Results: Significant differences in treatment duration were found between the study groups: 1.17 years for the group after urodynamic assessment versus 2.83 years for the group without urodynamics (p < 0.001 *). The majority of patients assessed urodynamically had an overactive bladder (52%). Conclusions: Urodynamic assessment and effective treatment of bladder dysfunction accelerate VUR resolution. Full article