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Effect of high and low calcium diets on stone forming risk during liberal oxalate intake.

PURPOSE: Recent studies suggest that a high calcium diet protects against calcium oxalate stone formation. We compared the effect of high and low calcium diets on urinary saturation of calcium oxalate during liberal oxalate intake. MATERIALS AND METHODS: A total of 10 healthy subjects (5 male, 5 female) participated in a 2-phase, randomized, crossover study comparing high (1,000 mg daily) and low (400 mg daily) calcium intake on a liberal oxalate diet (200 mg daily). During each phase subjects adhered to an instructed diet for 3 days followed by a controlled, metabolic diet for 4 days. Blood and 24-hour urine specimens collected on the last 2 days of each phase were analyzed for serum biochemistry studies and stone risk factors, respectively. RESULTS: Urinary calcium was higher (mean +/- SD 171 +/- 64 vs 124 +/- 49 mg daily, p = 0.002) and oxalate was lower (25 +/- 4.8 vs 27 +/- 4 mg daily, p = 0.02) on the high vs low calcium diet. Overall, the urinary relative saturation ratio of calcium oxalate was higher on the high compared with the low calcium diet (3.3 vs 2.5, p <0.0001) even after adjusting for confounding variables. CONCLUSIONS: In normal subjects urinary saturation of calcium oxalate was higher on a high calcium diet than a low calcium diet during liberal oxalate intake because the decrease in urinary oxalate did not overcome the effect of increased calcium. A high calcium diet during liberal oxalate intake may pose an increased risk of calcium oxalate stone formation.

Adult↗

Improved HPLC method for analysis of 1-hydroxypyrene in human urine specimens of cigarette smokers.

A method for the rapid and sensitive analysis of 1-hydroxypyrene in human urine was improved by high performance liquid chromatography (HPLC) with fluorescence detection. The limit of detection decreased approximately threefold when triethylamine was added to the extracts prior to the chromatographic determination. The sensitivity was increased mainly due to triethylamine blockage of the secondary-retention effects. The analysis time was approximately 20 min per run. This method was applied to the analysis of 1-hydroxypyrene in 125 urine specimens collected from cigarette smokers who had non-occupational exposure to polycyclic aromatic hydrocarbons. The average concentrations of urinary 1-hydroxypyrene were 0.04 micromol/mol creatinine in non-smokers, 0.20 micromol/mol creatinine in light smokers, 0.46 micromol/mol creatinine in medium smokers, and 1.16 micromol/mol creatinine in heavy cigarette smokers. The levels of urinary 1-hydroxypyrene correlated strongly with cigarette intake (r2 = 0.93, P < 0.001).

Adolescent↗

Screening for sexually transmitted infections in substance abuse treatment programs.

OBJECTIVES: We evaluated the prevalence of the sexually transmitted infections (STIs) chlamydia and gonorrhea in clients at a methadone maintenance program and a residential detoxification program. METHODS: We collected urine specimens for chlamydia and gonorrhea ligase chain reaction testing and assessed sexual, substance abuse and STI histories. RESULTS: Of 700 subject assessments, 490 occurred among detoxification clients and 210 in methadone maintenance. Chlamydia trachomatis was detected in 5/700 (0.9, 95% CI=0.1-1.8%) and Neisseria gonorrhoeae in none. All chlamydia infected subjects were recruited from the detoxification program. Subjects reported high risk sexual behavior: 17% reported commercial sex exchange, and 22% reported inconsistent condom use with multiple sexual partners during the prior 2 months. CONCLUSION: Based on prevalence in Boston, MA, universal screening for STI in substance abuse treatments programs is not warranted. However, routine screening for younger substance abusers and in communities with high prevalence should be considered.

Adult↗

Contingency management in outpatient methadone treatment: a meta-analysis.

A meta-analysis was conducted on contingency management interventions in outpatient methadone treatment settings. The outcome measure of interest was drug use during treatment, as detected through urinalysis. The results confirm that contingency management is effective in reducing supplemental drug use for these patients. The analysis of behavioral interventions yielded an overall effect size (r) of 0.25 based on 30 studies. Significant moderators of outcomes included type of reinforcement provided, time to reinforcement delivery, the drug targeted for behavioral change, number of urine specimens collected per week, and type of subject assignment. These factors represent important considerations for reducing drug use during treatment.

Behavior Therapy↗

Determination of the major urinary metabolite of flurazepam in man by high-performance liquid chromatography.

A high performance liquid chromatographic method for the determination of N-1-hydroxyethylflurazepam, the major urinary metabolite of flurazepam, in human urine is described. Urine specimens were incubated enzymatically to deconjugate N-1-hydroxyethylflurazepam glucuronide (metabolite) and were then extracted at pH 9.0 to extract the metabolite. The extracts were chromatographed on a microparticulate silica gel column using automatic sample injection, isocratic elution at ambient temperature and UV monitoring at 254 nm. The internal standard was 7 chloro-5(2'-chlorophenyl) 1,3-dihydro-1-2-dimethylaminoethyl-2H-1,4-benzodiazepine-2-one. The recovery from urine, in the 0.5-25.0 microgram/ml range, was 96.5 +/- 11.5% (S.D.), and the sensitivity limit was 0.5 microgram/ml. The method was found to be specific for N-1-hydroxyethylflurazepam in the presence of intact flurazepam and other possible urinary metabolites of flurazepam. The method was successfully applied to urine specimens collected from human subjects following the administration of 30-mg single oral doses of flurazepam dihydrochloride.

Adult↗

High-performance liquid chromatographic method for the determination of eclanamine and its N-desmethyl and N,N-didesmethyl metabolites in urine.

A high-performance liquid chromatographic method has been defined for the determination of eclanamine (free base of eclanamine maleate) and two of its metabolites, N-desmethyleclanamine and N,N-didesmethyleclanamine in urine. The method employs 10-ml urine samples, has a linear range from 5 to 500 ng/ml for the three compounds, and has a detection limit of 0.5 ng/ml for each compound. Sample preparation uses a cyanopropylsilane extraction column with washes of water, acetonitrile-water (30:70, v/v), and acetonitrile, and elution with 2% trifluoroacetic acid in acetonitrile. The eluate is evaporated to dryness, the residue dissolved in 1.0 ml acetonitrile-water (10:90, v/v) and 100 microliter are injected onto a Supelcosil LC-CN column. Eclanamine and its metabolites are eluted with an acetonitrile-water (35:65, v/v) eluent containing 0.01 M triethylamine and adjusted to pH 7.0 with phosphoric acid. The method has been validated by preparing and analyzing a series of fortified urines (range 2-500 ng/ml for each compound) on four separate days. Good linearity, precision, reproducibility, and specificity were obtained. Certification of the analytical method was accomplished by analyzing urine specimens collected from one volunteer administered a single oral dose of 45 mg eclanamine maleate. The data suggest that the metabolites of eclanamine have long elimination half-lives with levels still quantifiable in the 72-96 h collection interval.

Anilides↗

Finnish quality assurance programme for biological monitoring of organic solvents.

The FIOH quality assurance programme for organic solvents and their metabolites consists of analyses for 2,5-hexadione, phenol, mandelic acid, methylenedianiline, methylhippuric acid, trans,trans-muconic acid and trichloroacetic acid in urine, and creatinine and relative density for standardisation. Four times a year two levels of spiked urine or urine specimens collected from occupationally exposed workers are distributed to the participants in 22 countries. RSD and recovery were studied during 1997-2000. Average RSDs of all participants varied between 23 and 56% and were clearly dependent on the analytical method used and the concentration level of the samples. Since 1997 the target values have been determined in reference laboratories for five of the analytes. Lower RSDs (9-21%) and good recoveries were obtained for all analytes in these laboratories, indicating that good performance can be achieved even in the complex analyses performed in biological monitoring of exposure to industrial chemicals.

Environmental Monitoring↗

Interpretation of GC-MS opiate results in the presence of pholcodine.

Although the cross reactivity of pholcodine with opiate immunoassays has been well documented there is little published information the potential for pholcodine interference with chromatographic analyses. Wilson and Smith [Ann. Clin. Biochem. 36 (1999) 592] recently described the 'misidentification' of morphine in quality control specimens that had been spiked with pholcodine. This report describes a sensitive, rapid gas chromatography-mass spectrometry (GC-MS) method for the detection and quantitation of pholcodine and morphine, together with 6-monoacetylmorphine (6-MAM), codeine and dihydrocodeine in urine. This method was used to analyse urine specimens collected from volunteers given single and multiple doses of pholcodine to establish the significance this drug on the analytical results obtained when performing drug screening according to the proposed UK and EU legally defensible workplace drug testing guidelines. The maximum urinary free morphine concentration achieved following a single 10mg oral dose of pholcodine was 1.39 mg/l at 2-4h post dose. Following multiple 10mg oral doses of pholcodine the maximum urinary free morphine concentration was determined as 0.4 mg/l at 170 h after the final dose was administered. This apparent anomaly in the morphine concentrations obtained following single and multiple pholcodine doses can be explained in part by differences in the concentration of the specimens, and may be overcome by applying a correction factor for specimen dilution using their creatinine concentration. The data from this study suggests that even following one single 10mg dose of pholcodine, free morphine concentrations greater than both the proposed UK workplace drug testing guidelines threshold of 0.3mg/l total morphine and the proposed European Union threshold of 0.2mg/l total morphine can be achieved. This highlights the need for caution when interpreting confirmatory opiate data, especially in medicolegal and clinical cases, and in cases where the use of pholcodine is suspected.

Antitussive Agents↗

Comparative analysis of amplified fragment length polymorphism and pulsed field gel electrophoresis in a hospital outbreak and subsequent endemicity of ampicillin-resistant Enterococcus faecium.

Reliable molecular methods for determination of relatedness between bacterial isolates have become increasingly important to evaluate outbreaks and endemic situations with nosocomial pathogens. In the present study Simpson's index of diversity with calculated confidence intervals was used to compare amplified fragment length polymorphism (AFLP) and pulsed field gel electrophoresis (PFGE) analysis of a hospital outbreak of ampicillin-resistant Enterococcus faecium and subsequent endemicity. The outbreak, in a Norwegian tertiary hospital, of infections caused by these enterococci started in 1995 and increased in 1996 after which the situation turned endemic. The purpose of this study was to compare the two methods in this setting and to determine the length of time during an outbreak that these methods are sufficiently valid to be of value for hospital infection control efforts. One hundred and sixty clinical isolates from urine specimens collected during the period 1995-1999 were included. The findings indicate that PFGE and AFLP are equally discriminative and could in this setting be used for typing purposes over the whole 5-year period.

Ampicillin↗

Doping control analysis of intact rapid-acting insulin analogues in human urine by liquid chromatography-tandem mass spectrometry.

Insulin and related synthetic therapeutics have been prohibited by the World Anti-Doping Agency for athletes demonstrably not suffering from diabetes mellitus. The primary specimen for doping controls has been urine, but the renal excretion of intact human insulin as well as synthetic analogues such as the rapid-acting products Humalog LisPro, Novolog Aspart, and Apidra Glulisine has been reported negligible owing to metabolic degradation. Nevertheless, employing solid-phase extraction in combination with immunoaffinity purification followed by a top-down sequencing-based mass spectrometric approach, an assay was established allowing the identification of three intact rapid-acting synthetic insulins in doping control urine samples. A volume of 25 mL of urine was concentrated, insulin analogues were isolated from the concentrate by immunoaffinity chromatography, and the eluate was analyzed using microbore liquid chromatography/tandem mass spectrometry. Characteristic product ion spectra obtained from 5-fold protonated intact analytes as well as isolated insulin B-chains allowed the unambiguous identification of target analytes with detection limits of 0.05 ng/mL (9 fmol/mL). Moreover, assay validation demonstrated recoveries between 72 and 80% for Humalog LisPro, Novolog Aspart, and Apidra Glulisine, and assay precisions ranged from 9 to 16%. A reliable tool is provided that allows the qualitative determination of rapid-acting insulins in urine specimens collected for sports drug testing.

Amino Acid Sequence↗

Prevalence and risk factor analysis of microalbuminuria in Japanese general population: the Takahata study.

Microalbuminuria, an indicator of glomerular injury, is associated with increased risk of progressive renal deterioration, cardiovascular disease, and mortality. However, the prevalence of microalbuminuria in Japanese general population is less certain. Thus, we examined the prevalence of microalbuminuria and its associated risk factors in Japan. Subjects of this cross-sectional study were asymptomatic individuals over 40 years in Takahata, Japan. Urine albumin-creatinine ratio was calculated from a single-spot urine specimen collected in the morning. Creatinine clearance (CCr) was obtained by Cockcroft-Gault equation. Multivariate logistic regression analysis was used to determine which risk factors (i.e., age, hypertension, diabetes, obesity, and salt intake) might predict the presence of microalbuminuria. A total of 2321 subjects (mean age, 64 years; men, 1034; women, 1287) were entered into the final analysis. Among them, the prevalence of microalbuminuria, macroalbuminuria, and proteinuria by dipstick test (> or = 1+) were 317 (13.7%), 39 (1.7%), and 103 (4.4%), respectively. Age, hypertension, and diabetes were independently associated with microalbuminuria in men. In addition to the classical risk factors detected in men, estimated 24-h urinary sodium excretion and uric acid were also independently associated with microalbuminuria in women. Among the 668 subjects with renal insufficiency (CCr <60 ml/min/1.73 m(2)), the prevalence of microalbuminuria and macroalbuminuria were 119 (17.8%) and 18 (2.7%), respectively. In conclusion, microalbuminuria is prevalent across all age groups and is associated with lifestyle-related risk factors in Japanese general population. However, there are a substantial number of subjects with renal insufficiency accompanying no microalbuminuria.

Aged↗

Field evaluation of a second-generation cytometer UF-100 in diagnosis of acute urinary tract infections in adult patients.

AIMS: The authors evaluated the analytical performance of the Sysmex UF-100 cytometer vs. the diagnosis of urinary tract infections (UTI). METHODS: We considered 2010 subjects, aged between 18 and 78, 870 males and 1140 females. The majority (90.2%) of the samples were voided urine specimens collected by using the midstream technique. Each sample was subjected to microbiological evaluation (culture + residual antibacterial activity), dipstick tests, UF-100 examination and microscopic observation. In order to obtain a final diagnosis of UTI these laboratory results were taken into consideration together with clinical data and patients' characteristics. The analytical performance of the laboratory tests was obtained by adopting this diagnosis as standard practice. RESULTS: Out of the total 2010 subjects considered a clinical diagnosis of UTI was obtained in 529 cases (26.32%). The UF-100-based screening had sensitivity, 0.94; specificity, 0.93; positive predictive value, 0.83; negative predictive value, 0.98; and correctly classified incidence, 0.93. CONCLUSIONS: In our experience the results of the UF-100-based screening show a very good correlation with the diagnosis of acute UTI in adults patients.

Acute Disease↗

Urinary screening for Neisseria gonorrhoeae in asymptomatic individuals from Queensland, Australia: an evaluation using three nucleic acid amplification methods.

A comparison between three nucleic acid amplification methods (COBAS AMPLICOR NG, Abbott LCx NG and in-house cppB gene PCR) for the detection of Neisseria gonorrhoeae was undertaken using non-consecutive urine specimens collected from a high prevalence asymptomatic population (260 patients) from multiple sites in Queensland, Australia. Sensitivity, specificity, positive predictive value and negative predictive value were as follows: COBAS 97.9, 93.9, 78.0, 99.5%; LCx 95.7, 100, 100, 99.1%; cppB 97.9, 100, 100, 99.5%. The results provide further evidence that false-positive COBAS AMPLICOR NG PCR results are a significant problem in our population, and that the Abbott NG LCx is a suitable alternative for screening asymptomatic populations with a high prevalence of N. gonorrhoeae carriage.

DNA, Bacterial↗

Comparison of PCR/nucleic acid hybridization and EIA for the detection of Chlamydia trachomatis in different populations in a regional centre.

Culture on McCoy cell monolayers has been accepted as the reference method for the detection of Chlamydia trachomatis. Recent studies have shown that polymerase chain reaction (PCR)/nucleic acid hybridization based methods have increased sensitivity over culture while still retaining specificity. In situations where organism viability is of concern, due to factors such as transportation delays, culture is inappropriate. Regional laboratories therefore have not been able to utilize the reference method and have been forced to use less reliable methods. The aims of our study were to assess the feasibility of performing PCR to diagnose infections due to C. trachomatis in a regional laboratory using a new commercial kit--Amplicor (Roche Molecular Systems, Branchburg, NJ) and to compare the current enzyme-immunoassay (EIA) based-methods used in our laboratory (VIDAS [bioMerieux Vitek, Hazelwood MO] and IDEIA [Novo Nordisk Diagnostics, Cambridge, UK]) against PCR. Thirteen positive Amplicor specimens were found in 267 urine specimens collected from asymptomatic adolescent males and females. All 13 were confirmed positive using major outer membrane protein gene PCR (MOMP). VIDAS and IDEIA showed 100% correlation to each other but only detected 5/13 positives. Of 140 consecutive patients attending the regional sexual health clinic, 13 were Amplicor positive, 11/13 MOMP positive and 10/13 positive by VIDAS. Five of 254 patients attending the hospital antenatal clinic were positive by Amplicor, all being confirmed by MOMP. No PCR inhibition was detected in a random sample of 100 varied negative Amplicor tests using a modification of the Amplicor kit. No contamination was experienced. The Amplicor kit was shown to be suitable for use in the routine clinical laboratory with minimal disruption to workflow. For regional laboratories this kit should provide more accurate results than EIA based methods, particularly in the detection of asymptomatic persons.

Adolescent↗

Detection of Candida casts in experimental renal candidiasis: implications for the diagnosis and pathogenesis of upper urinary tract infection.

The distinction between upper versus lower urinary tract infection in patients with candiduria is a commonly encountered and therapeutically important diagnostic dilemma. Candida casts have been reported in the urine of several individual case reports of human renal candidiasis. The specificity of Candida casts would identify unequivocally a patient with upper urinary tract disease. Little is known, however, about the sensitivity and the formation of Candida casts. We therefore studied the diagnostic yield, methods for detection and pathogenesis of Candida cast formation in serially collected urine specimens from immunologically intact and granulocytopenic rabbit models of haematogenous disseminated candidiasis. Refractile blastoconidia and pseudohyphae of Candida encased in the granular matrix were seen on wet mounts while Candida stained a brilliant red in the fuschia pink tubular matrix on periodic acid Schiff (PAS) stained cytopathology filters. Among 24 rabbits with disseminated candidiasis, 11 (46%) had Candida casts detectable by wet mount and PAS-stained urine filters in comparison to none of 10 non-infected immunologically normal controls (P = 0.014). Fifteen (70%) of 21 episodes of Candida casts were detected within the first 3 days of infection, indicating possible utility in the early diagnosis of renal candidiasis. No Candida casts were detected in the urine of granulocytopenic rabbits, possibly due to the rapid destruction of tubules and abrogation of cast formation. This absence of detectable Candida in eight infected granulocytopenic rabbits differed significantly from that of 24 non-granulocytopenic infected rabbits, in which Candida casts were detected in 11 (46%) (P = 0.029). Candida cast formation occurred predominantly in the cortex. Histopathological examination demonstrated invasion of Candida into the glomerular tufts and peritubular capillaries, followed by development of Candida casts in the proximal and distal tubules, respectively. Detection of renal Candida casts may be a useful diagnostic marker in distinguishing upper versus lower urinary tract candidiasis.

Animals↗

A comparison of the OnTrak Testcup-5 to laboratory urinalysis among arrestees.

Research studies that collect urine specimens to measure recent drug use have traditionally sent the specimens to laboratories for analysis. A new method of urinalysis-instant urine testing-may offer a quicker, equally accurate alternative to laboratory assays. To date, however, no studies have explored the efficacy of instant urine technology with individuals under criminal justice supervision. To address this limitation, the authors administered a single instant urine test-the OnTrak Testcup-5-to 136 adult male arrestees surveyed through Maryland's Substance Abuse Need for Treatment among Arrestees (SANTA) study. The specimens were subsequently shipped to an independent laboratory for reanalysis. Analyses indicate high agreement between the two techniques for marijuana, cocaine, and heroin. Potential uses for the OnTrak Testcup-5 are discussed.

Adolescent↗

Congenital cytomegalovirus infection in an urban Canadian community.

congenital cytomegalovirus (CMV) infection was diagnosed in 64 (0.42%) of 15,212 infants born in Hamilton, Ontario, Canada, during a period of 44 consecutive months. Urine specimens collected within 48 hr of birth were screened for CMV by inoculation into cell cultures. Only four infants had clinical or laboratory findings in the first week of life that suggested cytomegalic inclusion disease; CMV infection was mild or inapparent in the remainder. Fourteen CMV-positive infants (21.9%) had birth weights of less than or equal to 2,500 g, including eight of 11 premature infants (less than or equal to 37 weeks of gestation). Only five CMV-positive infants were small for gestational age. Overall, there were no significant differences between CMV-positive and CMV-negative infants in mean gestational age or mean measurements of weight and head circumference at birth. Mothers of CMV-positive infants were predominantly younger, primiparous women of lower educational and economic status, and the number who were unmarried was about threefold greater than among mothers of uninfected infants.

Adolescent↗

Detection of specific bacterial antigen in urine of patients infected with Bacteroides fragilis.

An indirect enzyme-linked immunosorbent assay was developed to detect specific Bacteroides fragilis antigen(s) in human urine. Specimens collected within 72 hr of a positive culture were centrifuged, dialyzed, and treated with Tween 20, polyethylene glycol, and bovine serum albumin. Goat hyperimmune gamma-globulin to B. fragilis strain ATCC 23745 was added and incubated, and supernatants were tested for antibody activity to polysaccharide-protein antigen of the same organism. Mean +/- SD results, reported as percentage inhibition of control values and interpreted blindly, were as follows: 29 normal subjects, 9.8% +/- 6.0%; 22 patients with Enterobacteriaceae bacteremia, 6.0% +/- 5.1%; six patients with nonbacteremic infections due to B. fragilis, 22.3% +/- 10.3%; and nine patients with B. fragilis bacteremia, 28.7% +/- 10.2%. Three of six nonbacteremic patients and eight of nine bacteremic patients yielded values greater than 2 SD of control values. None of 22 patients with Enterobacteriaceae bacteremia was falsely positive (specificity, 100%).

Antigens, Bacterial↗