Pregnancy and ovulation detection in bison (Bison bison) assessed by means of urinary and fecal steroids.

Sexually mature bison (Bison bison) cows were tested for both pregnancy and ovulation by means of urinary steroid metabolites and fecal steroids. The accuracy of pregnancy diagnosis was determined among 18 bison cows, in approximately the third month of gestation, by means of urinary pregnanediol-3 delta-glucuronide (PdG), urinary estrone conjugates (E1C), and fecal total estrogens (TE). Urinary PdG was 100% accurate, urinary E1C was 89% accurate, and fecal TE was 100% accurate in predicting pregnancy. Fecal progesterone (P4) and TE, as well as urinary E1C and PdG concentrations all increased from conception (August) through January, but significant differences were not apparent until November. During the rutting season ovulation was detected by increases in either urinary PdG or fecal P4 concentrations. Both pregnancy and ovulation were detected in uncaptured bison with reasonable accuracy by means of urinary and fecal steroids or their metabolites.     

Randomized controlled trial of dietary intervention: association between level of urinary phenolics and anti-mutagenicity

We have undertaken a randomized trial to confirm the ability of a class of phenolics, flavonoids, to increase urinary anti-mutagenicity in smokers. Ninety heavy smokers were recruited and randomly assigned to three groups, who were given three different diets. One diet was rich in flavonoids, but not based on supplementation ('flavonoid'), one was a normal iso-caloric diet with an adequate administration of fruit and vegetables ('normal'), and one was based on supplementation of flavonoids in the form of green tea and soy products ('supplement'). The urinary anti-mutagenicity-as inhibiting effect of the urinary extracts on the mutations induced by MeIQx-was measured in Salmonella typhimurium YG1024 in the presence of liver S9 from male Sprague-Dawley rats treated with Aroclor 1254. The amount of total phenolics in the urinary extracts was measured by use of spectrometric analysis. We found that important dietary modifications can be achieved through special recipes and instructions given by a cook during an intensive course. The intervention was focused on increasing the flavonoid intake, and it was successful in that respect. In fact, differences in flavonoid intake were appreciated mainly between the first group (normal diet) and the other two (flavonoid-rich and supplemented diet), suggesting that dietary modification can be as effective as supplementation. However, both urinary anti-mutagenicity and the amounts of urinary phenolics did not change as a consequence of the trial. These results suggest that only a small fraction of urinary phenolics is influenced by dietary changes in the intake of flavonoids, and that most urinary anti-mutagens and phenolics are metabolites of dietary flavonoids, whose formation is more affected by the activity and diversity of bacterial flora in the colon than by the quantity and type of intake. A strong correlation was found between urinary phenolics and anti-mutagenicity in all the groups involved in the trial. Such correlation was not explained by dietary variables.     

Neo-regeneration of urinary bladder: a desired metaplasia of autologous membrane from rectosigmoid colon containing stem cells of intestinal crypts

The current management of diseases of urinary bladder requiring resection is by augmentation cystoplasty or transplantation of ureters. Transplantation of ureters is associated with morbidity and mortality. Ideal management will be by regenerating urinary bladder in vivo. Neo-regeneration of tissues and organs like abdominal wall, aponeurosis etc., has been attempted and patented. After neo-regeneration of mesoderm tissues and organs, regeneration of urinary bladder (developed from endoderm) was. In vivo surgical techniques were developed in dogs. It is known that the embryonic morphogenesis of urinary bladder is from uro-genital sinus of hind gut. A membrane, containing endoderm stem cells in crypts of recto-sigmoid colon, was surgically isolated and colonized with remnant of urinary bladder wall after extensive resection. Experimental study was performed in dogs, for 60 days to one and a half year. Regeneration of all the layers of tissues of the wall of urinary bladder was observed. The neo-regeneration phenomenon has been recognized as "desired metaplasia". The regenerated neo tissue/organ on histological examination and cystometry studies was found compatible with normal urinary bladder. The hypothesis, neo-regeneration and desired metaplasia, is discussed.     

Effect of calcium on intracellular pH

The effect of intracellular calcium on intracellular pH in the turtle urinary bladder was examined with phosphorus nuclear magnetic resonance. The turtle urinary bladder is capable of acidification in vitro and urinary acidification by this membrane is inhibited by an increase in intracellular calcium. Since calcium is capable of altering intracellular pH, it remains unclear whether the inhibition of urinary acidification is the result of an increase in intracellular pH. In the present study, intracellular calcium was increased by the cholinergic agent, carbachol, the ionophore A23187 and replacement of extracellular Na by sucrose. All agents decreased intracellular pH in the turtle bladder, thus suggesting that inhibition of urinary acidification by these agents is not due to an increase in intracellular pH.     

Morphological and conformational studies of Tamm-Horsfall urinary glycoprotein

We have studied the circular dichroic properties of normal and cystic fibrotic Tamm-Horsfall urinary glycoproteins, and the asialo-derivatives (ca. 80% removal of sialic acid with neuraminidase). There was no evidence of α-helicity in Tamm-Horsfall urinary glycoprotein, but the results do indicate a significant amount of β-structure. The circular dichroic spectra of normal and cystic fibrotic Tamm-Horsfall urinary glycoproteins and the asialo-derivatives were identical, thus suggesting that there is no major difference in the ordered secondary structure of Tamm-Horsfall urinary glycoprotein in cystic fibrosis (relative to normal Tamm-Horsfall urinary glycoprotein), and that sialic acid exerts no major effect on the β-structure. Also, the circular dichroic spectrum of Tamm-Horsfall urinary glycoprotein was not affected by Ca²⁺ at concentrations just below that required for gel formation. Electron microscopic studies reveal the presence of a supramolecular helical structure arising from subunit interactions. This structure was characterized by a repeat of 120–130 Å and a minimal helix diameter of ca. 40 Å, although this value varied depending on the number of interacting helices. The helical structure was observed for normal, cystic fibrotic, and asialo derivatives of Tamm-Horsfall urinary glycoproteins, and was independent of added Ca²⁺. Guanidine hydrochloride treatment, followed by dialysis, irreversibly destroyed this supra-molecular helical structure, but the β-structure was partially restored, as indicated by the circular dichroic spectrum. The Ca²⁺-mediated gel formation was found to be inhibited in asialo-Tamm-Horsfall urinary glycoprotein.     

Seasonal Variation in the Occurrence of Acute Urinary Retention

Objective: To determine whether acute urinary retention shows a seasonal variation. Methods: All acute urinary retentions observed in the emergency room of St. Anna Hospital, Ferrara, along an eight-year period (1991–1998) were considered. Diagnosis was made on the basis of history, physical examination, and bladder drainer by insertion of a catheter with registration of the retained urinary volume. The main determining cause of the acute urinary retention event was tentatively determined. Month and day of each event were categorized both by seasons and monthly intervals. For statistical analysis chi-square test for goodness of fit and partial Fourier series were used. Results: 1,133 acute urinary retention events were observed in 929 different subjects (871 males, 58 females). In 738 cases it was possible to define the main determining cause of the event. The seasonal distribution showed a higher frequency peak of events in summer and autumn both for total population and males subgroup. Analysis by determining cause of the event demonstrated a greater frequency of events in summer only in patients with prostatic hypertrophy. Chronobiologic evaluation showed a circannual pattern for acute urinary retention, both for total sample and male subgroup, with a significant peak in late summer (August–September). Analysis by main determining cause revealed a similar circannual pattern only for cases of acute urinary retention determined by prostatic hypertrophy. Conclusions: The present study shows that acute urinary retention exhibits a circannual distribution in its occurrence, particularly when caused by prostatic hypertrophy.     

Site of Action of Nitrofurantoin in Experimental Urinary Tract Infection

The effectiveness of nitrofurantoin in suppressing bacterial growth in the urinary tract was evaluated by using two different experimental models. Pyelonephritis was produced in rats by direct inoculation of 10(4)Escherichia coli in the medulla of left kidney. Ascending urinary tract infection was induced by inoculation into the urinary bladder of 10(7)Proteus mirabilis, after a partial cystectomy. Nitrofurantoin was shown to be effective in suppressing bladder bacteriuria, in preventing ascending pyelonephritis, and also in preventing bacterial multiplication in kidney tissue following direct inoculation.     

Relationship of urinary kallikrein excretion to urinary potassium excretion during furosemide administration with and without amiloride

The relationship of urinary kallikrein excretion to urine volume, and to urinary sodium and potassium excretions was studied in normal rats during furosemide diuresis and superimposed injection of amiloride, a K+-sparing diuretic. Continuous infusion of furosemide increased urinary kallikrein, sodium and potassium excretions and the urine volume. Amiloride injection during furosemide diuresis caused further increase in diuresis and natriuresis, but a prompt decrease in urinary kallikrein excretion to basal level, and potassium excretion to below the basal level. The significant correlation of urinary kallikrein excretion to urinary potassium excretion, but not to urine volume and urinary sodium excretion after amiloride injection suggests that the major determinant of urinary kallikrein excretion is renal potassium secretion through a mechanism that is affected by amiloride.     

Structure of the urinary bladder in the Pacific cod

The urinary bladder of the Pacific cod,Gadus macrocephalus was investigated. A pair of ureters was found to unite and form a sac-like protrusion on the posteroventral aspect of the gonad. This protrusion was confirmed as the urinary bladder based on its histology and the presence of ammonia and urea in the fluid within this structure. The urinary bladder consisted of a central lumen and two lateral expansions found on the right and left sections. The luminal epithelial cells of the Pacific cod urinary bladder, unlike those of other animals, were characterized by microvilli on the free surface, and the presence of a number of vesicles and mitochondria across the apical portion on the cells. This suggests that the luminal epithelium of the urinary bladder might be actively engaged in transportation of water or other materials from the urine. The Pacific cod urinary bladder may therefore be closely associated with osmoregulation by reabsorbing water from the urine. Ammonia-N and urea-N levels of 65–213 μg/dl and 1.5–3.5 mg/dl were measured in the fluid which filled the urinary bladder.     

Seasonal Variation in the Occurrence of Acute Urinary Retention

Objective: To determine whether acute urinary retention shows a seasonal variation.

Methods: All acute urinary retentions observed in the emergency room of St. Anna Hospital, Ferrara, along an eight-year period (1991-1998) were considered. Diagnosis was made on the basis of history, physical examination, and bladder drainer by insertion of a catheter with registration of the retained urinary volume. The main determining cause of the acute urinary retention event was tentatively determined. Month and day of each event were categorized both by seasons and monthly intervals. For statistical analysis chi-square test for goodness of fit and partial Fourier series were used.

Results: 1,133 acute urinary retention events were observed in 929 different subjects (871 males, 58 females). In 738 cases it was possible to define the main determining cause of the event. The seasonal distribution showed a higher frequency peak of events in summer and autumn both for total population and males subgroup. Analysis by determining cause of the event demonstrated a greater frequency of events in summer only in patients with prostatic hypertrophy. Chronobiologic evaluation showed a circannual pattern for acute urinary retention, both for total sample and male subgroup, with a significant peak in late summer (August-September). Analysis by main determining cause revealed a similar circannual pattern only for cases of acute urinary retention determined by prostatic hypertrophy.

Conclusions: The present study shows that acute urinary retention exhibits a circannual distribution in its occurrence, particularly when caused by prostatic hypertrophy.     

Purification and characterization of rat urinary renin

Rat urinary renin was purified by a procedure involving ammonium sulfate fractionation, pepstatin-aminohexyl-Sepharose 4B chromatography, ion exchange chromatography and gel filtration. The resulting preparation was essentially homogeneous, as assessed by polyacrylamide gel electrophoresis. The molecular weight of the preparation was estimated to be 39000 by SDS-gel electrophoresis and 40000 by gel filtration. The optimum pH determined with rat angiotensinogen was 7.0, and the Km was 3.6 microM. These properties agreed well with those of purified rat renal renin. The activity of urinary renin was specifically inhibited by anti-renin antibody. These results suggest that urinary renin may originate in the kidney.     

Renal synthesis of prostacyclin and thromboxane in healthy women: differential effects of a short-term saline loading

It is accepted that the urinary excretions of the stable metabolites of prostaglandin (PG)I2 and thromboxane(Tx) A2, 6-keto-PGF1alpha (6KPGF) and TxB2 respectively, provide an accurate estimate of both basal and stimulated renal synthesis of their precursors. The excretory profile of these metabolites has been evaluated in healthy women submitted to a short-term expansion in extracellular fluid volume. Salt retention (SR group, n=6) was induced by physiological saline (0.9% NaCl) i.v. infusions (2 L per day) over a period of 2 days. On the third day the increase in body weight was 0.92 +/- 0.27 kg (P<0.05). The results of the study have been compared to those previously obtained in normal balance of sodium and potassium (N group, n=20) and in induced salt depletion (SD group, n=14). A common study protocol was used. Basal values of plasma renin activity (PRA) and urinary aldosterone excretion were determined. Renal functional exploration [clearance (cl.) method] was performed during hypotonic polyuria (induced by oral water load) and subsequent moderate antidiuresis (induced by low-dose infusion of an antidiuretic hormone analogue). Urinary 6KPGF and TxB2 concentrations were estimated by RIA method and their urinary excretions were determined at both high and low urinary flow rates. The linear regressions of the urinary metabolite excretions vs. urinary flow rate were estimated by using the data obtained in both hypotonic polyuria and antidiuresis. Salt retention (SR vs. N group) was effective in decreasing the basal values of plasma renin activity and urinary aldosterone excretion. Moreover, during hypotonic polyuria it was effective in increasing the absolute and fractional excretions of sodium and chloride, in the absence of significant variations in mean arterial pressure and creatinine cl. Regarding urinary prostanoid excretions the following results were obtained. 1. Comparative data for hypotonic polyuria. In the SR vs. N group, the urinary excretion of 6KPGF was significantly higher, whereas that of TxB2 was not significantly different. In the SR vs. SD group, the urinary excretion of 6KPGF was not significantly different, whereas that of TxB2 was significantly lower. 2. Comparative data for the regression lines of the urinary prostanoid excretions vs. diuresis. In the SR vs. N group, the regression line slope for 6KPGF excretion was significantly higher, whereas that for TxB2 excretion was not significantly different. In the SR vs. SD group, the regression line slope for 6KPGF excretion was not significantly different, whereas that for TxB2 excretion was significantly lower. 3. Correlative data in the SR group during hypotonic polyuria. The plasma chloride concentration was positively correlated with urinary flow rate, absolute and fractional chloride excretions, and 6KPGF excretion but not with TxB2 excretion. In conclusion, functionally effective salt retention in healthy women induces a selective stimulation of renal synthesis of prostacyclin, unlike salt depletion, in which the synthesis of both PGI2 and TxA2 is upregulated.     

Spot determinations of urinary cortisol for the screening of Cushing's syndrome.

The usefulness of spot determination of urinary cortisol in the screening of Cushing's syndrome was evaluated by measuring the cortisol concentration in randomly sampled urine in 68 normal subjects and in 9 patients with Cushing's syndrome. The urinary cortisol concentration in the morning was significantly higher in patients with Cushing's syndrome but some overlap existed between normal subjects and patients with Cushing's syndrome. In contrast, there was a clear discrimination between two groups when urinary cortisol was measured in the late evening: urinary cortisol was lower than 75 micrograms per gram creatinine (microgram/gCr) in normal subjects but higher than 150 micrograms/gCr in patients with Cushing's syndrome. When 1 mg dexamethasone was administered at 2300 h in the evening, spot urinary cortisol the next morning was less than 80 micrograms/gCr in normal subjects while it was above 100 micrograms/gCr in patients with Cushing's syndrome. Dexamethasone-induced suppression of urinary cortisol in normal subjects lasted until late in the afternoon, which allows sampling of urine at any time in the morning and possibly in the afternoon. These results suggest the usefulness of spot determination of urinary cortisol in the screening of Cushings' syndrome.     

Determination of estradiol-17-sulfate in human urine by a direct radioimmunoassay: urinary levels throughout the menstrual cycle

The measurement of urinary estradiol-17-sulfate concentration by direct radioimmunoassay was established. The urinary estradiol-17-sulfate levels measured by the radioimmunoassay correlated well with those determined by high-performance liquid chromatography equipped with electrochemical detector. Estradiol-17-sulfate concentrations in early morning urine of six healthy adult men was 8.2 +/- 2.0 ng/mL, or 5.7 +/- 1.8 ng/mg creatinine. The urinary levels in women throughout the menstrual cycle showed a characteristic three-peak excretion pattern: the first and the second peaks appeared just after and three days before the urinary LH peak, and the third peak appeared a few days before menstruation.     

尿枸橼酸盐全自动酶法测定及临床应用

目的:建立全自动生化分析仪测定尿枸橼酸盐的方法,探讨其在枸橼酸钾治疗泌尿系结石患者中的应用。方法:采用酶偶联速率法,观察重复性、线性及回收实验,并测定36例泌尿系结石病人口服枸橼酸钾片前及服药后24小时尿枸橼酸、尿钾和尿钙。结果:该法重复性CV分别为2.26%和3.34%,枸橼酸浓度在5mmol/L的范围内呈线性,r为0.9981。服药后,尿枸橼酸明显升高(p<0.05),尿钙显著降低(p<0.05)。结论:使用全自动酶法测定尿枸橼酸盐重复性好,准确度高,口服枸橼酸钾能碱化尿液,降低尿钙,起到防治泌尿系结石的作用。     

The significance of ultrasonography in diagnosing and follow-up of cystic cystitis in children

Cystic cystitis is a separate form of urinary bladder inflammation, detected by cystoscopy in children with recurrent urinary infections. Cystoscopy is an invasive method, so the aim of this investigation was to determine the ultrasonographic characteristics of cystic cystitis and to assess the reliability of ultrasound in relation to cystoscopy in diagnosing cystic cystitis. The study included 115 girls with repeated urinary infections. Cystoscopy and ultrasonography was performed in all. According to the cystoscopic finding the subjects were divided into 4 groups. Lateral and posterior urinary bladder wall thickness was measured during ultrasonography. A statistically significant difference was found between all 4 groups, the method demonstrated a high degree of sensitivity (0.97) and specificity (0.91). Percentile calculations were determined for wall thickness. Ultrasonography can replace endoscopy in diagnosis and follow-up of cystic cystitis in children, with at least 50% fullness of the urinary bladder as a prerequisite.     

Isolation of tissue kallikrein in rat spleen by monoclonal antibody-affinity chromatography

Rat spleen kallikrein was identified and purified by DEAE-cellulose and monoclonal antibody-affinity chromatography. The purified enzyme has Tos-Arg-OMe esterase activity and kinin-releasing activity from a purified low-molecular-weight kininogen substrate. In the direct radioimmunoassay for tissue kallikrein, the splenic enzyme displays parallelism with standard curves of rat urinary kallikrein. The pH profiles of the Tos-Arg-OMe esterase activities of spleen and urinary kallikrein were identical with optima at 9.0 Rat spleen kallikrein was inhibited strongly by aprotinin and affinity-purified kallikrein antibody and weakly by soybean trypsin inhibitor. The IC₅₀ values were similar to those observed against rat urinary kallikrein. Neither the urinary nor the splenic enzyme was inhibited by lima bean trypsin inhibitor or preimmune serum immunoglobulins. Spleen kallikrein was labeled with [¹⁴]diisopropylphosphorofluoridate and visualized by fluorography on a sodium dodecyl sulfate-polyacrylamide gel. The electrophoretic mobility of the splenic enzyme was indistinguishable from that of urinary kallikrein A with an estimated Mr of approx. 38 000. With Western blot analyses using a rabbit anti-kallikrein antibody followed by ¹²⁵I-labeled protein A binding, the spleen and urinary kallikreins were again visualized at identical positions by autoradiography. The data show that there is a rat splenic tissue kallikrein which is indistinguishable from a renal kallikrein with respect to physicochemical properties, immunological character and susceptibility to inhibitors.     

Role of prostanoids in the control of renal function in normal potassium balance and in acute experimental potassium depletion. 4. Relation of extrarenal parameters, renal function parameters and urinary excretion of prostanoids

The renal function has been evaluated by clearance (cl.) method during hypotonic polyuria and successive moderate antidiuresis induced by a low dose of lysine-8-vasopressin; four 15 min and two 60 min cl. periods were performed, respectively. Glomerular filtration rate was estimated by creatinine cl.; the osmotic cl. (Cosm, CH2O), the absolute and fractional excretions of water, sodium, potassium and chloride were determined by usual methods. The urinary concentrations of PGE2, 6-keto-PGF1 alpha (6KPGF) and TxB2 were measured by RIA. The study protocol was applied in normal potassium balance and experimental potassium balance (KD), both in absence and presence of indomethacin. In KD groups with a potassium cumulative deficit of 198.4 +/- 22.2 meq (D3; n = 6) during polyuria significant correlations are consistent with the hypothesis that the lower the plasma potassium concentration is the higher the urinary chloride excretion and the inhibition of distal fractional chloride reabsorption. Moreover, by utilizing the polyuria and antidiuresis data pool, the effects of urine flow rate changes on PGE2 and 6KPGF urinary excretions are blunted as compared to normal potassium balance (n = 14). After indomethacin treatment (D3.I) the following functional relationships are disclosed: a) the lower the kaliemia is the lower the urinary chloride and potassium excretions and the higher the fractional isosmotic reabsorption; b) the lower the urinary potassium excretion is the lower the urinary chloride excretion. In both D3 and D3.I experimental groups the positive correlation between urinary chloride excretion and urinary potassium excretion is significant.     

Retrospective studies of urinary infections in the elderly

Retrospective analysis of the urinary infection course involved 55 elderly patients. The patients were divided into groups basing on the clinical diagnosis. These groups included patients with obstructive chronic pyelonephritis and the patients with lower urinary infections. It was found that reinfections occur in the elderly patients with both upper and lower urinary infections while recurrence is seen in case of the upper urinary infections. Bacteriuria is significantly more frequent in the obstructive chronic pyelonephritis than in other urinary infections. This form of bacteriuria is frequently accompanied by exacerbations than in bacteriuria of periodic origin. Arterial blood hypertension is frequent in the elderly with urinary infections but its incidence does not correlate with the localization of the infection. Decompensated renal failure in the course of the urinary infections in the elderly is nearly always combined with arterial blood hypertension.     

Resistance of proteinuric rats to furosemide: Urinary drug protein binding as a determinant of drug effect

The effect of drug binding to urinary proteins on the diuretic response to furosemide was assessed in normal and nephrotic rats. Nephrosis was induced by treating Sprague-Dawley rats with puromycin aminonucleoside. Binding of furosemide to urinary proteins was found to range from 60 to 95% depending on the concentration of urinary protein. The diuretic response to furosemide reaching the renal tubular lumen was inversely correlated with the degree of proteinuria, a finding that was independent of serum protein concentration of glomerular filtration rate. These data suggest that the binding of furosemide to urinary protein decreases the diuretic effect of furosemide and that drug-protein interactions of this type may also be important in modulating the activity of other lumenally-active drugs or endogenous substances exhibiting a high degree of protein binding. The binding of furosemide to urinary protein may explain the refractoriness of some patients with proteinuria to this agent.