Radioimmunoassay technology in mass drug screening: an evaluation of an absorbent paper disk transport system

These studies have demonstrated the feasibility of using urine-saturated paper disks in place of urine in the RIA system for drug abuse detection. Results with the disks are consistent with those using urine. A satisfactory procedure has been devised which provides reproducibility of results with no loss of sensitivity or specificity. Further, the procedure is essentially the same as the current procedure requiring urine except that a paper disk punched from a filter paper strip impregnated with urine is used. Complete flexibility is retained to switch from urine to disk. No new or additional equipment is required. It is envisioned that the urine would remain at the collection site and dried filter paper strips containing urine under test be shipped to toxicology laboratories. Sould the disk assay be positive, the urine specimen identified with that disk could then be shipped to the laboratory for confirmation by gas-liquid chromatography or other acceptable methods. The time and expense incurred in shipping large volumes of urine would thus be eliminated.     

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.     

Concentrations of specific epithelial growth factors in the urine of interstitial cystitis patients and controls

PURPOSE: Interstitial cystitis (IC) is a chronic bladder disease for which the etiology is unknown. Because the bladder epithelium is often abnormal in IC, we determined whether the levels of specific urine growth factors postulated to be important for bladder epithelial proliferation are altered in IC. MATERIALS AND METHODS: ELISAs were used to determine levels of epidermal growth factor (EGF), insulin-like growth factor 1 (IGF1), insulin-like growth factor binding protein 3 (IGFBP3), and heparin binding epidermal growth factor-like growth factor (HB-EGF) in urine specimens from women with IC, asymptomatic women without bladder disease, and women with bacterial cystitis. RESULTS: Urine HB-EGF levels were specifically and significantly decreased in IC patients as compared to asymptomatic controls or patients with bacterial cystitis, whether expressed as concentration (amount per volume of urine) or the amount relative to urine creatinine in each specimen. In contrast, urine EGF, IGF1, and IGFBP3 levels were all significantly elevated in IC patients compared to asymptomatic controls. Further, the amounts of urine EGF and IGF1 were also elevated in IC patients as compared to patients with bacterial cystitis, and urine IGFBP3 levels were significantly elevated when expressed per milligram of urine creatinine. CONCLUSIONS: These findings indicate that complex changes in the levels of urine epithelial cell growth factors (EGF, IGF1, and HB-EGF) and a growth factor binding protein (IGFBP3) are associated with IC. While EGF, IGF1, and IGFBP3 levels are either the same or increased in the urine of IC patients as compared to patients with bacterial cystitis or asymptomatic controls, HB-EGF levels are significantly decreased in the urine of IC patients. Understanding the reasons for these changes may lead to understanding the pathogenesis of this disorder.     

Integration of Processes to Treat Wastewater and Source-Separated Urine

Human urine contributes 80% of the total nitrogen and 40–50% of the total phosphate load to municipal wastewater. This study examines the impact of separate urine collection and treatment on wastewater treatment. An integrated wastewater and urine treatment process was defined, in which single high-rate ammonium removal over nitrite and anaerobic ammonium oxidation processes and struvite recovery are at the heart of the nutrient management. The model study demonstrated that if 50% or more of urine were collected and treated separately, integrated wastewater treatment with more compact and energy-efficient processes would be possible. The integrated wastewater and urine treatment is compared to an existing state-of-the-art treatment process. The main advantage of urine separation is not only a better effluent quality. Existing processes including tertiary treatment can already produce very good effluent quality with total effluent nitrogen and phosphate concentrations of 2.5 and 0.5?g/m3, respectively. The main advantage of urine separation is the production of this same good effluent quality with a remarkable saving in resources. With sufficient urine separation, generation of net primary energy is possible.     

The effect of probucol on low density lipoprotein oxidation and femoral atherosclerosis

Our knowledge of the traits possessed by extraintestinal isolates of Escherichia coli, necessary for growth and survival in urine, is limited. To identify such determinants, transposon (TnphoA'1,4) mutant libraries of a clinical isolate (CP9) were generated and screened for derivatives exhibiting decreased growth in urine in vitro, and for mutants with active lacZ fusions that were induced in urine relative to laboratory medium. Using this approach we identified two genes, guaA (CPA24) and argC (CPI-1), which were previously unrecognized as being important for growth in human urine. Unexpectedly, not only does CPA24 (guaA) not grow in human urine in vitro, but it is sensitive to its effects, undergoing a 2-3 log loss of viability over 6 h. By contrast, CPA24 neither grows nor is killed in M9 minimal medium and artificial urine. Therefore, we postulate that lack of guanine or its derivatives in urine, and the inability of CPA24 to synthesize these compounds de novo, prevents CPA24 from synthesizing other guanine (or derivatives)-dependent products that are critical for growth and survival in urine. Although it seems logical that decreased growth in urine in vitro should correlate with diminished urovirulence, this concept was tested by challenging mice with CPA24 in vivo in a mouse model of urinary tract infection (UTI). Indeed, CPA24 was found to be significantly less virulent compared with its wild-type parent CP9. CPI-1(argC) was identified because of the significant induction of its argC::lacZ fusion in urine. Subsequent testing in urine demonstrated that its growth was significantly diminished in all urine samples tested (four females, three males). Polyamine synthesis is dependent upon, in part, the arginine biosynthetic pathway. Therefore, we tested whether the induction of argC in urine and/or the decreased growth of CPI-1 was a result of low levels of polyamines or arginine in urine. The results suggest that low levels of arginine, but not polyamines, in human urine are responsible. When tested in vivo in the mouse model of UTI, CPI-1 was also found to be significantly less virulent than CP9. In summary, we have established that guaA and argC are the first genes, which we are aware of, that have been shown to contribute to the growth of E. coli in urine in vitro and both have diminished urovirulence in vivo. These results support the concept that urine can be used in vitro as a screening tool to identify urovirulence traits.     

A urine analysis method suitable for children's nappies

BACKGROUND: Urinary tract infection in infancy continues to be underdiagnosed, despite its association with renal scarring and thus hypertension, renal failure, and other sequelae. Low ascertainment of urinary tract infections reflects the many difficulties in establishing a diagnosis, some of which could be eliminated by a simple, reliable method for preliminary investigation of children's urine. AIM: To assess the accuracy of a new, simple method for testing urine for nitrite and leucocyte esterase, which could be applied to children in primary care. METHODS: An in vitro study was carried out to compare the results of conventional urine analysis with urine analysis on urine soaked on to panty-liners, and with the laboratory investigation. Two urine analysis stick types were used (Boehringer Mannheim Nephur sticks and Bayer Multistix 8SG) and two brands of panty-liners. Analysis examined evidence of agreement and bias for different methods in addition to sensitivity, specificity, and negative predictive values for urine analysis. RESULTS: Pressing urine analysis test sticks on to panty-liners soaked with urine achieved consistent results compared with the results of conventional dipstick urine analysis. At a prevalence of 21.8%, sensitivity and negative predictive values of urine analysis for laboratory confirmed urinary tract infection were 94% and 98%, respectively, for Boehringer sticks, and 76% and 93%, respectively, for Bayer sticks. At prevalences of 5% and 1% (prevalences that could be expected in primary care) Bayer sticks had negative predictive values of 98.7% and 99.7%, respectively, and Boehringer sticks had values of 99.6% and 99.9%, respectively. CONCLUSIONS: Testing urine on panty-liners is accurate compared with conventional urine analysis. It may be possible to apply this method to testing unwell children presenting in primary care to identify those who require microbiological urine culture to confirm or eliminate a diagnosis of urinary tract infection.     

Mathematical optimization of the technology for preparing tablets containing amidopyrine and butadione

A method is proposed for urine specific gravity determination by urine refractometric index reading. Only two-three drops are required. Abbe refractometer is used. The urine specific gravity is determined by the formula: (formula: see text), where y = urine specific gravity, x = the refraction read. The method could not be used in cases with glucosuria and ketoniria as well as in case of a considerable proteinemia. The author's results correlate with the results of some foreign authors.     

Interventional magnetic resonance. Initial clinical experience with a 1.5-tesla magnetic resonance system combined with c-arm fluoroscopy

In our studies on diurnal 6-sulphatoxymelatonin (aMT6s) rhythms in various species, we have sometimes obtained fluctuating patterns. In most of these, the volume of individual urine fractions was not accurately measured because of methodological problems. Here, we report a simple method to overcome these problems by using urinary creatinine to estimate urine volume. The benefit of this method is demonstrated in two representative examples of the diurnal aMT6s rhythms of rats, domestic pigs and humans. Because the human urine fractions were collected accurately, the qualitative pattern of the aMT6s rhythm was not altered by using urinary creatinine as a substitute for urine volume. The total creatinine excretion (urine volume x creatinine concentration) was constant within a small range and showed no diurnal rhythm. In rats and pigs, the highly variable aMT6s concentrations relative to urine volume throughout the 24-hr period were changed drastically by referring to creatinine. All aMT6s patterns became stable and qualitatively similar to those of the rest of the group. From these results it can be concluded that creatinine is an adequate substitute for urine volume and a beneficial parameter with which to overcome technical problems with urine collection from laboratory animals or unknown urine volumes in human studies.     

Urinary antibody level and survival in bacteriuric institutionalized older subjects

OBJECTIVE: In a previous study, elevated urine antibody levels in institutionalized subjects with asymptomatic bacteriuria were associated with decreased survival. This study was undertaken to confirm the previous observation in a prospective study in a larger cohort and to explore selected other variables that may be associated with survival. DESIGN: A prospective, 24-month, observational cohort study. SETTING: Three large nursing homes in Winnipeg, Manitoba. PARTICIPANTS: Permanent residents were identified by initial screening urine cultures and subjects with bacteriuria were enrolled. The median age of subjects was 76 years, 51% were women, and the median duration of residence before enrollment was 26 months. Subjects were highly functionally impaired. MEASUREMENTS: Monthly urine specimens were collected for culture, leukocyte count, and urine antibody. Serum specimens for antibody to uropathogens and IL6 were obtained at enrollment and every 6 months. Anthroporphometric tests of nutritional status and functional and mental status were also measured every 6 months. Residents were stratified as having elevated or not elevated urine antibody, based on the initial urine specimen. The mean urine antibody for all of each subject's specimens was also calculated, and subjects were stratified as low, intermediate, or elevated urine antibody. Outcomes measured included mortality, infection and antibiotic use, and functional, mental, and nutritional decline. RESULTS: Ninety-eight bacteriuric subjects were enrolled in the study; 34 (35%) had elevated urine antibody and 64 (65%) had not elevated urine antibody. The two groups did not differ in demographic features, co-morbidities, functional status, medication use, or infecting organisms. Survival was significantly (P < .001) poorer in the group with elevated urine antibody. At 24 months, 35% of subjects with an elevated urine antibody were alive compared with 75% with a low urine antibody level. This survival difference was consistent when the two groups were stratified by sex, institution, and presence of a chronic indwelling catheter. Subjects with elevated urine antibody had no evidence for accelerated functional or nutritional decline during the study period compared with the group with low urine antibody. These subjects did, however, have an increased incidence of episodes of symptomatic urinary infection and infections at non-urinary sites. CONCLUSIONS: Older, bacteriuric, institutionalized subjects with elevated urine antibody had decreased survival rates compared with subjects with lower urine antibody levels. There is no clinical evidence to support accelerated decline caused by chronic infection to explain this observation. Urine antibody may be a marker for immune dysregulation, which precedes death in older impaired subjects.     

Acidic urine pH is associated with elevated levels of free urinary benzidine and N-acetylbenzidine and urothelial cell DNA adducts in exposed workers

We evaluated the influence of urine pH on the proportion of urinary benzidine (BZ) and N-acetylbenzidine present in the free, unconjugated state and on exfoliated urothelial cell DNA adduct levels in 32 workers exposed to BZ in India. Postworkshift urine pH was inversely correlated with the proportions of BZ (r = -0.78; P < 0.0001) and N-acetylbenzidine (r = -0.67; P < 0.0001) present as free compounds. Furthermore, the average of each subject's pre- and postworkshift urine pH was negatively associated with the predominant urothelial DNA adduct (P = 0.0037, adjusted for urinary BZ and metabolites), which has been shown to cochromatograph with a N-(3'-phosphodeoxyguanosin-8-yl)-N'-acetylbenzidine adduct standard. Controlling for internal dose, individuals with urine pH < 6 had 10-fold higher DNA adduct levels compared to subjects with urine pH > or = 7. As reported previously, polymorphisms in NAT1, NAT2, and GSTM1 had no impact on DNA adduct levels. This is the first study to demonstrate that urine pH has a strong influence on the presence of free urinary aromatic amine compounds and on urothelial cell DNA adduct levels in exposed humans. Because there is evidence that acidic urine has a similar influence on aromatic amines derived from cigarette smoke, urine pH, which is influenced by diet, may be an important susceptibility factor for bladder cancer caused by tobacco in the general population.     

Measurement of kidney concentrating ability in children with nonrenal glycosuria]

In non-renal (diabetic) glucosuria we did not find any statistically real relations between the concentration of glucose in the urine and cryoscopically measured osmolality in children with healthy kidneys. The close negative correlation of the conductance of the urine to the concentration of glucose is not only to be explained by changes of the viscosity, but is an expression of an increased re-absorption of sodium as a result of a compensatory hyperaldosteronism. In renal insufficiency the electrolytic conductibility of the urine is lower than the borderline area of the normal, even when under influence of the glucose excretion the osmolality of the urine is still to be found normal. Thus also on the conditions of a considerable glucosuria we can further judge the concentrating ability of the kidney in diabetes mellitus with the help of the measurement of the conductance of the urine.     

Nocturnal polyuria and natriuresis in male patients with nocturia and lower urinary tract symptoms

PURPOSE: We investigated the circadian variation in urine output, plasma angiotensin II, aldosterone, atrial natriuretic peptide, arginine vasopressin and blood pressure. MATERIALS AND METHODS: We studied 17 elderly men with nocturia and lower urinary tract symptoms, and 10 age matched controls without nocturia. RESULTS: Of the 17 patients studied 11 had a lack of diurnal variation in urine output and increased nocturnal urine production associated with increased nocturnal sodium excretion, and 6 had a diurnal variation in urine output comparable to controls. CONCLUSIONS: Nocturia in a large proportion of elderly men with lower urinary tract symptoms is caused by nocturnal polyuria and natriuresis.     

Interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) enhance lipopolysaccharide binding to neutrophils via CD14

BACKGROUND: Microalbuminuria is an early marker of prognostic significance in diabetic renal disease. However, testing for microalbuminuria in a timed sample of urine using the double antibody radioimmunoassay (RIA) method is cumbersome and requires special laboratory facilities. Recently, a test strip for microalbuminuria, the Micral Test was available and we evaluated the performance of this test strip as a screening method for detection of microalbuminuria. METHODS: One hundred consecutive diabetic patients who were tested to be dipstick-negative (Albustix) for proteinuria were enrolled for the study. Micral Tests were performed on a paired first morning and random urine specimen from the same patient and the results compared with a timed 24-hour urine measurement of urine albumin excretion using the RIA method. RESULTS: Eighteen specimens were tested positive by the RIA method with a urinary albumin range of 32-177 mg/24 hours. With the Micral Test, the following sensitivity, specificity, positive and negative predictive values were obtained: 66.7%. 97.6%, 85.7% and 93.0% for the first morning urine specimens, and 77.8%, 91.5%, 66.7% and 94.9% for the random urine specimens. CONCLUSIONS: These results suggest that Micral Test with either the first morning or random urine specimen offers a simple, reliable, rapid and convenient method for screening of microalbuminuria in the diabetic patient.     

Significance of the antibacterial agent assay of urine for bacteriological diagnosis and control of chemotherapy of urinary tract infections (author's transl)

The disc agar-diffusion-test using Bacillus subtilis ATCC 6051 as test organism is a simple and rapid method for routine testing of antibacterial agents in urine specimens. The test records urine levels which are expected under medium dosage, and in many cases even lower concentrations of renal excreted antibiotics. Out of 5655 analysed urine samples 22% contain antibacterial substances. In urine specimens over which information was volunteered that either no chemotherapy had been administered or that more than a three day's interval free of therapy existed, inhibitory substances are found in 8% and 27% respectively. Urine specimens which are supposedly collected from patients under current chemotherapy do not show therapeutic relevant antibiotic levels in 26%. Between urine specimens with and without antibacterial activity there is no significant difference in the incidence of viable counts of 10-4-10-5/ml and 10-5/ml. From urine samples with antibacterial content increases in the numbers of multiple resistant strains of E. coli, Proteus spp., Pseudom. aerug. and Enterobacter spp. together with high numbers of Candida spp. are observed.     

Urinary bile acids in population screening for inapparent liver disease

BACKGROUND/AIMS: We performed this study in order to evaluate the diagnostic potential of bile acids in random samples of urine for detection of latent liver disease and to compare a radioimmunoassay for urine bile acids with an enzymatic method that detects bile acid sulfates. This was a prospective cohort study carried out at the VA Medical Center involving 151 adults who attended a Community Health Fair at the hospital and wanted to know if they had liver disease. METHODOLOGY: Urinary bile acids in random specimens of 5-10 ml urine were measured. Radioimmunoassay for primary bile acids and an enzymatic assay with or without sulfated bile acids, all corrected with creatinine for urine flow were performed. Serum primary bile acids were determined by radioimmunoassay. In addition, routine liver profile and clinical examination were carried out. RESULTS: In 78 of 151 subjects there was at least one recent liver profile to match with the urine bile acids. Of these 78, 52 subjects with normal urine bile acids had a normal liver profile. In 11 subjects abnormal urine bile acids were associated with an abnormal liver profile. Nine of these 11 subjects were anti HCV positive, one was HIV positive. Urine bile acids correctly predicted the outcome of routine liver tests in 89% of 78 subjects. In nine cases there was a discordance between urinary bile acids and the liver profile. Failure to correctly predict the liver profile using urine, was reduced from nine subjects to three when urine bile acids were obtained twice at separate intervals. Urine bile acids predicted the outcome of anti HCV testing in 37 subjects with similar accuracy as serum ALT or AST. Urine bile acids correlated with serum bile acids at r=0.96, 0.88 and 0.76 for the radioimmunoassay, enzymatic assay that included sulfated bile acids and enzymatic assay without the sulfates, respectively. CONCLUSION: Bile acids in a random sample of urine are useful for population screening for latent liver disease. Prediction of sub-clinical hepatitis C is comparable to that of serum ALT or AST. Inclusion of bile acid sulfates mildly increases the predictive value of urine. Urine bile acids highly correlate with serum bile acids, indicating their surrogate diagnostic value.     

Detection of urinary tract infections by reduction of nitroblue tetrazolium

BACKGROUND: Nitroblue tetrazolium (NBT) reduction to formazan has been used as a marker for nitric oxide synthase (NOS). Since inducible NOS activity is elevated in urine from patients with urinary tract infections (UTIs), we investigated the accuracy of NBT reduction as an early predictor of UTIs and quantified the relationship between inducible NOS and NBT. METHODS: Urine samples from 434 patients were screened for the presence of UTIs with leukocyte-esterase and nitrite dipsticks and with NBT reduction. The rapid screening results from each test were compared to urine culture results. In addition, NBT reduction parameters were measured in urine pellet at 595 nm after incubation with one of four factors: NOS cofactors, NOS inhibitors, NADH, or superoxide dismutase/catalase. RESULTS: As a urine screening test for UTIs, NBT reduction was more sensitive with a higher negative predictive accuracy than the nitrite dipstick. NBT reduction also was more specific with a higher positive predictive accuracy and negative predictive accuracy than the leukocyte-esterase dipstick. In infected urine pellet, both NADPH, a NOS cofactor, and NADH increased NBT reduction. Superoxide dismutase/catalase decreased NBT reduction. CONCLUSIONS: Although NOS may not be the only NBT reducing enzyme, rapid, visible reduction of NBT is induced in urine from patients with UTIs.     

MP84 expression in the urine specimens of acute rejection renal transplant recipients

MP84 is a novel protein synthesized in response to all cytokines. This antigen is expressed only in stimulated mesangial cells and decreased kidney sections, but not in the normal kidney sections (1,2). This study was performed to determine the excretion of MP84 in the urine of renal transplant recipients with acute rejection. Six persons with renal transplant acute rejection and 10 healthy persons were included. Two urine specimens from each person were collected. Dot-blot assay was performed. It was shown that 12 urine specimens from 6 persons with acute rejection revealed MP84 in the matrix dot-blot assay while there was no staining for MP84 in the urine specimens of healthy persons. This could be due to the immunological alteration during the acute rejection which could lead to autocrine and paracrine secretion of growth factors and then the excretion of MP84 in the urine. The mechanism of MP84 secretion is not clear.     

Immunochemical approaches to AGE-structures: characterization of anti-AGE antibodies

An extractionless method for determining aflatoxin M1 (AFM1), a major metabolite of aflatoxin B1 (AFB1), in human urine was developed. The biological fluid is injected directly into the chromatographic system after simple dilution and centrifugation. A pre-column, packed with a cation-exchange phase and coupled on-line to a column-switching liquid chromatography (LC) system, is used for sample pre-treatment and concentration. The analytes are non-selectively desorbed with the LC eluent and cleaned by means of a column-switching procedure. Pre-treatment and analysis were performed within 40 min. Average AFMI recovery reached 97% in the 10-100 ng/l range of urine. The detection limit of AFM1 in urine and milk was 2.5 ng/l for 1 ml of injected sample. A comparison with an immunoaffinity column clean-up and LC method was performed. The method was applied to determine AFM1 in the urine of AFB1 gavaged rats, and in the urine of both potentially exposed and supposedly unexposed workers. The method was also extended to milk.     

Six cases of upper urinary tract diseases including tumors and inflammatory lesions which suggest the significance of urine cytology in preoperative diagnosis

We present six cases of upper urinary tract diseases including tumors and inflammatory lesions in which the urine cytology rather than the radiological examinations was useful for their preoperative diagnoses. Three of the six cases had malignant diseases and the others had benign diseases. In all cases preoperative results of urine cytology were identical to histopathological findings of resected specimens; the cases with positive findings in urine cytology had ureter cancers and those with negative findings had benign diseases. Primary CIS of upper urinary tract was found in two of six cases, which is still uncommon in Japan. Since it is very difficult to make a preoperative diagnosis of primary CIS by radiological examinations, the present study showed that urine cytology is useful for its preoperative diagnosis. Recently endoscopic techniques for the diagnosis of upper urinary tract tumors are in clinical use. The ureteroscopic biopsy is recommended for the case in which the diagnosis using urine cytology is difficult.     

Effect of acute water loading on urinary excretion of prostaglandin E in hypertensive patients

To assess the relationship or urine flow to the urinary excretion of prostaglandin E (PGE), urinary excretion of PGE was measured before and after acute water loading (20 ml/kg orally) in patients with hypertension. Water loading promptly increased urinary excretion of PGE as well as urine flow rate and decreased urine osmolality (all p less than 0.001), but did not affect urinary excretions of sodium, potassium and creatinine, plasma renin activity and plasma aldosterone concentration. There was a significant positive correlation between urine flow rate and urinary PGE excretion rate (p less than 0.01). Urinary PGE concentration correlated negatively with urine flow rate when the flow was lower than 5 ml/min (p less than 0.01). Urinary PGE concentration correlated negatively with urine flow rate when the flow was lower than 5 ml/min (p less than 0.01), whereas it did not change when the urine flow rate was larger than 5 ml/min. These results may support the hypothesis that urinary excretion of PGE is determined mainly by urine flow rate in the situation of water diuresis.