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PubMed1999

Urine antibody tests: new insights into the dynamics of HIV-1 infection.

BACKGROUND: Noninvasive methodologies provide alternatives to diagnostic blood tests and have high patient acceptance, increased safety, and reduced costs. Such tests may supplement or replace blood diagnostic assays currently in use. METHODS: Using a licensed urine-based test for antibody to HIV-1, we performed 25 991 HIV-1 urine antibody enzyme immunoassay (EIA) screening tests [confirmable by HIV-1 Western blot (WB)] on paired urine and blood specimens obtained from high- and low-risk HIV-1 subjects collected at six sites representative of the US population. RESULTS: Using HIV-1 urine EIA tests confirmed by urine Western blot, a compartmentalized immune response (urine positive/serum negative) occurred in 0.24% of a cohort of 11 896 subjects. In the same cohort, specimens that were urine negative/serum positive occurred in 0.17% of subjects. In a second study of 25 991 subjects that included 859 high-risk individuals, the false-positive urine EIA frequency (urine WB negative or indeterminate) was 1.3%. This false-positive frequency in the high-risk cohort was attributed, in part, to an IgA antibody response. We tabulated urine and serum indeterminate reactivities and examined their possible causes. Data are presented showing that antibodies from a seroindeterminate HIV-1vau group O subject were reactive in urine EIA and urine WB tests. An analysis of the HIV-1vau strain group O env nucleotide sequence disclosed a high frequency of homology with human chromosome 7q31, a fragile site implicated in many human malignancies. CONCLUSIONS: These results demonstrate the utility of urine for alternative HIV-1 antibody testing and provide new insights into the pathogenesis of HIV-1 infection and into potential application of this approach in investigation of other microbial pathogens and toxic compounds.

Antibodies, Viral↗
PubMed1990

Assessment of proteinuria by using the protein/creatinine ratio of single-voided urine.

Quantitative 24-hour urine protein excretion is used in the diagnosis, the monitoring of therapeutic effects and the prognosis of renal disease. However, this method is time-consuming, cumbersome and often inaccurate. Many studies have shown that the single voided urine protein/creatinine (Pr/Cr) ratio relates well with 24-hour urine protein excretion and can be substituted for evaluating some conditions. In our study, 41 patient with renal disease (25 men, 16 women) with a mean age of 43 years (range, 20-79 years), not only collected 24-hour urine, but also collected single voided urine at four different times. There was an excellent correlation between 24-hour urine protein excretion and the single voided urine Pr/Cr ratio at various degrees of creatinine clearance and ranges of proteinuria. The highest correlation was found in urine specimens collected at 16:00 hours with a correlation coefficient (r) of 0.91. Other urine specimens also showed a good relation with a correlation coefficient of above 0.80. The creatinine clearance (Ccr) greater than or equal to 70 ml/min group showed a better relationship than the Ccr less than 70 ml/min group. The proteinuria more than 1.0 g/day group also showed a better relationship than the group with proteinuria of less than 1.0 g/day. However, these differences were not statistically significant. No significant differences between the different age groups or sexes, using the single voided urine Pr/Cr ratio to estimate the 24-hour urine protein excretion were found. It is concluded that the single voided urine Pr/Cr ratio correlates highly with 24-hour urine protein excretion and could be an alternative means for disease monitoring and screening.

Adult↗
PubMed1993

Gravimetric method for the dynamic measurement of urine flow.

The rate of urine formation is a primary index of renal function, but no techniques are currently available to accurately measure low rates of urine flow on a continuous basis, such as are normally found in rats. We developed a gravimetric method for the dynamic measurement of urine flow in anesthetized rats. Catheters were inserted directly into the ureters close to the renal pelves, and a siphon was created to collect all of the urine formed as rapidly as it was produced. Urine flow was determined by measuring the weight of the urine using a direct-reading analytical balance interfaced to a computer. Basal urine flow was measured at 2-sec intervals for 30 to 60 min. The dynamic response of urine flow to a rapid decrease in arterial pressure produced by a bolus intravenous injection of acetylcholine (0.5 micrograms) was also measured. Intrinsic drift, evaporative losses, and the responsiveness of the system to several fixed pump flows in the low physiologic range were evaluated in vitro. The gravimetric method described was able to continuously measure basal urine flows that averaged 37.3 +/- 12.4 microliters/min. Error due to drift and evaporation was negligible, totaling less than 1% of the measured urine flow. Acetylcholine-induced declines in arterial pressure were followed within 8 sec by a decline in urine flow. These data demonstrate that this new gravimetric method provides a simple, inexpensive, dynamic measurement of urine flow in the microliter/min range.

Acetylcholine↗
PubMed1982

Evaluation of Gram-stain screen and Micro-ID methods for direct identification of Enterobacteriaceae from urines.

A rapid method of urine screening and enterobacterial identification was evaluated. Results indicated that an average of 13.5 bacteria/oil immersion field (threshold value greater than or equal to 1) was observed in unsedimented urine of patients with significant bacteriuria, with an average of less than 1 bacterium/field in urines of patients without significant bacteriuria. In centrifuged urines, numbers of bacteria divided by amount of urine sedimented yielded similar results. Of 1758 urines studied, 136 yielded greater than or equal to 10(5) bacteria/ml, and 58 greater than 10(4) but less than 10(5) bacteria/ml, by conventional techniques. Gram-screening of unsedimented specimens gave sensitivity rates of 94.1%, specificity of 97.7%, and predictive positive and negative values of 78.5%, 99.5%, respectively; similar values were obtained with sedimented urines. Sensitivity rates of both screening methods for the 58 urines with greater than 10(4) but less than 10(5) bacteria/ml were 9.0%, 10.0%, respectively. Total correct enteric identification in 113 urines with positive screens and significant bacteriuria (greater than or equal to 10(5)/ml) was 82.3% and 90.3% with direct saline and broth Micro-ID methods, respectively. In 99 urines yielding pure or predominantly pure growth of 1 species of Enterobacteriaceae identification by direct saline and broth Micro-ID corresponded with isolated colony identification in 85.9%, 94.9% of cases, respectively. Gram-stain screening (together with back-up conventional plating in certain patient categories) and enterobacterial identification by direct broth Micro-ID, of urines with pure stains suggestive of greater than or equal to 10(5) Gram-negative rods/ml has been shown to be useful in laboratories without automated equipment for urine screening.

Bacteriological Techniques↗
PubMed1999

Urine colour as an index of hydration in critically ill patients.

In order to test the hypothesis that urine colour can be used as an index of hydration in critically ill patients, we selected 40 intensive care and high-dependency patients and correlated urine colour (scored on an eight-point scale) with various indices of hydration: urine:plasma sodium, osmolality and urea ratios, urine output and central venous pressure. In addition, we compared the colour-chart score with scores made by intensive care nurses (without the benefit of a colour chart) in order to test subjective assessment of urine colour. There were weak but statistically significant correlations between urine colour and urine output (Spearman's r = - 0.555) and between urine colour and urine:plasma sodium ratio (Spearman's r = - 0.459). Subjective assessment of urine colour appeared to be reliable. Thus, although urine colour does vary with hydration in the critically ill, assessment of urine colour adds little to the overall assessment of hydration in this group of patients.

Adolescent↗
PubMed1996

Evaluation of suprapubic bladder aspiration for detection of group B streptococcal antigen by latex agglutination in neonatal urine.

Our objective was to determine whether urine collection by suprapubic bladder aspiration (SBA) improves the specificity of the group B streptococcal (GBS) latex agglutination (LA) test by avoiding contamination of urine with GBS from perineal and rectal colonization that can result in a positive LA test in an uninfected infant when the urine is collected by bag. Part 1 consists of a retrospective review of the medical records of 113 infants who had urine collected by SBA for GBS LA testing as part of evaluation for possible sepsis. The sensitivity and specificity of the urine LA test was assessed by comparing it with blood culture results. In part 2, a prospective analysis was performed of 19 newborns who had rectal and vaginal/penile cultures as well as urine by SBA and bag for GBS cultures and LA. Results of LA testing on urine collected by both of these methods were compared with results of urine, perineal, and rectal cultures. In the retrospective review of GBS LA testing performed on 113 consecutive urine specimens collected by SBA from neonates being evaluated for suspected sepsis, the sensitivity and specificity were 67% and 89%, respectively, when compared with blood culture results. Twelve infants who had a positive LA test result but a sterile blood culture (BC-,LA+) were compared with 95 infants with both blood cultures and urine LA tests negative for GBS (BC-, LA-). BC-, LA+ infants were more likely than those with BC-, LA- to have an immature to total neutrophil (I/T) ratio > or = 0.16 at 12 and 24 hours (p = 0.04 and 0.02, respectively). In the prospective study, we found that a positive GBS LA test can be due to perineal contamination and possibly to gastrointestinal absorption of GBS antigen. No false positive LA test results occurred on urine obtained by SBA; however, use of this method failed to detect the one infant with GBS bacteremia. Because of suboptimal sensitivity and specificity, use of the GBS LA test on urine obtained either by SBA or bag cannot be recommended for diagnosis of early onset GBS disease.

Antigens, Bacterial↗
PubMed1999

Use of urine samples from healthy humans, nephritis patients or other animals as an alternative to foetal calf serum in the culture of Leishmania (L.) donovani in vitro.

The effect of supplementing in-vitro cultures of Leishmania donovani with urine was investigated. The parasites were isolated from Bangladeshi patients with visceral leishmaniasis. The urine samples used were collected from healthy human donors, patients with nephrotic syndrome, diabetic nephritis (DN) or diabetes mellitus, a dog and a cow. Promastigotes from blood-agar cultures were inoculated into RPMI-1640 basal medium with 10% heat-inactivated foetal calf serum (FCS) and/or 1%-20% urine. The parasites were then counted in a haemocytometer, on days 2, 4, 5, 6, 7, 8, 10, 12 and 14 post-inoculation. From day 4, the numbers of parasites/ml in cultures containing 5% healthy-human urine but no FCS were at least as high as those in cultures containing 10% FCS but no urine (P = 0.191). The wet weights of parasites harvested from mass cultures of the parasites in RPMI-1640 plus 5% healthy-human urine and in RPMI-1640 plus 10% FCS were practically the same. Multiplication of the parasites in the presence of 5% urine from a DN patient was significantly greater (P < 0.01) than that seen with other urine samples at the same concentration or with 10% FCS. The multiplication seen with 8% canine urine was almost the same as with 5% healthy-human urine. Parasites could be maintained in RPMI-1640 plus 5% healthy-human urine for at least 40 days, sub-culturing every 4 days. Urine may be a better and much cheaper stimulant of Leishmania multiplication in vitro than FCS.

Animals↗
PubMedDate not supplied

Gonococcal urethritis diagnosed from enzyme immunoassay of urine sediment.

First-catch urine specimens were obtained before clinical examination from 101 sexually active men who attended a sexually transmitted diseases clinic with a complaint of urethral discharge. Urethral swab specimens were used for preparation of smears and were then cultured on Martin-Lewis medium. Enzyme immunoassay (EIA) tests were performed on (1) uncentrifuged urine, (2) urine sediment, and (3) urine sediment diluted 1:6 with detergent buffer. Urethral cultures from 65 (64%) of the 101 men were positive for N. gonorrhoeae. EIA performed on urine sediment diluted 1:6 yielded the highest sensitivity: 98.5% (64/65). Sensitivity of EIA for uncentrifuged urine was only 66% (43/65). Specificity of all samples tested exceeded 97.2%. Overall agreement between results of EIA on diluted urine sediment and culture was 98% (99/101). Discordant culture and EIA results were unrelated to urine volume, time since prior urination, quantity of gonococcal growth on Martin-Lewis medium, duration of urine storage (less than 72 hours) before testing, or immunotype. EIA tests using urine sediment are highly sensitive and specific, and they offer an alternative means of diagnosing gonorrhea in men who refuse urethral manipulation. They also provide a means of screening men at high risk for gonorrhea who have submitted a urine specimen for other reasons.

Adult↗
PubMed1990

Urine flow rates in male and female urodynamic patients compared with the Liverpool nomograms.

The Liverpool male and female nomograms provide normal reference ranges for the maximum and average urine flow rates over a wide range of voided volumes. A known urine flow rate, in combination with the respective voided volume (and in men the age), can be converted into an equivalent centile ranking, using the equation for the relevant nomogram. Centile rankings allow the urine flow rates of specific male and female populations to be compared. The maximum and average urine flow rates of urodynamic patients (504 consecutive male and 168 consecutive female) were converted into centile rankings. These centile rankings were then compared with the respective urodynamic diagnoses. The median centile rankings for their maximum and average urine flow rates were 21 and 11 (male) respectively and 31 and 18 (female) (vs 50 for the normal populations). The low urine flow rates applied to all diagnostic groups, including those symptomatic men and women who were found to be urodynamically normal. Low urine flow rates may be one of a number of subtle manifestations of urological disease in these patients. Men with obstruction and women with voiding difficulties had the lowest urine flow rates. The 25th centile (men) and the 10th centile (women) appeared to be most appropriate lower limits of normality for both urine flow rates to identify those men more likely to be obstructed and those women at higher risk of voiding difficulties. Men and women with detrusor instability had the highest urine flow rates. Detrusor instability was present in 71% of men with centile rankings for the maximum urine flow rate over 50; 50% of women whose maximum urine flow rate centiles were over 90 had detrusor instability.

Adolescent↗
PubMed2005

Dairy farm effluent effects on urine patch nitrous oxide and carbon dioxide emissions.

Dairy farm effluent (DFE) comprises animal feces, urine, and wash-down water collected at the milking shed. This is collected daily during the milking season and sprayed onto grazed dairy pastures. Urine patches in grazed pastures make a significant contribution to anthropogenic N(2)O emissions. The DFE could potentially mitigate N(2)O emissions by influencing the N(2)O to dinitrogen (N(2)) ratio, since it contains water-soluble carbon (WSC). Alternatively, DFE may enhance N(2)O emissions from urine patches. The application of DFE may also provide a substrate for the production of CO(2) in pasture soils. The effects of DFE on the CO(2) and N(2)O emissions from urine patches are unknown. Thus a laboratory experiment was performed where repeated DFE applications were made to repacked soil cores. Dairy farm effluent was applied at 0, 7, or 14 d after urine deposition. The urine was applied once on Day 0. Urine contained (15)N-enriched urea. Measurements of N(2)O, N(2), and carbon dioxide (CO(2)) fluxes, soil pH, and soil inorganic N concentrations were made. After 43 d the DFE had not mitigated N(2)O fluxes from urine patches. A small increase in the N(2)O flux occurred from the urine-treated soils where DFE was applied 1 wk after urine deposition. The amount of WSC applied in the DFE proved to be insignificant compared with the amount of soil C released as CO(2) following urine application. The priming of soil C in urine patches has implications for the understanding of soil C processes in grazed pasture ecosystems and the budgeting of C within these ecosystems.

Animals↗
PubMed1991

[Anti-bacterial defense mechanisms of the urinary tract constructed from intestinal segments. Studies on cell population and phagocytotic activity of urinary leucocytes, and bacterial growth in urine].

Cell population of urinary leucocytes of 22 patients (intestine group) who underwent operations using intestinal segments for the urinary tract was compared with that of 26 complicated UTI patients without surgical intervention (control group). Eosinophils were recognized in 15.5 per cent of urinary leucocytes of the intestine group. However, in the control group, urine eosinophils were recognized only in 0.15 per cent. Although in sterile urines of the intestine group eosinophils were recognized in 30.8 per cent, in infected urines, the percentage of eosinophils decreased. Conversely, the percentage of neutrophils increased to 91.9 per cent. These findings suggest that neutrophils play an important role in infected urines of the intestine group as in urines of the control group. Significant differences were found in the values of urinary secretory IgA, IgG, IgM and urinary osmolarity. To evaluate the influence of these differences on the activity of phagocytosis of urinary leucocytes, the activity of phagocytosis of polymorphonuclear leucocytes (PMN), isolated from the peripheral blood, was investigated in immersion in urines of both groups. The mean rate of phagocytosis of E. coli in urines of both groups showed no statistically significant differences. However, urinary osmolarity of the intestine group was within the suitable range for phagocytosis and the activity of phagocytosis in urine was correlated with the value of IgG, which suggests that IgG has the opsonic effect. In contrast, the activity of phagocytosis in urine of the control group was strongly correlated with the value of urinary osmolarity. The growth of Providencia, Streptococcus, P. aeruginosa, whose frequency of isolation from urine of both groups was different in our previous study, and E. coli was studied in urine of the two groups. No significant difference in the growth of all bacteria was found, however. This finding suggests that the difference in the frequency of isolation of these bacteria from urine possibly depends on the adhesion of bacteria to intestinal epithelium.

Adolescent↗
PubMed1999

Tubular reabsorption and sodium excretion during urine reinfusion.

BACKGROUND: The mechanisms responsible for the natriuresis that follows urine reinfusion was investigated in rats by clearance and micropuncture techniques. METHODS: In each animal two urine reinfusion periods (R1 and R2) were performed and compared to a non-urine-reinfusion, saline infusion period (S) sandwiched between them. RESULTS: Switching from urine reinfusion to an equivalent rate of saline loading was followed by a fall in Na excretion from 1.9 +/- 0.5 to 0.5 +/- 0.2% of filtered load, p < 0.002. Urine osmolality rose, and urine to plasma inulin concentration ratio rose significantly from 73 +/- 14 to 147 +/- 21 (p < 0.002). The changes in GFR, SNGFR, absolute and percent proximal reabsorption could not account for these findings. A reduced Na excretion coupled to increased urine osmolality indicates enhanced transport along a segment responsible for the urinary concentrating mechanism. Thus the data can be interpreted then as due to enhanced reabsorption along the ascending limb of Henle's loop. These changes were reversed by reinstituting urine reinfusion after the S period. The consensual changes in Na+ and K+ excretion excluded an effect of urine reinfusion on the distal exchange site. There was a continuous fall in proximal reabsorption from R1 (76 +/- 3%) to S (69 +/- 3%) to R2 (62 +/- 5%) which was inversely correlated with the changes in hematocrit (R = 0.49, p < 0.026). This indicates that part of the late diuresis and natriuresis was due to volume expansion. An osmotic effect of reinfused urine solutes was suggested by a late rise in plasma osmolality, from 312 +/- 13 to 323 +/- 8 mOsm/kg. Osmotic diuresis could have exerted additive effects upon those of volume expansion, accounting for the late fall in proximal reabsorption. CONCLUSIONS: We conclude that the acute effects of urine reinfusion are due to changes in transport of solutes and permeability to water along distal tubular segments. The changes in plasma osmolality during the last period of the present acute experiments, suggest the possibility that solute retention may be linked to the chronic effects of urine reinfusion.

Animals↗
PubMed1998

[Some considerations on the avoidance of excess body burden in case of high dose 131I treatment: with special reference to the urination frequency].

In case of the treatment of metastatic thyroid cancer with radioiodine (131I), 3.7 GBq (100 mCi) or more is often repeatedly administered to the patient. Therefore, we must be aware of avoiding unnecessary exposure to the radioiodine except for the lesions to be treated. The administered radioiodine is excreted mainly through urinary system, resulting in an accumulation of the urine in the bladder for a certain period, which is highly concentrated with it. Based on this fact, we set one case having a particular pattern of the whole body retention curve. Then, two different modes of urination were considered; mode A indicates every 2 hour-urination x 6 times followed by every 6 hour-urination x 2 times, and mode B indicates every 6 hour-urination x 4 times. Focusing on the different amounts of urine in the bladder upon the different modes of urination, the radiation exposures from the urine to the neighboring organs, such as bladder wall, uterus, ovary and testis, and also to the whole body were calculated. As the results, it was found that the urination mode B would cause radiation exposure from the urine in the bladder twice as much as the urination mode A to the neighboring organs as well as to the whole body. This study will supply arguments for the necessity of frequent urination in the cases receiving radioiodine treatment for metastatic thyroid cancer.

Body Burden↗
PubMed1997

First-trimester urine free beta hCG, beta core, and total oestriol in pregnancies affected by Down's syndrome: implications for first-trimester screening with nuchal translucency and serum free beta hCG.

We have examined maternal urine concentrations of beta core, free beta human chorionic gonadotrophin (hCG), and total oestriol in 373 control pregnancies and 43 pregnancies affected by aneuploidy (including 22 cases of Down's syndrome) in an attempt to see if any of the analytes have a value in Down's syndrome screening between the tenth and 14th week of pregnancy. We have compared the performance of these analytes against nuchal translucency measurement combined with maternal serum free beta hCG at the same period of pregnancy. Our results show that levels of urine free beta hCG and beta core are increased in Down's syndrome with average multiple of the median levels of 1.81 and 2.91, respectively. Urine total oestriol was reduced (0.83) whilst maternal serum free beta hCG was increased (1.72). In trisomy 18 the levels of all analytes were reduced, although serum free beta hCG was the most discriminating. The spread of results in the control and the Down's group for urine beta core was more than three times than that for serum free beta hCG and with urine free beta hCG it was two times wider. In combination with maternal age, urine total oestriol had a 32 per cent detection rate at a fixed 5 per cent false-positive rate; urine beta core 34 per cent, urine free beta hCG 36 per cent, maternal serum free beta hCG 44 per cent, and nuchal translucency 82 per cent. In combination with nuchal translucency, urine total oestriol added an extra 1 per cent detection, urine beta core an extra 2 per cent, urine free beta hCG an extra 3 per cent, and serum free beta hCG an extra 5 per cent. It is unlikely that any of the urine markers will be of value in first-trimester screening. Optimal first-trimester screening programmes will rely for the foreseeable future on nuchal translucency, serum free beta hCG, and possibly pregnancy-associated plasma protein A.

Chorionic Gonadotropin, beta Subunit, Human↗
PubMed1980

Ectopic production of human chorionic gonadotropin (hCG) by neoplasms: the value of measurements of immunoreactive hCG in the urine as a screening procedure.

Immunoreactive human Chorionic Gonadotropin (hCG) was measured in serum and urine extracts from patients with malignant disease using a radioimmunoassay that detects efficiently hCG and its betasubunit. Of the 70 patients examined, 12 (17.1%) were positive for hCG in serum and 31 44.3%) in urine. Eleven patients who were positive in serum were also positive in urine; 20 patients (28.6%) were positive only in urine. Sephadex G-100 chromatography of urine from two serum-negative and urine-positive patients showed that the hCG immunoreactive material in the urine of these patients was mostly a molecular species smaller than hCG and hCG-beta. The nature of this molecule(s) is unknown and is called here metabolite(s) "X" of hCG-beta. The urine of 2 patients who where positive for hCG in both serum and urine contained considerable amount of metabolite(s) "X" as well as the native hCG-beta subunit, which was present also in the serum of these 2 patients. The metabolite(s) "X" was also shown by chromatography in the urine of a pregnant woman. It is concluded that the ectopic production of hCG is found more than twice as frequently in urine as compared to when serum alone is examined. The urine of serum-negative tumor patients can be positive for hCG because of the presence in it of the metabolite(s) "X" of hCG-beta or hCG which presumably circulates in the blood of these patients at non-detectable levels.

Chorionic Gonadotropin↗
PubMed1998

Blood lead-urine lead relationships in adults and children.

To determine the potential for using instead of blood as an indicator of lead exposure, especially in infants, lead concentrations and high-precision lead isotopic measurements have been compared in venous blood and "spot" urine (n > 260 from 182 different subjects) collected within the same 24-h period. Physiological conditions for the children and most of the adults were considered to be in a steady-state between body stores and lead in the environment. In the case of some adults, conditions were initially not steady-state because exposure conditions changed (for example, subjects moved to a country with lead of different isotopic composition.) There was a high correlation (r2 = ) between the blood and urine measurements of the isotope ratios but about 10% of measurements were outliers--the blood and urine measurements were further apart than was consistent with the measurement error that was generally obtained. The discrepancy was usually found to be associated with the urine measurement and was attributed to contamination during sampling. Weekly urine and monthly blood monitoring of an adult male over a 24-month period showed and excellent correlations, although the standard deviations were about an order of magnitude higher than the precision measured for replicate analyses of a single blood or urine sample. "Spot" urine analyses for two male subjects gave excellent agreement with 24-h urine samples. Standard deviations of the spot analyses were of similar order to those in the 24-month monitored subject. In cases where female adults from Eastern Europe migrated to Australia, there was generally a more rapid exchange of skeletal lead with Australian environmental lead in urine compared with blood. These data do not support a differential partitioning of endogenous lead into the plasma. At this stage, isotopic measurements of urine can be used as a proxy for isotopic measurements in blood. However, lead concentrations in blood and in urine are only weakly related. Concentrations of lead in urine cannot serve to predict concentrations of lead in blood, particularly at the lower range of exposures, for example, at blood concentrations less than 10 microgram/d1.

Adult↗
PubMed1988

Fibrin activatable urokinase (FA-UK): a latent form of UK found in urine related to a complex with an inhibitor/fibrin-interacting cofactor.

In order to investigate the binding of pro-urokinase (pro-UK) in urine to fibrin/Celite, the property which led to its discovery, the effect of fibrin on the plasminogen activator activity of urine was studied. The plasminogen activator activity in urine was found to be consistently about 2-fold higher when measured by fibrin plate assay than by amidolytic substrate (S-2444), when normalized against the UK reference standard. When the amidolytic activity measurement was preceded by incubation of urine with soluble fibrin, a 2-fold increase in amidolytic activity was also found. Fibrin similarly increased plasmin generation in urine enriched with Glu- or Lys-plasminogen as determined by synthetic substrate S-2251. The observed promoting effect was common to several forms of soluble fibrin and was dose dependent, whereas fibrinogen had little effect. The promoting effect of fibrin was not expressed in the presence of pro-UK or two-chain UK (TC-UK) in buffer and therefore was attributed to another constituent of urine. Since the activity was inhibited by antibodies to UK but not to t-PA, it was called fibrin activatable UK (FA-UK). Gel filtration (Sephacryl-200) of urine revealed FA-UK activity in fractions eluting at a molecular weight of approximately 100K. A 100 K band of activity was also consistently seen when concentrated urine was subjected to zymography. Treatment of concentrated urine with hydroxylamine (1 M) eradicated both these activities and was associated with an increase in baseline amidolytic activity in the urine sample indicative of the release of UK from an inhibitor complex. Moreover, passage of urine over insolubilized monoclonal antibody against UK-inhibitor (PAI-3) complexes resulted in loss of FA-UK activity and of the 100 K band on the zymogram suggesting that the complex responsible for FA-UK was related to a PAI-3 complex. Since the FA-UK activity appeared to bind to fibrin/Celite, attempts were made to investigate complexation with pro-UK. Unfortunately, due to the instability of pro-UK in urine, no reliable data were obtained. It was concluded that PAI-3 may serve as a fibrin-interacting co-factor of UK, and therefore may play a role in fibrinolysis.

Antibodies, Monoclonal↗
PubMed1992

An ephemeral sex pheromone in the urine of female house mice (Mus domesticus).

From previous research, the ultrasonic vocalizations of male mice (Mus domesticus) to female mouse urine were hypothesized to be learned as a result of classical conditioning during adult heterosexual encounters. According to this interpretation, a previously neutral conditioned stimulus in female urine comes to elicit vocalizations as a result of its association with some other unknown unconditioned stimulus associated with adult females. However, the research from which this hypothesis was derived utilized urine collected from females housed in metabolic cages. Three experiments further examined the classical conditioning hypothesis using two types of female urine: (i) metabolic-cage-collected urine and (ii) freshly voided urine. Experiment 1 demonstrated that, in contrast to vocalizations to metabolic-cage-collected urine, adult heterosexual experience was not necessary for males to vocalize to freshly voided female urine. In addition, unlike metabolic-cage-collected urine (Experiment 3), freshly voided urine remained a potent stimulus for eliciting vocalizations during repeated testing (Experiments 2 and 3). Finally, freshly voided urine appeared to cause a previously neutral stimulus (cotton swab) to acquire ultrasound eliciting properties (Experiment 2). We suggest from these findings that two chemosignals that elicit vocalizations from males may exist in female mouse urine: (i) a potent, but volatile or easily degraded, unconditioned stimulus to which males vocalize without sexual experience and (ii) a nonvolatile, chemically stable conditioned stimulus.

Analysis of Variance↗