Urine Samples In Order Research Articles

An updated approach to the evaluation of the urinary sediment.

Examination of the urinary sediment (U-sed) is an important non-invasive, rapid, and inexpensive tool for the diagnosis and surveillance over time of renal diseases. In this Educational Review, we describe first how to collect, prepare, and examine urine samples in order to obtain reliable results. Then, we describe the U-sed findings in isolated microscopic hematuria, glomerular diseases, acute interstitial nephritis, acute kidney injury, reactivation of the BK virus in kidney transplant recipients, and crystalluric genetic diseases.

Innovative molecular networking analysis of steroids and characterisation of the urinary steroidome

Steroids are cholesterol-derived biomolecules that play an essential role in biological processes. These substances used as growth promoters in animals are strictly regulated worldwide. Targeted assays are the conventional methods of monitoring steroid abuse, with limitations: only detect known metabolites. Metabolism leads to many potential compounds (isomers), which complicates the analysis. Thus, to overcome these limitations, non-targeted analysis offers new opportunities for a deeper understanding of metabolites related to steroid metabolism. Molecular networking (MN) appears to be an innovative strategy combining high-resolution mass spectrometry and specific data processing to study metabolic pathways. In the present study, two databases and networks of steroids were constructed to lay the foundations for the implementation of the GNPS-MN approach. Steroids of the same family were grouped together, nandrolone and testosterone were linked to other analogues. This network and associated database were then applied to a few urine samples in order to demonstrate the annotation capacity in steroidome study. The results show that MN strategy could be used to study steroid metabolism and highlight biomarkers.

Highly Efficient CeO2–CuCrO2 Composite Nanofibers Used for Electrochemical Detection of Dopamine in Biomedical Applications

Dopamine (DA) plays a crucial role in the functioning of the human central nervous system, participating in both physiological and psychological processes. It is an important research topic in biomedical science. However, we need to constantly monitor the concentration of dopamine in the body, and the sensors required for this usually require good sensitivity in order to achieve fast and accurate measurements. In this research project, a CeO2 and CuCrO2 composite nanofiber was prepared for the electrochemical detection of dopamine. Coaxial electrospinning techniques were used to prepare CeO2–CuCrO2 composite nanofibers. The characterization techniques of X-ray diffractometer (XRD), Raman, and X-ray photoelectron spectroscopy (XPS) were used to analyze the composite’s crystal structure, vibrational bonds, and elemental composition, while SEM and TEM were used to analyze the composite’s surface structure, morphology, and microstructure. The prepared nanofiber outer layer was found to have an average thickness of 70.96 nm, average fiber diameter of 192.49 nm, and an average grain size of about ~12.5 nm. The BET analysis was applied to obtain the specific surface area (25.03 m2/gm). The proposed nanofiber-decorated disposable screen-printed carbon electrode acted as a better electrochemical sensor for the detection of dopamine. Moreover, the electrocatalyst had a better limit of detection, 36 nM with a linear range of 10 to 100 μM, and its sensitivity was 6.731 μA μM−1 cm−2. In addition, the proposed electrocatalyst was successfully applied to real-time potential applications, namely, to the analysis of human urine samples in order to obtain better recovery results.

Urine Metabolites Enable Fast Detection of COVID-19 Using Mass Spectrometry.

The COVID-19 pandemic boosted the development of diagnostic tests to meet patient needs and provide accurate, sensitive, and fast disease detection. Despite rapid advancements, limitations related to turnaround time, varying performance metrics due to different sampling sites, illness duration, co-infections, and the need for particular reagents still exist. As an alternative diagnostic test, we present urine analysis through flow-injection-tandem mass spectrometry (FIA-MS/MS) as a powerful approach for COVID-19 diagnosis, targeting the detection of amino acids and acylcarnitines. We adapted a method that is widely used for newborn screening tests on dried blood for urine samples in order to detect metabolites related to COVID-19 infection. We analyzed samples from 246 volunteers with diagnostic confirmation via PCR. Urine samples were self-collected, diluted, and analyzed with a run time of 4 min. A Lasso statistical classifier was built using 75/25% data for training/validation sets and achieved high diagnostic performances: 97/90% sensitivity, 95/100% specificity, and 95/97.2% accuracy. Additionally, we predicted on two withheld sets composed of suspected hospitalized/symptomatic COVID-19-PCR negative patients and patients out of the optimal time-frame collection for PCR diagnosis, with promising results. Altogether, we show that the benchmarked FIA-MS/MS method is promising for COVID-19 screening and diagnosis, and is also potentially useful after the peak viral load has passed.

Prevalence of drug abusers in patients undergoing elective surgeries at the Cairo University Teaching Hospital; Prospective cohort study

ABSTRACT Background and Aim : Drug abusers are presenting a challenge to the anaesthetist because of the added potential risks involved in the administration of anaesthesia to this subset of patients. In this study, we aimed at screening all patients scheduled for elective orthopaedic, general, vascular, and plastic at the Cairo University Teaching Hospital during a set period of time for the most commonly abused drugs in Egypt. Methods : All patients included in the study were consented for taking part in this study. Each patient was asked to answer a form and submit a urine sample in order to be screened for the most commonly abused drugs in Egypt (hashish, tramadol, benzodiazepine, and morphine). Patients were then followed up both intra and post-operatively. Incidence of drug abuse among those patients was set as primary outcome, intraoperative hemodynamic, Richmond Agitation-Sedation Scale (RASS) scores as well as clinical recovery scores (CRS) were set as secondary outcomes. Results : Out of a total of 1106 patients screened during the study period, only 500 met inclusion criteria and consented to be part of the study. Results showed evidence of drug abuse among 14.4% of study patients, with frequencies of drugs abused as follows: hashish (11.4%); tramadol (5%); benzodiazepines (1.6%); and morphine (0.4%). Most patients that showed evidence of drug abuse were among those scheduled for orthopaedic surgeries, and a higher percentage of them were males. Conclusions : Within the study group, cannabinoids were the most frequent substance of abuse, followed by tramadol and then other opioids.

Graphene supported poly(3-aminophenylboronic acid) surface via constant potential electrolysis for facile and sensitive paracetamol determination

The present work demonstrates the first example of an electro-sensor platform based on disposable graphite surface that was coated electrochemically with poly(3-aminophenylboronic acid)-graphene for highly sensitive and selective determination of paracetamol. Fabrication of this sensing platform was realized using constant potential electrolysis with 3-aminophenylboronic acid monomer and graphene in a single step preparation. Optimization of the electrochemical polymerization conditions was done with different electrolysis durations and various concentrations of the monomer and graphene. Coated electrodes were then characterized by using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX). Electrochemical determination studies performed with differential pulse voltammetry (DPV) presented that the electro-sensor platform had a good linear paracetamol concentration range of 0.15 μM to 100 μM and a very low detection limit of 0.028 μM (R2 = 0.9941, n = 3). The effect of a possible and important interferent, dopamine was also examined. The results proved the simultaneous determination of these two analytes with a good separation and high sensitivity. These results were compared with the results of the bare electrode: 15-fold and 13-fold improvements were achieved at the coated electrode for the electrooxidation of dopamine and paracetamol, respectively. In addition, this robust platform showed a good long-term stability. At last, recovery studies were done in urine samples in order to present the possible use of the electro-sensor in clinical studies. The outputs highlighted the high accuracy of the system by giving good recoveries (94.0–103.7%).

Daily Urinary Sodium Excretion Monitoring in Critical Care Setting: A Simple Method for an Early Detection of Acute Kidney Injury.

Introduction:Making an early diagnosis of acute kidney injury (AKI) is crucial. Classical biomarkers are not capable of early detection of AKI, but novel biomarkers that do have this capability are expensive and not universally available. This prospective study attempts to mitigate these limitations through the evaluation of daily urine analysis on patient admitted to a critical care unit in order to detect early AKI.Methods:Daily urinary indices were measured on every patient admitted to the intensive care unit (ICU) from the time of admission until his/her discharge from the ICU or death. This renal monitoring consisted of daily blood and spot morning urine samples in order to measure creatinine, urea, sodium, chloride and potassium in order to calculate the fractional excretion of sodium (FENa), chloride, urea and potassium. The data collected on these patients in the previous days was analyzed to determine whether or not there was a significant statistical difference in the urinary indices one day before the clinical diagnosis of AKI (day – 1) and 2 days before the diagnosis (day – 2). The statistical test applied was a single rank test, using as a limit of significance a value of P < 0.05.Results:Of the 203 patients included, 61 developed AKI. A statistical significant difference was documented only in the value of urinary sodium (UNa) and FENa between day-1 (one day before AKI clinical diagnosis) and day-2 (two days before AKI clinical diagnosis).Conclusion:Daily monitoring of UNa and FENa detected a significant change in their basal values 24 hours before clinical diagnosis of AKI was made.

The key points in the pre-analytical procedures of blood and urine samples in metabolomics studies.

Metabolomics provides measurement of numerous metabolites in human samples, which can be a useful tool in clinical research. Blood and urine are regarded as preferred subjects of study because of their minimally invasive collection and simple preprocessing methods. Adhering to standard operating procedures is an essential factor in ensuring excellent sample quality and reliable results. In this review, we summarize the studies about the impacts of various preprocessing factors on metabolomics studies involving clinical blood and urine samples in order to provide guidance for sample collection and preprocessing. Clinical information is important for sample grouping and data analysis which deserves attention before sample collection. Plasma and serum as well as urine samples are appropriate for metabolomics analysis. Collection tubes, hemolysis, delay at room temperature, and freeze-thaw cycles may affect metabolic profiles of blood samples. Collection time, time between sampling and examination, contamination, normalization strategies, and storage conditions may alter analysis results of urine samples. Taking these collection and preprocessing factors into account, this review provides suggestions of standard sample preprocessing.

MoS2 nanostructured materials for electrode modification in the development of a laccase based amperometric biosensor for non-invasive dopamine detection

A novel laccase-based amperometric biosensor was developed through the modification of carbon paper electrodes with layered two-dimensional molybdenum disulfide (MoS2) presented in two different morphologies, flowers (MoS2-F) and ribbons (MoS2-R). Two laccase isoforms from Pycnoporus sanguineus CS43 fungi (LacI and LacII) were evaluated for the first time as a biological recognition element, and their performance was compared with commercial laccase from Trametes versicolor (TvL). The best results were observed with the MoS2-R morphology at a concentration of 2 mg/mL Biosensors modified with MoS2-R and with the three enzymes, showed two linear ranges of detection, from 0.1 to 0.5 µM and from 1 to 5 µM. Biosensors modified with LacII exhibited a high sensitivity of 340.3 nA/µM; and low limit of detection (LOD) of 10 nM, which are the best values reported in the literature for dopamine detection by laccase based biosensor. Optimized electrodes modified with enzymes LacII and TvL were tested in a synthetic urine sample in order to evaluate their practical effectiveness, resulting in LOD of 0.67 and 2.67 µM respectively, which are below the average concentrations of dopamine normally found in real urine samples (around 2 µM).

Semiquantitative, fully automated urine test strip analysis.

ObjectivesUrinalysis is one of the most frequently ordered diagnostic laboratory tests. In order to reduce workload and costs, rapid screening tests such as urine test strip analyses are applied. The aim of this study was to evaluate the analytical performance of the UC‐3500 as well as the diagnostic performance in comparison with reference methods.Design and methodsWe measured within‐run and between‐run imprecision based on quantitative reflectance values. 347 prospectively included urine specimens were investigated for the presence of glucose, protein, albumin, leukocyte esterase, and hemoglobin peroxidase activity, and ordinal scale results were compared to an automated urine particle analyzer (UF‐5000, Sysmex, Kobe, Japan) and wet chemistry (Roche Cobas 8000, Mannheim, Germany).ResultsWithin‐run and between‐run imprecision results based on reflectance data for both the 9 and 11 parameter test strips ranged from 0.07% to 1.36% for the low‐level control and from 0.37% to 6.13% for the high‐level control, depending on the parameter. Regarding diagnostic performance, the sensitivity/specificity for glucose, protein, albumin, leukocyte esterase, and hemoglobin peroxidase was 100/60%, 94.2/88.2%, 81.8/89.2%, 81.7/92.8%, and 85.1/88.6%, respectively; the negative predictive value was 100%, 83.3%, 89.1%, 94.6%, and 96.1%. The Spearman correlation coefficients of the UC‐3500 vs reference methods ranged from 0.915 to 0.967, depending on the parameter.ConclusionThis fully automated urine test strip analyzer overall shows a satisfying performance and can reliably screen out negative urine samples in order to focus on further characterization of positive samples in the following steps of the workflow.

Sexually Transmitted Infections: A Novel Screening Strategy for Improving Women's Health in Vulnerable Populations.

Background: Migrant women are one of the most vulnerable population to health problems and well-being. This study aimed at implementing a counseling and preventive strategy for sexually transmitted infections (STIs) in undocumented migrant women in Milan, Italy. Methods: Women (ages 18–65) were enrolled at the NAGA Centre (2012–2013) and asked for a urine sample in order to carry out molecular detection of Human papillomavirus (HPV), Chlamydia trachomatis (Ct), Trichomonas vaginalis (Tv), Neisseria gonorrhoeae (Ng)-DNA. Socio-demographic and sexual behavior information were collected. All HPV/Ct+ women were offered Pap tests and/or were prescribed antibiotic treatment. Results: 537/757 women participated in the study (acceptability rate: 70.9%). Most of the women were from Latin America (45.6%) and Eastern Europe (30.7%); >60% of them had stable partners, did not use contraception and had had at least one pregnancy. The prevalence rates of HPV, Ct, Tv and Ng infections were 24.2%, 7.8%, 4.8% and 0%, respectively. In all, 43.2% of the positive women agreed to undergo a gynecological examination and accepted suitable treatment. Conclusions: This study shows an overall high prevalence of STIs in undocumented migrant women in Milan. The screening strategy based on counseling and urine testing contributed to the successfully high acceptability rate. More appropriate health services that adequately address all aspects of women’s health are required.

Diaper Slit Perineal Bag Urine Sampling.

Many indications justifiably drive the pediatrician to request a urine sample in order to assess the health status of the incontinent baby or infant. Urine collection by means of an adhesive perineal bag is the most widely used in children who do not control urine emission, despite its inaccurate reflection of bacteriuria. We suggest a novel technique to immediately visualize the micturition event upon occurrence, in attempt to shorten waiting times and exhausting checking and rechecking opaque diapers in order to determine whether the desired sample has finally been delivered. The perineal urine collection bag is pulled through a vertical slit created in the front of the disposable diaper. The bag is now visible at all times. In general, waiting times appear shortened since the implementation of this practice. Since the adoption of this technique at a number of our clinics, we have not experienced bag displacement as was previously experienced. Forgoing the need to manipulate the diaper and repeatedly awakening irritable infants was gladly welcomed by parents. Widespread adoption of the diaper slit technique seems a promising procedure enhancing comfort and facilitating the process of obtaining a urine sample in the incontinent child. Assumptions that this procedure actually shortens waiting times and raises the success rates of sample retrieval remain to be proven by formal comparative trials.

A nyugat-nílusi vírus kimutatása humán betegmintákból: nyomon követéses vizsgálatok a 2015. évi szezonális időszakban

West Nile virus, a mosquito-borne viral zoonosis is responsible for human infections in Hungary. Laboratory diagnosis is based on serological tests, however the application of molecular methods has been appreciated. The aim of the study was to investigate blood, cerebrospinal-fluid and urine samples of acutely ill patients and to follow-up PCR positive cases to ascertain the length of virus excretion. Clinical specimens were examined by indirect-immunofluorescent, haemagglutination-inhibition, two PCR tests and Sanger-sequencing. Virus isolation in case of two patients was successful. A follow-up study could be carried out in case of 5 patients. Viral nucleic acid was detectable in urine even for several weeks after symptom onset and viral RNA was present at higher concentration compared with other samples. PCR analysis of urine could provide useful epidemiological and diagnostic information. Therefore, it is recommended to collect urine samples in order to supplement the serological diagnosis. Orv Hetil. 2017; 158(20): 791-796.

Assessment of 1H NMR-based metabolomics analysis for normalization of urinary metals against creatinine

BackgroundProton nuclear magnetic resonance (1H NMR, or NMR) spectroscopy and inductively coupled plasma-mass spectrometry (ICP-MS) are commonly used for metabolomics and metal analysis in urine samples. However, creatinine quantification by NMR for the purpose of normalization of urinary metals has not been validated. We assessed the validity of using NMR analysis for creatinine quantification in human urine samples in order to allow normalization of urinary metal concentrations. MethodsNMR and ICP-MS techniques were used to measure metabolite and metal concentrations in urine samples from 10 healthy subjects. For metabolite analysis, two magnetic field strengths (600 and 700MHz) were utilized. In addition, creatinine concentrations were determined by using the Jaffe method. ResultsCreatinine levels were strongly correlated (R2=0.99) between NMR and Jaffe methods. The NMR spectra were deconvoluted with a target database containing 151 metabolites that are present in urine. A total of 50 metabolites showed good correlation (R2=0.7–1.0) at 600 and 700MHz. Metal concentrations determined after NMR-measured creatinine normalization were comparable to previous reports. ConclusionsNMR analysis provided robust urinary creatinine quantification, and was sufficient for normalization of urinary metal concentrations. We found that NMR-measured creatinine-normalized urinary metal concentrations in our control subjects were similar to general population levels in Canada and the United Kingdom.

Presencia de Streptococcus bovis en muestras de orina de sujetos con síntomas de infección del aparato urinario

Given the relevance of proper clinical validation of Streptococcus bovis, we here consider revising its presence in urine samples in order to determine its relative frequency and the pattern of antibiotic susceptibility. The susceptibility to antibiotics of 91 isolates of S. bovis from urine samples was retrospectively reviewed over a period of 4 years (2012-2015). The mean age of patients was 55 years, 81% of whom were women and 37.4% were hospitalized patients suffering from urological diseases (61%). Susceptibility to penicillin, vancomycin and teicoplanin was 97.8%. Due to the fact that S. bovis can be infrequent in urine isolates and given its presence in patients suffering from urological diseases, further pathogenic studies, showing the true ability of this group of bacteria to produce disease, are required.

Human biomonitoring of bisphenol A exposure in an Austrian population

AbstractBisphenol A (BPA) is an industrial chemical widely used as a monomer in polycarbonate, epoxy resin and thermal paper production, and as a stabiliser in polyvinyl chloride (PVC) production. Thus, BPA is present in a wide range of common consumer products. Due to contact with BPA-containing products, humans are regularly exposed via different routes. BPA is an endocrine active substance and can potentially lead to different adverse health outcomes. In the present study, total BPA levels were analysed by on-line SPE-HPLC-MS/MS in spontaneous urine samples in order to investigate the exposure to an Austrian population aged 6-15 and 18-81 years. In the total study population (n=594), total BPA were found in 15.7% of all samples, with concentrations up to 17 μg/l (median: n.d., 95th percentile: 3.7 μg/l). These results show generally low BPA levels among the investigated study population. However, children and adolescents exhibited higher exposure levels. Daily BPA intakes were estimated and compared to acceptable exposure levels, whereat no exceedances could be identified. Although BPA exposure was shown to be low in the Austrian population, the wide distribution and the ongoing discussion related to possible negative health effects even at low doses indicate need for further monitoring and investigations.

Bile acid profiling and quantification in biofluids using ultra-performance liquid chromatography tandem mass spectrometry.

Bile acids are important end products of cholesterol metabolism. While they have been identified as key factors in lipid emulsification and absorption due to their detergent properties, bile acids have also been shown to act as signaling molecules and intermediates between the host and the gut microbiota. To further the investigation of bile acid functions in humans, an advanced platform for high throughput analysis is essential. Herein, we describe the development and application of a 15 min UPLC procedure for the separation of bile acid species from human biofluid samples requiring minimal sample preparation. High resolution time-of-flight mass spectrometry was applied for profiling applications, elucidating rich bile acid profiles in both normal and disease state plasma. In parallel, a second mode of detection was developed utilizing tandem mass spectrometry for sensitive and quantitative targeted analysis of 145 bile acid (BA) species including primary, secondary, and tertiary bile acids. The latter system was validated by testing the linearity (lower limit of quantification, LLOQ, 0.25-10 nM and upper limit of quantification, ULOQ, 2.5-5 μM), precision (≈6.5%), and accuracy (81.2-118.9%) on inter- and intraday analysis achieving good recovery of bile acids (serum/plasma 88% and urine 93%). The ultra performance liquid chromatography-mass spectrometry (UPLC-MS)/MS targeted method was successfully applied to plasma, serum, and urine samples in order to compare the bile acid pool compositional difference between preprandial and postprandial states, demonstrating the utility of such analysis on human biofluids.

Berberine ameliorates nonbacterial prostatitis via multi-target metabolic network regulation.

Metabolomics has been increasingly applied to discovering biomarkers and identifying perturbed pathways. Berberine has been shown to exhibit anti-inflammatory, antioxidant, and anticancer properties, but its mechanisms for treating nonbacterial prostatitis (NBP) remain unclear completely. We developed the untargeted metabolomics approach based on UPLC-Q-TOF-HDMS to profile the metabolite changes in urine samples in order to discover novel potential biomarkers to clarify mechanisms of berberine in treating a rat model of capsaicin-induced nonbacterial prostatitis (NBP). The changes in metabolic profiling were restored to their base-line values after berberine treatment according to the principal component analysis (PCA) score plots. Fourteen different potential biomarkers and five acutely perturbed metabolic pathways contributing to the treatment of NBP were discovered and identified. Specifically, the berberine-treated rats are located closer to the normal group, indicating that the NBP-induced disturbances to the metabolic profile were partially reversed by berberine treatment. After treatment with berberine, the relative contents of 12 potential biomarkers were effectively regulated, which suggested that the therapeutic effects of berberine on NBP may involve regulating disturbances to the metabolism. Our results show that the protective effect of berberine occurs in part through a reversal of the NBP-caused disturbances.

Quantitation of methadone and metabolite in patients under maintenance treatment.

Gas chromatography-mass spectrometry quantitative method was developed to monitor concentrations of methadone and its metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in plasma and urine of patients. The developed method was simple, accurate and reproducible to quantify methadone and EDDP in plasma and urine samples in the concentration range of 15-1,000 and 50-2,000 ng/mL, respectively. The proposed analytical method was applied to plasma and urine samples obtained from 96 patients undergoing methadone maintenance treatment (MMT) with daily methadone doses of 2-120 mg/day. Urinary methadone excretion was observed to be significantly affected by pH, in which the ratio of methadone to EDDP was two times higher in acidic urine (P = 0.029). The findings of this study further enhance the guidelines for monitoring of methadone treatment among outpatients. Methadone-to-EDDP ratio in urine was found to be consistent at 24 and 4 h, hence suggesting the possibility that outpatients may be monitored with single urine sample in order to check for compliance. This study which provides data on peak concentrations of methadone and EDDP as well as the ratio of both compounds has added to the body of knowledge regarding pharmacokinetic properties of methadone among heroin-dependent patients under MMT.

Employment-based abstinence reinforcement following inpatient detoxification in HIV-positive opioid and/or cocaine-dependent patients.

Employment-based reinforcement interventions have been used to promote abstinence from drugs among chronically unemployed injection drug users. The current study used an employment-based reinforcement intervention to promote opioid and cocaine abstinence among opioid and/or cocaine-dependent, HIV-positive participants who had recently completed a brief inpatient detoxification. Participants (n = 46) were randomly assigned to an abstinence and work group that was required to provide negative urine samples in order to enter the workplace and to earn incentives for work (n = 16), a work-only group that was permitted to enter the workplace and to earn incentives independent of drug use (n = 15), and a no-voucher control group that did not receive any incentives for working (n = 15) over a 26-week period. The primary outcome was urinalysis-confirmed opioid, cocaine, and combined opioid/cocaine abstinence. Participants were 78% male and 89% African American. Results showed no significant between-groups differences in urinalysis-verified drug abstinence or HIV risk behaviors during the 6-month intervention. The work-only group had significantly greater workplace attendance, and worked more minutes per day when compared to the no-voucher group. Several features of the study design, including the lack of an induction period, setting the threshold for entering the workplace too high by requiring immediate abstinence from several drugs, and increasing the risk of relapse by providing a brief detoxification that was not supported by any continued pharmacological intervention, likely prevented the workplace from becoming established as a reinforcer that could be used to promote drug abstinence. However, increases in workplace attendance have important implications for adult training programs.