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Urine Samples (urine + sample)
Kinds of Urine Samples Selected AbstractsDetermination of Diclofenac in Urine Samples by Molecularly-Imprinted Solid-Phase Extraction and Adsorptive Differential Pulse VoltammetryELECTROANALYSIS, Issue 15 2007Laura Fernández-Llano Abstract A molecularly imprinted polymer for diclofenac (DCF) was prepared by thermal polymerization over silica beads using 2-(dimethylamino)ethyl-methacrylate as functional monomer. After silica elimination by HF treatment, the polymer was applied to the selective solid-phase extraction of the drug from urine followed by its quantification by adsorptive differential pulse voltammetry. Results indicate that the drug could be selectively extracted from the sample and quantified at clinically relevant concentrations (,g/mL). [source] Longitudinal study of urinary albumin excretion in young diabetic patients,-Wessex Diabetic Nephropathy ProjectDIABETIC MEDICINE, Issue 5 2001S. Twyman Summary Aims This study was established to follow changes in albumin/creatinine ratio (ACR) and to determine the prevalence and degree of progression of microalbuminuria (MA) or of clinical proteinuria (CP) in children with Type 1 diabetes. The study has investigated subjects for up to 12 years in establishing the correlation between MA and gender, age, duration of diabetes and glycated haemoglobin (HbA1c). The study has defined clinical cut-offs for MA in daytime clinic urine samples in young diabetic subjects. Methods Three hundred and sixty-one patients were involved in the study, with 221 (61.2%) having over six sets of data. Urine samples were collected at routine annual clinic visits and analysed without prior freezing for ACR. Blood samples were taken for HbA1c measurement. Data including sex, age and duration of diabetes were recorded. Results A random clinic ACR of <,4.5 mg/mmol (males) and 5.2 mg/mmol (females) creatinine was used as the ,clinical cut-off' to define the presence of MA. The presence of MA was independent of HbA1c and duration of diabetes but appeared be associated with the adolescent years (> 10 years). There was little evidence of progression from normoalbuminuria to MA, or from MA to CP. Of patients aged 10,18 years, 30.9% of males and 40.4% of females had one or more episodes of MA. Conclusions Persistent MA and random episodes of MA or CP may be associated with the adolescent years but not with duration of diabetes. Further study will reveal if the substantial increases in ACR sometimes seen during adolescence are predictive of diabetic nephropathy. Clinical cut-offs of <,4.5 and <,5.2 mg/mmol creatinine for males and females, respectively, are suggested for the interpretation of changes in ACR in random urine samples in young people with Type 1 diabetes. [source] DNA damage in leukocytes of workers occupationally exposed to arsenic in copper smeltersENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 2 2005Jadwiga Palus Abstract Inorganic arsenic (i-As) is a known human carcinogen; however, humans continue to be exposed to i-As in drinking water and in certain occupational settings. In this study, we used the Comet assay to evaluate DNA damage in the somatic cells of workers from three Polish copper smelters who were occupationally exposed to i-As. Blood samples were collected from 72 male workers and 83 unexposed male controls and used for the detection of DNA damage, oxidative DNA damage, and DNA damage after a 3-hr incubation in culture. Urine samples were collected to assess the level of exposure. The mean concentration of arsenic metabolites in urine [the sum of arsenite (AsIII), arsenate (AsV), monomethylarsenate (MMA) and dimethylarsenate (DMA)] and the concentrations of DMA (the main metabolite in urine) were higher in workers than in controls, but the differences were not statistically significant. By contrast, the level of DNA damage, expressed as the median tail moment, was significantly higher in the leukocytes of workers than in the controls. Comet assays conducted with formamidopyrimidine glycosylase (FPG) digestion to detect oxidative DNA damage indicated that oxidative lesions were present in leukocytes from both the exposed and control groups, but the levels of damage were significantly higher among the workers. Incubation of the cells in culture resulted in a significant reduction in the levels of DNA damage, especially among leukocytes from the workers, suggesting that the DNA damage was subject to repair. Our findings indicate that copper smelter workers have increased levels of DNA damage in somatic cells, suggesting a potential health risk for the workers. Although i-As was present in air samples from the smelters and in urine samples from workers, no clear association could be made between i-As exposure and the DNA damage. Environ. Mol. Mutagen., 2005. © 2005 Wiley-Liss, Inc. [source] A survey on two years of medication regulation in horse races in IranEQUINE VETERINARY JOURNAL, Issue 2 2010S. LOTFOLLAHZADEH Summary Reasons for performing study: The present survey evaluated the use of prohibited substances cases in the first 2 years of medication regulation in horseracing in Iran so that the impact of these regulations on the level of positive cases over the period could be assessed. Objectives: To determine the prevalence of positive tests for prohibited substances in horse races during 2 years of a drugs testing programme in Iran. Methods: A total of 656 horses that were winners or second in races were tested during the 2 year study. In the first year 354 horses (209 males and 145 females) and in the second year 302 horses (155 males and 147 females) were tested. In the 2 years, 306 were found to be positive. Urine samples were taken from candidate horses and sent to the Central Doping Laboratory. Blood samples were taken from those horses where a urine sample could not be taken within one hour. Detection and measurement of prohibited substances were carried out by ELISA, GC and HPLC using standard methods. Results: Thirty-two percent of males were positive for prohibited substances, which was not significantly different from the percentage of females (25.5%). In the second year, of the 302 horses tested for prohibited substances, 33.5% of males were positive, again similar to females (33.3%). Almost 83% of horses tested positive for prohibited substances once in the first year, 15% tested positive twice and 2% tested positive 3 times. In the second year 78% tested positive once, 15% tested positive twice and 7% tested positive 3 times. Morphine was the most used prohibited substance and was detected 42 times during the survey, followed by caffeine and phenylbutazone. Morphine was also the most used drug in combination with other drugs in both years. Conclusions: Morphine and caffeine were the most popular prohibited substances found in the measurements. As these substances were found in the environment and food stuffs, their presence in the samples may be due to unintentional feeding of contaminated materials (bread, hay and chocolate). [source] Disability payments, drug use and representative payees: an analysis of the relationshipsADDICTION, Issue 7 2003James A. Swartz ABSTRACT Aims This study attempted to determine: if US federal cash disability payments increase the use of cocaine or opiates among those requalifying for supplemental security income (SSI) disability benefits compared with those who lost benefits; if drug use peaks at the beginning of the month after the receipt of the disability cash disbursement; and if money management by representative payees of requalifying SSI recipients suppresses drug use. Design A multi-site, prospective, 2 year longitudinal design was used with follow-up interviews conducted every 6 months. Urine samples were collected at the final three follow-up interviews. Setting Data were collected in Chicago, IL, Los Angeles, CA, and Seattle, WA, USA. Participants This study used a randomly selected sample of 740 former recipients of SSI who had received disability benefits for drug addiction and alcoholism (DA&A) in 1996, were between the ages of 21 and 59 years, had not received concurrent social security disability insurance and provided testable urine samples and complete self-report data for at least one follow-up interview. Measurements Independent variables included demographics, SSI status at follow-up, representative payee status, drug treatment participation and income. Time of drug testing was operationalized as the first 10 days of the month versus the last 20,21 days based on when the urine sample was collected. The dependent variables were cocaine and opiate use, determined by urinalysis results. Findings Participants were 28% more likely to test positive for cocaine use in the first 10 days of the month than later in the month. This effect was general across all subjects and was not restricted to those receiving SSI benefits. No such effect was found for opiate use. Receiving SSI benefits did not increase cocaine or opiate use generally, nor did having a representative payee suppress use. Conclusions The findings do not support the contentions that federal cash benefits appreciably increase drug use or that representative payees discourage use, at least when use is defined dichotomously. The ,check effect' for cocaine use appears to be general and not confined to those receiving federal cash benefits. The lack of a ,check effect' for opiate use is probably the result of the difference between a relatively steady state of opiate use associated with addiction and a binge pattern of cocaine use triggered by suddenly flush resources. [source] The relationship of magnesium intake to serum and urinary calcium and magnesium levels in Trinidadian stone formersINTERNATIONAL JOURNAL OF UROLOGY, Issue 3 2005TREVOR I ANATOL Abstract, Background:, The present study was undertaken to investigate the relationship between the dietary intake of magnesium and the serum and urinary levels of calcium and magnesium in a group of Trinidadian stone formers. Methods:, A group of 102 confirmed stone formers presenting to urological clinics were interviewed using a questionnaire designed to obtain a semi-quantitative estimate of their oral magnesium intake. Patients were invited to give blood samples for serum calcium and magnesium levels and to provide 24-h urine specimens for the measurement of urinary levels of these minerals, as well as total urinary volumes. A group of 102 controls was subjected to a similar interview and blood and urinary testing. Chi-square tests and Student's t -tests were used to examine group demographic differences. The Mann,Whitney test investigated differences in biochemical indices. Binary logistic regression was used to identify predictors of stone formation. Results:, Blood samples were obtained from 60 patients and 98 controls. Urine samples were returned by 34 patients and 97 controls. Only 10 stones were retrieved from patients. Patients had a significantly lower magnesium intake, but higher median serum and urinary calcium levels, and higher serum calcium to magnesium ratios than controls. Independent variables capable of predicting stone formation included total magnesium intake and serum and urinary calcium levels. Conclusions:, Increased serum and urinary calcium levels, calcium to magnesium ratios, and a low magnesium intake were predictive of stone formation in this Trinidadian population. [source] Age-related differences in susceptibility to cisplatin-induced renal toxicity,,JOURNAL OF APPLIED TOXICOLOGY, Issue 2 2010P. Espandiari Abstract Limited experimental models exist to assess drug toxicity in pediatric populations. We recently reported how a multi-age rat model could be used for pre-clinical studies of comparative drug toxicity in pediatric populations. The objective of this study was to expand the utility of this animal model, which previously demonstrated an age-dependent sensitivity to the classic nephrotoxic compound, gentamicin, to another nephrotoxicant, namely cisplatin (Cis). Sprague,Dawley rats (10, 25, 40 and 80 days old) were injected with a single dose of Cis (0, 1, 3 or 6,mg,kg,1 i.p.). Urine samples were collected prior and up to 72,h after treatment in animals that were , 25 days old. Several serum, urinary and ,omic' injury biomarkers as well as renal histopathology lesions were evaluated. Statistically significant changes were noted with different injury biomarkers in different age groups. The order of age-related Cis-induced nephrotoxicity was different than our previous study with gentamicin: 80 > 40 > 10 > 25 day-old vs 10 , 80 > 40 > 25-day-old rats, respectively. The increased levels of kidney injury molecule-1 (Kim-1: urinary protein/tissue mRNA) provided evidence of early Cis-induced nephrotoxicity in the most sensitive age group (80 days old). Levels of Kim-1 tissue mRNA and urinary protein were significantly correlated to each other and to the severity of renal histopathology lesions. These data indicate that the multi-age rat model can be used to demonstrate different age-related sensitivities to renal injury using mechanistically distinct nephrotoxicants, which is reflected in measurements of a variety of metabolite, gene transcript and protein biomarkers. Published in 2009 by John Wiley & Sons, Ltd. [source] Quantification of urinary 8-iso-prostaglandin F2, using liquid chromatography,tandem mass spectrometry during cardiac valve surgeryJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 4 2010Yun-Hui Teng Abstract Oxidative stress is an unavoidable event during many complex surgical procedures. 8-iso-prostaglandin F2, (8-iso-PGF2,) is a reliable biomarker for the evaluation of oxidative stress in vivo. The aim of this study is to develop simple and accurate liquid chromatography,tandem mass spectrometry (LC-MS/MS) methods for the detection of urinary 8-iso-PGF2, in samples collected from patients who received a cardiopulmonary bypass (CPB) during cardiac valve surgery. Urine samples of 14 patients with cardiac valve diseases were collected before, during, and after CPB. The level of 8-iso-PGF2, was detected via selected-reaction monitoring triple quadrupole MS/MS and the result was compared with 12 healthy volunteers. The method's detection limit (3S/N) was 0.25,pg for 8-iso-PGF2,, with a linear working range of 0.25,20,ng/ml. For patients with cardiac valve disease, the 8-iso-PGF2, levels before the bypasses were the same as those of healthy individuals (P>0.05) and the 8-iso-PGF2, levels during and after CPB were significantly higher than those before the bypasses (P<0.05). In conclusion, we present a simple and specific protocol for LC-MS/MS quantification of urinary 8-iso-PGF2, collected during CPB. Using this technique, it would be feasible to assess the levels of oxidative stress during cardiac surgery and thereby helpful for the management of oxidative injury. J. Clin. Lab. Anal. 24:237,245, 2010. © 2010 Wiley-Liss, Inc. [source] Measurement of pulmonary surfactant disaturated-phosphatidylcholine synthesis in human infants using deuterium incorporation from body waterJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 7 2005Paola E. Cogo Abstract The aim of the study was to determine surfactant palmitate disaturated-phosphatidylcholine (DSPC-PA) synthesis in vivo in humans by the incorporation of deuterium from total body water into DSPC-PA under steady state condition. We studied three newborns and one infant (body weight (BW) 4.6 ± 2.9 kg, gestational age 37.5 ± 2 weeks, age 9 ± 9 days) and four preterm newborns (BW 1.3 ± 0.6 kg, gestational age 30.3 ± 2.5 weeks, postnatal age 8.8 ± 9.2 h). All infants were mechanically ventilated during the study and the four preterm infants received exogenous surfactant at the start of the study. We administered 0.44 g 2H2O/kg BW as a bolus intravenously, followed by 0.0125 g 2H2O/kg BW every 6 h to maintain deuterium enrichment at plateau over 2 days. Urine samples and tracheal aspirates (TA) were obtained prior to dosing and every 6 h thereafter. Isotopic enrichment curves of DSPC-PA from sequential TA and urine deuterium enrichments were analyzed by Gas Chromatography-Isotope Ratio,Mass Spectrometry (GC-IRMS) and normalized for Vienna Standard Mean Ocean Water. Enrichment data were used to measure DSPC-PA fractional synthesis rate (FSR) from the linear portion of the DSPC-PA enrichment rise over time, relative to plateau enrichment of urine deuterium. Secretion time (ST) was defined as the time lag between the start of the study and the appearance of DSPC-PA deuterium enrichment in TA. Data were given as mean ± SD. All study infants reached deuterium-steady state in urine. DSPC-PA FSR was 6.5 ± 2.8%/day (range 2.6,10.2). FSR for infants who did not receive exogenous surfactant was 5.7 ± 3.5%/day (range 2.6,9.9%/day) and 7.3 ± 2.1%/day (range 5.1,10.2%/day) in the preterms, whereas DSPC-PA ST was 10 ± 10 h and 31 ± 10 h respectively. Surfactant DSPC-PA synthesis can be measured in humans by the incorporation of deuterium from body water. This study is a simpler and less invasive method compared to previously published methods on surfactant kinetics by means of stable isotopes. Copyright © 2005 John Wiley & Sons, Ltd. [source] Prevalence of human papillomaviruses in urine samples of male patients infected with HIV-1 in Sao Paulo, Brazil,JOURNAL OF MEDICAL VIROLOGY, Issue 12 2009Fernando A.M. Costa Abstract Human papillomavirus is a DNA virus that includes 118 genotypes. HPV16 is responsible for 80% of cervical cancer in women. Men are important reservoirs and major transmitters of HPV to their partners. The aim of this study was to detect HPV DNA and to determine the prevalence of HPV types 6, 11, 16, and 18 in urine samples of men infected with HIV-1. This study included 223 patients infected with HIV-1 from the Center of Reference on HIV/AIDS (CRT-SP) and an outpatient clinic of HIV. Urine samples were collected and after DNA extraction real-time PCR was performed for detection of HPV DNA. Positive samples were then tested by conventional PCR using type-specific primers for the four HPV types. A total of 223 men infected with HIV-1 were tested, 81% of whom were on HAART. Four (5.8%) were positive for HPV6, 18 (26.1%) were positive for HPV11, 22 (31.9%) were positive for HPV16 and five (7.2%) were positive for HPV18 by conventional PCR. Twenty (29%) patients had other HPV types and five patients (1.5%) had multiple types. The mean T CD4+cells count was 517 and 441,cells/mm3 (P,=,0.30), in HPV negative and positive men, respectively. The HIV viral load was higher in the HPV negative group than for in the men with HPV (P,=,0.0002). A 30.9% prevalence of HPV was found in asymptomatic urine samples of men infected with HIV-1. This study suggests that urine may be a useful specimen for HPV screening. J. Med. Virol. 81:2007,2011, 2009. © 2009 Wiley-Liss, Inc. [source] Rapid screening of beta-adrenergic agents and related compounds in human urine for anti-doping purpose using capillary electrophoresis with dynamic coatingJOURNAL OF SEPARATION SCIENCE, JSS, Issue 20 2009Monica Mazzarino Abstract This paper presents a capillary electrophoresis method, developed for the detection, in human urine, of beta-adrenergic agents and phenolalkylamines. The electrophoretic separation is achieved in less than 10 min and is based on the use of CEofix kit, for the dynamic capillary coating. The effects of accelerator buffer pH and separation voltage were investigated. The optimum buffer pH was found to be 2.5 for beta2-agonists and 6.2 for beta-blockers and phenoalkylamines with a separation voltage of 15 kV. Urine samples spiked with the compounds here studied were treated according to the standard procedure (SPE and evaporation to dryness) and analyzed by CE interfaced with an UV diode-array, set at 195 and 210 nm. The quantitative validation results, obtained analyzing samples at three different concentrations, show a good precision of peak areas that do not exceed 5% for intra-day assays and 10% for inter-day assays. Good linearity (r2 > 0.995) was obtained within the 50,500 ng/mL concentration range. The qualitative validation data show a relative migration times (MTs) variation lower than 1%. The analytes were clearly distinguishable in urine, with LOD and LOQ in the range of 10,80 and 40,100 ng/mL, respectively. [source] Day-to-Day Variation of the Urine Protein: Creatinine Ratio in Female Dogs with Stable Glomerular Proteinuria Caused by X-Linked Hereditary NephropathyJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 3 2007Mary B. Nabity Background:Interpretation of serial urine protein: creatinine (UPC) values is confounded by a lack of data regarding random biologic variation of UPC values in dogs with stable glomerular proteinuria. Hypothesis:That there is minimal day-to-day variability in the UPC of dogs with unchanging proteinuria and the number of measurements needed to reliably estimate UPC varies with the magnitude of proteinuria. Animals:Forty-eight heterozygous (carrier) female dogs with X-linked hereditary nephropathy (XLHN) causing stable proteinuria. Methods:Urine samples were obtained daily by cystocentesis for 3 consecutive days on 183 occasions (549 samples). The UPC was measured for each sample with a single dry-film chemistry auto-analyzer. Data were analyzed retrospectively by a power of the mean model because the variance of UPC values within the 3-day evaluation periods increased as the magnitude of proteinuria increased. Results:To demonstrate a significant difference (P < .05) between serial values in these proteinuric dogs, the UPC must change by at least 35% at high UPC values (near 12) and 80% at low UPC values (near 0.5). One measurement is adequate to reliably estimate the UPC when UPC < 4, but 2,5 determinations are necessary at higher UPC values. Conclusions and Clinical Importance: These guidelines for interpretation of serial UPC values in female dogs with XLHN may also be helpful for interpretation of UPC values in dogs with other glomerulopathies. [source] Evaluation of erythrocyte dysmorphism by light microscopy with lowering of the condenser lens: A simple and efficient methodNEPHROLOGY, Issue 2 2010GYL EANES BARROS SILVA ABSTRACT: Aim: To demonstrate that the evaluation of erythrocyte dysmorphism by light microscopy with lowering of the condenser lens (LMLC) is useful to identify patients with a haematuria of glomerular or non-glomerular origin. Methods: A comparative double-blind study between phase contrast microscopy (PCM) and LMLC is reported to evaluate the efficacy of these techniques. Urine samples of 39 patients followed up for 9 months were analyzed, and classified as glomerular and non-glomerular haematuria. The different microscopic techniques were compared using receiver,operator curve (ROC) analysis and area under curve (AUC). Reproducibility was assessed by coefficient of variation (CV). Results: Specific cut-offs were set for each method according to their best rate of specificity and sensitivity as follows: 30% for phase contrast microscopy and 40% for standard LMLC, reaching in the first method the rate of 95% and 100% of sensitivity and specificity, respectively, and in the second method the rate of 90% and 100% of sensitivity and specificity, respectively. In ROC analysis, AUC for PCM was 0.99 and AUC for LMLC was 0.96. The CV was very similar in glomerular haematuria group for PCM (35%) and LMLC (35.3%). Conclusion: LMLC proved to be effective in contributing to the direction of investigation of haematuria, toward the nephrological or urological side. This method can substitute PCM when this equipment is not available. [source] 1H NMR spectroscopic method for diagnosis of malabsorption syndrome: a pilot studyNMR IN BIOMEDICINE, Issue 2 2004Lakshmi Bala Abstract Despite its well-documented limitations, colorimetry has been commonly used for the d -xylose test in the diagnosis of malabsorption syndrome (MAS). With a possibility of overcoming its limitations, the use of 1H NMR spectroscopy for D -xylose test is explored herein. Urine samples from 35 adults with suspected MAS were obtained before and after oral ingestion of D -xylose. The diagnosis of MAS was based on fecal fat (72,h excretion using Van de Kamer's technique, normal <,7,g/24,h and/or Sudan III stain of spot stool specimen, normal,10 droplets/high power field) and/or endoscopic duodenal biopsy. Urinary excretion of D -xylose over 5,h after consumption of 5,g D -xylose, using both colorimetry and NMR was compared (normal,1,g/5,g/5,h). In vitro experiments on the standard specimens of D -xylose were also performed independently using both methods. Colorimetry showed a lower value for the quantity of D -xylose excreted in urine than NMR [median 0.73 (0.17,1.89,g) vs 1.37 (0.17,3.23,g), respectively; p<0.0001, Wilcoxon's signed ranks test]. Colorimetry and NMR correctly diagnosed 11/12 and 10/12 (p=N.S.) patients with MAS and 14/23 and 20/23 (p<0.05) without MAS, respectively. Sensitivity and specificity of colorimetry and NMR were 91.6 and 60.7% vs 83.3 and 86.9%, respectively. In in vitro experiments, the values obtained for standard xylose using NMR showed a maximum error of 7%, whereas the colorimetric method showed 20%. The NMR method is simple and may be more accurate for the D -xylose absorption test. Colorimetry was found to be inferior as compared with NMR due to its low specificity. Copyright © 2004 John Wiley & Sons, Ltd. [source] Urinary leukotriene E4 levels in children with allergic rhinitis treated with specific immunotherapy and anti-IgE (Omalizumab)PEDIATRIC ALLERGY AND IMMUNOLOGY, Issue 5 2003Matthias Volkmar Kopp Recently, we were able to demonstrate that Omalizumab, a humanized monoclonal anti-IgE antibody, reduces in vitro leukotriene (LT) release of peripheral leukocytes stimulated with allergen in children with allergic rhinitis undergoing allergen immunotherapy. The aim of this study was to investigate the effect of anti-IgE in combination with specific immunotherapy (SIT) on urinary leukotriene E4 (LTE4) levels. Children and adolescents with sensitization to birch and grass pollens and suffering from seasonal allergic rhinitis were included in a phase III, placebo-controlled, multicenter clinical study. Within the four-arm study, patients were randomized to receive SIT for either birch or grass pollen and to receive either subcutaneous anti-IgE or placebo for 24 weeks during the pollen season. From a total population of 225 children, we collected three urine samples in a subgroup of 19 children [n = 12 boys (63%); mean age 11.8 years; range 7.2,17.5 years; Group A (n = 10): SIT (grass or birch) + anti-IgE; Group B (n = 9): SIT (grass or birch) + placebo]. Urine samples were collected before, during and at the end of treatment. Endogenous urinary LTE4 was separated by high-performance liquid chromatography (HPLC) and determined by enzyme immunoassay with a specific antibody. No differences in urinary LTE4 concentrations were observed between the anti-IgE and the placebo groups before (A: 35.2; B: 36.5 nmol/mol creatinine), during (A: 27.0; B: 29.3) and after treatment (A: 28.9; B: 26.5 nmol/mol creatinine). We conclude that urinary LTE4 levels are not helpful in monitoring patients treated with anti-IgE and SIT. [source] Leukotriene synthesis during respiratory syncytial virus bronchiolitis: Influence of age and atopy,PEDIATRIC PULMONOLOGY, Issue 4 2005Giovanni Piedimonte MD Abstract Respiratory syncytial virus (RSV) infection is the most common cause of bronchiolitis in infants and an important risk factor for the development of recurrent wheezing and asthma. Cysteinyl leukotrienes were implicated in the pathophysiology of these diseases, and are being targeted for their diagnosis and therapy. We measured urinary leukotriene E4 (LTE4) in infants with RSV bronchiolitis in comparison with controls without respiratory infection, and investigated whether medical and family history, age, and passive exposure to tobacco smoke are related to urinary leukotriene excretion. We studied 33 infants with bronchiolitis and 25 controls, 1,12 months of age. Demographic and historical data were obtained from informed-consent forms and questionnaires completed by the parents. RSV was detected in nasal secretions by enzyme-linked immunoassay. Urine samples were collected on day of admission and were analyzed for LTE4 with an enzyme-linked immunoassay. Urinary LTE4 was 8-fold higher in infants with bronchiolitis than in controls. Leukotriene excretion was significantly higher in infected infants <6 months of age with a medical history of eczema or dry cough and/or family history of asthma. Multivariate analysis revealed that eczema and dry cough are independently associated with high LTE4 excretion during bronchiolitis. Exposure to tobacco smoke did not affect urinary LTE4. Our study shows that leukotriene synthesis during bronchiolitis is particularly elevated in younger infants with an atopic/asthmatic background. Urinary LTE4 may become a valuable, noninvasive marker for the identification of patients who will benefit most from therapy with leukotriene modifiers for management of bronchiolitis. Pediatr Pulmonol. © 2005 Wiley-Liss, Inc. [source] Rapid screening of clenbuterol in urine samples by desorption electrospray ionization tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 12 2008Ziqing Lin Rapid screening of clenbuterol in urine was performed by combining desorption electrospray ionization (DESI) and tandem mass spectrometry (MS/MS). Optimization experiments were carried out including the selection of substrates, spray solutions, nebulizing gas pressures, high-voltage power supplies and flow rates of spray solution. The limit of detection (LOD), defined as the lowest quantity that can be detected, was 5.0 pg for the pure compound. Using DESI coupled with solid-phase extraction (SPE), the linear response range was from 10 to 400,ng/mL (R2,=,0.993) and the concentration LOD for urine sample was 2.0,ng/mL. The analysis for one spiked urine sample was achieved within 4,min. In addition to the fast analysis speed, MS/MS provided structural information for the confirmation of clenbuterol. Urine samples from different people were investigated and the recoveries were within 100,±,20%. The developed method can potentially be used for screening of clenbuterol in doping control. Copyright © 2008 John Wiley & Sons, Ltd. [source] Determination of urinary S -phenylmercapturic acid, a specific metabolite of benzene, by liquid chromatography/single quadrupole mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 9 2005Luciano Maestri A high-performance liquid chromatography/single quadrupole mass spectrometry (LC/MS) method is described for the determination of urinary S -phenylmercapturic acid (S-PMA), a specific metabolite of benzene. Urine samples were spiked with [13C6]S-PMA (used as the internal standard) and acidified; then they were purified by solid-phase extraction (SPE) on C18 cartridges. Analyses were conducted on a reversed-phase column by gradient runs with 1% aqueous acetic acid/methanol mixtures at different proportions as the mobile phase. The detector was used in electrospray negative ion mode (ESI,), the ions m/z 238 for S-PMA and 244 for [13C6]S-PMA being recorded simultaneously. The detection limit (for a signal-to-noise ratio,=,3) was 0.2,,g/L, thus allowing for the measurement of background excretion of S-PMA in the general population. The use of the internal standard allowed us to obtain good precision (CV% values <3%) and a linear calibration curve within the range of interest for monitoring occupational exposure to benzene (up to 500,,g/L). The method was applied to assay the metabolite concentration in a group of 299 workers (68 smokers and 231 non-smokers) occupationally exposed to relatively low levels of benzene (environmental concentration,=,0.4,220,,g/m3, mean 11.4,,g/m3) and 236 non-exposed subjects (134 smokers and 102 non-smokers). The results clearly showed that smoking must be taken into account for the correct interpretation of the results of S-PMA measurements for the assessment of work-related benzene exposure. When only non-smokers were selected, the mean excretion of S-PMA was significantly higher in workers exposed to benzene (1.2,±,0.9,,g/g creatinine) than in the control group (0.7,±,0.6,,g/g creatinine) (p,<,0.001), thus confirming the role of S-PMA as a biomarker of benzene on a group basis, even for relatively low exposure degrees. Copyright © 2005 John Wiley & Sons, Ltd. [source] Simultaneous determination of t,t -muconic, S -phenylmercapturic and S -benzylmercapturic acids in urine by a rapid and sensitive liquid chromatography/electrospray tandem mass spectrometry methodRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 17 2004Anna Barbieri We describe a rapid and sensitive high-performance liquid chromatography/electrospray tandem mass spectrometry (HPLC/ESI-MS/MS) method for simultaneous determination of the most relevant metabolites of benzene and toluene, t,t- muconic acid (t,t -MA), S -phenylmercapturic acid (S-PMA), and S -benzylmercapturic acid (S-BMA). Urine samples were purified before analysis by solid-phase microextraction (SPE) on SAX cartridges with 50,mg sorbent mass. The developed method fulfils all the standard requirements of precision and accuracy. Calibration curves were linear within the concentration range of the standards (0,80,,g/Lurine for t,t -MA, and 0,25,,g/Lurine for S-PMA and S-BMA), and had correlation coefficients ,0.997. Limits of detection were 6.0,,g/L for t,t -MA, 0.3,,g/L for S-PMA, and 0.4,,g/L for S-BMA. The method was used to determine t,t -MA, S-PMA and S-BMA levels in urine of 31 gasoline-station workers, with personal monitoring data obtained from radial symmetry passive diffusive samplers. In the context of mean work-shift exposures of 75.9,,g/m3 (range 9.4,220.2) for benzene and 331.9,,g/m3 (78.2,932.1) for toluene, metabolite concentrations in end-of-shift urine samples ranged from 23.5,275.3,,g/gcreatinine for t,t -MA, non-detectable to 0.9,,g/gcreatinine for S-PMA, and 3.8,74.8,,g/gcreatinine for S-BMA. No significant correlation was found between the environmental concentrations and urinary metabolites (p,>,0.05 for all cases); the ratios of benzene metabolites could be influenced by exposure levels and co-exposure to xylenes and toluene. The high throughput of this procedure should facilitate exploration of the metabolic effects of benzene-related co-exposure to toluene and alkylbenzenes in large populations of subjects exposed to gasoline. Copyright © 2004 John Wiley & Sons, Ltd. [source] Urine NGAL and IL-18 are Predictive Biomarkers for Delayed Graft Function Following Kidney TransplantationAMERICAN JOURNAL OF TRANSPLANTATION, Issue 7 2006C. R. Parikh Delayed graft function (DGF) due to tubule cell injury frequently complicates deceased donor kidney transplants. We tested whether urinary neutrophil gelatinase-associated lipocalin (NGAL) and interleukin-18 (IL-18) represent early biomarkers for DGF (defined as dialysis requirement within the first week after transplantation). Urine samples collected on day 0 from recipients of living donor kidneys (n = 23), deceased donor kidneys with prompt graft function (n = 20) and deceased donor kidneys with DGF (n = 10) were analyzed in a double blind fashion by ELISA for NGAL and IL-18. In patients with DGF, peak postoperative serum creatinine requiring dialysis typically occurred 2,4 days after transplant. Urine NGAL and IL-18 values were significantly different in the three groups on day 0, with maximally elevated levels noted in the DGF group (p < 0.0001). The receiver,operating characteristic curve for prediction of DGF based on urine NGAL or IL-18 at day 0 showed an area under the curve of 0.9 for both biomarkers. By multivariate analysis, both urine NGAL and IL-18 on day 0 predicted the trend in serum creatinine in the posttransplant period after adjusting for effects of age, gender, race, urine output and cold ischemia time (p < 0.01). Our results indicate that urine NGAL and IL-18 represent early, predictive biomarkers of DGF. [source] Methotrexate catabolism to 7-hydroxymethotrexate in rheumatoid arthritis alters drug efficacy and retention and is reduced by folic acid supplementationARTHRITIS & RHEUMATISM, Issue 8 2009Joseph E. Baggott Objective To assess the catabolism of methotrexate (MTX) to 7-hydroxy-MTX (7-OH-MTX) in patients with rheumatoid arthritis as well as the effect of folic acid and folinic acid on this catabolism. Methods Urinary excretion of MTX and its catabolite, 7-OH-MTX, was measured in 2 24-hour urine specimens collected after MTX therapy. Urine samples were collected from patients after the sixth and seventh weekly doses of MTX. MTX and 7-OH-MTX concentrations were determined by high-performance liquid chromatography mass spectrometry. Swelling and pain/tenderness indices were used to measure symptoms before and at 6 and 7 weeks of therapy. Patients received either folic acid or folinic acid supplements (1 mg/day) from week 6 to week 7. Results Folic acid inhibited aldehyde oxidase (AO), the enzyme that produces 7-OH-MTX, but folinic acid did not. Excretion of 7-OH-MTX (determined as a percentage of the dose of MTX or as mg 7-OH-MTX/gm creatinine) was not normally distributed (n = 39). Patients with marked improvement in swelling and pain/tenderness indices had a lower mean 7-OH-MTX excretion level (P < 0.05). Patients who received folic acid supplements had decreased 7-OH-MTX excretion (P = 0.03). Relatively high 7-OH-MTX excretion was correlated with relatively high MTX excretion and with relatively low MTX retention in vivo (P < 0.05) (n = 35). Conclusion Our findings of a non-normal distribution of 7-OH-MTX excretion suggest that there are at least 2 phenotypes for this catabolism. Decreased 7-OH-MTX formation suggests folic acid inhibition of AO and a better clinical response, while increased 7-OH-MTX formation may interfere with MTX polyglutamylation and binding to enzymes and, therefore, may increase MTX excretion and decrease MTX retention and efficacy in vivo. [source] Relative lung and systemic bioavailability of sodium cromoglycate inhaled products using urinary drug excretion post inhalationBIOPHARMACEUTICS AND DRUG DISPOSITION, Issue 4 2002Osama Aswania Abstract The relative lung and systemic bioavailability of sodium cromoglycate following inhalation by different methods have been determined using a urinary excretion pharmacokinetic method. On three separate randomised study days, 7 days apart, subjects inhaled (i) 4×5 mg from an Intal® metered dose inhaler (MDI), (ii) 4×5 mg from an MDI attached to a large volume spacer (MDI+SP) and (iii) 20 mg from an Intal Spinhaler® (DPI). Urine samples were provided at 0, 0.5, 1, 2, 5 and 24 h post dose. The mean (S.D.) amount of sodium cromoglycate excreted in the urine during the first 30 min post inhalation was 38.1 (27.5), 222.3 (120.3) and 133.1 (92.2) ,g following MDI, MDI+SP and DPI, respectively. The mean ratio (90% confidence interval) of these amounts excreted in the urine over the first 30 min for MDI+SP vs MDI, DPI vs MDI and MDI+SP vs DPI was 801.0 (358.0, 1244; p<0.002)%, 457.0 (244.0, 670.0; p<0.02)% and 262.4 (110.2, 414.5)%, respectively. Similarly for the 24 h cumulative amount of sodium cromoglycate excreted over the 24 h post inhalation the ratios were 375.4 (232.9, 517.9; p<0.005)%, 287.5 (183.4, 391.6; p<0.02)% and 211.4 (88.3, 334.5)%, respectively. The results highlight better lung deposition of sodium cromoglycate from a metered dose inhaler attached to a large volume spacer. Copyright © 2002 John Wiley & Sons, Ltd. [source] Matrix metalloproteinases (MMPs) in bladder cancer: the induction of MMP9 by epidermal growth factor and its detection in urineBJU INTERNATIONAL, Issue 1 2003J.E. Nutt OBJECTIVES To investigate the matrix metalloproteinases (MMPs) 2 and 9 in bladder cancer cell lines stimulated with epidermal growth factor (EGF), and to investigate the presence of gelatinases in the urine of patients with bladder tumours, in relation to the stage and grade of tumour and the EGF receptor (EGFR) status. PATIENTS, SUBJECTS AND METHODS Conditioned media from cultured tumour cells were analysed by zymography. Urine samples from 28 patients with transitional cell carcinoma and 12 normal volunteers were also analysed. Western blotting was used to verify the bands of gelatinolytic activity. The EGFR status of the tumours was assessed by immunohistochemistry. RESULTS MMP9 was induced by EGF in the RT112 but not the RT4 bladder tumour cell line, whereas MMP2 production was unaffected by EGF. Gelatin zymography of urine samples from patients with bladder tumours showed high levels of MMP activity, with 78% positive for MMP9 and 28% positive for MMP2. The total gelatinolytic and MMP9 activity were significantly higher in patients with high-stage invasive tumours than in those with superficial tumours (P < 0.05), and were higher than in normal controls. Gelatinolytic activity at 130 and 200 kDa in urine was identified as MMP9 and MMP2. There was no significant relationship of urinary MMP9 activity to EGFR status of the tumour. CONCLUSION EGF induces MMP9 but not MMP2 in bladder cells. Analysis of urinary gelatinases is a useful noninvasive technique and both total gelatinase and MMP9 activity are associated with high stages of bladder tumours. [source] Incidence and clinical outcome of cytomegalovirus transmission via breast milk in preterm infants ,31 weeksACTA PAEDIATRICA, Issue 2 2009Horst Buxmann Abstract Aim: To evaluate incidence, timing and clinical relevance of acquired human cytomegalovirus (HCMV) infection in preterm infants. Methods: The prospective longitudinal study included preterm infants ,31 weeks. Congenital HCMV infection was excluded by negative HCMV culture from urine or by HCMV-PCR-negative umbilical cord blood. Infants from HCMV-IgG-positive mothers received thawed frozen breast milk until 33 weeks. Urine samples were obtained weekly for HCMV culture. Data were collected regarding clinical course and milk-intake. Results: Twenty-nine mothers (29/48, 60%) of 35 infants were HCMV-IgG-positive. Five of 35 infants (14%) excreted HCMV in urine. Three of five children remained asymptomatic. One child developed a respirator-dependent HCMV pneumonia, the other child an upper airway infection and a transient thrombocytopenia. HCMV infected children had a significant longer hospital stay (median 96 vs. 73 days, p = 0.025) and received more formula milk (89 vs. 44 mL/kg/day, p = 0.04). Mothers of infected children had significantly higher HCMV-IgG levels than those of non-infected children (mean 1557 vs. 921 AU/mL, p = 0.048). Nineteen of 48 mothers (40%) with 23 infants were HCMV-IgG-negative. These children remained HCMV negative. Conclusion: Feeding preterm infants ,31 weeks of HCMV-IgG-positive mothers with thawed frozen breast milk until 33 completed weeks does not prevent symptomatic HCMV infection in all cases. These infections can be associated with a prolonged hospital stay. [source] Endocrinological and endometrial factors in recurrent miscarriageBJOG : AN INTERNATIONAL JOURNAL OF OBSTETRICS & GYNAECOLOGY, Issue 12 2000T. C. Li Consultant Gynaecologist Objective To investigate the endocrinological and endometrial factors in women with unexplained recurrent miscarriage Design Prospective, case study Setting Recurrent miscarriage clinic, Jessop Hospital for Women, Sheffield Participants One hundred and forty-four women with unexplained recurrent (, 3) miscarriages Methods A blood sample was obtained in early follicular phase (day 3,5) to measure follicle stimulating hormone, luteinising hormone, prolactin, androgens and thyroid function; daily blood/urine samples were obtained from mid-follicular phase to measure luteinising hormone until the luteinising hormone surge was identified; endometrial biopsy and a further blood sample for progesterone measurement were obtained in the mid-luteal phase. A transvaginal ultrasonography was performed to evaluate ovarian morphology. Results Hypersecretion of luteinising hormone or ultrasonographic features of polycystic ovarian disease was present in 8% and 7.8% of women, respectively. The free androgen index was elevated in 14.6% of subjects. In the mid-luteal phase, low progesterone level was found in 17.4% and delayed endometrial development was noted in 27.1% of women. Although women with recurrent miscarriage women and delayed endometrium had significantly lower progesterone levels than those with normal endometrial development, only 8/24 had mid-luteal progesterone levels below 30 nmol/L. Recurrent miscarriage was not associated with hyperprolactinaemia or abnormal thyroid function test. Conclusions Endocrinological and endometrial abnormalities are present in about a quarter of women with unexplained recurrent miscarriage. [source] Finnegan neonatal abstinence scoring system: normal values for first 3 days and weeks 5,6 in non-addicted infantsADDICTION, Issue 3 2010Urs Zimmermann-Baer ABSTRACT Objective The neonatal abstinence scoring system proposed by Finnegan is used widely in neonatal units to initiate and to guide therapy in babies of opiate-dependent mothers. The purpose of this study was to assess the variability of the scores in newborns and infants not exposed to opiates during the first 3 days of life and during 3 consecutive days in weeks 5 or 6. Patients and methods Healthy neonates born after 34 completed weeks of gestation, whose parents denied opiate consumption and gave informed consent, were included in this observational study. Infants with signs or symptoms of disease or with feeding problems were excluded. A modified scoring system was used every 8 hours during 72 hours by trained nurses; 102 neonates were observed for the first 3 days of life and 26 neonates in weeks 5,6. A meconium sample and a urine sample at weeks 5,6 were stored from all infants to be analysed for drugs when the baby scored high. Given a non-Gaussian distribution the scores were represented as percentiles. Results During the first 3 days of life median scores remained stable at 2 but the variability increased, with the 95th percentile rising from 5.5 on day 1 to 7 on day 2. At weeks 5,6 median values were higher during daytime (50th percentile = 5, 95th percentile = 8) than night-time (50th percentile = 2, 95th percentile = 6, P = 0.02). Conclusion Scores increase from days 1,3 to weeks 5,6 and show day,night cycles with 5,6 weeks. Values above 8 can be considered pathological. This data may help to raise suspicion of narcotic withdrawal and to guide therapy. [source] Separation and Detection of Narcotic Drugs on a Microchip Using Micellar Electrokinetic Chromatography and ElectrochemiluminescenceELECTROANALYSIS, Issue 6 2008Yan Du Abstract A new approach for fast and sensitive electrochemiluminescence (ECL) detection of narcotic drugs on a microchip after separation by micellar electrokinetic chromatography (MEKC) is presented, taking the cocaine and its hydrolysate ecgonine as the test analytes. The mixture of hydrophilic BMIMBF4 ionic liquid (IL) and sodium dodecyl sulfate (SDS) was used directly as the buffer of MEKC with less noisy baselines, lower electrophoretic current and satisfactory separation performance. This developed microchip MEKC,ECL system was successfully applied to the determination of two very similar narcotics, heroin and codeine, within 100s in urine sample and was demonstrated as a promising method in clinical and forensic analysis. [source] Direct chiral analysis of primary amine drugs in human urine by single drop microextraction in-line coupled to CEELECTROPHORESIS, Issue 16 2009Kihwan Choi Abstract Three-phase single drop microextraction (SDME) was in-line coupled to chiral CE of weakly basic amine compounds including amphetamine. SDME was used for the matrix isolation and sample preconcentration in order to directly analyze urine samples with the minimal pretreatment of adding NaOH. A small drop of an acidic aqueous acceptor phase covered with a thin layer of octanol was formed at the tip of a capillary by simple manipulation of the liquid handling functions of a commercial CE instrument. While the saline matrix of the urine sample was blocked by the octanol layer, the basic analytes in a basic aqueous donor phase were concentrated into the acidic acceptor drop through the octanol layer by the driving force of the pH difference between the two aqueous phases. The enantiomers of the enriched amines were resolved by using (+)-(18-crown-6)-tetracarboxylic acid as a chiral selector for the subsequent CE separation. From 10,min SDME with the agitation of the donor phase by a small stirrer retrofit to the CE instrument, enrichment factors were about a 1000-fold, yielding the LOD of 0.5,ng/mL for amphetamine. This low LOD value as well as the convenience of in-line coupled SDME make the proposed scheme well suited for the demanding chiral analysis of amphetamine-type stimulants. [source] Enantioselective analysis of pheniramine in urine by charged CD-mediated CZE provided with a fiber-based DAD and an on-line sample pretreatment by capillary ITPELECTROPHORESIS, Issue 15 2007Jozef Marák Abstract Application potentialities of CZE on-line coupled with capillary ITP and DAD to the identification and determination of trace concentration levels (,g/L) of pheniramine (PHM) enantiomers and their metabolites present in complex ionic matrices of biological origin (urine) are shown. An enhanced (enantio)selectivity of the CZE separation system obtained by the addition of carboxyethyl-,-CD (CE-,-CD) to the carrier electrolyte provided CZE conditions for a reliable identification of similar/identical DAD spectra of structurally related compounds (PHM enantiomers and their metabolites) in clinical urine samples differing in qualitative and quantitative composition of sample matrix constituents. A high sample loadability (a 30,,L sample injection volume), partial sample clean-up (removing macroconstituents from the sample), and preconcentration of the analytes in ITP stage resulted in the decrease of concentration LOD for PHM enantiomers in urine to 5.2 and 6.8,,g/L (2.2×10,8 and 2.8×10,8,mol/L), without using any sample pretreatment technique. The background correction and smoothing procedure applied to the raw DAD spectra provided analytically relevant DAD spectra of PHM enantiomers and their metabolites also when they were present in urine sample (30,,L injection volumes of ten-times diluted urine sample) at a 9×10,8,mol/L concentration. DAD spectra of PHM enantiomers present in urine samples matched their reference spectra with reasonable certainties. DAD spectra of PHM metabolites were compared with the reference spectra of PHM enantiomers and a good match was found which indicates the similarities in the structures of enantiomers and their metabolites detected in the urine samples. This fact allows performing the quantitative analyses of PHM metabolites in the urine samples by applying the calibration parameters of PHM enantiomers also for PHM metabolites and the results show the possibilities of using the ITP,CZE,DAD combination for the direct analysis of PHM enantiomers and/or their metabolites in urine without any sample pretreatment. ITP,CZE,DAD method with oppositely charged selector is suggested to use in clinical research as it provides favorable performance parameters including sensitivity, linearity, precision, recovery, and robustness with minimal demands on sample preparation. [source] Poly(methacrylic acid-ethylene glycol dimethacrylate) monolith in-tube solid-phase microextraction applied to simultaneous analysis of some amphetamine derivatives in urine by capillary zone electrophoresisELECTROPHORESIS, Issue 16 2005Fang Wei Abstract A method based on in-tube solid-phase microextraction and capillary zone electrophoresis (CZE) was proposed for simultaneously determining four amphetamines (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, and 3,4-methylenedioxymethamphetamine) in urine. A poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column, which can provide sufficient extraction efficiency, was introduced for the extraction of amphetamines from urine samples. The hydrophobic main chains and acidic pendant groups of the monolithic column make it a superior material for extraction of basic analytes from aqueous matrix. After extraction, the samples were analyzed by CZE. The best separation was achieved using a buffer composed of 0.1,M disodium hydrogen phosphate (adjusted to pH,4.5 with 1,M hydrochloric acid) and 20% methanol v/v, with a temperature and voltage of 25°C and 20,kV, respectively. By applying electrokinetic injection with field-amplified sample stacking, detection limits of 25,34,µg/L were achieved. Excellent method of reproducibility was found over a linear range of 0.1,5,mg/L. Determination of these analytes from abusers' urine sample was also demonstrated. [source] | ||||||||||||||||||||||||||||||||||