Significant Increase In Urinary Levels Research Articles

Polycystic Ovary Syndrome Accompanied by Hyperandrogenemia or Metabolic Syndrome Triggers Glomerular Podocyte Injury

Objective: To determine whether the urinary excretion of podocyte degradation products varies according to PCOS phenotype and metabolic syndrome (MetS). Methods: The concentrations of podocalyxin (PDX) and nephrin, chronic markers of podocyte damage, and neutrophil gelatinase-associated lipocalin (NGAL), a marker of acute glomerular damage, were analyzed in the morning urine samples of 50 PCOS patients and 50 healthy controls matched by age and BMI. Albuminuria was assessed by calculating the urine albumin–creatinine ratio (uACR). Results: The PDX, nephrin and NGAL concentrations of PCOS participants were significantly higher than those of the control group. While PDX, nephrin and NGAL levels of classic phenotypes were similar, they were higher than ovulatory and non-hyperandrogenic phenotypes. Significant increases in urinary levels of each podocyte protein were detected in PCOS patients with MetS compared to patients without MetS. A positive significant correlation between podocyte proteins and BMI, systolic blood pressure, testosterone, glucose, HOMA-IR and uACR. After adjusting for age and BMI, podocyte proteins were an independent risk factor for microalbuminuria. The incidence of microalbuminuria in PCOS increased 6-fold compared to controls. The frequency of microalbuminuria was higher in classical phenotypes than in ovulatory phenotype. The frequency of microalbuminuria in PCOS patients with MetS was 6.5 times higher than in PCOS patients without MetS. Conclusions: In PCOS accompanied by hyperandrogenemia or metabolic syndrome, leakage of acute and chronic podocyte breakdown products into the urine becomes more pronounced.

EXOSOMAL MICRORNA-26A RESPONSE TO TGF-B1 STRESS IN HYPERTENSION

Objective: In a previous work we established an exosomal miRNA signature associated to urinary albumin excretion (UAE) in hypertension, which included microRNA (miRNA) miR-26a, and obtaining cell signaling (MAPK, calcium, p53), TGF-ß and VEGF pathways as the most regulated biological processes. The aim of this work was to validate the exosomal miR-26a changes and its potential association with TGF-ß in human podocytes. Design and method: This prospective study analysed the miR-26a levels in 52 hypertensive patients, 24 with UAE (162,8 ± 168.2 mg/g creatinine; mean age 52.7 ± 8.4 years) and 28 normoalbuminuric (mean age 54.5 ± 5.6 years). Exosomes were isolated by differential ultracentrifugation from urine and plasma and miRNA levels were calculated by qRT-PCR. Urinary levels of TGF-ß1 were quantified by ELISA. Finally, human podocytes cultures were stimulated with recombinant human TGF-ß1 at several concentrations (5 ng/mL, 10 ng/mL and 15 ng/mL) for up 24 h to assess miR-26a changes in exosomal and cellular fractions. Results: We found a decrease in the levels of miR-26-5p in plasma and urinary exosomes of patients con increased UAE (p < 0.05 and p < 0.01, respectively). In addition, miR-26a-5p correlated inversely with UAE levels, both in urinary and plasma exosomes (r = -0.49, p < 0.01; r = -0.53, p < 0.01). Our patients also showed a significant increase in urinary levels of TGF-ß1 (2-fold increase, p < 0.05) in hypertensive patients with UAE. Finally, we analysed the effect in vitro of TGF-ß1 stress. Significant changes in scaffolding of podocytes, such as an increase of vimentin levels, has been also observed. Finally, significant down-regulated miR-26a expression in exosomes from podocytes after TGF-ß1 stimulation (5 – 15 ng / mL): 2.23, 1.96 and 1.7-fold decrease, with p < 0.001 and p < 0.05, respectively. No significant changes were obtained at intracellular level. Conclusions: Our results show that miR-26a-5p is decreased in urinary and plasma exosomes of microalbuminuric patients. In addition, exosomal miR-26a podocyte release is associated with TGF-ß1 pathway, key molecule in the development and maintenance structure of the glomerular filtration barrier and fibrosis.

Gentamicin induced acute renal damage and its evaluation using urinary biomarkers in rats.

Consistent, sensitive biomarkers of acute kidney injury in animal models and humans have historically represented a poorly met need for investigators and clinicians. Detection of early kidney damage using urinary biomarkers is essential to assess the adversity in preclinical toxicology studies, which will help in reducing attrition of lead candidates in drug development. This study was undertaken to evaluate recently identified urinary biomarkers use in identifying acute kidney injury compared to traditional serum markers in experimentally induced nephrotoxicity in male Sprague Dawley (SD) rats. Gentamicin induced nephrotoxicity in Sprague Dawley rats is commonly detected using serum markers and histological evaluation of kidneys. Gentamicin, an aminoglycoside was administered at 30 and 100 mg/kg/day dose (subcutaneous) for seven consecutive days to induce nephrotoxicity. On day 4 and day 8 post treatment, serum and urine samples from these rats were analyzed for traditional serum/urine and novel urinary biomarkers and microscopic evaluation of kidneys. On Day 4, no statistically significant change in serum BUN and creatinine level, but increase in urinary microalbumin (mALB) and urinary protein (UP) noticed in both doses of Gentamicin treated rats. On Day 8 significant increase in serum blood urea nitrogen (BUN), serum creatinine, UP and urinary mALB at 100 mg/kg/day, increase in total protein and decrease in albumin in 30 and 100 mg/kg/day and decrease in BUN and creatinine at 100 mg/kg of Gentamicin treated rats. The BUN and creatinine levels or fold change was comparable between control and 30 mg/kg of Gentamicin on Day 8, however, there was 5.6 and 3.4 fold change in BUN and Creatinine level noticed at 100 mg/kg/day of Gentamicin. On Day 4 and 8, significant increase in urinary levels of Clusterin was noted with animals administered both doses of Gentamicin. Similarly, significant increase in urinary levels of kidney injury molecule 1 (Kim-1), Cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) were noticed with animals administered Gentamicin at 100 mg/kg/day on both Day 4 and 8. All these markers have shown dose-dependent change. Histological changes seen on Day 4 and Day 8 were of minimal to mild and moderate to severe in nature at both doses, respectively. The results demonstrated the sensitiveness and accuracy of detecting acute renal damage with novel urinary biomarkers, and their use in diagnosing early kidney damage. This helps in adversity assessment in animal toxicology studies and advocating right treatment to patients who have early renal injury which otherwise can only be diagnosed by elevated levels of traditional biomarkers in blood only after >30% of kidneys is damaged.

Urinary biomarkers for early detection of platinum based drugs induced nephrotoxicity

BackgroundNephrotoxicity is a major hazard complicating the use of platinum based drugs (PBD), which can hinder using higher doses protocols to maximize the therapeutic gain. Shortage of serum creatinine level as an accurate biomarker for acute kidney injuries (AKI) necessitates searching for novel biomarkers with better sensitivity and specificity in patients on PBD.MethodsIn a prospective cohort design, 132 patients receiving PBD were selected for the study. AKI was diagnosed by continuous follow up of serum creatinine level according to Kidney Disease: Improving Global Outcomes (KDIGO) guidelines 2012. Serum creatinine and urinary biomarkers (KIM-1, NGAL and cystatin C) was measured in the day of treatment and for 3 days after PBD cycle.ResultsAKI occurred in 35 patients (26.52% of patients). KIM-1, Cystatin C, and NGAL showed significant increase in samples collected in the day of AKI in comparison to their corresponding basal levels (P < 0.0001). In addition, significant increase in urinary levels of the biomarkers in samples collected 1 day before AKI in comparison to their basal levels (P < 0.0001, P < 0.0001, and P = 0.013 for KIM-1, NGAL and Cystatin C respectively). Furthermore KIM-1 data showed a significant increase 2 days before serum creatinine rise in comparison to the corresponding KIM-1 levels in patients who developed AKI (P = 0.001).ConclusionsUrinary KIM-1, Cystatin C and NGAL can predict PBD induced AKI in earlier stages than serum createnine. KIM-1 is the most sensitive biomarker for early detection of AKI in patients receiving PBD.

Elevation in and persistence of multiple urinary biomarkers indicative of oxidative DNA stress and inflammation: Toxicological implications of maleic acid consumption using a rat model.

Maleic acid (MA), an intermediate reagent used in many industrial products, instigated public health concerns in Taiwan when it was used to adulterate an array of starch-based delicacies to improve texture and storage time. Established studies reported that exposure to high concentrations of MA induce renal injury; little is known whether oxidative stress is induced at a relative low dose. This study aims to investigate the effect of oral single dose exposure of MA on the status of oxidative stress and inflammation. Single dose of MA at 0, 6 and 60 mg/kg (control, low- and high-dose groups, respectively) were orally administered to adult male and female rats. Urine samples were collected and analyzed to measure 8-hydroxy-2’-deoxyguanosine (8-OHdG), 8-iso-prostaglandin F2α (8-IsoPGF2α), 8-nitroguanine (8-NO2Gua) and N-acetyl-S-(tetrahydro-5-hydroxy-2-pentyl-3-furanyl)-L-cysteine (HNE-MA) using LC-MS/MS. Results revealed that oral consumption of MA induced oxidative DNA damage and lipid peroxidation, as demonstrated by the statistically significant increases in urinary levels of 8-NO2Gua, 8-OHdG, and 8-isoPGF2α, in high-dosed male rats within 12 h of oral gavage (p < 0.05). Additionally, increases in concentration of these biomarkers persist for days after consumption; male rats appear to be more sensitive to oxidative burden compared to their counterparts. The aforementioned findings could help elucidate the mechanisms through which nephrotoxicity occur.

Perspectives on using a multiplex human kidney safety biomarker panel to detect cisplatin-induced tubular toxicity in male and female Cynomolgus monkeys

Multiplex biomarker panel assays would enable early de-risking of discovery compound related kidney safety liabilities. The objective of this study was to evaluate the usefulness of the Myriad RBM Human KidneyMAP (Multi-Analyte Profile)® v.1.0 panel to detect experimental nephrotoxicity in Cynomolgus monkeys following a single intravenous administration of cisplatin (2.5mg/kg). Urine samples were collected at baseline on day −2; at approximately 4hr post-dose on day 1; and on days 4, 9, 15 and/or 20. Blood samples were collected at predose on day −2; at 4hr post-dose on day 1; and on days 2, 5, 10 and/or 21. Changes in toxicokinetic and biochemistry parameters in plasma, qualitative/quantitative urinalysis parameters, and urinary kidney safety biomarkers were assessed. Kidney tissues were collected on days 2, 5, 10 and 21 for routine microscopy. Cisplatin-induced tubular alterations were characterized by acute and progressive cortical tubular degeneration/necrosis, regeneration, tubular dilation and proteinaceous cast in the absence of statistically significant changes in traditional plasma biochemistry and urinalysis parameters. When normalized to urinary creatinine, cisplatin-induced significant increases in urinary levels of kidney injury molecule 1 (females on day 4), increases in calbindin D28k (males and females on day 4), decreases in Tamm-Horsfall glycoprotein (males on days 1, 4 and 9), and increases in clusterin (females and males on days 15 and 20, respectively), when compared to concurrent controls. This study revealed the usefulness of the Human KidneyMAP® multiplex panel when measuring changes in urine-based biomarkers to reliably detect cisplatin-induced acute/progressive cortical tubular injury in male and female Cynomolgus monkeys.

Biomarkers of Organophosphate Pesticides and Attention-Deficit/ Hyperactivity Disorder in Children: A Case-Control Study

Children are frequently exposed to significant hazard for organophosphate poisoning (OP) in their environment; since the brain is growing during this period and it is extra liable to neurotoxicants. The aim of this work was to determine the relationship between urinary concentrations of dialkylphosphates (DAP) metabolites, which are biomarkers of OP pesticide exposure, and attention-deficit/hyperactivity disorder (ADHD) in Egyptian children ages seven to twelve years. A case-control study was conducted at Alzhraa University hospital. We recruited 40 children diagnosed as ADHD according to “the diagnostic and statistical manual of mental disorder (DSM-5) criteria” by psychiatrist. Control group of 40 healthy children who did not have any psychiatric or neurologic disorders, age and sex matched. Exposure was determined by analyzing six urinary metabolites of DAP. We investigated the association between ADHD subtypes, coincidence of epilepsy and OP exposure. Significant increase in urinary level of all DAP metabolites for children with ADHD in comparison to control group except Diethyl-dithio-phosphate. ADHD children with epilepsy had statistically significant elevated level of urinary Dimethyl-dithio-phosphate, Diethyl- phosphate and Dimethyl-phosphate metabolites in comparison to those without epilepsy. By logistic regression analysis, Children with high level of urinary Dimethyldithio- phosphate (OR=2.29), Diethyl-phosphate (OR=2.40), Dimethyl-phosphate (OR =2.02), and Dimethyl-thiophosphate (OR=1.82) have two fold increased risk of developing ADHD than those having lower concentration of these metabolites. Our findings back up the theory that existing environmental concentration of organophosphate pesticide exposure may lead to the development of ADHD and increase the risk of epilepsy in ADHD children.

Urinary excretion of 9α,11β-prostaglandin F2 and leukotriene E4 in patients with exercise-induced bronchoconstriction.

Increased levels of mast cell-derived eicosanoids, such as prostaglandin (PG) D2 and cysteinyl leukotrienes (CysLTs), have been reported in patients with exercise-induced bronchoconstriction (EIB), suggesting that mast cell activation is involved in the mechanism of EIB. However, it is still controversial since these results have not been reproduced in other studies. The aim of this study was to evaluate the role of PGD2 and LTE4 in adult asthma with EIB, as measuring urinary levels of their metabolites-9α,11β-PGF2 and LTE4 before and after an exercise challenge test. Eight patients with asthma and EIB and five normal controls without EIB were enrolled. Exercise challenge tests comprised of 6 min of treadmill exercise or free running were performed in all study subjects, and urine samples before and 1 h after the challenge were collected. Urinary levels of 9α,11β-PGF2 and LTE4 were measured by enzyme immunoassay (EIA). No significant differences were observed in 9α,11β-PGF2 and LTE4 levels before/after the exercise challenge between patients with EIB and normal controls. No significant increases in urinary levels of 9α,11β-PGF2 or LTE4 were detected during the exercise challenge in patients with EIB and normal controls. No significant correlations were observed between the percent decrease in forced expiratory volume in 1 s (FEV1) or percent changes in 9α,11β-PGF2 and LTE4 levels after the exercise challenge. Urinary 9α,11β-PGF2 and LTE4 levels did not increase after an exercise challenge in patients with EIB, suggesting that urinary excretion of 9α,11β-PGF2 and LTE4 may not be a good marker of mast cell activation in patients with EIB.

Leptin and urinary leukotriene E4 and 9α,11β-prostaglandin F2 release after exercise challenge

BackgroundLeptin-related effects on inflammation and bronchial hyperresponsiveness (BHR) in the human airway have not been demonstrated. ObjectivesTo investigate the relationship between the levels of serum leptin and BHR and urinary leukotriene E4 (LTE4) and 9α,11β-prostaglandin F2 (9α,11β-PGF2) release after exercise challenge in asthmatic children. MethodsEighty-six prepubertal children between 6 and 10 years old were enrolled and divided into 4 groups: 19 obese asthmatic children, 25 normal-weight asthmatic children, 21 obese nonasthmatic children, and 21 healthy controls. We measured serum leptin levels and urinary LTE4 and 9α,11β-PGF2 levels in children before and 30 minutes after the exercise challenge. ResultsSerum leptin levels were significantly higher in obese asthmatic children compared with normal-weight asthmatic children. Significant increases in urinary levels of LTE4 and 9α,11β-PGF2 were observed in obese asthmatic children after the exercise challenge. Although smaller than in obese asthmatic children, significant increases in the urinary levels of LTE4 and 9α,11β-PGF2 were also observed in the normal-weight.Asthmatic children Logarithmic serum leptin values were significantly associated with the logarithmic maximum percentage change in forced expiratory volume in 1 second, the logarithmic urinary LTE4 change, and the logarithmic urinary 9α,11β-PGF2 change from baseline to after exercise in both obese and normal-weight asthmatic children. ConclusionThe serum levels of leptin were significantly associated with BHR and urinary LTE4 and 9α,11β-PGF2 release induced by exercise challenge in asthmatic children.

Disparities in arsenic exposure among children and adolescents in the United States

Chronic arsenic exposure in developing children is associated with cognitive delays, reduced IQ, poor memory, and a range of negative health outcomes later in life. In this study, we examined demographic and socioeconomic disparities in arsenic exposure among children and adolescents in the U.S. Subjects aged 6–19 years were selected from the 2005–2006 and 2007–2008 cycles of the National Health and Nutrition Examination Survey. Exposure to arsenic was determined based on the urinary concentrations of total arsenic (tAs) and speciated arsenic. Multivariate logistic regression analysis was performed using SAS 9.3 to assess the association of demographic and socioeconomic variables with arsenic exposure. A statistically significant increase in urinary levels of tAs and dimethylarsinic acid was found in both African-American children and other races, as compared with Whites, as well as in those who recently consumed fish and shellfish. The racial disparity in arsenic exposure observed in this study may be related to genetic factors governing arsenic metabolism and excretion patterns in urine, as well as to socioeconomic status, dietary options, or residential proximity to industry. Further studies are needed to identify factors that contribute to such racial disparities in arsenic exposure.

A randomized targeted amino acid therapy with behaviourally at-risk adopted children

Increasing numbers of children are at-risk for behavioural and emotional disorders, a phenomenon contributing to increased use of pharmacological interventions for paediatric clients. Adverse side effects and other risks associated with pharmacological approaches have helped fuel interest in nutritional interventions for behaviourally at-risk children. The current randomized clinical trial evaluates the efficacy of a neurochemical intervention involving the glutamine and glutamate analogue L-theanine and 5-hydroxytryptophan, the precursor for serotonin, with children adopted from traumatic backgrounds. Results include significant increases in urinary levels of the biomarkers for serotonin and gamma-aminobutyric acid, coupled with significant decreases in parent reports of the children's behaviour problems. While further research is needed, these initial findings are encouraging and are consistent with a growing number of studies indicating the efficacy of nutritional approaches to help behaviourally at-risk children.

Urinary Metabolites of a Tobacco-Specific Lung Carcinogen in Nonsmoking Hospitality Workers

Exposure of nonsmokers to environmental tobacco smoke results in increased risk for cancer and other diseases. In spite of this finding, some restaurants and bars continue to permit smoking. This study examined the uptake of nicotine and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a potent lung carcinogen, in nonsmokers who work in restaurants and bars that permitted smoking. Urine samples were collected for 24 hours on working and nonworking days and were analysed for total NNAL [the sum of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its glucuronides (NNAL-Glucs)], metabolites of NNK. In addition, urine samples were analysed for total nicotine (nicotine plus nicotine glucuronide), and total cotinine (cotinine plus cotinine-N-glucuronide). The results showed significant increases in urinary levels of total NNAL, total nicotine, and total cotinine on working days compared with nonworking days. The results of this study show that smoke exposure in bars and restaurants may have important health effects on nonsmoking employees, elicited by the increase in carcinogen levels.

GC/MS analysis of urinary excretion of 9alpha,11beta-PGF2 in acute and exercise-induced asthma in children.

9Alpha,11beta-prostaglandin (PG) F2 is an initial metabolite of PGD2 which has a potent bronchoconstrictive activity. We measured the urinary levels of 9alpha,11beta-PGF2 in asthmatic children to investigate its role in not only acute asthmatic attack in a time course study but also in exercise-induced asthma (EIA). In the acute asthma study, 30 asthmatic children were examined. Urine samples were collected on the first, third, and sixth days. Urinary levels of 9alpha,11beta-PGF2 were measured with gas chromatography mass spectrometry using the electron impact method. In the exercise challenge study, 14 children with EIA and 14 children without EIA were studied. Urine samples were collected before exercise challenge, and at 1 h, and 5 h after exercise challenge. Urinary levels of 9alpha,11beta-PGF2 were measured. Elevated urinary levels of 9alpha,11beta-PGF2, which were observed on the first day when treatment was started in the hospital, were gradually decreased on the third day (P < 0.05), and on the sixth day (P < 0.01). A significant correlation between urinary levels of 9alpha,11beta-PGF2 and symptom scores (P < 0.005) was observed on the first day. In EIA, there was a significant increase in urinary levels of 9alpha,11beta-PGF2 at 1 h (P < 0.01) and at 5 h (P < 0.01) after exercise challenge, but not in the children without EIA. 9Alpha,11beta-PGF2 may be involved in the pathogenesis of acute and exercise-induced asthma in children.