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The effect of osmotic and ion-osmotic stresses on the blood and urine composition and urine flow of rainbow trout (Salmo gairdneri).

Changes in urine and plasma concentrations (sodium, potassium, magnesium, calcium and total osmotic) and urine production were determined in fish exposed to various concentrations of an ionically active substance, sodium chloride, and a non-electrolyte, mannitol, as well as freshwater. Responses occurred for the most part over a short crisis period preceding establishment of new stable conditions. It was shown that plasma homeostasis was not maintained in response to changing ion-osmotic and osmotic gradients. Urinary osmotic and ionic concentrations were unaffected and urine production was shown to be inversely related to the external concentration. It is suggested that ionic shifts between body compartments are an important aspect of ion-osmotic adaptation.

Animals↗

Modifiers of calcium oxalate crystallization found in urine. II. Studies on their mode of action in an artificial urine.

The relative potencies of various modifiers of the crystallization of calcium oxalate (CaOx) were determined under "whole urine equivalent" conditions using a batch crystallizer. The system was used to measure changes in the degree of agglomeration of CaOx crystals produced spontaneously at a level of supersaturation within the range found in the urines of recurrent CaOx stone-formers. The modifiers tested included RNA, heparin, chondroitin-4-sulphate, pyrophosphate (at pH 5, 6 and 7), Tamm-Horsfall mucoprotein and a greater than 10,000 dalton fraction of macromolecules obtained from pooled normal urine by dialysis. The effects of these urinary constituents were also measured on the zeta potential produced at the surface of CaOx crystals. In the case of pyrophosphate, there was a clear correlation between the degree of inhibition of agglomeration and the zeta potential observed on the crystal surface indicating that, for this ion, repulsive electrostatic forces dominate the tendency for CaOx crystals to agglomerate. For the other modifiers tested, the tendency to agglomerate appeared to be dependent on the balance between the positive viscous binding ("sticky") forces and the negative electrostatic forces produced by these macromolecules on the CaOx crystal surface.

Calcium Oxalate↗

Modifiers of calcium oxalate crystallization found in urine. I. Studies with a continuous crystallizer using an artificial urine.

Various modifiers of the rate of crystallization of calcium oxalate (CaOx) known to be present in urine were studied in a continuous crystallizer system under conditions approximating those in whole urine. Of the small ions examined, magnesium and citrate were only weak inhibitors of the crystal growth rate and degree of agglomeration of CaOx within the urinary range of concentration of these ions. At and beyond the upper limit of the normal, however, both ions became much more active mainly through their ability to complex oxalate and calcium ions respectively. Pyrophosphate showed similar characteristics although its ability to act as an inhibitor at high concentrations was not due to complexation of calcium ions but to adsorption onto the crystal surface of the freshly generated CaOx crystals. The polyanionic inhibitors, chondroitin-4-sulphate, heparin and ribonucleic acid (RNA), also acted as adsorption inhibitors of growth and agglomeration. Of all the inhibitors tested RNA was the most active within the concentration range found in normal urine. Under the conditions of ionic strength employed, it was not possible to measure the effect of Tamm-Horsfall mucoprotein satisfactorily since it polymerized to form particles sufficiently large to interfere with the true crystal counts of CaOx in the test system.

Calcium Oxalate↗

Evaluation of full-scanning GC/ion trap MS analysis of NIDA drugs-of-abuse urine testing in urine.

Analytical methods developed for the Finnigan MAT ITS40 gas chromatograph-ion trap mass spectrometer (GC/MS) were evaluated for the confirmation of drugs-of-abuse in urine. The specific drugs evaluated are those listed by the National Institute on Drug Abuse (NIDA): 11-nor-9-carboxy-delta 9-tetrahydrocannabinol (9-carboxy-THC), benzoylecgonine (BE), codeine and morphine, phencyclidine (PCP), amphetamine, and methamphetamine. Drugs were extracted from urine using solid-phase columns, separated by capillary gas chromatography, and analyzed by ion trap mass spectrometry following electron impact ionization. All drugs except PCP were derivatized prior to analysis. The full scan limits of detection (LOD), quantitation (LOQ), and linearity were 2.5, 5.0, and 1000 ng/mL, respectively, for 9-carboxy-THC; 37, 75, and 5000 ng/mL for BE; 50, 100, and 2500 ng/mL for the opiates; 0.25, 0.50, and 500 ng/mL for PCP; and 50, 100, and 5000 ng/mL for the amphetamines. The limits of detection (LOD) and limits of quantitation (LOQ) met the minimum criteria for the signal-to-noise (S/N) ratio and spectral match criteria for drug identification. Absolute LODs and LOQs (in ng/mL) for the ITS40 based on single ion monitoring of blank urines were: 0.8 and 2.0 for 9-carboxy-THC; 8.9 and 25 for BE; 3.3 and 9.6 for codeine; 6.2 and 16.7 for morphine; 0.25 and 0.32 for PCP; 0.7 and 2.0 for amphetamine; and 2.4 and 5.7, for methamphetamine, respectively. The coefficient of variation ranged from 5 to 10%, and analytical recoveries were in the range of 90-114%. The ion trap mass spectrometer permits full scan identification of drugs while maintaining analytical LOQ that are below NIDA guidelines, and has equivalent or better detection limits to quadrupole analyzers for high sensitivity applications.

Evaluation Studies as Topic↗

Urine and urine-derived compounds induce c-fos mRNA expression in accessory olfactory bulb.

Soiled bedding from male mice induced c-fos mRNA expression in the accessory olfactory bulb (AOB) and main olfactory bulb (MOB) of female mice. The increase observed in the AOB, but not the MOB, was dependent on the presence of the vomeronasal organ (VNO). Male urine alone also increased c-fos mRNA expression in the AOB. The urine-derived compounds dehydro-exo-brevicomin (DHB) and sec-butyl-dihydrothiazole (SBT) in combination with major urinary protein (MUP) induced significantly greater c-fos mRNA expression in the AOB than in the MOB. The results indicate that compounds derived from male urine are detected at the AOB and suggest that specific urinary compounds play an important role in AOB-mediated reproductive events.

Alpha-Globulins↗

Monitoring for polyomavirus BK And JC in urine: comparison of quantitative polymerase chain reaction with urine cytology.

Clinical monitoring for polyomavirus infection is becoming common, but the optimal detection technique remains undefined. We compared the relative efficacy of exfoliative cytology and polymerase chain reaction (PCR) for detecting viruria in 100 urine samples. Definite decoy cells were seen in 8% and probable decoy cells in 5% of specimens. PCR showed BK virus (BKV) DNA in all these and in an additional 18% of samples. Using decoy cells as a marker of polyomavirus viruria cytology has a sensitivity of 41.9% and negative predictive value of 82.8%. The specificity and positive predictive value for viruria (not viral nephropathy) are 100%. False-negative results occurred in samples with suboptimal cellularity, vaginal contamination, and a low viral load. One patient with a false-negative urine cytology developed BKV nephropathy on follow-up. Compared with PCR, urine cytology is a less sensitive technique, which requires morphologically intact cells, and cannot distinguish BKV from JC virus.

BK Virus↗

[Purple urine bag syndrome (PUBS) associated with strong alkaline urine].

Mechanisms for purple discoloration of the plastic urine bag in purple urine bag syndrome (PUBS) were investigated. Activities of bacterial indoxyl sulfatase catalyzing the conversion of indoxyl sulfate to indigo (or indirubin) were detected in strong alkaline liquid media but not in normal ones. These enzyme activities were particularly high in simple and combined cultures of Proteus mirabilis and/or Klebsiella pneumoniae. These results suggest that occurrence of PUBS is associated with strong alkaline urine as well as urinary tract infections induced by some species of bacteria with indoxyl sulfatase.

Aged↗

[Bacterial contamination of infant urine samples obtained from filter papers used in neuroblastoma-screening tests. (6) Protective effects of preaddition of chlorhexidine digluconate in filter papers or containers for urine on bacterial break-down of creatinine, vanillylmandelic acid and homovanillic acid].

The effects of chlorhexidine digluconate preadded to urine samples for a neuroblastoma-screening test in preventing the break-down of creatinine (Cre), vanillylmandelic acid (VMA), and homovanillic acid (HVA) by creatinine-cleaving bacteria and the influence of the added disinfectant on the HPLC determination of urinary Cre, VMA, vanillillactic acid (VLA) and HVA. Laboratory or field experiments showed that preadded disinfectant (0.02% volume) nearly completely inhibited the growth of the bacteria and satisfactorily protected urinary Cre, VMA, and HVA from bacterial decomposition. Chlorhexidine digluconate was comparable to benzalkonium chloride in its inhibitory effects on the growth and activities of creatinine-cleaving bacteria. Unlike benzalkonium chloride, chlorhexidine digluconate added to urine samples gave no troubles in subsequent analytical processes. Addition of the disinfectant (0.02% volume) to standard Cre, VMA, VLA, and HVA solution did not affect retention times and sensitivities in HPLC analyses. To estimate influences of the added disinfectant on the serial HPLC determinations of VMA, VLA, and HVA on a large number of urine samples (40, 80, 120, 160, 200, or 5, 500), HPLC analyses were performed on standard VMA, VLA, and HVA solution with repeated injection of the disinfectant in great amounts into a column. The results showed no appreciable changes in the retention times and sensitivities of VMA, VLA, and HVA, and almost complete elution of the disinfectant retained on the column were possible with water-methanol (1:1, 2:8) or methanol washing.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Infections↗

A urine collector for separating the initial, midstream and final portions of voided urine in infancy and early childhood.

The object of this study was to design a simple and non-invasive method for the collection of uncontaminated urine samples in children aged up to 4 years. We developed a disposable plastic midstream urine collector for use in combination with a specially designed chair which was simple to use and cost effective. In this study we compared the initial portion, midstream and final portion during a single voiding in 40 children and found significant differences between initial and other fractions of the urine sample.

Child, Preschool↗

Effects of urine acidification on plasma and urine phencyclidine levels in overdosage.

A gas chromatography--mass fragmentography--electron impact (GC-MS-EI) assay of phencyclidine (PCP) was adapted for human plasma and urine. This assay is specific for PCP and very sensitive (approximately 1 ng/ml). Patients with the putative diagnosis of PCP overdosage were studied to correlate plasma and urinary levels with clinical state. Urinary PCP levels were enhanced in an acid urine, which suggests that acidification of the urine is an adjunct in the therapy of PCP overdosage.

Chromatography, Gas↗

A rapid and sensitive method for quantitation of nucleosides in human urine using liquid chromatography/mass spectrometry with direct urine injection.

Oxidized nucleosides are biochemical markers for tumors, aging, and neurodegenerative diseases. However, during the last decade, the analytical methods for nucleosides by gas chromatography/mass spectrometry (GC/MS) and high-performance liquid chromatography (HPLC) with single-parameter detectors like electron-capture detection (ECD) have not been sufficiently rapid or reliable to detect nucleosides in urine and to analyze clinical samples. It has been reported (Dudley et al., Rapid Commun Mass Spectrom. 2000; 14: 1200) that liquid chromatography with electrospray mass spectrometry (LC/ESI-MS) is more specific and sensitive for analysis of nucleosides than HPLC with conventional detectors; however, this method required complex extraction steps. In the present work a direct LC/ESI-MS method for nucleosides without extraction of urine samples has been developed. Analysis of nucleosides using positive-ion mode with selected reaction monitoring effectively eliminated potential interferences from endogenous constituents of the urine. This highly selective and sensitive method made it possible to analyze urinary nucleosides with a lower limit of quantitation of 0.2 nmol/mL. The method has been validated, with both excellent linearity and reproducibility, in the calibration range from 0.2-400 nmol/mL. The correlation coefficients of the calibration curves were higher than 0.987. The coefficients of variation were in the range 0.03-14.92% (inter-day) and 0.54-14.39% (intra-day), respectively.

Chromatography, High Pressure Liquid↗

Cotinine concentrations in amniotic fluid and urine of smoking, passive smoking and non-smoking pregnant women at term and in the urine of their neonates on 1st day of life.

Cotinine was measured in the amniotic fluid and urine of 31 pregnant women and in the urine of their offspring. Amniotic fluid cotinine was 8 times higher in active and 2.5 times higher in passive smokers than in non-smokers. In general, amniotic fluid cotinine was considerably higher than urinary cotinine both in active and in passive smokers. Estimation of cotinine both in amniotic fluid immediately before delivery and in urine of the newborn on the 1st day of life aids in assessing the degree of prenatal exposure to tobacco smoke.

Amniotic Fluid↗

Quantitation of the niacin metabolites 1-methylnicotinamide and l-methyl-2-pyridone-5-carboxamide in random spot urine samples, by ion-pairing reverse-phase HPLC with UV detection, and the implications for the use of spot urine samples in the assessment of niacin status.

A simple ion-pairing reverse-phase HPLC method, with UV diode array detection, was developed and validated for quantitation of the urinary niacin metabolites 1-methylnicotinamide and l-methyl-2-pyridone-5-carboxamide in a single run. Urine samples were purified using a polymer-based mixed mode anion exchange reverse-phase cartridge. Analysis was performed on a reverse-phase C18 column, using a methanol gradient elution system, containing phosphate buffer pH 7.0, 1-heptanesulphonic acid as the ion-pairing agent and trimethylamine as a modifier. The assay was applied to the measurement of the niacin status of two subjects using spot urine samples. The samples were collected over 4 consecutive days and at four time points during 1 day. Status, expressed as the concentration ratios (2-PYR or 1-MN)/creatinine and 2-PYR/l-MN, varied within and between days and was least for fasting samples. This work illustrates the potential of spot urine sampling for niacin status assessment, but highlights the need for further validation prior to its use in field nutritional surveys.

Chromatography, High Pressure Liquid↗

Determination of free catecholamines in human urine by direct injection of urine into a liquid chromatographic column-switching system with fluorimetric detection.

An ion-exchange chromatographic method combined with ion exclusion was developed for the determination of free catecholamines in human urine. Catecholamines were separated by ion exclusion from most acidic and neutral impurities by filtration through an anion-exchange column with a hydrophilic matrix (Asahipak ES-502N) and the excluded catecholamines were separated by ion-exchange chromatography on a column of weakly acidic ion exchanger with a hydrophilic matrix (Asahipak ES-502C), connected in series to the Asahipak ES-502N column with a four-way automatic valve. A sodium succinate-borate buffer of pH 6.7 (0.035 mol of succinic acid, 0.0075 mol of borate and 0.5 mmol of ethylenediaminetetraacetate were dissolved in 1 kg of water and the pH of the solution was adjusted to 6.7 with sodium hydroxide) was used as the mobile phase, and the temperature of both columns was kept at 30 degrees C. The catecholamines in the eluate were determined fluorimetrically by post-column derivatization with glycylglycine. A diluted urine sample was injected directly onto the first column. The first column was back-flushed with the mobile phase for 52.5 min after the elution of the catecholamines from the first to the second column. Then the columns were washed with the mobile phase for 10 min in the normal direction before the next sample was injected into the first column. Samples could be analysed every 70 min and 5 pmol/ml of epinephrine, 5 pmol/ml of norepinephrine and 25 pmol/ml of dopamine in human urine could be determined.

Catecholamines↗

Six spectroscopic methods for detection of oxidants in urine: implication in differentiation of normal and adulterated urine.

Six separate methods to detect oxidants in urine were developed. The presence of the oxidants was established by initial oxidation of ferrous to ferric ion and detecting the ferric by chromogenic oxidation or complex formation. The reagents for chromogenic oxidation were N,N-dimethylamoino-1,4-phenylenediamine (DMPDA), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS), and 2-amino-p-cresol (APC), and the reagents for the chromogenic complex were xylenol orange (XO), 8-hydroxy-7-iodo-5-quinolinesulfonic acid (HIQSA), and 4,5-dihydroxy-1,3-benzene-di-sulfonic acid (HBSA). All methods showed comparable results when tested for ferric, chromate, permanganate, oxychloride, hydrogen peroxide, oxone, tert-butylhydroperoxide, and cumenehydroperoxide at a concentration of 1.0 mmol/L in water (CV < 7%). The nitrite results are comparable only with DMPDA and APC. Periodate responded to the highest oxidation number (ON = 8) by chromogenic oxidation but lowest (ON = 2) by the chromogenic complex. The iodate responded only to the chromogenic oxidation with ON = 6. The linearity of the procedures was established by chromate in water. The linear concentrations were 0.09-12.00 mE/L for DMPDA, ABTS, APC, and HBSA and 0.09-6.00 mE/L for XO and HIQSA. In all methods, the correlation coefficients were > or = 0.9991 and precisions were within +/- 5.6%. The methods were used to test oxidants in 238 urine specimens. The chromate at 3.0 mE/L in water was used as standard. The correlation coefficients of 0.9600-0.9853 and the ANOVA test (F = 0.90, F(critical) = 2.22 at P(0.48)) indicated that the methods correlated well. The median concentration of oxidants in the specimens was 0.21 mE/L with an average and standard deviation of 0.62 +/- 1.19 (range 0.04-8.83 mE/L). When Grubbs' statistical test was applied to the specimen results, no specimen was found to be outlier or could be considered as adulterated. The Grubbs' test also revealed that the threshold concentration to identify urine adulteration was 29 mE/L at confidence level of 99%.

Chromogenic Compounds↗

Quantitation of renal antigen excretion in the urine of normal children and of children with various renal diseases. I. Quantitation of renal antigens in random urine samples.

This study reports a serologic method for the measurement of kidney-derived antigens in the urine of healthy children and of children with renal diseases. Two hundred twenty patients were studied. Four groups were recognized: group A, patients with no evidence of renal disease; group B, patients with past history of active urinary tract infection; group C, patients with active urinary tract infection; group D, patients with other renal diseases. Urinary renal antigen concentration was tested by the complement fixation method, in which titers of antigens in the urine were compared with a standard human renal antigen extract. The distribution of renal antigen concentrations in group C differed significantly (P(X2Y less than 0.001) from the other three groups. About 85% of patients in groups A, B, and D had levels below 0.6 mg/ml, whereas in group C only 53% of patients had similar concentrations. After factoring the results by the urinary concentration of creatinine, 85% of patients in group C had antigen levels above 0.6 mg/ml as opposed to 24%, 44%, and 27% in groups A, B, and D, respectively. The results of the study are consistent with the assumption that the rate of discharge of renal antigenic material in the urine is accelerated in certain renal diseases.

Antigens↗

Can a two-glass urine test or leucocyte esterase test of first-void urine improve syndromic management of male urethritis in southern Thailand?

The goal of this study was to determine whether a urine two-glass test or a leucocyte esterase (LE) test of first-void urine (FVU) improve the sensitivity or specificity of the World Health Organization (WHO) algorithm for the syndromic management of men with urethritis in southern Thailand. A secondary aim was to determine whether infection with Trichomonas vaginalis was sufficiently common to include treatment for it in a syndromic management protocol. One hundred and twenty-nine men with symptoms of urethritis seen at 2 STD clinics in Songkla Province, Thailand were enrolled. Symptoms and signs of each man were recorded and a urethral swab collected for microscopy and culture for Neisseria gonorrhoeae. A two-glass urine test and an LE test of an FVU specimen were performed. The FVU was tested by polymerase chain reaction (PCR) for N. gonorrhoeae, Chlamydia trachomatis and T. vaginalis. Dysuria was a symptom in 78% of men. A urethral discharge was a symptom in 68% but was evident on examination in 95% of the men. The prevalences of infection were 32.6% for N. gonorrhoeae, 23.3% for C. trachomatis, 1.6% for T. vaginalis and 51.9% for any infection. The sensitivities and specificities of urethral discharge on examination, two-glass test and LE test of FVU as indicators of infection with either or both of N. gonorrhoeae or C. trachomatis were 97% and 8%; 57% and 83%; and 59% and 78% respectively. Combinations of urethral discharge on examination and one of the other indicators were more specific but much less sensitive than the presence of discharge alone. Culture for N. gonorrhoeae was found to be only 43% sensitive compared with an expanded gold standard involving a PCR test. Our analysis demonstrates that neither the two-glass test nor the LE test of FVU were useful in improving on the WHO algorithm for management of men with urethritis. T. vaginalis was not common enough to include in a first-line syndromic management protocol for male urethritis. We recommend that, in southern Thailand, men with symptoms of urethritis in whom a urethral discharge is present on examination be offered immediate treatment for both N. gonorrhoeae and C. trachomatis as per the WHO algorithm.

Animals↗

Lead levels in urine of never-smoking adult women in non-polluted areas in Japan, with references to cadmium levels in urine.

The present study was initiated to assess current level of general population exposure to lead (Pb) in terms of Pb in urine (Pb-U). For this purpose, spot urine samples were collected from 2,332 never-smoking adult women in 10 areas all-over Japan, and were subjected to analysis for Pb-U by graphite furnace atomic absorption (GFAA). Data on cadmium in urine (Cd-U) as measured by GFAA were cited from a previous publication). The grand geometric means (GM) for Pb-U as observed, as corrected for creatinine (cr) and as corrected for a specific gravity of 1.016 were 1.06 microg/l, 1.28 microg/g cr and 1.14 microg/l, respectively, with small intra- and inter-variations with geometric standard deviations of about 2 or less. Arithmetic means of the 10 GMs (one each for the 10 areas) were 1.05 microg/l, 1.19 microg/g cr and 1.18 microg/l, respectively. The levels observed appeared to be lower than levels reported for other populations in the world. Chronological comparison within Japan suggested a gradual decrease in Pb-U in past over 20 yr. No close correlation was detected between Pb-U and Cd-U.

Adult↗