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We systematically characterized gene expression, inflammation and neovascularization in patients with interstitial cystitis/bladder pain syndrome to obtain biological evidence supporting diagnosis and classification.We sequenced RNA obtained from bladder mucosal biopsies of 33 patients with 3 subtypes of interstitial cystitis/bladder pain syndrome, including Hunner lesions in 12, no Hunner lesions in 11 but with glomerulations and neither Hunner lesions nor glomerulations in 10, and 9 controls. Differentially expressed genes of each subtype were searched to identify subtype specific biological pathways and candidate genes important for pathogenesis. Candidate genes were validated by quantitative polymerase chain reaction and immunohistochemistry. Digital immunohistochemical quantification was performed to assess subepithelial lymphoplasmacytic cell and microvessel density. Relationships between candidate gene over expression and symptom severity were explored.Patients with Hunner lesions showed a distinct gene expression profile associated with significant up-regulation of biological processes involving immune responses and infection, and an increase in subepithelial lymphoplasmacytic cell and microvessel density. Over expression of 2 candidate genes, VEGF and BAFF, correlated with symptom severity. Meanwhile, the gene expression profiles of patients with the 2 subtypes without Hunner lesions were similar to those of controls. No difference in biological pathways or subepithelial lymphoplasmacytic cell and microvessel density were detected between these 2 subtypes and controls.Interstitial cystitis/bladder pain syndrome with Hunner lesions shows distinct genomic and histological features associated with immune responses and infection. In addition, VEGF and BAFF are potential disease biomarkers and therapeutic targets. This subtype should be considered separate from the syndrome.

The objective of this multicenter cross-sectional study was to investigate the relationship of nocturnal polyuria in patients with common lifestyle related diseases and overactive bladder, with special attention to hypertension.After baseline assessment, patients recorded 24-hour urinary frequency/volume, blood pressure and heart rate for 3 days. They were stratified into 4 groups based on mean blood pressure, including no hypertension, and controllable, untreated and uncontrolled hypertension, respectively.The 2,353 eligible patients, who had urinary urgency once or more per week and 1 or more nocturnal toilet visits, were enrolled from 543 sites in Japan. Of these patients complete data, including the 24-hour frequency volume chart, were collected from 1,271. Multivariable analyses showed a statistically significant association of nocturnal polyuria with increasing age (OR 1.04, 95% CI 1.02-1.05, p0.001) and gender (women vs men OR 0.75, 95% CI 0.59-0.96, p = 0.02), and for controllable (OR 1.10, 95% CI 0.83-1.460), untreated (OR 2.62, 95% CI 1.55-4.45) and uncontrolled (OR 1.15, 95% CI 0.81-1.62) hypertension vs no hypertension (p = 0.005). However, when assessed separately in men and women, hypertension and heart rate were significantly associated with nocturnal polyuria in women alone (p = 0.01 and 0.03, respectively). Lower urinary tract symptoms suggestive of benign prostatic hyperplasia were significantly associated with nocturnal polyuria in men alone (p0.001).The current study demonstrates that nocturnal polyuria was significantly associated with age, male gender, and untreated hypertension in patients with lifestyle related diseases and overactive bladder. The association between hypertension and nocturnal polyuria was significant in women alone.

We evaluated aging related bladder dysfunctions and biological changes in the bladder and dorsal root ganglia in rats. We also investigated whether long-term caloric restriction may have preventive effects on these changes.Male Fischer 344 rats were divided into a young group (age 6 months) and an old group (age 25 to 28 months), each with free access to normal food, and an old group (age 25 to 28 months) with food restricted to 3 days per week. Conscious cystometry, cDNA microarray analysis, immunohistochemistry and oxidative stress measurements of the bladder and dorsal root ganglia were performed.The old group with free access to normal food showed higher threshold pressure, more nonvoiding contractions and lower bladder compliance than the young group with free access to food. Old rats with free access showed greater post-void residual volume and lower voiding efficiency than old rats with caloric restriction and young rats. In the old group with free access 83 genes in the bladder and 48 in the L6 dorsal root ganglia were up-regulated compared with old rats with caloric restriction and young rats. These genes were mostly related to immune and inflammatory responses. Immunohistochemistry showed stronger expression of the immune response protease Gzm (granzyme) B and the collagenase Mmp13 (matrix metalloproteinase-13) in the bladder of old rats with free access vs old rats with caloric restriction and young rats. The level of malondialdehyde, an oxidative stress marker, was higher in the bladder of old rats with free access than in young rats but there was no difference between old rats with caloric restriction and young rats with free access to food.In rats aging leads to storage and voiding dysfunctions associated with immune and inflammatory related responses in the bladder and dorsal root ganglia, and with increased oxidative stress in the bladder. Caloric restriction reduced these aging related changes.

We investigated the relationships among nocturnal polyuria, sleep apnea and body fluid volume to elucidate the pathophysiology of nocturia in sleep apnea syndrome.We enrolled 104 consecutive patients who underwent polysomnography for suspected sleep apnea syndrome. Self-assessed symptom questionnaires were administered to evaluate sleep disorder and lower urinary tract symptoms, including nocturia. Voiding frequency and voided volume were recorded using a 24-hour frequency-volume chart. Body fluid composition was estimated in the morning and at night using bioelectric impedance analysis. Frequency-volume chart data were analyzed in 22 patients after continuous positive airway pressure therapy.Patients with nocturnal polyuria showed a higher apnea-hypopnea index (33.9 vs 24.2, p = 0.03) and a larger circadian change in extracellular fluid adjusted to lean body mass (0.22 vs -0.19, p = 0.019) than those without nocturnal polyuria. These relations were more evident in patients 65 years old or older than in those 64 years or younger. A multivariate linear regression model showed an independent relationship of nocturnal polyuria with the apnea-hypopnea index and the circadian change in extracellular fluid adjusted to lean body mass (p = 0.0012 and 0.022, respectively). Continuous positive airway pressure therapy significantly improved nocturnal polyuria and nocturia only in patients with nocturnal polyuria.This study identified sleep apnea and the circadian change in extracellular fluid as independent factors for nocturnal polyuria.

Purpose Bladder function is often impaired with aging. In other organs caloric restriction has had a prophylactic effect on the biological changes associated with aging. We tested the hypothesis that long-term caloric restriction can prevent age related impaired bladder function in the rat. Materials and Methods Fisher 344 male rats were divided into 3 groups, including 16 young rats at age 6 months with free access to normal food, 15 old rats at age 25 to 28 months with free access to normal food and 16 old rats at age 25 to 28 months fed with normal food 3 days per week since age 6 weeks (caloric restriction). We performed frequency volume measurements, in vitro organ bath functional studies using full-thickness longitudinal detrusor strips, evaluation of muscarinic and purinergic receptor mRNA expression, and histological examination with Masson trichrome staining of bladder tissue. Results Frequency volume changes did not significantly differ among the 3 groups. The old group fed normal food showed weaker contractile responses to carbachol and electrical field stimulation (especially in the cholinergic component), lower M 3 receptor mRNA expression and higher collagen deposition compared to the young group. These age related bladder changes were milder in the old group with caloric restriction than in the old group fed normal food. Conclusions This study suggests that in the rat long-term caloric restriction has a preventive effect against age related functional and morphological bladder changes. These changes include impaired detrusor contractility that may be related to decreased expression of M 3 receptors and to bladder wall fibrosis.

Imidafenacin and fesoterodine are used to treat overactive bladder. Imidafenacin, fesoterodine and its active metabolite 5-hydroxymethyl tolterodine are muscarinic receptor antagonists. It is believed that these agents act on afferent nerves in addition to smooth muscle. We investigated the effects of imidafenacin and 5-hydroxymethyl tolterodine on single unit afferent activity of mechanosensitive capsaicin sensitive and insensitive primary bladder afferent nerve fibers in rats.Female Sprague Dawley® rats were anesthetized. Single unit afferent activity was recorded from the L6 dorsal roots and classified by conduction velocity as that of Aδ or C fibers. After measuring control single afferent activity during constant filling cystometry the procedure was repeated with intravenous administration of imidafenacin (0.3 to 30 μg/kg) or 5-hydroxymethyl tolterodine (0.01 to 1 mg/kg) at cumulative doses with or without intravesical capsaicin or oxotremorine-M instillation.A total of 139 single unit afferent fibers were isolated from 111 rats, including 19 Aδ and 120 C fibers. Neither imidafenacin nor 5-hydroxymethyl tolterodine significantly affected the overall single unit afferent activity of Aδ or C fibers. Based on capsaicin sensitivity C fibers were divided into capsaicin sensitive and insensitive groups. Each antimuscarinic inhibited the single unit afferent activity of capsaicin sensitive C fibers but not of capsaicin insensitive C fibers at the highest dose. Moreover, oxotremorine-M facilitated single unit afferent activity in a proportion of C fibers. The facilitated single unit afferent activity was significantly attenuated by the highest dose of imidafenacin.These findings demonstrate that imidafenacin and 5-hydroxymethyl tolterodine can selectively inhibit capsaicin sensitive C fibers among mechanosensitive bladder afferents by antagonizing bladder muscarinic receptors.

FAAH degrades endocannabinoids and fatty acid amides. FAAH inhibition reduces micturition frequency and counteracts bladder overactivity in rats. We studied the effects of the peripherally active selective FAAH inhibitor URB937, and the CB1 and CB2 receptor antagonists rimonabant and SR144528, respectively, on single unit afferent activity of primary bladder afferents in rats.Female Sprague Dawley® rats were anesthetized. Single unit afferent activity of Aδ or C-fibers from the L6 dorsal roots was recorded during bladder filling before and after URB937 administration with or without rimonabant or SR144528. Drugs (1 mg/kg) were given intravenously. FAAH, CB1 and CB2 expression, and expression of the sensory marker CGRP in the L6 dorsal root ganglion were compared by immunofluorescence.A total of 102 single afferent fibers (48 Aδ and 54 C-fibers) were isolated from 57 rats. URB937 decreased single unit afferent activity of C-fibers to a mean ± SEM of 78% ± 9% and of Aδ-fibers to a mean of 67% ± 7% while increasing bladder compliance to a mean of 116% ± 3%. The effects of URB937 on single unit afferent activity and bladder compliance were counteracted by rimonabant or SR144528. Rimonabant increased single unit afferent activity of each fiber type but SR144528 affected only Aδ-fiber activity. CGRP positive L6 dorsal root ganglion neurons showed strong FAAH, CB1 and CB2 staining.To our knowledge we report for the first time that inhibiting peripheral FAAH depresses the Aδ and C-fiber activity of primary bladder afferents via CB1 and CB2 receptors. CB antagonists alone exerted facilitatory effects on single unit afferent activity during bladder filling in rats. The endocannabinoid system may be involved in physiological control of micturition as regulators of afferent signals.

We assayed mRNA expression of the TRP family of channels and ASIC1 in bladder tissue from patients with interstitial cystitis.Bladder biopsies of 1) nonclassic interstitial cystitis, 2) nonulcerative portions of classic interstitial cystitis, 3) ulcerative portions of classic interstitial cystitis and 4) noncancerous portions of bladder cancer as the control were placed immediately in ice-cold RNAlater® and subjected to real-time reverse transcriptase-polymerase chain reaction. We compared the mRNA expression of TRP channels, ASIC1, NGF, CXCL9 and UPK3A with that of controls, and correlated expression with symptom severity.We analyzed specimens from 17 patients with nonclassic interstitial cystitis, 22 with classic interstitial cystitis and 11 controls. In nonclassic interstitial cystitis samples TRPV2 and NGF showed significantly increased expression. In classic interstitial cystitis samples nonulcerative portions demonstrated a significant increase in the expression of TRPA1, TRPM2 and 8, TRPV1 and 2, ASIC1, NGF and CXCL9, and a significant decrease in UPK3A and TRPV4. Ulcerative portions showed similar changes for TRPM2, TRPV1, 2 and 4, CXCL9 and UPK3A. Increased expression of TRPM2, first noted in interstitial cystitis tissue, was the most pronounced one of the TRP family. All symptom measures correlated with TRPM2 and TRPV2 expression, and partially with that of the other genes.This study showed increased expression of the genes involved in pronociceptive inflammatory reactions in interstitial cystitis, including TRPV1, 2 and 4, ASIC1, NGF and CXCL9, and to our knowledge TRPM2 for the first time. The different expression patterns suggest distinct pathophysiologies for classic and nonclassic interstitial cystitis. The genes and their products are potential candidates for use as biomarkers or novel therapy targets.

It was previously thought that renal cell carcinoma with a diameter of 3 cm or less has low potential to cause distant metastasis. However, metastasis develops in a small number of cases, which cannot be ignored. We investigated the clinicopathological characters of small renal cell carcinoma with metastasis to further understand this condition.From January 1983 to February 2009, 165 cases of sporadic renal cell carcinoma 3 cm or less were treated at our department. Bilateral and von Hippel-Lindau disease were excluded from study. Clinicopathological parameters and outcome data were collected on each patient and analyzed.Histologically the 165 cases of primary renal cell carcinoma 3 cm or less included 151 of clear cell, 10 of papillary and 4 of chromophobe renal cell carcinoma, of which 4 had sarcomatoid differentiation, 6 had perinephric and/or sinus invasion and 20 had microvascular invasion. Overall we identified 10 metastatic cases (6.06%), of which 5 were synchronous. Univariate analysis revealed that age 60 years or greater (p = 0.0139), symptoms (p = 0.0054) and microvascular invasion (p0.0001) were significant risk factors. Multivariate analysis showed that only microvascular invasion was a significant risk factor (p = 0.00062). Perinephric and/or sinus fat invasion was not a significant risk factor.Metastasis also develops in small renal cell carcinoma cases. Results suggest that microvascular invasion is a significant risk factor and patients with microvascular invasion should be followed more carefully.

We investigated the genes responsible for ulcerative interstitial cystitis by DNA microarray analysis and quantitative real-time polymerase chain reaction.Bladder urothelial tissues were taken from a site apart from the ulcerative lesion in 9 patients with ulcerative interstitial cystitis and from a normal-looking area in 9 controls, including 7 with bladder carcinoma and 2 with benign prostatic hyperplasia. Total RNA was extracted from bladder samples and gene expression was compared between these 2 groups using Whole Human Genome DNA microarray 44K (Agilent Technologies, Santa Clara, California). Microarray data were analyzed by GeneSpring GX software and Ingenuity Pathway Analysis. Chosen genes were confirmed for altered transcription by quantitative real-time polymerase chain reaction.We identified 564 probes that were significantly expressed in mRNA more than 4-fold vs those in controls using volcano plot analysis (p0.001). Further network Ingenuity Pathway Analysis of these genes showed the top 3 functions, including 1) cell-to-cell signaling and interaction, and hematological system development and function, 2) inflammatory disease and 3) cellular development. Quantitative real-time polymerase chain reaction confirmed increased mRNA expression of several genes in the bladder samples of patients with ulcerative interstitial cystitis, including CXCR3 binding chemokines (CXCL9, 10 and 11) and TNFSF14 (LIGHT).Our study using DNA microarray analysis followed by quantitative real-time polymerase chain reaction reveals over expression of genes related to immune and inflammatory responses, including T-helper type 1 related chemokines, and cytokines such as CXCR3 binding chemokines and TNFSF14. These genes may be potential interstitial cystitis biomarkers.

Interstitial cystitis remains a poorly understood urological condition characterized by chronic pelvic pain and increased urinary frequency in the absence of any known etiology. Urothelial dysfunction and other abnormalities are presumed to be involved in the disease. Uroplakins that are expressed by urothelial cells are thought to have an important role as major barrier proteins on the apical surface of the urothelium.Gene expression of uroplakin Ia, Ib, II, III and III-delta4 was quantitatively measured in bladder biopsy samples from 29 patients with interstitial cystitis and 16 control subjects using real-time reverse transcriptase-polymerase chain reaction.The mRNA levels of the uroplakin Ia, Ib and II genes were relatively low and uroplakin III was relatively high in interstitial cystitis bladders compared to normal controls, although not significantly. Uroplakin III-delta4, a splicing variant of uroplakin III, was significantly up-regulated in interstitial cystitis samples (p0.001). When patients with interstitial cystitis were divided into those with and without ulcerative changes, the uroplakin III and III-delta4 genes were significantly up-regulated only in patients with nonulcerative interstitial cystitis. Even more interesting was the finding that up-regulation of uroplakin III-delta4 was much more prominent than that of uroplakin III, that is 26.5 vs 5.6-fold compared to the median values of normal subjects.Although the clinical implications of the over expression of uroplakin III and III-delta4 in nonulcerative interstitial cystitis bladders remains to be clarified, from the diagnostic viewpoint uroplakin III-delta4 is a potential marker for identifying nonulcerative interstitial cystitis.

We studied the relationship of nocturia with the International Prostate Symptom Score in men with lower urinary tract symptoms due to benign prostatic hyperplasia (BPH).Baseline symptom score and the change in the score by treatment were analyzed in 219 consecutive Japanese men with BPH using basic statistics, and correlation, cluster and principal component analyses.The average nocturia score was the middle of that of the 7 symptoms at baseline and by far the highest after treatment. Hence, the score change was the smallest. Analysis of baseline scores demonstrated that all symptoms were the initial component in principal component analysis with individual symptom scores correlating with the total symptom score and quality of life index. However, the correlation was least for nocturia. Observations were similar when analyzing the score change by treatment. These results indicate that the nocturia score is least specific to symptoms associated with BPH or least sensitive to the therapeutic effect on symptoms. This finding may be related to the high nocturia score in the age matched control population.The nocturia score of the International Prostate Symptom Score behaves differently in the symptom complex of BPH in Japanese men, partially because it is most confounded by the aging factor.

Few studies are available on imaging the urethral sphincter. We performed transvaginal ultrasonography to examine the muscle in continent and incontinent women.Transvaginal ultrasonography was performed using a 7.5 MHz. transrectal probe in 19 continent and 69 incontinent supine women. Incontinent cases were classified as urge (14) and stress (55), and included urethral hypermobility (22) and intrinsic sphincter deficiency (33). Cadaver specimens were also examined to confirm the anatomy of ultrasound images.Transvaginal ultrasonography showed the urethra as a round structure consisting of a relatively high echogenic central zone and a relatively low echogenic peripheral zone. Matching ultrasound images with the anatomy of cadaver specimens indicated that the peripheral zone represents rhabdosphincter most reliably at the lateral aspects. Thus, its thickness was measured at the 9 o'clock position of the mid urethra. Mean thickness plus or minus standard error of mean was significantly decreased in stress incontinence compared with continence and urge incontinence (2.14 +/- 0.04, 2.78 +/- 0.08 and 2.87 +/- 0.11 mm., respectively, p0. 001), while in intrinsic sphincter deficiency it was thinner than in hypermobility (2.00 +/- 0.05 versus 2.35 +/- 0.06 mm., p0.01). There was no difference in central zone thickness according to continence status.Urethral structures may be visualized by transvaginal ultrasonography. The rhabdosphincter image was thinner in stress incontinence, especially in intrinsic sphincter deficiency. Transvaginal ultrasonography may be a useful clinical test for examining the morphology of the urethral sphincter muscle in women.

The objective of the current study is to investigate the relationship between bcl-2 and p53 expression and prognosis in prostatic carcinoma.One hundred and forty-six needle core biopsy specimens obtained before any treatment were used for immunohistochemical detection of bcl-2 and p53 positivity. The relationship between these proteins was assessed by serial sections. Associations with Gleason score, clinical stage and patient survival were studied. Additionally, multivariate analysis by Cox proportional hazard model was used to determine the prognostic significance of these proteins.Bcl-2 positivity was found in 20% and p53 in 27% of 146 prostatic carcinomas. Both bcl-2 and p53 positivity were found only in 5%. They were expressed almost reciprocally in the tumors. P53 positivity correlated with high Gleason grade tumors. Bcl-2 positivity correlated with high T stage. Both bcl-2 and p53 positivity correlated with poor survival and short progression-free period. Multivariate analysis revealed that bcl-2 positivity was an independent prognostic indicator (p = 0.001).Bcl-2 and p53 were almost independently expressed in prostatic cancer. Both correlated with malignant phenotypes of prostatic cancer. The combined data from this staining further improved the ability to predict the patient prognosis.

A nationwide Japanese survey of 210 cases from 116 collaborating institutions demonstrated a dramatic increase in incidentally detected adrenal tumor from 4 cases in 1980 to 54 in 1988. The most common diagnosis was nonfunctioning cortical adenoma (69 cases), followed by pheochromocytoma (49). A total of 14 malignant tumors (6.7%) and 16 functioning benign cortical lesions was also found. Histological study revealed that functioning tumors were small and found in relatively young female patients, nonfunctioning cortical adenomas were of similar size but found in older patients, and all malignant tumors were larger than 6.5cm. in diameter and occurred in patients 58 years or younger. These results indicate an increase in the occurrence of incidental adrenal tumor and the complexity of pathology.

A survey of renal cell carcinoma detected incidentally (incidental renal cancer) in 116 collaborating institutions was conducted nationwide in Japan between 1980 and 1988. The analysis of 1,428 cases demonstrated a dramatic increase in the frequency of incidental renal cancer from 20 cases in 1980 to 338 cases in 1988. Detection occurred at routine health examinations in 32.8% of the cases and during examinations for unrelated diseases in the remainder. The chief method of detection was ultrasonography (68.2%), followed by computerized tomography (21.8%) and excretory urography (6.4%). Tumor extension was within the renal capsule in 76.5% of the cases and the mean tumor diameter was 5.4 cm. The 5-year survival rate was 76.5% but it was better in cases detected by ultrasonography compared with other methods. These results were compatible with previous reports and suggest that incidental renal cancer detection may improve the prognosis of renal cancer as a whole.

The inhibitory effect of oral administration of alpha-difluoromethylornithine (DFMO) on urinary bladder carcinogenesis induced by N-butyl-N-(4-hydroxybutyl)-nitrosamine (BHBN) was explored. Since DFMO in the drinking water at 0.5% and 0.2% had been demonstrated to inhibit tumorigenesis, lower doses (0.2%, 0.1%, 0.03% and 0.01%) of DFMO were examined in the present study. After six-week treatment with the drinking water containing 0.05% BHBN, water containing DFMO at 0.2%, 0.1%, 0.03%, 0.01% or 0% was administered during the subsequent 34 weeks. Incidence of bladder carcinoma was 15/35 (43%), 14/35 (40%), 21/35 (60%), 20/35 (57%) and 27/35 (77%) in the 0.2%, 0.1%, 0.03%, 0.01% and 0% DFMO groups, respectively. Stastistical analysis indicated significant tumor suppression in the 0.2% and 0.1% DFMO groups. Tumor multiplicity and size were not affected by DFMO treatment. No untoward effects were demonstrated in body weight gain or examination of pertinent organs. Our data indicate that bladder carcinogenesis induced by six-week exposure to 0.05% BHBN is significantly inhibited by daily administration of DFMO at the level of 0.1% or higher in drinking water.