Can Bladder Wall Thickness Predict Videourodynamic Findings in Children with Spina Bifida?

PURPOSE: We investigated whether bladder wall thickness measured at specifically defined bladder volumes could predict videourodynamic findings in children with spina bifida. MATERIALS AND METHODS: We prospectively investigated patients with spina bifida on intermittent catheterization who underwent ultrasound examination simultaneously with videourodynamics. We evaluated the association between bladder wall thickness measured at maximum cystometric capacity and parameters including age, maximum detrusor pressure during filling or at leak and bladder compliance. Differences in bladder wall thickness measured at each percent maximum cystometric capacity were compared between patients with and without unfavorable videourodynamic findings. Maximum detrusor pressure 40 cm H2O or greater during filling or at leak, bladder compliance less than 10 ml/cm H2O, detrusor overactivity, bladder trabeculation and vesicoureteral reflux were defined as unfavorable videourodynamic findings. RESULTS: A total of 23 males and 30 females with spina bifida (median age 7.8 years) underwent measurement of bladder wall thickness at maximum cystometric capacity. Mean ± SD bladder wall thickness measured at maximum cystometric capacity was 1.7 ± 0.5 mm. Only age had a weak correlation with bladder wall thickness measured at maximum cystometric capacity (p <0.05). In 31 patients bladder wall thickness was measured at each percent maximum cystometric capacity. Regarding unfavorable videourodynamic findings, there were no significant differences in bladder wall thickness measured at each percent maximum cystometric capacity, except for bladder trabeculation. CONCLUSIONS: Even if bladder wall thickness is measured at specifically defined bladder volumes, it cannot predict videourodynamic findings other than bladder trabeculation in children with spina bifida.

Winter is associated with failure in the alarm treatment of nocturnal enuresis.

OBJECTIVE: To assess whether the winter season in Japan is associated with failure in the alarm treatment of nocturnal enuresis (NE). PATIENTS AND METHODS: Consecutive patients with NE referred to our center between June 2009 and May 2010 were treated with the enuresis alarm (EA). The EA was used for 16 weeks with each child. Patients were divided into a success group and a treatment failure group. Pretreatment variables were collected, including age, sex, night-time urine volume, severity of enuresis, presence of nocturnal polyuria, presence of daytime incontinence, and treatment initiation season. These variables and initial success rates were retrospectively compared between the two groups. Chi-square, Student t tests, and multivariate regression analysis were used for statistical analysis. RESULTS: A total of 67 children with NE were evaluated, 37 (55%) in the success group and 30 (45%) in the failure group. None of the pretreatment variables differed significantly between groups except for season; winter season initiation was an independent risk factor for failure in multivariate regression analysis. CONCLUSIONS: Winter was associated with failure in the EA treatment. We recommend that EA be introduced in the summer season in Japan to achieve an optimal success rate.

Association between graves' disease and renal coloboma syndrome: a case report.

Renal coloboma syndrome is an autosomal dominant condition characterized by renal lesions and optic nerve abnormalities. We report an 11-yr-old Japanese girl with familial renal coloboma syndrome, who also had Graves' disease. Four affected family members had a previously reported heterozygous mutation (c.76dupG, p.Val26Glyfs*28) in the PAX2 gene. We hypothesized that PAX2 mutations may increase the risk of autoimmune diseases through alterations of human ß-defensin 1 expression.

The value of finding a closed internal ring on laparoscopy in unilateral nonpalpable testis.

PURPOSE: There is an ongoing debate about whether initial laparoscopy is as efficient as initial extra-abdominal exploration for unilateral nonpalpable testis. The aim of this study was to clarify whether the finding of a closed internal ring on laparoscopy accurately predicts extra-abdominal findings in unilateral nonpalpable testis. METHODS: Both laparoscopy and subsequent extra-abdominal exploration were prospectively performed in consecutive patients with unilateral nonpalpable testis. Laparoscopic findings were reviewed with respect to the status of the vas deferens, spermatic vessels, and the appearance of the internal ring. Except for patients with an abdominal testis, all patients underwent subsequent open exploration regardless of the laparoscopic findings. Open explorations were primarily performed by scrotal incision. During open exploration, any testicular remnants were removed and evaluated histologically. RESULTS: A total of 100 patients with unilateral nonpalpable testis were evaluated, of whom 15 had an abdominal testis. Of the 85 patients without an abdominal testis, an open internal ring was recognized in only 8. Of the remaining 77 with a closed internal ring, subsequent open exploration revealed testicular nubbins in all patients (100%), regardless of the laparoscopic findings of the vessels and vas. Nine nubbins were associated with intra-abdominal vanishing vessels above the closed internal ring. All nubbins were confirmed by histological findings, and the incidence of viable germ cell elements was 7.4%. CONCLUSIONS: When laparoscopy demonstrates no abdominal testis in patients with unilateral nonpalpable testis, the finding of a closed internal ring on laparoscopy is always associated with the presence of an extra-abdominal nubbin, regardless of the status of the vessels and the vas at the internal ring. Subsequent extra-abdominal exploration can be obviated in this circumstance if nubbin excisions are skipped. Initial diagnostic laparoscopy for unilateral nonpalpable testis provides not only the intra-abdominal findings, but also exact prediction of the extra-abdominal findings in patients with a closed internal ring.

Bladder augmentation using tissue-engineered autologous oral mucosal epithelial cell sheets grafted on demucosalized gastric flaps.

BACKGROUND: At present, autologous intestinal segments are often used for bladder reconstruction. However, the gastrointestinal mucosa often causes various complications. METHODS: Oral mucosal tissues were obtained from the buccal cavity of beagle dogs. Primary oral mucosal epithelial cells were cultured on temperature-responsive culture dishes with a mitomycin C-treated 3T3 feeder layer for 2 weeks. Cultured epithelial cells were harvested as contiguous sheets by reducing the temperature to 20°C. The study consisted of three groups. In group 1, oral mucosal epithelial cell sheets were autografted on demucosalized gastric flaps. Next, the gastric flaps with the oral mucosal epithelial cell sheets were used for bladder reconstruction. Bladder reconstruction was once immediately and then 5 days after epithelial cell sheet grafting in groups 2 and 3, respectively. Three weeks after bladder reconstruction, the gastric flaps with the oral mucosal epithelial cell sheets were examined by immunohistology. RESULTS: Flaps grafted with oral mucosal epithelial cell sheets showed epithelial regeneration in groups 1 and 3. Regenerated epithelia were stratified and similar to native oral mucosa. However, the regenerated epithelium was absent from the reconstructive segment, and urothelial ingrowth was observed in group 2. Macroscopically, all reconstructive segments showed contracture. CONCLUSIONS: We successfully performed a bladder reconstruction using oral mucosal epithelial cell sheet-grafted flaps that exhibited epithelial regeneration. Further study should consider shrinkage prevention.

The role of the retrograde colonic enema in children with spina bifida: is it inferior to the antegrade continence enema?

OBJECTIVES: The aim of this study was to evaluate the efficacy of the retrograde colonic enema relative to the Malone antegrade continence enema. METHODS: We retrospectively investigated 25 children with spina bifida and fecal incontinence. Thirteen children had started retrograde colonic enema and twelve had started Malone antegrade continence enema. Fecal continence, water volume, time to washout, procedure frequency, pain during procedure, performance independence and demographical data were compared between the two groups. RESULTS: Fecal continence was achieved for 10 of 13 (76.9%) in the retrograde group and 9 of 12 (75.0%) in the antegrade group. In the antegrade group 8 of 12 (66.7%) performed procedure independently, while 3 of 13 (23.1%) did so in the retrograde group. Achievement of fecal continence did not differ between the groups, but procedure independence was significantly better in the antegrade group. CONCLUSIONS: Our results suggest that retrograde colonic enema was not inferior to Malone antegrade continence enema on fecal continence. We recommend considering retrograde colonic enema prior to introduction of Malone antegrade continence enema in children with spina bifida.

Scrotal nubbins associated with blind-ending spermatic vessels and a normal vas deferens on laparoscopy.

OBJECTIVES: To determine whether a scrotal nubbin is present in children with unilateral non-palpable testis when diagnostic laparoscopy demonstrates blind-ending vessels and a normal vas deferens entering a closed internal ring. METHODS: Eighty consecutive patients with a unilateral nonpalpable testis were retrospectively reviewed. Patients underwent initial diagnostic laparoscopy, and, if needed, subsequent inguinal exploration was performed. On inguinal exploration, any testicular remnant or nubbin-like tissue was removed and evaluated histologically. Patients with a patent processus vaginalis were excluded from this analysis. RESULTS: Overall, 60 of the 80 patients had neither an abdominal testis nor a patent processus vaginalis. Of these 60, 34 patients had both a vas deferens and spermatic vessels entering a closed internal ring, and all of these underwent inguinal exploration. A total of 17 patients had both a blind-ending vas deferens and blind-ending spermatic vessels; no inguinal exploration was attempted. In nine patients, laparoscopy revealed blind-ending vessels with a normal vas deferens entering the closed internal ring. Of these nine, six underwent inguinal exploration, and a scrotal nubbins was found in three. At histological examination, hemosiderin deposit and calcification were seen in the nubbin tissue. No viable germ cell was detected in these specimens. CONCLUSIONS: A laparoscopic finding of blind-ending vessels above the closed internal ring does not mean intra-abdominal vanished testis, regardless of the appearance of the vas deferens.

Predicting early recurrent urinary tract infection in pretoilet trained children with vesicoureteral reflux.

PURPOSE: We identified independent factors predicting recurrent urinary tract infection within 1 year after the first urinary tract infection in pretoilet trained children with vesicoureteral reflux. MATERIALS AND METHODS: We retrospectively reviewed the records of infants younger than 2 years with primary vesicoureteral reflux and a history of febrile urinary tract infection. Patients were divided into 2 groups based on the presence or absence of recurrent febrile urinary tract infection. Analysis included age, gender, reflux laterality and grade, abnormalities on dimercapto-succinic acid renal scan and prophylactic antibiotic type. Univariate and multivariate analyses were performed to identify risk factors for recurrent febrile urinary tract infection. RESULTS: From 2004 to 2007, 78 children met study inclusion criteria. Mean age at the first urinary tract infection was 4 months (range 1 week to 16 months). None of the males were circumcised. Of 78 children 25 (32%) had a recurrent febrile urinary tract infection during 1 year of followup. Univariate analysis showed that bilateral reflux, high grade reflux (IV-V) and abnormal dimercapto-succinic acid scan were statistically significant predictors of early recurrent urinary tract infection (p <0.05). However, on multivariate analysis only an abnormal dimercapto-succinic acid scan showed a significant association with early recurrent urinary tract infection (OR 8.01, 95% CI 2.10-30.51, p = 0.002). CONCLUSIONS: Abnormal dimercapto-succinic acid renal scan is an important predictor of early recurrent urinary tract infection in pretoilet trained children with vesicoureteral reflux. Whether the explanation lies in congenital or infection related damage, in this patient subgroup careful clinical followup or early surgical management for reflux should be considered.

The significance of 99mtechnetium dimercapto-succinic acid renal scan in children with spina bifida during long-term followup.

PURPOSE: (99m)Technetium dimercapto-succinic acid renal scans are ideal for demonstrating renal scarring in children with spina bifida. However, doubt persists about the need for routine application. We assessed the associations among abnormal (99m)technetium dimercapto-succinic acid renal scans, vesicoureteral reflux and urodynamic findings in patients with spina bifida during long-term followup. MATERIALS AND METHODS: We retrospectively reviewed the records of 64 patients with spina bifida followed at our center. All patients were older than 10 years (mean 15.8, range 10 to 23). Dimercapto-succinic acid renal scans were considered abnormal with differential function of less than 40% or focal defects. Patient age, gender, previous febrile urinary tract infections, positive vesicoureteral reflux history, timing of clean intermittent catheterization initiation and the latest urodynamic findings were noted. Patients were grouped based on normal/abnormal scan results. Statistical analysis included univariate and multivariate regression analyses and chi-square tests. RESULTS: A total of 16 patients (25%) had abnormal scans. Mean patient age, male-to-female ratio, leak point pressure, bladder compliance and timing of clean intermittent catheterization initiation did not differ between groups. Rates of previous febrile urinary tract infections differed significantly (11 of 16 in the abnormal group vs 9 of 48 in the normal group, p <0.01), as did positive reflux history (100% vs 31%, p <0.01). No patient with a negative reflux history had an abnormal dimercapto-succinic acid renal scan. Multivariate analysis identified previous febrile urinary tract infections as a significant risk factor for an abnormal scan. CONCLUSIONS: A positive vesicoureteral reflux history and febrile urinary tract infections were associated with abnormal dimercapto-succinic acid renal scan in followup of patients older than 10 years with spina bifida. Thus, these factors are indicators of proactive evaluation of renal function using dimercapto-succinic acid renal scanning.

Asymmetric development of tumor-like cysts in a child with autosomal dominant polycystic kidney disease.

We report a case of asymmetric development of tumor-like cysts in a child with autosomal dominant polycystic kidney disease (ADPKD). The preliminary considerations were renal cell carcinoma and Wilms' tumor. After open biopsy, the patient was diagnosed with ADPKD with atypical renal cyst development. The development pattern of renal cysts in children with ADPKD is varied, and open biopsy is sometimes needed.

Endocrine disruptors and hypospadias: role of genistein and the fungicide vinclozolin.

OBJECTIVES: The phytoestrogen (plant estrogen) genistein, present in soy products, is of interest because in utero exposure to genistein can cause hypospadias in our mouse model and maternal consumption of soy is prevalent in human populations. Another compound of interest is the fungicide vinclozolin, which also causes hypospadias in the mouse and rat and can occur concurrently with genistein in the diet as a residue on exposed foods. A study in the United Kingdom found no relationship between a maternal organic vegetarian diet and hypospadias frequency, but women who consumed nonorganic vegetarian diets had a greater percentage of sons with hypospadias. Because nonorganic diets can include residues of pesticides such as vinclozolin, we sought to assess the interaction of realistic daily exposures to genistein and vinclozolin and their effects on the incidence of hypospadias. METHODS: Pregnant mice were fed a soy-free diet and orally gavaged from gestational days 13 to 17 with 0.17 mg/kg/day of genistein, 10 mg/kg/day of vinclozolin, or genistein and vinclozolin together at the same doses, all in 100 microL of corn oil. The controls received the corn oil vehicle. The male fetuses were examined at gestational day 19 for hypospadias, both macroscopically and histologically. RESULTS: We identified no hypospadias in the corn oil group. The incidence of hypospadias was 25% with genistein alone, 42% with vinclozolin alone, and 41% with genistein and vinclozolin together. CONCLUSIONS: These findings support the idea that exposure to these compounds during gestation could contribute to the development of hypospadias.

Multiple human isoforms of drug transporters contribute to the hepatic and renal transport of olmesartan, a selective antagonist of the angiotensin II AT1-receptor.

Olmesartan, a novel angiotensin II AT1-receptor antagonist, is excreted into both bile and urine, with minimal metabolism. Because olmesartan is a hydrophilic anionic compound, some transporters could be involved in its hepatic and renal clearance. In this study, we characterized the role of human drug transporters in the pharmacokinetics of olmesartan and determined the contribution of each transporter to the overall clearance of olmesartan. Olmesartan was significantly taken up into human embryonic kidney 293 cells expressing organic anion-transporting polypeptide (OATP) 1B1, OATP1B3, organic anion transporter (OAT) 1, and OAT3. We also observed its saturable uptake into human hepatocytes and kidney slices. Estimated from the relative activity factor method and application of specific inhibitors, the relative contributions of OATP1B1 and OATP1B3 to the uptake of olmesartan in human hepatocytes were almost the same, whereas OAT3 was predominantly involved in its uptake in kidney slices. The vectorial transport of olmesartan was observed in OATP1B1/multidrug resistance-associated protein (MRP) 2 double transfectants, but not in OATP1B1/multidrug resistance (MDR) 1 and OATP1B1/breast cancer resistance protein (BCRP) transfectants. ATP-dependent transport into membrane vesicles expressing human MRP2 and MRP4 was clearly observed, with K(m) values of 14.9 and 26.2 microM, respectively, whereas the urinary excretion of olmesartan in Mrp4-knockout mice was not different from that of control mice. We also investigated the transcellular transport of olmesartan medoxomil, a prodrug of olmesartan. Vectorial basal-to-apical transport was observed in OATP1B1/MRP2, OATP1B1/MDR1 double, and OATP1B1/BCRP double transfectants, suggesting the possible involvement of MRP2, MDR1, and BCRP in the limit of intestinal absorption of olmesartan medoxomil. From these results, we suggest that multiple transporters make a significant contribution to the pharmacokinetics of olmesartan and its prodrug.

Species difference in the inhibitory effect of nonsteroidal anti-inflammatory drugs on the uptake of methotrexate by human kidney slices.

Simultaneous use of nonsteroidal anti-inflammatory drugs (NSAIDs), probenecid, and other drugs has been reported to delay the plasma elimination of methotrexate in patients. Previously, we have reported that inhibition of the uptake process cannot explain such drug-drug interactions using rats. The present study quantitatively evaluated the possible role of the transporters in such drug-drug interactions using human kidney slices and membrane vesicles expressing human ATP-binding cassette (ABC) transporters. The uptake of methotrexate by human kidney slices was saturable with a K(m) of 45 to 49 microM. Saturable uptake of methotrexate by human kidney slices was markedly inhibited by p-aminohippurate and benzylpenicillin, but only weakly by 5-methyltetrahydrofolate. These transport characteristics are similar to those of a basolateral organic anion transporter (OAT) 3/SLC22A8. NSAIDs and probenecid inhibited the uptake of methotrexate by human kidney slices, and, in particular, salicylate, indomethacin, phenylbutazone, and probenecid were predicted to exhibit significant inhibition at clinically observed plasma concentrations. Among ABC transporters, such as BCRP/ABCG2, multidrug resistance-associated protein (MRP) 2/ABCC2, and MRP4/ABCC4, which are candidates for the luminal efflux of methotrexate, ATP-dependent uptake of methotrexate by MRP4-expressing membrane vesicles was most potently inhibited by NSAIDs. Salicylate and indomethacin were predicted to inhibit MRP4 at clinical plasma concentrations. Diclofenac-glucuronide significantly inhibited MRP2-mediated transport of methotrexate in a concentration-dependent manner, whereas naproxen-glucuronide had no effect. Inhibition of renal uptake (via OAT3) and efflux processes (via MRP2 and MRP4) explains the possible sites of drug-drug interaction for methotrexate with probenecid and some NSAIDs, including their glucuronides.

Progesterone receptors in the developing genital tubercle: implications for the endocrine disruptor hypothesis as the etiology of hypospadias.

PURPOSE: In fetal mice genital tubercles the ontogenetic expression of progesterone receptors and the effect of in utero estrogen and testosterone exposure were investigated. MATERIALS AND METHODS: To evaluate ontogenetic progesterone receptor expression genital tubercles from untreated fetuses at gestational days 12, 14, 16 and 18, and newborn pups were prepared for real-time reverse transcriptase-polymerase chain reaction or immunohistochemistry. To evaluate estrogen and testosterone effects pregnant dams were gavaged once daily with corn oil (vehicle), ethinyl estradiol or testosterone propionate from gestational days 12 through 17. At gestational day 19 the genital tubercles of delivered fetuses were harvested for morphological examination and then pooled for real-time reverse transcriptase-polymerase chain reaction. RESULTS: Progesterone receptor protein was first detected at gestational day 12 in the urethral plate and mesenchyma. At later stages staining intensity increased with a greater progesterone receptor signal, especially in the urethra. Progesterone receptor mRNA expression showed different increasing patterns in each sex until birth. However, no difference was noted between male and female genital tubercles in terms of the distribution and quantity of progesterone receptor expression. In utero ethinyl estradiol led to 8.2, 9.7 and 5.2-fold increases in progesterone receptor mRNA in females and in males with and without hypospadias, respectively. Testosterone propionate significantly decreased progesterone receptor mRNA levels in females and males. CONCLUSIONS: Progesterone receptors are expressed in developing genital tubercles, suggesting a direct role of progesterone in normal genital tubercle patterning. Their increasing expression until birth also implies increasing sensitivity of the genital tubercles to the effects of estrogenic and progestogenic endocrine disruptors during fetal life. Ethinyl estradiol and testosterone propionate lead to opposing effects on progesterone receptor expression, in addition to their opposing morphological effects on the genital tubercles. These findings expand our knowledge of genital tubercle morphogenesis and provide important information for understanding the effects of endocrine disruptors.

Estrogen receptor-alpha and beta are differentially distributed, expressed and activated in the fetal genital tubercle.

PURPOSE: We examined the ontogenic and sex specific expression of estrogen receptor-alpha and beta in mouse genital tubercles and assessed the effects of in utero estrogen exposure on these parameters. MATERIALS AND METHODS: Expression of the 2 genes was detected in mouse genital tubercles from fetuses collected on gestational days 12, 14, 16 and 18, and from newborns using immunohistochemistry and quantitative polymerase chain reaction. Pregnant dams were exposed to ethinyl estradiol or corn oil as the control. RESULTS: Estrogen receptor-alpha and beta proteins first appeared on gestational days 12 and 14, respectively. The 2 proteins were expressed in the urethral plate and mesenchyma. Staining intensity was more prominent in the mesenchyma for estrogen receptor-alpha and in the urethral plate for estrogen receptor-beta. Female genital tubercles expressed more estrogen receptor-alpha than male genital tubercles (p <0.01), while estrogen receptor-alpha expression increased gradually in the 2 sexes until birth. Estrogen receptor-beta expression did not differ between males and females, and it showed no notable variation during fetal life. Ethinyl estradiol led to a 2.1 and 3.8-fold increase in estrogen receptor-alpha expression in females and in males with hypospadias (p = 0.002 and 0.04, respectively). Estrogen receptor-beta expression did not change in response to ethinyl estradiol. CONCLUSIONS: This study provides in vivo evidence that estrogen receptor-alpha expression in the genital tubercles of each sex increases until parturition but estrogen receptor-beta expression does not, implying genital tubercle sensitivity to estrogen increases during fetal life. Exogenous administration of estrogens results in a response of increased expression of estrogen receptor-alpha but not of estrogen receptor-beta. These differential findings for estrogen receptor-alpha and beta imply that the 2 receptors may have different roles in normal or anomalous genital tubercle development.

Urothelial sonic hedgehog signaling plays an important role in bladder smooth muscle formation.

During bladder development, primitive mesenchyme differentiates into smooth muscle (SM) under the influence of urothelium. The gene(s) responsible for this process have not been elucidated. We propose that the Sonic hedgehog (Shh) signaling pathway is critical in bladder SM formation. Herein, we examine the role of the Shh-signaling pathway during SM differentiation in the embryonic mouse bladder. Genes in the Shh pathway and SM expression in mouse embryonic (E) bladders (E12.5, 13.5, and 14.5) were examined by immunohistochemistry (IHC), in situ hybridization, and reverse transcription polymerase chain reaction (RT-PCR). To examine the effects of disrupting Shh signaling, bladder tissues were isolated at E12.5 and E14.5, that is, before and after bladder SM induction. The embryonic bladders were cultured on membranes floating on medium with and without 10 muM of cyclopamine, an Shh inhibitor. After 3 days, SM expression was examined by assessing the following: SM alpha-actin (SMAA), SM gamma-actin (SMGA), SM-myosin heavy chain (SM-MHC), Patched, GLI1, bone morphogenic protein 4 (BMP4), and proliferating cell nuclear antigen (PCNA) by IHC and RT-PCR. SM-related genes and proteins were not expressed in E12.5 mouse embryonic bladder before SM differentiation, but were expressed by E13.5 when SM differentiation was initiated. Shh was expressed in the urothelium in E12.5 bladders. Shh-related gene expression at E12.5 was significantly higher than at E14.5. In cyclopamine-exposed cultures of E12.5 tissue, SMAA, SMGA, GLI1, and BMP4 gene expression was significantly decreased compared with controls, but PCNA gene expression did not change. In cyclopamine-exposed E14.5 cultures, SMGA and SM-MHC gene expression did not change compared with controls. Using an in vitro embryonic bladder culture model, we were able to define the kinetics of SM- and Shh-related gene expression. Cyclopamine inhibited detrusor SM actin induction, but did not inhibit SM-MHC induction. SMAA and SMGA genes appear to be induced by Shh-signaling pathways, but the SM-MHC gene is not. Based on Shh expression by urothelium and the effects of Shh inhibition on bladder SM induction, we hypothesize that urothelial-derived Shh orchestrates induction of SM in the fetal mouse bladder.

Characterization of the uptake of organic anion transporter (OAT) 1 and OAT3 substrates by human kidney slices.

The activities of renal multispecific organic anion transporters (OATs) 1 and 3 have not been fully evaluated in human kidneys. In the present study, the uptake of some organic anions was characterized in kidney slices from human intact renal cortical tissues: hOAT1 and hOAT3 substrates [p-aminohippurate (PAH) and 2,4-dichlorophenoxyacetate (2,4-D)] and hOAT3 substrates [benzylpenicillin (PCG), dehydroepiandrosterone sulfate (DHEAS), and estrone sulfate (ES)]. Despite large inter-batch differences, hOAT1 and hOAT3 mRNA levels correlated well, and there was a good correlation between the uptake of PAH and PCG by kidney slices. The uptake of organic anions by kidney slices was saturable with Km values of 31 to 48 microM for PAH, 0.73 to 4.9 microM for 2,4-D, 14 to 90 microM for PCG, and 9.2 to 11 microM for ES. These parameters were comparable with those for hOAT1 and/or hOAT3. The uptake of DHEAS consists of two saturable components with Km values of 2.2 to 3.9 and 1300 microM, and the Km value of the high-affinity component was close to that for hOAT3. Furthermore, PAH more potently inhibited the uptake of 2,4-D than that of PCG and DHEAS. PCG had a weaker effect on the uptake of PAH and 2,4-D than expected from its Km value. Taken together, it is likely that the uptake of PAH and 2,4-D is due to OAT1, and the uptake of PCG and ES and part of DHEAS uptake are due to OAT3 in human kidney slices. Human kidney slices are useful tools for characterizing the renal uptake of drugs.

Serum response factor, its cofactors, and epithelial-mesenchymal signaling in urinary bladder smooth muscle formation.

Little is known about the mechanism of bladder smooth muscle differentiation. We hypothesize that epithelial-mesenchymal signaling induces the expression of smooth muscle proteins in bladder mesenchyme resulting in smooth muscle differentiation. We confirmed that smooth muscle differentiation in the mouse urinary bladder occurs first at gestational day 14 (E14) based upon immunohistochemical localization of smooth muscle alpha-actin (SMAA). To investigate murine bladder smooth muscle differentiation and epithlelial-mesenchymal signaling in the developing bladder, we analyzed gene expression profiles of intact embryonic murine bladders and separated epithelial and mesenchymal components at embryonic days E13, E14, E15, E16, and postnatal day 1 (P1). Using cDNA microarray, we identified regulators of vascular smooth muscle differentiation in bladder mesenchyme, including serum response factor (SRF) and its cofactors, ELK1 and SRF accessory protein (SAP)1, as well as two SRF-associated pathways, angiotension receptor II and transforming growth factor- beta2. Immunohistochemistry showed diffuse expression of SRF in the bladder at E12 with localization of expression to the peripheral mesenchyme at E13 and E14. Our results suggest that bladder smooth muscle differentiation may share a similar gene expression program as occurs during vascular smooth muscle differentiation. The unique structure of the urinary bladder makes it an ideal model for studies of smooth muscle differentiation and epithelial-mesenchymal signaling.

Transplantable urothelial cell sheets harvested noninvasively from temperature-responsive culture surfaces by reducing temperature.

Augmentation cystoplasty using gastrointestinal flaps may induce severe complications such as lithiasis, urinary tract infection, and electrolyte imbalance. The use of viable, contiguous urothelial cell sheets cultured in vitro should enable us to avoid these complications. Transplantable urothelial cell sheets were obtained by utilizing a temperature-responsive cell culture method, and then examined by immunostaining and electron microscopy. Canine urothelium was produced on the surfaces of temperature-responsive culture dishes covalently bonded with the thermally sensitive polymer, poly(N-isopropylacrylamide). Stratified urothelial cell sheets were cultured and then harvested intact without enzymatic treatment from these dishes by reducing the temperature. Histological structure and cell-to-cell junctions were compared between these urothelial cell sheets and those harvested with dispase. All urothelial cell sheets were harvested from the bonded surfaces by reducing the culture temperature without the need for dispase. Electron microscopy revealed well-developed microridge, microvilli, and cell junction complexes. Conversely, these same cell features were destroyed by dispase treatment. Immunoblotting revealed that dispase fragmented occludin, whereas it remained unchanged in the intact urothelial cell sheets. Novel urothelial cell sheets obtained by culture on temperature-responsive culture surfaces were successfully harvested much less destructively than with dispase. This technology should prove useful in urinary tract tissue engineering in the near future.

Factors associated with failure of extracorporeal shock-wave lithotripsy for ureteral stones using Dornier lithotripter U/50.

BACKGROUND: In the present series of 170 patients who underwent extracorporeal shock-wave lithotripsy (SWL) treatment for ureteral stones, the authors determine which patients with ureteral stones had an unsuccessful outcome. METHODS: The records of 170 patients with ureteral stones who were treated with SWL using the Dornier lithotriptor U/50 (EMSE 140) between January 1998 and December 1999 were retrospectively investigated. One hundred and thirty-one patients were treated with SWL alone (single session, n = 98; multiple session, n = 33) and 39 patients required auxiliary treatment due to failure of SWL (33 with transurethral ureterolithotripsy (TUL), one with open lithotomy, and five with residual fragments who were followed up). These two groups were compared using multivariate logistic regression analysis. RESULTS: Lower ureteral stones and stones more than 12 mm in diameter were associated with a poor outcome of SWL. There were no significant differences in age, gender, number of stones, JJ stent placement, and degree of ureteral obstruction due to the stone between the two groups. The odds ratios of lower ureteral stones and stones > or = 12 mm were 4.18 and 2.57, respectively. CONCLUSION: Patients with distal ureteral stones and/or stones more than 12 mm in diameter were difficult to treat successfully with SWL. Alternatives such as TUL should possibly be considered as a first-line therapy for these stones.