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PubMed2003

Leptin measurement in urine in children and its relationship to other growth peptides in serum and urine.

OBJECTIVE: Leptin has been implicated in the interaction between nutrition, energy balance and sexual maturation in humans. A non-invasive method of measuring leptin would greatly facilitate longitudinal studies of changes in leptin in normal children. The aim of this study was to evaluate the use of urinary leptin as a surrogate for serum leptin measurements. DESIGN: We have modified and validated a serum immunoradiometric assay (IRMA) kit for the measurement of leptin in urine, and subsequently investigated the relationship between urinary leptin and other growth-related proteins. METHODS: Cross-sectional study: urinary leptin, measured in the first morning urine voided and expressed as ng excreted overnight, and serum concentrations of leptin, IGF-I, IGF-II, IGFBP-3 and IGFBP-1 were determined in a cohort of 188 healthy schoolchildren aged 5-19 years (88 males, 100 females). Height, weight and pubertal status were assessed in all children. Longitudinal study: urinary levels of leptin, IGF-I and GH were measured daily in two adults (one male, one female) over a period of 6 weeks. RESULTS: The detection limit of this modified assay was 0.59 ng/L. The intra- and interassay coefficients of variation range was 4-8% and 4-9%, respectively. The recovery of recombinant leptin added to urine was 98-108%, and the assay had a recovery rate for serial dilution in the range of 106-112%. Urinary leptin correlated significantly with serum leptin (r = +0.65, P < 0.01). Urinary leptin showed similar changes through puberty to those of serum leptin, with levels rising in females throughout puberty, whereas in males levels peaked at G2/G3 then decreased. BMI SDS was the main determinant of urinary leptin, as it was for serum leptin. In the cross-sectional study urinary leptin correlated significantly with serum IGF-I (r = +0.41, P = 0.001), IGF-II (r = +0.19, P = 0.05), IGFBP-3 (r = +0.29, P = 0.001) and IGFBP-1 (r = -0.25, P = 0.001). In the adult study, leptin was also detected in urine with similar night-to-night variability to that found for IGF-I and GH. CONCLUSION: Urinary leptin is a valid marker of serum leptin concentrations, and therefore this non-invasive assay would be a useful tool for longitudinal assessment of changes in leptin in children.

Adolescent↗
PubMed2003

Comparing ambulatory spot urine protein/creatinine ratios and 24-h urine protein measurements in normal pregnancies.

OBJECTIVE: To determine whether a single specimen protein/creatinine ratio correlated with a 24-h urine protein measurement in ambulatory women with normal pregnancies. METHODS: A prospective cohort was evaluated of healthy women performing a 24-h urine collection and spot urine measurement for protein and creatinine in each trimester and the postpartum period. Data were analyzed to establish the correlation of values and the reliability of cut-off values. RESULTS: Fifty-eight women were enrolled in the study. Thirty-three completed at least one measurement. Three (5.1%) developed pre-eclampsia. With the use of linear regression, the spot protein/creatinine ratio correlated with the 24-h protein measurement in the first trimester and postpartum periods only (p < 0.001 and p < 0.043, respectively). It did not correlate in the second or third trimester (p = 0.637 and p = 0.290, respectively). CONCLUSION: In this population, the protein/creatinine ratio correlated only with the 24-h urine protein measurement in the first trimester and postpartum periods. In the periods of pregnancy where physiological changes are most prevalent, the ratio does not predict total urinary protein reliably.

Adult↗
PubMedDate not supplied

Nitrite adulteration of workplace urine drug-testing specimens. I. Sources and associated concentrations of nitrite in urine and distinction between natural sources and adulteration.

The active ingredient in the commercial workplace urine drug-testing adulterant, Klear, was previously determined to be nitrite ion. Nitrite adulteration compromises the confirmation of some drugs, notably the marijuana metabolite. A previously reported bisulfite step overcomes some nitrite adulteration, but it cannot do so in every case, which leaves the laboratory to report the specimen as not suitable for testing. Unlike many other adulterants, nitrite is found in normal urine at low concentrations. In order to defend a report of nitrite adulteration, it is necessary to provide evidence that the amount of nitrite in a workplace urine specimen could not arise by normal means. The objectives of this study were to identify all sources of nitrite in urine and the range of concentrations associated with these sources and to determine if nitrite adulteration can be supported based upon a quantitative result. The scientific literature was reviewed for internal and external sources of nitrite and their concentration ranges and are reported. The following specimens were obtained and nitrite concentrations measured by a spectrophotometric method: clinical specimens nitrite positive by test strip (< 15 micrograms/mL); specimens culture positive for nitrate-reducing microorganisms (< 36 micrograms/mL); specimens from patients on medications that may metabolize to nitrite (< 6 micrograms/mL); and drug-test specimens, both negative (< 130 micrograms/mL) and others that appeared to be adulterated with nitrite (range 1910-12,200 micrograms/mL, mean 5910). The literature and the nitrite measurements of this study indicate a substantial difference between concentrations from natural sources compared with adulteration. A quantitative measurement of nitrite by a well-structured assay can provide scientifically valid and forensically defensible proof of adulteration with a nitrite-containing substance.

Drug Contamination↗
PubMed1995

Determination of amounts of polyamines excreted in urine: demonstration of N1,N8-diacetylspermidine and N1,N12-diacetylspermine as components commonly occurring in normal human urine.

An analytical system developed for fractionating free and monoacetylated polyamines [Hiramatsu, K. et al. (1994) J. Biochem. 115, 584-589] was proved useful also in detecting diacetylpolyamines, namely N1,N8-diacetylspermidine (diAcSpd) and N1,N12-diacetylspermine (diAcSpm). Detection limits were 0.9 and 0.6 pmol (S/N = 5) for diAcSpd and diAcSpm, respectively. Analytical recovery and within-run variation were also satisfactory. Human urine samples were found to contain diAcSpd and diAcSpm. These polyamines were identified on the basis of the following observations: (i) their retention times were coincident with those of authentic samples; (ii) they were deacetylated to N8-acetylspermidine and monoacetyl- and free spermine, respectively, by acetylpolyamine amidohydrolase; and (iii) they were practically inert to direct oxidation by bacterial polyamine oxidase as were authentic samples. The amounts of eleven polyamine species including diAcSpd and diAcSpm in urine samples from 52 healthy persons were determined. Mean values for the major polyamine components were consistent with those reported by others. Although the amounts of diAcSpd and diAcSpm were very small, comprising only 1.4 and 0.46% of total polyamines, respectively, these two compounds were found to be always present in healthy human urine as regular constituents. Moreover, variation in their content among individuals was small, suggesting that excretion of these components in urine is strictly regulated.

Biogenic Polyamines↗
PubMed2001

Detection of Legionella pneumophila antigen in urine samples by the BinaxNOW immunochromatographic assay and comparison with both Binax Legionella Urinary Enzyme Immunoassay (EIA) and Biotest Legionella Urin Antigen EIA.

The new BinaxNOW Immunochromatographic (ICT) Assay for the detection of Legionella pneumophila antigens was used to test 535 urine specimens from patients with and without Legionnaires' disease. The specificity, calculated by testing 112 samples from patients with pneumonia of aetiologies other than Legionella infection, and 167 urine specimens from urinary tract infections, was found to be 97.1% if the manufacturer's guidelines were followed. However, it was determined that the 'false positive' results characterised by very weak bands could be discounted by re-examination of the results at 60 min, yielding a specificity of 100%. With this minor modification of the procedure applied to examination of urine samples from 117 patients with legionellosis confirmed by isolation of L. pneumophila and 70 patients who had seroconverted to L. pneumophila serogroup 1, sensitivity was calculated to be 79.7%. In comparison, the sensitivities of the Binax Urinary Antigen Enzyme Immunoassay (EIA) and Biotest Urin Antigen EIA were estimated to be 79.1 and 83.4%, respectively. Eleven cases (5.9%) were positive by BinaxNOW assay but negative by Binax or Biotest EIA, or both. The sensitivities of all assays increased to c. 94% if only diagnosis of cases confirmed by isolation of serogroup 1 L. pneumophila was considered, although the sensitivity for infections caused by L. pneumophila serogroup 1 monoclonal antibody (MAb) subgroup Bellingham was significantly lower than for other MAb subgroups. The Biotest EIA recognised 10 (45%) of the 22 cases not caused by L. pneumophila serogroup 1, whereas the two Binax kits detected only three each. The ICT assay BinaxNOW can be recommended as a rapid specific test for the diagnosis of Legionnaires' diseases caused by L. pneumophila serogroup 1, although very weak bands should be interpreted cautiously.

Antibodies, Monoclonal↗
PubMed2006

The value of urine specific gravity in detecting diabetes insipidus in a patient with uncontrolled diabetes mellitus: urine specific gravity in differential diagnosis.

When a patient with diabetes mellitus presents with worsening polyuria and polydipsia, what is a sensible, cost-effective approach? We report the unique coincidence of type 2 diabetes mellitus and diabetes insipidus. A 46-year-old woman with poorly controlled type 2 diabetes complained of polyuria with a daily output of 5 L. Although urinalysis demonstrated significant glucosuria, diabetes insipidus was suspected owing to a low urine specific gravity (1.008). The low specific gravity persisted during a water deprivation test. Ultimately, diabetes insipidus was confirmed when urine specific gravity and urine osmolality normalized following desmopressin administration. This case emphasizes the importance of accurately interpreting the urine specific gravity in patients with polyuria and diabetes mellitus to detect diabetes insipidus.

Diabetes Insipidus↗
PubMed1988

Screening for proteinuria in a rheumatology clinic: comparison of dipstick testing, 24 hour urine quantitative protein, and protein/creatinine ratio in random urine samples.

Measurements of protein/creatinine ratio in 'spot' urine samples were compared with measurements of 24 hour quantitative proteinuria and side room 'dipstick' testing in 104 samples from 90 patients presenting consecutively to a rheumatology unit. Linear regression analysis showed a highly significant correlation between the random urinary protein/creatinine ratio and total protein excretion in 24 hour urine samples (r = 0.92, p less than 0.001, y = 6.55x + 0.04). Although an approximation of 24 hour urinary protein excretion could have been made from the regression line: 24 hour urine protein = 6.55 x protein/creatinine ratio + 0.04 (g/l), there was a wide scatter of values, particularly in patients with greater than 1 g/24 h urinary protein excretion. Nevertheless, significant proteinuria (greater than 300 mg/24 h) could have been confirmed or excluded with a sensitivity and specificity of 97% by adopting random protein/creatinine values of less than 0.04 as 'normal'. Specificity and sensitivity could have been increased to 100%, however, by excluding patients with values lying between 0.01 and 0.10 as all the false negatives (n = 3) and false positives (n = 3) lay within this range. In comparison, dipstick testing, although 100% sensitive, had a poor specificity due to the high false positive rate (40/83 (48%] in patients with 1+ to 3+ readings. Assessment of random urinary protein/creatinine ratio may obviate the need for 24 hour urine collections in the initial assessment of suspected proteinuria. A wider application of this technique seems indicated in view of the obvious advantages in terms of cost, time, and patient convenience.

Creatinine↗
PubMed1994

Clearance half life of mercury in urine after the cessation of long term occupational exposure: influence of a chelating agent (DMPS) on excretion of mercury in urine.

The elimination of mercury (Hg) in urine was investigated in 12 former chloro-alkali workers exposed to metallic Hg vapour for two to 18 (median five) years. Morning urine samples were taken on several (median 9) occasions after change of employment or retirement. The median follow up time was 28 months. The decrease in concentration of Hg in urine (U-Hg) was well characterised by a one compartment model. Three different regression methods were used; non-linear least squares regression (NLSR), weighted non-linear least squares regression (WNLSR), and linear least squares regression (LLSR) after log transformation of the U-Hg data. The median half life from the WNLSR method was 55 days. There were no large differences in the half life estimates given by the WNLSR or the NLSR methods, but for five subjects the LLSR method gave poor fits. There was a non-significant tendency towards longer half lives with higher initial U-Hg. About three years after the cessation of occupational exposure a mobilisation test with 2,3-dimercaptopropane-1-sulphonate (DMPS) was performed on seven subjects. Excretion of Hg, copper (Cu), and zinc (Zn) in urine was estimated before and after the ingestion of 300 mg of DMPS. Treatment with DMPS increased 24 hour urinary excretion by a factor of 7.6 for Hg, 12 for Cu, and 1.5 for Zn. The relative increase in U-Hg was not significantly higher than that obtained in a previous study of an occupationally unexposed group. A major proportion (62%) of Hg excreted during 24 hours after DMPS appeared in the first six hours.

Adult↗
PubMed1980

Quantification of polar glucocorticosteroids in the urine of pregnant and nonpregnant women: a comparison with 6 alpha-hydroxylated metabolites of cortisol in neonatal urine and amniotic fluid.

6 alpha-Hydroxy metabolites of cortisol were determined in the urine of pregnant (36-40 weeks of gestation) and nonpregnant women and in amniotic fluid from nearly fullterm pregnant women because relatively large amounts of these compounds are excreted in the urine of 2-day-old infants (> 200 micrograms/day). The corticosteroids analyzed by high pressure liquid chromatography and gas chromatography-mass spectrometry were 6 alpha-hydroxy derivatives of (allo)tetrahydrocortisone (3 alpha, 17 alpha, 21-trihydroxy-5 epsilon-pregnan-11,20-dione), (allo)tetrahydrocortisol (3 alpha, 11 beta, 17 alpha, 21-tetrahydroxy-5 epsilon-pregnan-20-one), and alpha- and beta-cortolone (3 alpha, 17 alpha, 20 epsilon, 21-tetrahydroxy-5 beta-pregnan-11-one). All of these compounds were found in the urine samples from both groups of women and in the amniotic fluid samples in contrast to those found in the urine samples from the neonates where 6 alpha-hydroxy compounds of (allo)tetrahydrocortisol and allotetrahydrocortisone were not positively identified because of insufficient yields. The pregnant women excreted significantly larger amounts of 6 alpha-hydroxy metabolites of cortisol (approximately 600 micrograms/day) than the control women (approximately 90 micrograms/day), and the rate of urinary excretion of these 6 alpha-hydroxy compounds was 7.82 and 1.30 micrograms/kg . day, respectively, for these groups of women compared to 54.3 micrograms/kg . day for the neonates. The precursors of these metabolites within the fetal body originated largely from the maternal circulation, and, therefore, the 6 alpha-hydroxy metabolites of cortisol excreted by the mother refer mainly to fetal metabolism and to a lesser extent, to the fetal secretion of cortisol.

Adult↗
PubMed2001

Safely reducing manual urine microscopy analyses by combining urine flow cytometer and strip results.

We aimed to reduce the number of manual urine microscopy examinations safely by cross-interpretation of the Sysmex UF-100 (TOA Medical Electronics, Kobe, Japan) and urine strip results such that microscopy would be performed if there was discordance between the UF-100 and urine strip results. We also evaluated the usefulness of the optional UF-100 expert software. We performed 2 studies: study 1 to establish review rules for eventual microscopic examination; study 2, a validation study. Our review rates were 40% and 48% and those of UF-100 software were 16% and 32% for the 2 studies. Our false-positive and false-negative results, among the samples not flagged for microscopic review, were acceptably low. We did not find a good correlation between the microscopic classification of RBC morphologic features and the classification given by the UF-100. Since incorporation of the automated urine strip reader and the UF-100 in routine use, our manual microscopy has been reduced to less than 40%.

Adolescent↗
PubMed1990

Preanalytical handling of stored urine samples, and measurement of beta 2-microglobulin, orosomucoid, albumin, transferrin and immunoglobulin G in urine by enzyme-linked immunosorbent assays (ELISA).

Urinary beta 2-microglobulin, orosomucoid, albumin, transferrin and IgG were measured by enzyme-linked immunosorbent assays (ELISA). In urine samples stored at -20 degrees C three of these proteins decreased during the period of freezing. After 1 week at -20 degrees C urinary transferrin decreased by 81%, IgG by 39% and albumin by 26% of the pre-freezing values; however, addition of Tween-20 restored these values. In previously stored urine samples with bovine albumin, the decreased value after freezing at -20 degrees C could be increased by changing the thawing procedure and including addition of Tween-20. Urine samples thawed at room temperature just before analysis decreased by 80% for transferrin, 57% for IgG, 30% for albumin and 26% for beta 2-microglobulin compared with samples thawed at 37 degrees C, had Tween-20 added and then were kept for a few days at room temperature before analysis. Furthermore, previously frozen urine samples that were thawed at 37 degrees C, had Tween-20 added and then were stored at room temperature did not show significant changes in any of the protein results measured the day after thawing and 35 days later. Orosomucoid seemed to be less variable as regards the effect of freezing and thawing procedure.

Adult↗
PubMed1991

[A latex immunologic method of analysis of low-molecular proteins and albumin in urine. II. Beta 2-microglobulin, retinol-binding protein and albumin levels in the urine of general Polish population and workers exposed to cadmium].

The aim of the work was to collect data about beta 2-microglobulin (beta 2M), retinol-binding protein (RBP) and albumin (Alb) levels in subjects not exposed to nephrotoxic agents in Poland. The data are necessary to interpret determination results in persons occupationally exposed to these chemicals. Concentrations of beta 2-microglobulin, retinol-binding protein and albumin in urine were determined using latex-immunological method. Examination of levels of low molecular proteins and albumin was made on population from agricultural regions of Poland (the Suwałki region) and on textile plant workers from Lódź not exposed to nephrotoxic agents. It was found that upper normal level of low molecular proteins in urine was 300 micrograms/l and of albumin 60 mg/l. The data obtained indicate that sex and age play an important role in beta 2-M and RBP excretion. In the workers of an alkaline accumulator plant who were occupationally exposed to cadmium, a statistically increased excretion of low molecular proteins in urine was observed compared to the controls from Lódź. In 20% of the subjects beta 2-M and RBP concentration in urine were higher than the accepted upper normal levels. Collected data make one aware of necessity to study the above determinations which lead to an early detection of renal function disorders.

Age Factors↗
PubMed1990

[Study on influence of cigarette smoking on the mutagenicity of urine. I. Influence of cigarette smoking on the mutagenicity of urine in healthy smokers and bladder cancer patients].

We studied the mutagenicity of urine of healthy smokers and smokers with bladder tumors by the Ames test. The 41 healthy smokers studied showed a significant increase in urinary mutagenic activity compared to the 24 passive smokers and the 22 non-smokers, but there was no significant difference between the passive smokers and non-smokers. In the 5 healthy subjects, the urinary mutagenic activity increased in accordance with the increase of tobacco consumption. In the 32 healthy subjects and 22 smokers with bladder tumors, the time-course changes of the mutagenicity of urine after smoking were investigated by testing the urine every 2 hours. In the healthy subjects, urinary mutagenic activity was increased up to 4 hours after cigarette smoking and decreased to the level noted before smoking after the 6th hour. By contrast, in the 22 smokers with bladder tumors, the urinary mutagenic activity remained high even after the 6th hour, and only decreased to the level seen before smoking after the 8th hour. The influence of smoking on the mutagenicity of urine tended to protract in smokers with bladder tumors in comparison to healthy smokers. There was no correlation between urinary mutagenic activity and tumor status and recurrence rate of the bladder tumors.

Adult↗
PubMed1983

Isolation and characterization of N-acetylglucosamine 6-sulfate from the urine of a patient with Sanfilippo type D syndrome and its occurrence in normal urine.

N-Acetylglucosamine 6-sulfate (GlcNAc6S) has been isolated from human urine and shown to be present at levels of approximately 0.02 and 14 mg/mmole creatinine in urine from normal individuals and a mucopolysaccharidosis type IIID (MPS IIID) patient respectively. We propose that the greater than 500-fold elevation of GlcNAc6S in urine from the MPS IIID patient indicates that this sulfated monosaccharide is also a substrate for the sulfatase deficient in MPS IIID patients. We further propose that part, if not all, of the GlcNAc6S found in urine may be produced from the cleavage by beta-N-acetylhexosaminidase A of non-reducing end beta-linked GlcNAc6S residues of keratan sulfate and/or sulfated glycoproteins.

Acetylglucosamine↗
PubMed1993

Can overnight urine replace 24-hour urine collection to measure urinary calcium in epidemiologic studies?

The present study investigated whether overnight urine collection can replace 24-hour urine collection to measure urinary calcium (Ca) excretion in healthy individuals. One hundred healthy men (age 25-74 years) on their usual diet participated in the present study as part of an ongoing epidemiological population-based survey. Two separate bags (daytime and overnight bags) were given together with instructions to collect a complete 24-hour urine sample divided in to the daytime and the overnight specimen. Urinary Ca was analyzed by atomic absorption spectrophotometry and creatinine by the picric acid colorimetric method. Overnight urinary Ca was correlated to daytime (r = 0.618, p < 0.0001) and 24-hour urinary Ca (r = 0.891, p < 0.0001). In the 17 men found to have hypercalciuria (24-hour urinary Ca > or = 7.5 mmol), the overnight urinary Ca averaged 4.44 +/- 0.29 mmol/12 h (mean +/- SEM, range 2.35-6.38 mmol/12 h), indicating that an overnight urinary Ca of < 2.35 mmol/12 h (< 60% of the overnight urinary Ca distribution) ruled out the possibility of finding hypercalciuria in 24-hour urine. Fourteen of the seventeen hypercalciuric men had overnight urinary Ca in the upper 20% of the distribution (> or = 3.25 mmol/12 h). Hypercalciuria was found in 14 of the 19 men (73.7%) with an overnight urinary Ca of > or = 3.25 mmol/12 h, but only in 3 of the 81 men (3.7%) with an overnight urinary Ca of < 3.25 mmol/12 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1998

Measures of urine concentration in maternal urine screening for Down syndrome.

A study was carried out to assess eight methods of normalizing the level of urinary beta-core human chorionic gonadotropin (hCG) for variable urine concentration. We compared the standard approach--creatinine determination by the Jaffe method--with high performance liquid chromatography (HPLC) measurement of creatinine, osmolarity and optical density at five wavelengths. Urine samples were included from a total of 472 women with unaffected singleton pregnancies at 15 weeks' gestation. The median beta-core hCG value was determined for each decile group when the results were ranked in turn according to the different measures of urine concentration. Creatinine using the Jaffe method had a much stronger relationship with median beta-core hCG than the other measures. Linear regression across the decile groups gave an R2 value for Jaffe of 0.85 compared with HPLC of 0.53, osmolarity of 0.52, optical density at 405 nm of 0.72, at 450 nm of 0.57, at 490 nm of 0.33, at 570 nm of 0.34 and at 630 nm of 0.33. We conclude that when screening with urinary beta-core hCG measuring creatinine appears to be an adequate method of allowing for variable urine concentration.

Chorionic Gonadotropin, beta Subunit, Human↗
PubMed1988

Hormone studies in Myxine glutinosa: failure of calcitonin to influence urine flow or plasma and urine electrolyte balance.

Hagfish, Myxine glutinosa, were used in a series of experiments to determine a possible role for calcitonin in plasma and urine electrolyte balance. Individual animals were anaesthetised and implanted with polythene cannulae for monitoring blood pressure and for infusion/injection. Urine was collected by manual palpation of the body surface whilst blood samples were withdrawn from the lateral sinus. Following injection of either 1.25 or 3.75 micrograms kg body wt-1 calcitonin, there was no significant change in either the plasma or urine composition of sodium, potassium, calcium, or magnesium ion levels. These results indicate that calcitonin may not be involved in electrolyte homeostasis in Myxine.

Animals↗
PubMed1977

Cow's urine poisoning in Nigeria: cardiorespiratory effects of cow's urine in dogs.

"Cow's urine" concoction (CUPR) is a traditional remedy for convulsive seizures in Nigeria. Its administration has been associated with severe poisoning, sometimes with a fatal outcome in Nigerian children. Recently, we showed that several of the components of the concoction are toxic. We have demonstrated in the present study certain cardio-respiratory effects of cow's urine concoction in dogs. On the cardiovascular system, an initial bradycardia followed by tachycardia and a biphasic effect on blood pressue characterized by a fall followed by a rise were demonstrated. Progressive hypotension following repreated administration of CUPR was also shown in all experimental dogs. On the respiratory system, a short period of respiratory arrest and/or respiratory depression, followed by tachyponoea with associated hypoventilation of the lungs, were observed. The possible explanations for the observed cardiorespiratory effects of the concoction were discussed. On the basis of our findings, an hypothesis is advanced for the possible mechanisms of the neurological sequelae and/or death following cow's urine poisoning.

Animals↗