Effects of phosphodiesterase-5 inhibitors on Leydig cell secretory function in oligoasthenospermic infertile men: a randomized trial.

OBJECTIVE: to evaluate the effects of phosphodiesterase-5 inhibitors (PDE5-i) on Leydig cell secretory function (LCSF). PATIENTS AND METHODS: in all, 75 men with oligoasthenospermia were treated daily for 12 weeks with either vardenafil (23 men, group A), sildenafil (25 men, group B) or l-carnitine (26 men, group C); a further group of 22 men with oligoasthenospermia (group D) received no treatment. Serum levels of insulin-like-3 peptide (INSL3) were evaluated before and after the end of the treatment in each of groups A, B and C, respectively. Serum INSL3 levels were measured in each participant of group D before and after the 12-week experimental period. RESULTS: within group A and B, the peripheral serum mean INSL3 concentration, sperm concentration, percentage of motile spermatozoa, and percentage of morphologically normal spermatozoa were significantly greater after PDE5-i treatment than before. CONCLUSION: we suggest that PDE5-i enhances LCSF, as the mean INSL3 concentration was significantly greater after PDE5-i administration than before, within groups A and B. This enhancement in LCSF might contribute to the increase in sperm concentration and sperm motility after administration of PDE5-i.

Apoptotic action of botulinum toxin on detrusor muscle in rats.

AIMS: To examine botulinum toxin A (BTX-A)-induced apoptosis in the detrusor muscle in male rats. METHODS: The bladder wall and prostate in adult male Sprague-Dawley rats were injected with BTX-A (10 U at each site). Tissues were harvested after 2 weeks, then the effects of BTX-A on proliferative and apoptotic indices were determined using proliferative cell nuclear antigen and TUNEL staining, respectively. In addition, real-time polymerase chain reaction was used to measure mRNA expressions of the apoptosis-related gene, bcl-2, and BAX in detrusor muscle. RESULTS: Although bladder weight was significantly increased in the BTX-A group compared to controls, no significant changes in prostate weight were identified. While significant increases in proliferative cells were apparent in the vesical mucosa after staining, no significant changes were noted in apoptotic or proliferative cells in detrusor muscle or prostate. In addition, no significant changes between the BTX-A group and controls were seen for mRNA expressions of bcl-2/beta-actin or BAX/beta-actin in detrusor muscle. CONCLUSIONS: The lack of significant changes in cellular dynamics of the detrusor muscle in the current study suggests that BTX-A injection into detrusor muscle for overactive bladder may not induce bladder atrophy.

Post-fertilization effects of chronic renal failure in male rats.

We evaluated the potential for growth and intrauterine development of embryos generated from the fertilization of oocytes with spermatozoa recovered from animals with chronic renal failure (CRF). Group A included sham-operated rats (n = 28), group B1 involved CRF rats that had undergone erythropoietin plus bromocryptine treatment (n = 28), and group B2 included CRF rats that had received normal saline. Embryos derived from the in vitro fertilization of oocytes with spermatozoa recovered from rats of group A or group B1 or group B2 were transferred to female recipients. We induced CRF in a group of rats (group B; n = 56; the total kidney volume was reduced to one-sixth with two operations). One week after the second operation, the rats of group B were randomly divided into group B1 (they subsequently received bromocryptine plus erythropoietin) and group B2 (they received injections of saline). Nine weeks after the second operation, the fertility of each male rat was assessed by mating tests and in vitro fertilization of oocytes. The mean litter size was significantly smaller in the subpopulation of fertile animals in group B2 than in the fertile rats of group B1 and in the fertile rats of group B1 than in the fertile rats of group A. Per cent of transferred blastocysts that developed into alive offspring were significantly lower in group B2 than in group B1 and in group B1 than in group A. Epididymal spermatozoa demonstrated a significantly larger DNA-oxidative damage in group B2 than in group B1 and in group B1 than in group A. These findings demonstrate that sperm-DNA damage because of CRF development is accompanied by a defect in the development of embryos generated in vitro. We may suggest that bromocryptine and erythropoietin protecting sperm DNA from oxidative damage improve reproductive potential in rats with CRF.

Effect of the rho-kinase inhibitor hydroxyfasudil on bladder overactivity: an experimental rat model.

OBJECTIVES: To investigate the effects of the rho-kinase inhibitor hydroxyfasudil on bladder overactivity in cyclophosphamide (CYP)-induced cystitis. METHODS: Female Sprague-Dawley rats received a single intraperitoneal injection of CYP (200 mg/kg). Four days later, bladder function was evaluated by: (i) monitoring micturition behavior in metabolic cages between hydroxyfasudil- and vehicle-treated animals; (ii) measuring changes in continuous cystometrograms in response to intravenous hydroxyfasudil under anesthesia; and (iii) conducting a functional study examining the effect of hydroxyfasudil on the concentration-response curves to carbachol in bladder tissue strips. RESULTS: Intraperitoneal injection of hydroxyfasudil (10 mg/kg) significantly increased both the average and maximal voided volumes. Hydroxyfasudil significantly decreased the maximal detrusor pressure, whereas the intercontraction interval was not significantly affected. After administration of 0.1, 0.3, 1, and 3 microM hydroxyfasudil, the maximal contraction of the concentration-response curves to carbachol was significantly reduced to 74.5 +/- 4.2%, 55.2 +/- 5.6%, 29.4 +/- 5.6%, and 21.6 +/- 8.2% of the control values, respectively. CONCLUSIONS: The present findings indicate that hydroxyfasudil might be a new treatment option for CYP-induced detrusor overactivity.

Effects of long-term estrogen treatment on micturition behavior and the sensory neurons of the urinary bladder in old female rats.

OBJECTIVES: The aim of this study was to examine the effect of estrogen treatment on the micturition characteristics and sensory neurons of the bladder in old female rats. METHODS: Thirteen-month-old female Wistar rats were divided into two groups: a control group and a group treated with estradiol for 12 weeks. After the treatment, we investigated the changes of micturition behavior under the condition of polyuria, and the immunoreactivity of P2X(3), calcitonin gene-related peptide, and substance P in the dorsal root ganglia (L(1)-L(2), L(6)-S(1)). RESULTS: Our data showed that estrogen induced a significant increase in maximum micturition volume and average micturition volume in the estrogen-treated rats. On the other hand, there was no significant difference in the proportion of the immunoreactivity of P2X(3), calcitonin gene-related peptide, and substance P between the two groups. CONCLUSIONS: Although it is known that estrogen facilitates an increase in the nerve's regenerative response, the present study reveals no significant change in the sensory neurons. However, the increase in micturition volume indicated that long-term estrogen treatment is beneficial for the improvement of the storage problems of the micturition cycle.

Fibroepithelial polyp of distal ureter with periodic prolapse into bladder.

Ureteric fibroepithelial polyp is a rare disease; it is of mesodermal origin and exhibits benign characteristics. Hydronephrosis occurs in rare cases, and it is generally accepted that it may result in an obstruction without causing alterations of renal function. In many cases it is difficult to differentiate from transitional cell carcinomas. Nowadays, endoscopic evaluation is the means of treatment and management. In our case study we report a patient with a long fibroepithelial polyp of the distal ureter prolapsing into the bladder in a periodic pattern. Cystoscopy revealed that movement of the polyp was moving forward and backward in the right ureteric orifice. Cold-cut biopsy established the diagnosis. The patient underwent ureteroscopic excision and remains asymptomatic a year later.

Genetic and epigenetic risks of intracytoplasmic sperm injection method.

Pregnancies achieved by assisted reproduction technologies, particularly by intracytoplasmic sperm injection (ICSI) procedures, are susceptible to genetic risks inherent to the male population treated with ICSI and additional risks inherent to this innovative procedure. The documented, as well as the theoretical, risks are discussed in the present review study. These risks mainly represent that consequences of the genetic abnormalities underlying male subfertility (or infertility) and might become stimulators for the development of novel approaches and applications in the treatment of infertility. In addition, risks with a polygenic background appearing at birth as congenital anomalies and other theoretical or stochastic risks are discussed. Recent data suggest that assisted reproductive technology might also affect epigenetic characteristics of the male gamete, the female gamete, or might have an impact on early embryogenesis. It might be also associated with an increased risk for genomic imprinting abnormalities.

[The effect of botulinum toxin injection into the bladder for overactive bladder: two case].

Two cases of intractable overactive bladder which were treated by intradetrusor botulinum-A toxin (BTX-A) are presented. Case 1: A 53-year-old woman suffered from adhesive arachinoiditis. She had severe detrusor hyperreflexia and urge urinary incontinence despite a high dose of anticholinergic medication. Under cystoscopic control, a total of 300 units of BTX-A were injected into the detrusor muscle at 30 sites (10 units per ml per site), sparing the trigone area using a 27G flexible injection needle. This procedure was done under both caudal anesthesia with 10 ml 1% lidocaine and bladder mucosa anesthesia by instilling 40 ml 2% lidocaine. At a 12-week followup she was completely continent. Case 2: A 63-year-old man with intractable detrusor overactivity of idiopathic origin was treated using the above technique. Urge urinary incontinence decreased 4 weeks after the BTX-A injections. However, postvoid residual urine volume was increased, and intermittent self-catheterization was needed. Intradetrusor BTX-A dramatically increases the bladder capacity and decreases the frequency of urge urinary incontinence. However, this treatment caused urinary retention in Case 2. We believe that the dosage amount of BTX-A should be changed case by case.

Expression of heat shock protein 70 and its mRNAs during ischemia-reperfusion in the rat prostate.

We investigated the expression of heat shock protein (HSP) 70 and its mRNAs during ischemia-reperfusion in the rat prostate. Eight-week-old rats were divided into six groups: a control group, a 30-min ischemia group, and 30-min ischemia+30-min, 60-min, 1-day, and 1-week reperfusion groups (groups A, B, C, D, E, and F, respectively). In vivo real-time blood flow and HSP 70-1 and 70-2 mRNAs and proteins in the prostate were measured using laser Doppler flow meter, real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) methods, respectively. Clamping of the aorta decreased blood flow to 10% of the basal level. The expressions of HSP 70-1/2 mRNAs increased in groups B, C, and D, and decreased in groups E and F. The expression of HSP 70 proteins was increased after a short interval of increase in their mRNAs. Our data indicated that the expressions of HSP 70 proteins and their mRNAs are dramatically changed during ischemia-reperfusion in the rat prostate.

Preventive effect of long-chain fatty alcohol on ischemia--reperfusion injury in the rat bladder.

We attempted in the present study to clarify the preventive effects of cyclohexenonic long-chain fatty alcohol on ischemia-reperfusion injury in the rat bladder. Rat bladders were exposed to 30 min of ischemia and a subsequent 30 min of reperfusion with or without several doses of cyclohexenonic long-chain fatty alcohol (0.5, 2, 8 mg/kg). Muscle-bath studies were performed, and malonaldehyde concentrations were measured in the bladder. Bladder dysfunction and lipid peroxidation caused by ischemia-reperfusion were prevented by cyclohexenonic long-chain fatty alcohol in a dose-dependent manner. Our data indicate that cyclohexenonic long-chain fatty alcohol can prevent the production of free radicals and ischemia-reperfusion injury in the bladder.

Effect of cyclohexenonic long-chain fatty alcohol on rat overactive bladder induced by bladder neck obstruction.

We attempted to clarify the preventive effects of cyclohexenonic long-chain fatty alcohol on detrusor overactivity induced by mild bladder neck obstruction. Bladder neck obstruction was created by partial ligation of the urethra. Female Sprague-Dawley rats were divided into three groups: those with bladder neck obstruction treated without long-chain fatty alcohol, those with bladder neck obstruction with long-chain fatty alcohol (8 mg/kg, i.p., every day) and the sham-operated control group (A, B, and C groups, respectively). Six weeks after the induction of bladder neck obstruction, voiding behavior was observed in the metabolic cage, and a cystometrogram was performed in the experimental animals. Furthermore, Hematoxylin and Eosin, Azan-Mallory, and Bodian stainings were performed in these bladders. Bladder weight, voiding behaviors and a cystometry indicated that rats in the A group showed detrusor overactivity, which was improved by treatment with long-chain fatty alcohol. The proportion of connective tissue and the density of bundles of neurofibers in the bladder of the A group was significantly less than that in the other bladders. Mild bladder neck obstruction induces detrusor overactivity, which is improved by treatment with long-chain fatty alcohol.

Effects of long-chain fatty alcohol on peripheral nerve conduction and bladder function in diabetic rats.

Diabetic cystopathy as manifested by an enlarged bladder is mainly caused by peripheral neuropathy. Long-chain fatty alcohol, which has been isolated from the Far-Eastern traditional medicinal plant, Hygrophilia erecta, Hochr., has been found to possess some neurotrophic activities on the central neuron. Cyclohexenonic long-chain fatty alcohol (FA) used in this study were synthesized in order to improve the efficiency of the molecules. The effects of this compound on peripheral nerves, however, have not yet been studied. To get more information, we evaluated the effects of this compound on peripheral nerves in streptozotocin-induced diabetic rats in terms of nerve conduction velocity and bladder function. Three experiments were performed 8 weeks after the administration of streptozotocin to 8-week-old rats: (i) motor sciatic nerve conduction (MNCV), (ii) monitoring micturition behavior in the metabolic cage, and (iii) cystometrogram under urethane anesthesia (CMG). Half of the diabetic rats were treated with FA (8 mg/kg/day, i.p.). The difference in MNCV between control rats (49.0 +/- 2.2 m/s) and untreated diabetic rats (42.4 +/- 0.5 m/s) after 8 weeks reached significance (p = 0.0183). FA-administrated diabetic rats showed an improved MNCV (45.8 +/- 1.2 m/s). We also identified a significant improvement of bladder function in these animals. The diabetic rats had a much higher maximal micturition volume per 24 hours (4.9 +/- 0.4 ml) than control animals (1.5 +/- 0.1 ml). However, the diabetic rats treated with FA had a maximal micturition volume of only 3.7 +/- 0.3 ml. Likewise, the diabetic rats had a CMG bladder capacity of 0.90 +/- 0.14 ml while the diabetic rats treated with FA had a capacity of 0.54 +/- 0.07 ml. These results indicate that cyclohexenonic long-chain fatty alcohol has a beneficial effect on peripheral neuropathy and cystopathy in streptozotocin-induced diabetic rats.

Expression of HSP 70 and its mRNAS during ischemia-reperfusion in the rat bladder.

HSP 70 is an important protein that repairs damaged tissue after injury. In the present study, we investigated the expression of HSP 70 and its mRNAs during ischemia-reperfusion in the rat bladder. Rat abdominal aorta was clamped with a small clip to induce ischemia-reperfusion injury in the bladder dome. Male Wistar rats, 8 weeks old, were divided into six groups: controls, 30-min ischemia, 30-min ischemia and 30-, 60-minute, 1- and 7-day reperfusion, groups A, B, C, D, E, and F, respectively. In functional studies, contractile responses to carbachol were measured in these groups. The expression of HSP 70-1/2 mRNAs was quantified using a real-time PCR method, and that of HSP 70 proteins was measured using ELISA in the bladders. In the functional study, Emax values of carbachol to bladders in the A, B, C, D, E and F groups were 9.3 +/- 1.3, 7.9 +/- 1.7, 4.3 +/- 0.8, 4.2 +/- 0.7, 4.5 +/- 0.6, and 8.1 +/- 1.2 g/mm2, respectively. In the control group, the expression of HSP 70-1/2 mRNA was detected, and the expression of HSP 70-1 mRNAs was significantly higher than that of HSP 70-2 mRNAs in each group. The expression of HSP 70-1 mRNA increased in groups B and C, but decreased in groups D, E, and F. The expression of HSP 70-2 mRNA in group C was significantly higher than that of groups A, D, E, and F. The expression of HSP 70-1/2 mRNAs after 1 day or 1 week of reperfusion was similar to control levels. The expression of HSP 70 proteins was increased shortly after the expression of their mRNAs. The expression of HSP 70 after 1 day or 1 week of reperfusion was almost identical to control levels. Our data indicate that contractile responses of the bladder were decreased by ischemia reperfusion, and that expression of HSP 70 and its mRNAs appeared to increase after a short period of the insult.

Effects of estrogen on age-related changes in muscarinic responsiveness of the urinary bladder and lumbosacral dorsal root ganglion cells in female rats.

The aim of this study was to investigate estrogen's effect on age-related changes in bladder function. Female Wistar rats were divided into three groups that included young rats (3-month-old) (YR), old rats (13-month-old) (OR), and old rats given subcutaneous treatments of estradiol for 6 weeks (OR + E). The groups were evaluated for (i) micturition behavior, (ii) changes of detrusor contractility and frequency of detrusor contraction in response to muscarinic stimulation in cystometrograms, (iii) messenger RNA (mRNA) expression of the muscarinic receptor subtype in the detrusor muscle, as measured by real-time polymerase chain reaction, and (iv) the immunoreactivity of P2X(3), CGRP, and substance P in the lumbosacral dorsal root ganglia. There were no significant changes for the average micturition volumes or micturition frequencies seen among the three groups. Moreover, there were no significant differences in the proportion of the immunoreactivities of P2X(3), CGRP, and substance P in afferent neurons among these three groups. However, an intravenous administration of muscarine significantly increased the frequency during continuous cystometrograms in the OR and OR + E groups. In these groups, there was a significant increase in the expression of the M(2) receptor mRNAs as compared to YR. It appears that the up-regulation of the M(2) receptor may lead to a decreased intercontraction interval by muscarinic stimuli. OR + E rats showed a significant increase in bladder weight as compared to the OR group. The muscarine-stimulated contractility of the detrusor in the cystometrogram also exhibited a significant increase in the OR + E group as compared to the OR group, which resulted from estrogen-induced functional hypertrophy of the detrusor muscle. These findings suggest that as little as 6 weeks of estrogen treatment is capable of improving the detrusor contractility, although the treatment contributes little to the storage phase of the micturition cycle.

Effects of long-term estradiol treatment on the contractile response to muscarine and muscarinic receptor subtypes in the bladder of aged female rats.

In this study, we tried to elucidate the effect of estrogen treatment on the detrusor contractile response to muscarine and muscarine receptor subtypes of the bladder in 13-month-old female Wistar rats. The rats were divided into two groups, controls and rats treated with estradiol for 12 weeks. After the treatment phase, we monitored micturition behavior in addition to performing cystometrograms after the administration of muscarine, and real-time polymerase chain reaction for mRNA expression of the muscarinic receptor subtypes in the detrusor muscle. Our data indicated that there was a significant increase in the maximum micturition volume in the estradioltreated rats. The urodynamic results indicated significant changes in the maximum detrusor pressure following the administration of muscarine in the estradiol-treated rats, in contrast to the controls for which no significant changes were observed. Furthermore, M(3) receptor mRNAs in the detrusor muscle were significantly decreased in the estradiol-treated rats as compared to the control rats, while there were no differences noted for the M2 receptor mRNAs. Our data demonstrates that long-term estradiol treatment might be capable of increasing the potential detrusor contractility, and thus, estradiol might be a therapeutic agent that can be used to target the M3 receptors during the treatment of detrusor instability.

Significance of Fas expression alteration during tumor progression of renal cell carcinoma.

BACKGROUND: In order to characterize the alteration of apoptotic regulatory molecule expression during tumor progression of renal cell carcinoma (RCC), we compared the expression between tumor and normal tissues, and evaluated the relationship of the expression in tumors with pathological and clinical characteristics. METHODS: Competitive reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) allowed the determination of Fas and bcl-2 mRNA and protein expression in surgically resected tumor and normal tissue of 50 RCC. RESULTS: The mRNA expression of Fas and bcl-2 in RCC was significantly reduced compared to that in normal tissues. An IHC analysis was supportive of the RT-PCR results. In terms of relationships with pathological and clinical characteristics, the mRNA and protein expression of Fas in high-stage or high-grade tumors was significantly higher than that in low-stage or low-grade tumors. Moreover, a statistically poor prognosis was observed in tumor cases expressing a high amount of mRNA. In bcl-2 analysis, the mRNA and protein expression was significantly reduced in clear cell tumors compared to chromophobe cell tumors. CONCLUSION: It is suggested that the reduced expression of Fas and bcl-2 in RCC compared with the expression in normal kidney is a prominent alteration of apoptotic regulatory molecules. The alteration of the up-regulated Fas expression might be characterized during the tumor progression stage. It is also suggested that the effect of alteration of bcl-2 expression might be minimal during the tumor progression stage because of the reduced expression in tumors of the clear cell type, which is the most dominant cell type in RCC.

Prostatic meningeal carcinomatosis with low serum level of prostate-specific antigen.

Less than 1% of patients with metastatic prostate cancer present with abnormally low serum levels of prostate-specific antigen (PSA). Furthermore, cases of prostatic metastases to the meninges with a low serum level of PSA are extremely rare. The present report describes a patient with prostatic meningeal carcinomatosis and low serum PSA levels.

'Successful' chemo- and radiotherapy prior to radical cystectomy does not necessarily correlate with clinical course in small cell carcinoma of the bladder.

We report 2 cases in which chemo- and radiotherapy induced pathological remission prior to radical cystectomy in small cell carcinoma of the bladder. However, one of these cases suffered a local recurrence 8 months after surgery.

Effectiveness of an anti-inflammatory drug, loxoprofen, for patients with nocturia.

AIM: There is increasing evidence that non-steroidal anti-inflammatory drugs are effective for the treatment of nocturia. In this study, we attempted to investigate the role of loxoprofen sodium (loxoprofen) in the therapeutic management of patients with nocturia. METHODS: Fifteen benign protastatic hyperplasia and/or overactive bladder patients (13 males and 2 females, 71.1 +/- 1.5 years old) with three or more voids per night were involved. These patients had received standard drug therapy. Although these patients had received standard drug therapy for more than half a year, they had still three or more episodes of nocturia. The patients took a single dose of 60 mg of loxoprofen at night prior to sleep. Before and 1 week after the initiation of this therapy, the effects of this treatment were assessed by frequency volume chart and a questionnaire. RESULTS: In the questionnaire, seven patients answered as excellent, six patients demonstrated improvement of their symptoms, two patients did not show a significant change in their symptoms and no patients demonstrated a deterioration in the symptoms. In frequency volume chart, total void per day, total void per night, total urine volume per day, total night urine volume per day and single voided volume in the night before and after this treatment were 9.97 +/- 0.81 and 8.99 +/- 0.74 per day, 3.82 +/- 0.25 and 1.82 +/- 0.27 per night, 1349 +/- 81 and 1258 +/- 91 mL per day, 567 +/- 46 and 325 +/- 51 mL per night, and 143 +/- 13 and 149 +/- 10 mL, respectively. CONCLUSION: Loxoprofen can be effective and useful for patients with nocturia. Our data suggest that the main mechanism of this effect is to decrease urine production during a night's sleep.