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Oxalate is overestimated in alkaline urines collected during administration of bicarbonate with no specimen pH adjustment.

We compared measurements of daily urine oxalate excretion in urines collected at the prevailing urine pH with measurements of urine oxalate excretion in urines collected into 20 mL of 6 mol/L HCl. We studied eight healthy adults fed constant diets. Urines were collected during control conditions and, in each subject, during the administration of NaCl, KCl, NaHCO3, or KHCO3, 90 mmol/day. Daily urine oxalate excretion calculated for collections made in acid averaged 271 (SD 79) mumol/day and did not vary with any of the salt supplements. When urines were collected at ambient urine pH (average 5.94, SD 0.23) during control conditions, and during the administration of NaCl or KCl, urine oxalate excretion averaged 263 (SD 88) mumol/day, a value not different from that for collections in acid. However, when urine was collected with no pH adjustment during NaHCO3 or KHCO3 administration (average pH 6.90, SD 0.14), apparent urine oxalate excretion averaged 398 (SD 132) mumol/day, significantly (P less than 0.025) exceeding the mean observed when urines were collected in acid. Moreover, the percentage increase in apparent oxalate excretion increased with urinary pH. These observations reinforce recommendations that urine specimens for measurement of oxalate be collected in acid to avoid the increase in apparent oxalate content that occurs during collection of alkaline urines. This increase presumably results from the well-known in vitro nonenzymatic conversion of ascorbate to oxalate.

Adult↗

Timing of specimen collection is crucial in urine screening of drug dependent mothers and newborns.

We compared results of urine drug analysis with clinical data and history to test the usefulness of peripartum drug screening and to establish guidelines for optimal testing. Urine from 28 mothers and 52 babies was analysed. Drugs not suspected by history were found in 10 mothers and six babies. Results assisted in the management of neonatal withdrawal in three babies. Drugs suspected by history were not found in 11/22 mothers and 23/35 babies. About half of these results were associated with delayed urine collection. In 12/28 mothers, drugs administered in hospital could have confused interpretation of screen results. We conclude that urine drug screening without strict protocols for specimen collection is of limited usefulness for management of drug abuse in pregnancy and neonatal drug withdrawal. We favour testing of maternal urine obtained before drugs are administered in hospital. Neonatal urine, if used, should be collected in the first day of life.

Drug Evaluation, Preclinical↗

Program monitoring for clinical practice. Specimen positivity across urine collection methods.

Urinalysis is a primary component of methadone programming. A clinic-based study was conducted to determine whether unsupervised urine collection is as reliable as other methods for detecting unauthorized drug use among methadone treatment clients. Drug positivity rates compared over a 30-week period revealed no significant differences as a function of collection method. Results confirm the efficacy of unsupervised urine collection using a heat sensitive strip or temperature indicator. Study findings also offer evidence to support clinical practice designed to ensure client privacy and dignity while maintaining standards for effective program monitoring.

Adult↗

Response of CEDIA amphetamines assay after a single dose of bitter orange.

Bitter orange has recently been substituted as an ingredient in many "ephedra-free" dietary supplements used for weight loss. The primary active ingredient in bitter orange is synephrine. Previous reports have documented false-positive results from ephedrine with urine amphetamine assays. Because of the similarity in chemical structure of ephedrine and synephrine, it is hypothesized that ingestion of a bitter orange supplement may have the potential to cause false-positive results with urine amphetamine assays. The purpose of this study was to determine the response of the CEDIA Amphetamines Assay after ingestion of bitter orange. Six healthy adult male volunteers were administered a single oral dose of Nature's Way Bitter Orange, a 900-mg dietary supplement extract standardized to 6% synephrine. Urine specimens were collected at baseline and 3 and 6 hours post-administration. Additional urine specimens were collected from 1 subject at 9, 12, and 15 hours after administration. All specimens were analyzed by the CEDIA Amphetamines Assay. Urine specific gravity and pH also were measured. All urine specimens demonstrated a negative response to the CEDIA Amphetamines Assay. Urine specific gravity ranged from 1.007 to 1.028, and pH ranged from 5.0 to 7.0; thus, reducing the possibility that the negative results were caused by diluted specimens or reduced excretion of synephrine into alkaline urine. This information will be of value when health care providers or those who interpret drug screens are asked to provide consultation regarding the interference of bitter orange supplements with the CEDIA Amphetamines Assay. A single-dose of Nature's Way Bitter Orange was not found to cause a false-positive response to the CEDIA Amphetamines Assay in 6 healthy adult male volunteers.

Administration, Oral↗

Relationship of upper and lower urinary tract infection and bacterial invasion of uroepithelium to antibody-coated bacteria test results in female dogs.

Urinary tract infection was demonstrated in 12 female dogs via bacteriologic culture of a specimen of bladder urine collected by antepubic cystocentesis. Escherichia coli was isolated in pure culture from the urine of 9 dogs. Urine specimens from 2 dogs contained E coli and alpha-streptococci and from 1 dog contained Streptococcus zymogenes in pure culture. In 6 dogs, urinary tract infection was limited to the urinary bladder, whereas 6 dogs had unilateral or bilateral culture-positive renal pelvic urine as well (specimens collected by percutaneous nephropyelostomy). An antibody-coated bacteria (ACB) test was conducted on a portion of the bladder urine specimen from each dog, and the urinary tissues from these 12 dogs and from 6 healthy, noninfected female dogs were examined at necropsy. Tissues were given a subjective score based on the severity of the lesions seen microscopically. Histologic scores, bacterial cultural results, and ACB test results were examined for significance. A significant difference was found in the histologic scores between infected and noninfected dogs (P less than 0.025), but comparisons among histologic scores, cultural results, and ACB test results were not significant among infected dogs. The ACB test could neither be used to localize bacterial infection within the urinary tract nor could it be used to indicate the presence of bacterial invasion of the uroepithelium in dogs.

Animals↗

Fimch antiserum inhibits the adherence of Escherichia coli to cells collected by voided urine specimens of diabetic women.

PURPOSE: With the increasing problem of resistance in pathogenic microorganisms the development of nonantimicrobial therapies is important. Diabetes mellitus (DM) is associated with an increased incidence of urinary tract infections. The majority of Escherichia coli strains, which is the most prevalent uropathogen, have type 1 fimbriae that bind to uroplakin in the bladder, as mediated by the adhesin FimH. A vaccine is being developed based on FimH adhesion. MATERIALS AND METHODS: The sequence of FimH adhesion of 29 E. coli strains isolated from women with DM was determined. For adherence experiments we used E. coli isolated from women with DM and a T24 bladder cell line as well as the 2 well-defined type 1 fimbriated E. coli strains Ctrl 39 and NU14, and uroepithelial cells from women with DM. RESULTS: The fimH sequence of E. coli strains isolated from women with DM was highly homologous to the known fimH sequence of E. coli from patients without DM. Adherence assays in a T24 bladder cell line showed that adherence of these E. coli strains from women with DM could be inhibited by pre-incubation with antiserum raised against the chaperone-adhesin complex FimC-FimH. AntiFimCH antiserum also inhibited the adherence of the 2 well-defined E. coli strains expressing type 1 fimbriae, NU14 and Ctrl 39, but not of the FimH mutant strain NU14 H-, to uroepithelial cells from women with DM. CONCLUSIONS: These findings suggest that a vaccine based on FimH adhesin of type 1 fimbriated E. coli is a potential method of preventing urinary tract infection in women with DM.

Adhesins, Escherichia coli↗

Comparison of neonatal thyroid-stimulating hormone levels and indicators of iodine deficiency in school children.

OBJECTIVES: To compare thyroid-stimulating hormone (TSH) levels in neonatal cord blood between study sites in Bangladesh, Guatemala and the United States. Also, to compare neonatal TSH results with indicators of iodine deficiency in school children. DESIGN: Consecutive births and, in school children, cross-sectional surveys. SETTING: Savar, Bangladesh; San Pedro Sacatepequez, Guatemala; and Atlanta, United States. SUBJECTS: In each study site, cord blood was spotted on to filter paper and TSH levels determined using a sensitive monoclonal assay. In the USA, heel stick blood specimens from newborns spotted on to filter paper were also obtained as well as exposure to iodine-containing antiseptics during the birthing process. Urine specimens were collected from mothers of newborns and tested for iodine concentration. School children in the same areas were surveyed for thyroid size by palpation and ultrasonography, and urine specimens collected for iodine concentration. RESULTS: Between 141 and 243 cord blood specimens were collected from each study site. The prevalence of elevated cord blood TSH levels (> 5 mUl(-1)) was high in all study sites, from 58% to 84%. All sites would be categorised as having 'severe' iodine deficiency based on WHO/UNICEF/ICCIDD criteria. Iodine-containing antiseptics were used during 98% of the births in the USA but not in Bangladesh or Guatemala. The neonatal TSH classification indicated more severe iodine deficiency levels than classifications based on urinary iodine and goitre in school children. CONCLUSIONS: In the USA, elevated TSH levels may be partially attributed to use of beta-iodine-containing antiseptics prior to birth. We recommend the cautious interpretation of TSH results in newborns for the assessment of iodine deficiency disorders when iodine-containing antiseptics are used during the birthing process.

Adolescent↗

Monitoring cocaine use in substance-abuse-treatment patients by sweat and urine testing.

Sweat and urine specimens were collected from 44 methadone-maintenance patients to evaluate the use of sweat testing to monitor cocaine use. Paired sweat patches that were applied and removed weekly (on Tuesdays) were compared with 3-5 consecutive urine specimens collected Mondays, Wednesdays, and Fridays. All patches (N = 930) were extracted in 2.5 mL of solvent and analyzed by ELISA immunoassay (cutoff concentration 10 ng/mL); a subset of patches (N = 591) was also analyzed by gas chromatography-mass spectrometry (GC-MS) for cocaine, benzoylecgonine (BZE), and ecgonine methyl ester (EME) (cutoff concentration 5 ng/mL). Urine specimens were subjected to qualitative analysis by EMIT (cutoff 300 ng/mL) and subsets were analyzed by TDx (semiquantitative, LOD 30 ng/mL) and by GC-MS for cocaine (LOD 5 ng/mL). Results were evaluated to (1) determine the relative amounts of cocaine and its metabolites in sweat; (2) assess replicability in duplicate patches; (3) compare ELISA and GC-MS results for cocaine in sweat; and (4) compare the detection of cocaine use by sweat and urine testing. Cocaine was detected by GC-MS in 99% of ELISA-positive sweat patches; median concentrations of cocaine, BZE, and EME were 378, 78.7, and 74 ng/mL, respectively. Agreement in duplicate patches was approximately 90% by ELISA analysis. The sensitivity, specificity, and efficiency of sweat ELISA cocaine results as compared with sweat GC-MS results were 93.6%, 91.3%, and 93.2%, respectively. The sensitivity, specificity, and efficiency between ELISA sweat patch and EMIT urine results were 97.6%, 60.5%, and 77.7%, respectively. These results support the use of sweat patches for monitoring cocaine use, though further evaluation is needed.

Adolescent↗

Endogenous sex hormones, prolactin, and breast cancer in premenopausal women.

Forty-one women with breast cancer and 119 controls participated in a case-control study of the relation of endogenous sex hormones to breast carcinoma in premenopausal women. During the follicular phase of the menstrual cycle, one overnight urine specimen was collected. During the luteal phase, urine and blood specimens were obtained. 17 beta-Estradiol, sex hormone-binding globulin, progesterone, and prolactin were measured in plasma, whereas estrogen metabolites (estrone, estradiol, and estriol) and pregnanediol were assessed in the urine. Breast cancer was associated with high-plasma estradiol and prolactin and with low progesterone. Similar but weaker associations were observed for urinary estrogens and pregnanediol in the luteal phase.

Adult↗

Are true-negative nitrite results affected by a two-hour delay in dipstick/pad analysis of urine in incontinence pads?

OBJECTIVES: To determine if true-negative nitrite results of a urine dipstick pressed into an incontinence pad (dipstick/pad method) are affected by a 2-hour delay in analysis. DESIGN: A quantitative study. SETTING AND SUBJECTS: Clean-catch urine specimens from a convenience sample of clinic patients, staff, and long-term care facility residents. INSTRUMENT: Changes from negative to positive for each group of urine specimens were evaluated using percentages within the groups. METHODS: Urine specimens were collected and a portion was cultured. Urine specimens negative for a urinary tract infection were included in the study. A portion of the specimen was poured into an incontinence pad. Initial nitrite results were determined using a dipstick pressed into an incontinence pad. Pads with true-negative dipstick/pad nitrite results were tested 2 hours later in the same manner. Urine culture results determined groups: mixed colonies; 50,000 to 75,000 colony-forming units per milliliter of a single type of uropathogen; greater than or equal to 50,000 cfu/mL diphtheroids; no significant growth; and no growth. RESULTS: Of the 443 urine cultures negative for a urinary tract infection, 441 initial dipstick/pad nitrite results were negative. Two initial true-negative nitrite dipstick/pad results, or 0.5%, changed from negative to positive over the 2-hour period: 1 in the "mixed colonies" group (0.4%) and 1 in the "no growth" group (0.7%). CONCLUSIONS: Results of this study indicate that true-negative nitrite results of a dipstick pressed into urine in an incontinence pad do not appear to be affected by a 2-hour delay in analysis.

Adolescent↗

A method comparison study to assess the reliability of urine collection pads as a means of obtaining urine specimens from non-toilet-trained children for microbiological examination.

BACKGROUND: In young, non-toilet-trained children, the collection of a urine sample for microbiology can be challenging, with the application of a urine bag being the main method of collection. However, recent research has shown that absorbent pads can be used yielding similar results to bag specimens. However, weaknesses in study design erode confidence in research findings. Therefore, improvements in research design are required to fully evaluate the reliability of pad collection. AIMS OF THE STUDY: This pilot study sought to test the feasibility of a technique for the collection of concurrent bag/pad urine samples from non-toilet-trained children, and to assess the reliability of urine pads over bags as a collection method for urine specimens for microbiological evaluation. DESIGN: A pilot, method comparison study. METHODS: Twenty concurrent bag and pad specimens were collected from non-toilet-trained children, following parental consent. Urine specimens were analysed for presence or absence of white cell count (WBC), and bacterial growth, using standard laboratory methods. DATA ANALYSIS: The Kappa (kappa) statistics and confidence interval (CI) estimation were used to assess agreement between the two collection methods. RESULTS: Despite concurrent samples there was a lack of agreement between bag and pad specimens on both main outcome measures. Agreement between bag and pad specimens for the presence of WBC yielded a kappa=0.10 (95% CI: 0.19, 0.39), indicating poor agreement, while a kappa of 0.5 (95% CI: 0.12, 0.88) was calculated for the degree of agreement in bacterial growth reflecting moderate agreement. Differences in proportions of the presence of WBC between bag and pad did not quite reach significance at the 5% level 0.2 (95% CI: 0.00, 0.42, P=0.062). For cultures the difference was calculated as 0.15 (95% CI: 0.05, 0.35, P=0.125). CONCLUSION: The pilot study demonstrates that concurrent urine samples can be obtained without difficulty. Despite poor to moderate agreement on outcome measures the level of agreement is greater than reported in those other studies, that use non-current methods of urine collection, suggesting an advantage of the concurrent technique. It is recommended that larger scale studies be undertaken using the concurrent collection technique to assess reliability of these findings.

Child, Preschool↗

Identification of hydrocodone in human urine following controlled codeine administration.

Allegations of illicit hydrocodone use have been made against individuals who were taking physician-prescribed oral codeine but denied hydrocodone use. Drug detection was based on positive urine opiate immunoassay results with subsequent confirmation of hydrocodone by gas chromatography-mass spectrometry (GC-MS). In these cases, low concentrations of hydrocodone (approximately 100 ng/mL) were detected in urine specimens containing high concentrations of codeine (> 5000 ng/mL). Although hydrocodone has been reported to be a minor metabolite of codeine in humans, there has been little study of this unusual metabolic pathway. We investigated the occurrence of hydrocodone excretion in urine specimens of subjects who were administered codeine. In a controlled study, two African-American and three Caucasian male subjects were orally administered 60 mg/70 kg/day and 120 mg/70 kg/day of codeine sulfate on separate days. Urine specimens were collected prior to and for approximately 30-40 h following drug administration. In a second case study, a postoperative patient self-administered 960 mg/day (240 mg four times per day) of physician-prescribed oral codeine phosphate, and urine specimens were collected on the third day of the dosing regimen. Samples from both studies were extracted on copolymeric solid-phase columns and analyzed by GC-MS. In the controlled study, codeine was detected in the first post-drug-administration specimen from all subjects. Peak concentrations appeared at 2-5 h and ranged from 1475 to 61,695 ng/mL. Codeine was detected at concentrations above the 10-ng/mL limit of quantitation for the assay throughout the 40-h collection period. Hydrocodone was initially detected at 6-11 h following codeine administration and peaked at 10-18 h (32-135 ng/mL). Detection times for hydrocodone following oral codeine administration ranged from 6 h to the end of the collection period. Confirmation of hydrocodone in a urine specimen was always accompanied by codeine detection. Codeine and hydrocodone were detected in all specimens collected from the postoperative patient, and concentrations ranged from 2099 to 4020 and 47 to 129 ng/mL, respectively. Analyses of the codeine formulations administered to subjects revealed no hydrocodone present at the limit of detection of the assay (10 ng/mL). These data confirm that hydrocodone can be produced as a minor metabolite of codeine in humans and may be excreted in urine at concentrations as high as 11% of parent drug concentration. Consequently, the detection of minor amounts of hydrocodone in urine containing high concentrations of codeine should not be interpreted as evidence of hydrocodone abuse.

Administration, Oral↗

Optical isomer analysis of 3,4-methylene-dioxyamphetamine analogues and their stereoselective disposition in rats.

Identification of the optical activity and simultaneous analysis of racemates (+/-) of three hallucinogens, 3,4-methylenedioxy-amphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA), and the urinary excretion of their optical isomers in rats was performed by high-performance liquid chromatographic analysis. Analysis of optical enantiomers of three N-alkyl MDA derivatives was performed within 50 min using two different detectors, polarimetry (OR) and ultraviolet spectroscopy (UV). The OR detector proved suitable for identification of the optically active forms, whereas the UV detector was suitable for simultaneous analysis of the enantiomers in urine. After the administration of each of the three N-alkylated derivatives, rat urine specimens were collected over four intervals, 0-4, 4-12, 12-20, and 20-24 h. After the administration of 30 mg/kg of racemic MDA and MDMA, somewhat less of the S(+)-forms of unchanged MDA and MDMA than of the R(-)-forms in each urine specimen were detected, which gave R/S ratios greater than 1.00 (p < 0.01). Conversely, after the administration of 30 mg/kg of racemic MDEA, more of the S(+)-form than the R(-)-form was found in the urine, thus giving R/S ratios less than 1.00 (p < 0.01). The percentage of the dose excreted up to 24 h was approximately 29.4% of the administered dose for MDA [S(+) 13.40% and R(-) 15.98%], 5.8% for MDMA [S(+) 1.96% and R(-) 3.79%], and 7.3% for MDEA [S(+) 3.89% and R(-) 3.43%]. Urinary excretion of optical isomers of N-dealkylated MDA from MDMA and MDEA origin were the opposite of those of the unchanged forms, and their R/S ratios up to 24 h were 0.48 to 0.72 (p < 0.01) and 1.31 to 1.50 (p < 0.01), respectively. The urinary excretion rates up to 24 h were approximately 4.3% for N-dealkylated MDA from MDMA origin [S(+) 2.72% and R(-) 1.63%] and 0.8% for N-dealkylated MDA from MDEA origin [S(+) 0.36% and R(-) 0.47%]. The total percent of unchanged forms and N-dealkylated MDA was approximately 10.1% for MDMA [S(+) 4.68% and R(-) 5.42%] and 8.2% for MDEA [S(+) 4.25% and R(-) 3.91%]. The total R/S ratio of MDMA was found to be 1.95 (p < 0.01), whereas that of MDEA was 0.88 (p < 0.01). The total R/S ratio of MDA was 1.20 (p < 0.01 ), which was comparable with that of MDMA. These three R/S ratios did not conform to the theoretical values for three N-alkyl derivatives used and neither did the R/S ratios of urine specimens collected at the four interval. These results suggested the stereoselective disposition of three N-alkyl MDA analogues in rat. The method would be suitable for the forensic chemistry and toxicology analysis of specimens obtained from hallucinogen abusers.

3,4-Methylenedioxyamphetamine↗