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Effect of urine component on leukocyte chemiluminescence response.

In the urine, the function of polymorphonuclear leukocytes (PMNs) is thought to be impaired because of the high osmolality and low pH along with a high concentration of inorganic salts. We investigated the effect of the properties of urine and its components on the chemiluminescence (CL) response of PMNs. This was helped by using an artificial urine. The CL response was measured by automatic luminometer following stimulation of phorbol myristate acetate. We found the CL response of PMNs to be significantly suppressed at a pH of 6 or 5, but not suppressed at a pH of 7 or 8. The CL response was significantly reduced when the osmolality was increased to 580 or 800 mOsm/kg H2O by the addition of urea as compared to the response in the standard artificial urine at an osmolality of 425 mOsm/kg H2O. A change in the osmolality by the addition of mannitol only minimally influenced the CL responses. In addition, the CL response was significantly impaired by both low and high concentrations of sodium at 12 and 300 mEq/L as compared to 77.5 mEq/L in the standard artificial urine. Potassium significantly reduced the CL response in a concentration-dependent manner in the range of from 4 to 31 mEq/L as compared to 52.3 mEq/L in the standard artificial urine. A high concentration of calcium at 8.1 or 9.9 mg/dL reduced CL response as compared to 3.9 mg/dL, whereas CL response was not reduced by the change in the concentration of magnesium. A high concentration of creatinine significantly reduced the CL response as compared to the standard artificial urine. We conclude that the function of PMNs in urine is reduced mainly by urine pH, concentration of urea, sodium, potassium, and creatinine. We suggest that reversal of these change in urine may restore functions of PMNs to clear bacteria in patients with urinary tract infections.

Bacteriuria↗

[The effects of human urine on the adhesion of calcium oxalate crystal to renal tubular cells].

PURPOSE: We examined the effects of human urine on the adhesion of calcium oxalate monohydrate (COM) crystals to Madin-Darby canine kidney (MDCK) cells in vitro. METHODS: Quantitative assay of COM crystal adhesion to MDCK cells: MDCK cells were exposed to COM crystal suspension for 5 minutes. Various urine samples were added in the COM crystal suspension. The adherent COM crystals on the MDCK cells were dissolved by 5 N hydrochloric acid. Calcium concentration of the solution was measured by atomic absorption analysis to quantify the volume of adherent COM crystals. This assay was applied for the experiments as follows. (1) Effect of human urine on COM crystal adhesion. (2) To investigate whether human urine inhibited COM crystal adhesion by acting on the crystal surface or on the cell surface. (3) Isolation of the substance from human urine that inhibits COM crystal adhesion. (4) Comparison of the inhibitory activities of the urine between from stone formers and healthy controls. RESULTS AND CONCLUSION: (1) Human urine had a strong inhibitory effect on COM crystal adhesion to MDCK cells. (2) The capacity of human urine to inhibit adhesion of COM crystals to MDCK cells was shown to be mediated by their ability to act on the crystal surface. (3) We isolated a macromolecular fraction (MW 60,000) that had strong capacity to inhibit cellular adhesion of COM crystals through ion exchange and gel filtration chromatography. (4) Nine urine samples from stone formers demonstrated extreme low inhibitory activities, which supposed us that the inhibitory capacity of the urine against crystal adhesion might be one of the risk factors in kidney stone formation.

Animals↗

Intra-individual variability in nighttime urine production and functional bladder capacity estimated by home recordings in patients with nocturnal enuresis.

PURPOSE: We evaluated the intra-individual variability and reproducibility of nighttime urine production on wet nights and functional bladder capacity estimated by long-term home recordings of monosymptomatic nocturnal enuresis. In particular, the intention was to evaluate the validity of 1 versus 2 weeks of recording when estimating urine volume on wet nights and 1 versus 2 weekends of recording when estimating functional bladder capacity. MATERIALS AND METHODS: We analyzed 120, 2-week home recordings of nighttime urine volume from patients with monosymptomatic nocturnal enuresis 6 to 16 years old (mean age 9.1) with at least 3 wet nights per week. Most patients were nonresponders or partial responders to desmopressin. Nighttime urine volume was estimated by weighing diapers before and after sleep, and measuring morning urine volume. Of the home recordings 62 included frequency volume charts for 2 weekends, which were evaluated for functional bladder capacity defined as the largest voided volume observed. RESULTS: No significant overall week-to-week differences were observed in average urine volume on wet nights and functional bladder capacity. There was a large intra-individual variability in all measured variables, which was most pronounced for functional bladder capacity and least pronounced for urine volume on wet nights. With regard to repeatability, the limits of agreement of urine volume on wet nights were -32% and 36% (95% confidence interval) as opposed to -54% and 48% for functional bladder capacity. CONCLUSIONS: In this study intra-individual week-to-week estimates of average urine volume on wet nights demonstrated acceptable variability and repeatability in contrast to functional bladder capacity. A reliable estimate of urine volume on wet nights could be obtained by 7 nights of home recording, whereas 4 days of daytime recording were necessary when estimating functional bladder capacity. Similar studies of patients who respond to desmopressin are needed.

Adolescent↗

An improved standardized procedure for urine mutagenicity testing.

The aim of the present study was to optimize the procedures for urinary mutagenicity testing in order to lower the baseline variation in mutagenic activity found in urine from unexposed subjects and to increase the sensitivity of the method. This was accomplished by using urine from nonsmokers and smokers as well as chemically spiked nonsmokers' urine. Diet was standardized. The number of mutants per ml of urine calculated from the linear portion of the dose-response curve was used as a measure of mutagenicity. The parameters investigated were (i) the total volume of urine per resin volume, (ii) the flow rate, (iii) the pH, (iv) the ionic strength of the urine, and (v) elimination of histidine. XAD-2 and C18 Sep-Pak resins recovered mutagens in smokers' urine and in chemically spiked urine with the same efficiency when an optimized procedure was adopted. The optimized procedure using a maximum volume of 50 ml acidified urine per Sep-Pak cartridge, or equal amount of XAD-2 resin, gave well over ten times greater recovery of mutagens from smokers' urine than in earlier reports. Histidine was effectively eliminated, and the background variation was also lowered.

Humans↗

[Quantitative urine microscopic examination using disposable counting chamber for diagnosis of urinary tract infection].

Routine urinalysis is performed as a screening test for urinary tract infection (UTI) in out-patients or in-patients. We assessed the usefulness of microscopic examination of unspun and unstained urine using a disposable slide with counting chambers (Kova Slide 10 grid, Miles-Sankyo) for diagnosis of significant bacteriuria. 173 fresh urine samples were obtained from 173 subjects (89 male and 84 (female), including 117 inpatients, aged from 0 to 96 years. Urine samples were examined for bacteriuria by the standard culture method and counting chamber method. Significant bacteriuria was defined as 10(5)/ml or more of bacilli for midstream urine and urine collected by bags and 10(4)/ml or more for urine collected by catheterization and from indwelling catheters. Urine leukocytes were also counted on disposable slide. The rapid dipstick test (N-multistix-SG-10, Miles-Sankyo) of leukocyte esterase activity and nitrite were measured in the urine specimens read by a photometer (Clinitek-10, Miles-Sankyo). Significant bacteriuria was diagnosed by standard culture method in 67 urine samples. Close correlation was obtained between bacterial counts determined by the bacterial culture and counting chamber method (Spearman's correlation coefficient p < 0.001). Sensitivity and negative predictive value for significant bacteriuria were 94.0 and 95.1%, respectively, when bacteriuria or pyuria was present in the counting chamber. Dipstick test had a sensitivity and negative predictive value of 86.6 and 89.9%, respectively, when either leukocyte esterase activity of + or more, or nitrite of + was found. In out-patients, both sensitivity and negative predictive value were as high as 100% in counting chamber method. Thus, we can conclude that urine microscopy on disposable counting chambers is a very sensitive, simple, time-saving and lost-effective method for diagnosis of UTI.

Adolescent↗

Detection of Chlamydia trachomatis in first-void urine from men and women as an alternative to swabs.

BACKGROUND: Ideally, an effective preventive strategy for the control of Chlamydia trachomatis infection should take into account the following attributes: rapid and simple specimen collection, low cost and noninvasive test processing. Therefore, we compared the performance profile of urine-based detection of C. trachomatis antigen in first-void urine with that of testing urethral and endocervical samples in men and women. MATERIAL AND METHODS: Urethral and endocervical samples and first-void urine from 285 men and 192 women attending the Sexually Transmitted Diseases Outpatient Clinic at the Medical University in Plovdiv, Bulgaria were tested using direct immunofluorescence assay (DFA) (MicroTrak, Syva, Palo Alto, CA, USA). RESULTS: Seventy (25%) of all men tested were positive for C. trachomatis antigen in either urethral or urine samples. 65 men (93%) had both a positive urethral and urine sample, three men (4%) had only a positive urethral sample and two (3%) had only a positive urine sample. Thirty-five women (18%) had C. trachomatis infection. Twenty-six women (74%) had both a positive endocervical and urethral sample, 6 (17%) had only a positive endocervical sample and 3 (8%) had only a positive urethral sample. All women with positive urethral samples tested positive on their urine samples. Two of the women with a negative urethral sample and a positive endocervical sample had a positive urine sample. CONCLUSIONS: These results show that using direct immunofluorescence assay on first-void urine samples is a reliable noninvasive method which can replace urethral swabs in the diagnosis of C. trachomatis infection in symptomatic men. Urine-based strategies are also an acceptable alternative for the diagnosis of C. trachomatis infection in symptomatic women when it is not possible to obtain an urogenital sample.

Adult↗

Comparison of first-voided urine specimens with endocervical swab specimens for enzyme-linked immunosorbent assay detection of Chlamydia trachomatis in women.

OBJECTIVE: To compare first-voided urine specimens with paired endocervical swab specimens from women to determine the role of urine in complementing or replacing swab specimens for the detection of the chlamydial antigen. DESIGN: For 18 months, both endocervical swab specimens (the criterion standard) and urine specimens were tested for the chlamydial antigen, using an enzyme-linked immunosorbent assay (Chlamydiazyme, Abbott Laboratories, North Chicago, Ill). Positive results were confirmed using a blocking reagent (Abbott Laboratories) and/or a direct fluorescent antibody test (Micro-Trak, Syva, Palo Alto, Calif). A low level of chlamydial antigen (below the enzyme-linked immunosorbent assay threshold recommended by the manufacturer) was also looked for and, when found, was confirmed by the direct fluorescent antibody test. SETTING: Prenatal and family practice clinics in a 500-bed community hospital. PATIENTS: Specimens were collected from 489 random asymptomatic or symptomatic women. MAIN OUTCOME MEASURE: The detection of the chlamydial antigen from endocervical swab specimens was compared with the detection from first-voided urine specimens. RESULTS: Acceptable swab and urine specimens were obtained from 300 (61.3%) of the patients. The antigen of Chlamydia trachomatis was confirmed in 20 (6.7%) of the 300 women. Of the infected patients, the antigen was detected in both swab and urine specimens for nine patients (45%), only in the swab specimens for eight (40%), and only in the urine specimens for three (15%). Testing urine in addition to endocervical swab specimens allowed for the detection of 18% more chlamydial infections, whereas confirming the presence of the antigen below the enzyme-linked immunosorbent assay cutoff resulted in the detection of 54% more infections. CONCLUSIONS: Collecting multiple specimens and testing for low levels of chlamydial antigen may significantly improve the detection of chlamydial infections in women. First-voided urine may be an appropriate complementary specimen to endocervical swab specimens, but urine by itself does not allow for the adequate detection of the chlamydial antigen in women.

Antigens, Bacterial↗

The role of urine sugar in diabetic management.

The concentration of reducing sugar in the urine is commonly used in the management of diabetes in children. Supplemental doses of regular insulin are administered in response to the concentration of urine sugar according to a protocol termed the "sliding scale." This practice assumes that the concentration of sugar in urine is a good indicator of the plasma glucose concentration. This assumption was tested by comparing urine sugar concentrations in first and second voided urines with the plasma glucose concentrations in 220 children with diabetes. The correlation was good (r = .92) for both the first and second voided urine specimens. Thus, urine sugar concentrations in general define the level of plasma glucose. The large standard deviation of the plasma glucose at each concentration of urine sugar, however, limits the usefulness of urine sugar as an accurate reflection of the coincident plasma glucose concentration. The urine sugar concentration, although useful for the general management of diabetes, provides significant risk when used to guide frequent adjustments in insulin administration. Therefore, the "sliding scale" should not be used in the treatment of children with diabetes.

Adolescent↗

Low-molecular-weight variants of osteopontin generated by serine proteinases in urine of patients with kidney stones.

Osteopontin (OPN) is a multifunctional glycosylated phosphoprotein found in body fluids, including urine, and has been implicated in urinary stone formation. We tested the hypothesis that OPN levels in urine of patients with kidney stones differed from normal individuals. To quantify OPN levels in the urine, we developed an ELISA using a combination of a mouse monoclonal and rabbit polyclonal antibodies raised against a recombinant glutathione-S-transferase-human OPN fusion protein. In a group of 34 patients diagnosed with kidney stones compared with a control group of 23 normal individuals, we found that OPN levels in urine of the patient and control groups ranged from 0.01 to 2.7 micrograms/ml, with no significant difference in their medians (P > 0.8, Mann-Whitney test). OPN in urine was qualitatively assessed by Western blotting using a biotinylated monoclonal antibody to detect various molecular forms. The urine of most individuals contained OPN species within in the 55- to 66-kDa electrophoretic mobility range. However, a significantly higher proportion of individuals in the patient group (13 of 34) was found to have aberrant urine OPN species (< or = 40 kDa) compared to 2 of 23 for the control group (P < 0.03, chi 2 test). Mixing experiments indicated that urine samples with aberrant OPN contain proteases inhibitable with phenylmethylsulfonyl fluoride. Such proteases could break down normal urine OPN in vitro. Therefore, urine from a high frequency of kidney stone patients contains serine proteases that contribute to proteolytic cleavage of OPN.

Adult↗

Serum and urine beta-2-microglobulin in hemophagocytic syndrome.

BACKGROUND: Previous reports suggesting a correlation between lymphoproliferative disease and serum levels of beta-2-microglobulin (beta-2M) and in vitro data indicating a role of cytokines in the production of beta-2M prompted a study of serum and urine beta-2M concentration in patients with hemophagocytic syndrome (HPS), because no data were previously available for HPS, a pathologic state associated with excessive cytokines secreted from activated reactive/malignant lymphocytes and histiocytes. METHODS: Serum and urine beta-2M levels were measured in six children with HPS during active and convalescent phase and in other diseases. RESULTS: Serum and urine beta-2M levels during active phase HPS were significantly high not only in serum (median, 7.5 mg/l; range, 2.3-16.0 mg/l; P < 0.01), but also in urine (median, > 31,650 micrograms/gram Creatinine (gCr); range, 8179-236,333 micrograms/gCr; P < 0.01), compared with levels during convalescent phase HPS (median, 2.0 mg/l; range, 0.9-2.5 mg/l in serum and median, 338 micrograms/gCr; range, 223-585 micrograms/gCr in urine) and in control subjects with diseases such as acute lymphocytic leukemia (median, 2.3 mg/l; range, 1.0-2.8 mg/l in serum and median, 327 micrograms/gCr; range, 48-2684 micrograms/gCr in urine), infectious mononucleosis (median, 2.9 mg/l; range, 2.5-5.5 mg/l in serum and median, 348 micrograms/gCr; range, 80-1325 micrograms/gCr in urine), and Kawasaki disease (median, 2.8 mg/l; range, 1.5-3.3 mg/l in serum and median, 1016 micrograms/gCr; range, 214-4500 micrograms/gCr in urine). Noteworthy was the observation that urine beta-2M levels correlated closely with HPS disease activity. CONCLUSIONS: Urine beta-2M appears to be a useful marker for assessing disease activity in patients with HPS.

Adolescent↗

Sensitive detection of transitional cell carcinoma of the bladder by microsatellite analysis of cells exfoliated in urine.

Transitional cell carcinoma (TCC) is the most common bladder tumor. Urine cytology can identify most high-grade tumors but sensitivity is lower if one includes lesions of all grades. Microsatellite marker alterations have been found in many tumor types including bladder cancer and have been used to detect cancer cells in body fluids including urine. The aim of our study is to further evaluate feasibility and sensitivity of microsatellite analysis to detect bladder cancer cells in urine. We studied 55 individuals: 21 with symptoms suggestive of bladder cancer, 23 patients with previous history of TCC and 11 healthy subjects. Genomic DNA was extracted from blood lymphocytes, urine sediment, bladder washings and tumor or normal bladder mucosa. Twenty highly informative microsatellite markers were analyzed for loss of heterozigosity (LOH) and microsatellite instability (MIN) by polymerase chain reaction. Microsatellite analysis of urine identified 33 of 34 (97%) patients with either primary or tumor recurrence, whereas urine cytology identified 27 of 34 (79%) patients (p = 0.0001). Detection of microsatellite abnormalities improved the sensitivity of detecting low-grade and/or stage bladder tumor: from 75-95% for grades G1-G2 and from 75-100% for pTis-pTa tumors. Bladder washings from 25 patients were also analyzed, and in all cases results were identical to those obtained from voided urine. None of the 16 patients without evidence of TCC showed LOH and/or MIN in urine samples or bladder washings. Interestingly, in a patient with persistent bladder mucosa abnormalities, microsatellite alterations were demonstrated 8 months before the histopathologic diagnosis of tumor recurrence. These results further indicate that microsatellite marker analysis is more sensitive than conventional urine cytology in detecting bladder cancer cells in urine and represents a potential clinical tool for monitoring patients with low-grade/stage TCC.

Adult↗

Analysis of an expanded width of albumin fraction by cellulose acetate membrane electrophoresis in IgA nephropathy urine before treatment.

Cellulose acetate membrane electrophoresis with colloidal silver stain re-vealed that the width of the albumin fraction in IgA nephropathy (IgAN) urine before treatment was significantly expanded. This phenomenon was not shown in IgAN urine after treatment or in non-IgAN urine. There was a reverse correlation between the width of the albumin fraction and the albumin con-centration in IgAN urine. By immuno-fixation, Tamm-Horsfall protein (THP) was located in the same position as the albumin band in IgAN urine before treatment; however, in the urine of a healthy subject it was located in the same position as alpha(1)-globulin. By ELISA, the THP-albumin complex concentration in IgAN urine before treatment was significantly higher than in the other two diseases. The width of the albumin fraction and the sodium ion concentra-tion of the urine were significantly correlated. The THP/albumin ratio in IgAN urine before treatment was significantly higher than in the other two groups. This suggests that the characteristic expanded width of albumin found by immunofixation indicates a THP-albumin complex, and that the sodium concentration of urine is involved in the formation of this complex.

Albuminuria↗

High-performance liquid chromatographic procedures for the analysis of carboplatin in human plasma and urine.

Specific, sensitive and reproducible high-performance liquid chromatographic procedures were developed for the quantitative analysis of carboplatin in human plasma and urine. Plasma and urine were ultrafiltered with an Amicon Centrifree micropartition system, and samples were injected onto a LiChrosorb diol column. The mobile phase was CH3CN/H2O (92:8, v/v) for plasma and CH3CN/0.015% H3PO4 (89:11, v/v) for urine. The effluents were monitored at 229 nm. Carboplatin eluted by 10 min. The detector response was linear from 0.5 (plasma) or 5 (urine) to 500 micrograms/ml. The lower limit of quantification was 1.0 micrograms/ml plasma and 5.0 micrograms/ml urine. Constituents in plasma and urine, and possible degradation products (cyclobutane mono- and dicarboxylic acids) did not interfere. Within-day precision was less than 4% for plasma and 9% and 4% for urine concentrations of 40 and 401 micrograms/ml, respectively. Within-day accuracy was 96% or greater for both matrices. Carboplatin was not bound to the Centrifree membrane and recovery was 94% for plasma and 96% for urine. The storage stability of carboplatin in water, plasma, plasma ultrafiltrate, and urine and the extent of binding to human plasma proteins were evaluated. The percentage of carboplatin reversibly bound to plasma proteins was minimal (less than or equal to 10%) over a range of 1-50 micrograms/ml. In human plasma at 37 degrees C the drug was stable for about 2 h, but then degraded with a half-life of 32 h. Carboplatin had limited stability in water, plasma, and urine stored at -25 degrees C. Biological samples, therefore, should be stored frozen and analyzed within a week of collection to obtain valid results.

Antineoplastic Agents↗

Urine interleukin-6 and interleukin-8 in children with acute pyelonephritis, in relation to DMSA scintigraphy in the acute phase and at 1-year follow-up.

The relationship between urine interleukin-6 (IL-6) and interleukin-8 (IL-8)/creatinine quotients and 99mTc-dimercaptosuccinic acid (DMSA) scintigraphy, performed within 10 days of acute first-time pyelonephritis and after 1 year, was studied in 41 children. The urine IL-6 and IL-8/creatinine quotients were also related to the urine N-acetyl-beta-D-glucosaminidase (NAG) and albumin/creatinine quotients. Presence of DMSA uptake defects, reflecting local inflammation, in children in the acute phase of pyelonephritis, were associated with elevated urine IL-6/creatinine quotients (median 27 pg/mumol); in children without DMSA changes there was no increase in quotients (median non-detectable) (P < 0.05). Persistent DMSA changes at the 1-year follow-up, probably reflecting renal scarring, were only seen in children with increased urine IL-6/creatinine quotients in the acute phase (P < 0.01). No correlation was found between urine IL-8 and DMSA uptake defects. Vesicoureteral reflux (VUR) at 6-8 weeks did not correlate with the urine cytokine levels in the acute phase. The urine excretion of NAG and albumin, reflecting renal dysfunction, was associated with values of both urine IL-6 and IL-8/creatinine quotients, but not with DMSA defects or VUR. Thus, the initial urine IL-6/creatinine quotients might be used as an indicator of risk for persistent renal damage in acute pyelonephritis.

Acetylglucosaminidase↗

Utility of urine reagent strip in screening women with incontinence for urinary tract infection.

The aim of this study was to determine the utility of urine reagent strips in screening women with urinary incontinence for urinary tract infections. Subjects were all women presenting with urinary incontinence. A urine specimen was screened for urinary tract infection with a urine reagent strip and urine culture. Using the urine culture result as the gold standard, the sensitivity, specificity, and positive and negative predictive values of the urine reagent strips were calculated. A total of 265 pairs of reagent strips and urine cultures were evaluated. Thirty-one cultures grew out of a single organism; nine of these had a positive urine reagent strip, yielding a sensitivity of 29%. The specificity of the urine reagent strip was 99%, the positive predictive value was 82% and the negative predictive value was 92%. For women presenting with urinary incontinence, the sensitivity of a urine reagent test for diagnosing urinary tract infection was low.

Bacteriological Techniques↗

Evaluation of a commercial enzyme immunoassay for HIV screening in urine.

A cross-sectional study was conducted to evaluate the utility of a commercial enzyme immunoassay (EIA) as a screening test for detecting HIV-1 antibody in urine in a population at risk for HIV infection in Catalonia, Spain. Paired urine and serum samples were collected consecutively from 99 patients who attended two drug-dependency treatment centres and 151 patients who attended a sexually transmitted diseases (STD) clinic in Barcelona. Antibodies against HIV in urine samples were detected using the Calypte HIV-1 Urine EIA (Calypte Biomedical Corporation, Berkeley, CA, USA) and confirmed by urine-based Western blot (WB) analysis. Sera were analysed using Bioelisa HIV-1+2 EIA (Biokit Laboratories, Barcelona, Spain), and the results were verified using serum-based WB analysis. Results of both urine and serum testing were available for 246 of 250 participants. For 52 individuals the results of both urine and serum testing were positive and for five the results were discordant (2 with urine-negative/serum-positive results and 3 with urine-positive/serum-negative results). The respective sensitivity and specificity values obtained for the urine EIA were 100% and 96.2% for intravenous drug users (IDUs) and 80% and 99.3% for persons attending the STD clinic. According to the 1997 UNAIDS/WHO strategy I recommendations, these values are acceptable for surveillance purposes, particularly in populations with a high prevalence of HIV infection.

Cross-Sectional Studies↗

A variant form of arylsulfatase A in human urine derived directly from the renal pelvis: kinetic and immunological characterization.

Arylsulfatase A (aryl-sulfate sulfohydrolase, EC 3.1.6.1) was examined in voided and in nephrostomic urine. A variant form of the enzyme was found in nephrostomic urine, in addition to the minor form, which is the sole component of arylsulfatase A in voided urine. The nephrostomic enzyme differed from the voided urine enzyme with respect to the kinetic parameters, the isoelectric point, heat stability and immunological reactivity. The isoelectric points of the voided urine and nephrostomic enzymes were 4.7 and 5.3, respectively. The nephrostomic enzyme was more heat-labile at 62.5 degrees C than the voided urine enzyme. Although the Km values of the two enzymes with nitrocatechol sulfate as substrate were almost the same, the V value of the nephrostomic enzyme was approx. one-hundredth that of the voided urine enzyme. The molecular weight (almost 130 000) did not differ between the voided urine and nephrostomic enzymes. It was demonstrated by various methods, using IgG antibody against the purified voided urine enzyme, that the nephrostomic enzyme was antigenically distinct from the voided urine enzyme.

Cerebroside-Sulfatase↗

Analysis of rat urine proteins and allergens by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting.

BACKGROUND: In rats, urine has been identified as a major source of the allergens that cause laboratory animal allergy, an important occupational health problem. METHODS AND RESULTS: Urinary proteins and allergens of Wistar rats were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Proteins excreted by male and female rats during puberty were similar and of low molecular weight. In adulthood, moderate increases in diffuse staining of 26 and 21 kd occurred in female urine. In males the 17 kd protein increased dramatically and the 23 and 21 kd proteins moderately. The urine excretion of high-molecular-weight proteins (75 to 63 kd) increased with age in males (females not studied). Immunoblot studies with six sera showed allergens in urine of male and female rats of all ages, three of which were present in all urine (75, 68, and 21 kd). Three allergens (17, 16, and 15 kd) in female urine may be allergenically similar to the 17 kd allergen in adult male urine. Adult male urine allergens were studied further with sera from 83 rat-hypersensitive subjects. Major allergens were identified at 23, 21, and 17 kd, and all sera had IgE to one or more of these proteins. Twenty-seven percent had IgE to 68 and 63 kd allergens. Minor allergens were identified at 75, 51, and 44 kd. CONCLUSIONS: Rat urine is an important source of the major allergens associated with rat hypersensitivity. Age and sex markedly influence the protein and allergenic constituents of rat urine.

Aging↗