Epigenetic Regulation of Filaggrin Gene Expression in Human Epidermal Keratinocytes

BACKGROUND: Loss-of-function mutations in the filaggrin gene (FLG), which encodes an epidermal protein crucial for the formation of a functional skin barrier, have been identified as a major predisposing factor in the etiopathogenesis of atopic dermatitis (AD). Recent reports of relatively low frequencies of FLG-null mutations among specific ethnic groups with AD necessitated analysis of the epigenetic regulation which may control FLG expression without altering its DNA sequence.OBJECTIVE: The study aimed to identify DNA methylation-dependent regulation of FLG expression.METHODS: Quantitative polymerase chain reaction was performed to determine the restoration of FLG mRNA expression in normal human epidermal keratinocyte (NHEK) cells after treatment with epigenetic modulating agents. Bisulfite genomic sequencing and pyrosequencing analyses of the FLG promoter region were conducted to identify the citical CpG sites relevant to FLG expression. We performed small-scale pilot study for epidermal tissues obtained from Korean patients with severe AD.RESULTS: We here show that DNA methylation in the FLG with non-CpG island promoter is responsible for the transcriptional regulation of FLG in undifferentiated NHEK cells. The methylation frequencies in a single CpG site of the FLG promoter were significantly higher in lesional epidermis than those in matched nonlesional epidermis of subjects with severe AD.CONCLUSION: Our in vitro and clinical studies point to this unique CpG site as a potential DNA methylation marker of FLG, which can be a promising therapeutic target in the complications of filaggrin-related skin barrier dysfunction as well as in AD.

The Internalization of Bacillus Calmette-Guerin (BCG) in Bladder Cancer Cells May be Inhibited by Human beta-defensin 3

PURPOSE: To investigate whether secretion of human beta-defensin 3 (HBD-3) is induced by bacillus Calmette-Guerin (BCG) and to determine whether HBD-3 affects BCG internalization in bladder cancer cells. MATERIALS AND METHODS: RTPCR analysis was used to determine whether HBD-3 mRNA increases after incubation with BCG. HBD-3 proteins in 5637 and T24 human bladder cancer cell lines were assayed by ELISA. The internalization rate was evaluated by double immunofluorescence assay and confocal microscopy to test the optimal dose of HBD-3 for BCG internalization. We also investigated the difference in internalization rates and cell viability between recombinant HBD-3 protein, anti-HBD-3 antibody, and HBD-3 plus anti-HBD-3 antibody pretreatments. RESULTS: BCG induced HBD-3 mRNA expression and HBD-3 production dose and time-dependently in bladder cancer cells and affected BCG internalization. Pretreatment with recombinant HBD-3 protein lowered internalization of BCG dose-dependently. Moreover, anti-HBD-3 antibody prevented the effect of HBD-3 on BCG internalization in bladder cancer cells. The internalization rate of BCG pretreated with anti-HBD-3 antibody was higher than that in the control. The BCG internalization rate in cells pretreated with anti-HBD-3 antibody plus recombinant HBD-3 protein was higher than that in the control. BCG decreased bladder cancer cell viability, and anti-HBD-3 antibody prevented the inhibitory role of HBD-3 on the anti-proliferative effects of M. bovis BCG in bladder cancer cells. CONCLUSIONS: Bladder cancer cells produce HBD-3 when they are infected by BCG to defend themselves against BCG internalization, which plays an important role during the initiation and propagation of the immunotherapeutic response in bladder cancer cells.

The association of 5-alpha reductase type 2 (SRD5A2) gene polymorphisms with prostate cancer in a Korean population

PURPOSE: Steroid 5-alpha reductase type 2 (SRD5A2) modifies testosterone to dihydrotestosterone (DHT) in the prostate. Single-nucleotide polymorphisms (SNPs) of the SRD5A2 gene might affect DHT. We sought to understand the relationship of SRD5A2 SNPs to prostate cancer in the Korean population. MATERIALS AND METHODS: Twenty-six common SNPs in the SRD5A2 gene were assessed in 272 prostate cancer cases and 173 controls. Single-locus analyses were conducted by using conditional logistic regression. Additionally, we performed a haplotype analysis for the SRD5A2 SNPs tested. RESULTS: Among the 20 SNPs and 4 haplotypes, there were no statistically significant results in the prostate cancer patients and the controls. In the logistic analysis of SRD5A2 polymorphisms with prostate-specific antigen (PSA) criteria, two SNPs (rs508562, rs11675297) and haplotype 1 displayed significant results (odds ratio [OR], 1.76; p=0.05; OR, 1.88-2.02; p=0.01-0.04; OR, 0.59; p=0.02, respectively). rs508562, rs11675297, rs2208532, and haplotype 1 (OR, 1.49; p=0.05; OR, 2.02; p=0.05; OR, 2.01; p=0.04; OR, 0.56-0.64, p=0.03-0.04, respectively) had significant associations with Gleason score. rs508562, rs11675297, and haplotype 1 (OR, 1.41-2.34; p=0.004-0.05; OR, 1.74-1.82; p=0.03-0.05; OR, 0.42-0.67; p=0.0005-0.03, respectively) were significantly associated with clinical stage. CONCLUSIONS: We conclude that there was no significant association between SRD5A2 SNPs and the risk of prostate cancer in the Korean population. However, we found that some SNPs and 1 haplotype influenced PSA level, Gleason score, and clinical stage.

Nicotine in High Concentration Causes Contraction of Isolated Strips of Rabbit Corpus Cavernosum

It is well known that cigarette smoke can cause erectile dysfunction by affecting the penile vascular system. However, the exact effects of nicotine on the corpus cavernosum remains poorly understood. Nicotine has been reported to cause relaxation of the corpus cavernosum; it has also been reported to cause both contraction and relaxation. Therefore, high concentrations of nicotine were studied in strips from the rabbit corpus cavernosum to better understand its effects. The proximal penile corpus cavernosal strips from male rabbits weighing approximately 4 kg were used in organ bath studies. Nicotine in high concentrations (10(-5)~10(-4) M) produced dose-dependent contractions of the corpus cavernosal strips. The incubation with 10(-5) M hexamethonium (nicotinic receptor antagonist) significantly inhibited the magnitude of the nicotine associated contractions. The nicotine-induced contractions were not only significantly inhibited by pretreatment with 10(-5) M indomethacin (nonspecific cyclooxygenase inhibitor) and with 10(-6) M NS-398 (selective cyclooxygenase inhibitor), but also with 10(-6) M Y-27632 (Rho kinase inhibitor). Ozagrel (thromboxane A2 synthase inhibitor) and SQ-29548 (highly selective TP receptor antagonist) pretreatments significantly reduced the nicotine-induced contractile amplitude of the strips. High concentrations of nicotine caused contraction of isolated rabbit corpus cavernosal strips. This contraction appeared to be mediated by activation of nicotinic receptors. Rho-kinase and cyclooxygenase pathways, especially cyclooxygenase-2 and thromboxane A2, might play a pivotal role in the mechanism associated with nicotine-induced contraction of the rabbit corpus cavernosum.

Predicting Recurrence and Progression of Non-Muscle-Invasive Bladder Cancer in Korean Patients: A Comparison of the EORTC and CUETO Models

PURPOSE: This study aimed to confirm the utility of the European Organization for Research and Treatment of Cancer (EORTC) and the Spanish Urological Club for Oncological Treatment (CUETO) scoring systems and to determine which model is preferred as a prognostic model in Korean patients with non-muscle-invasive bladder cancer. MATERIALS AND METHODS: Between 1985 and 2011, 531 patients who were treated by transurethral resection of bladder cancer were retrospectively analyzed by use of the EORTC and CUETO models. Statistically, we performed Kaplan-Meier survival analysis; calculated Harrell's concordance index, receiver operating characteristic (ROC) curve, and cutoff values; and performed univariate and multivariate Cox proportional hazards regression analyses. RESULTS: For risk of recurrence, with the use of the EORTC model, all groups had statistically significant differences except between the group with a score of 0 and the group with a score of 1-4. With the use of the CUETO model, all groups differed significantly. For risk of progression, with the use of the EORTC model, significant differences were observed between all groups except between the group with a score of 2-6 and the group with a score of 7-13. With the use of the CUETO model, a significant difference was observed between the group with a score of 0 and the other groups. The concordance index of the EORTC and CUETO models was 0.759 and 0.836 for recurrence and 0.704 and 0.745 for progression, respectively. The area under the ROC curve for the EORTC and CUETO models was 0.832 and 0.894 for recurrence and 0.722 and 0.724 for progression, respectively. CONCLUSIONS: Both scoring systems, especially the CUETO model, showed value in predicting recurrence and progression in Korean patients, which will help in individualizing treatment and follow-up schedules.

Febrile Urinary Tract Infection After Prostate Biopsy and Quinolone Resistance

PURPOSE: Complications after prostate biopsy have increased and various causes have been reported. Growing evidence of increasing quinolone resistance is of particular concern. In the current retrospective study, we evaluated the incidence of infectious complications after prostate biopsy and identified the risk factors. MATERIALS AND METHODS: The study population included 1,195 patients who underwent a prostate biopsy between January 2007 and December 2012 at Chung-Ang University Hospital. Cases of febrile UTI that occurred within 7 days were investigated. Clinical information included age, prostate-specific antigen, prostate volume, hypertension, diabetes, body mass index, and biopsy done in the quinolone-resistance era. Patients received quinolone (250 mg intravenously) before and after the procedure, and quinolone (250 mg) was orally administered twice daily for 3 days. We used univariate and multivariate analysis to investigate the predictive factors for febrile UTI. RESULTS: Febrile UTI developed in 39 cases (3.1%). Core numbers increased from 2007 (8 cores) to 2012 (12 cores) and quinolone-resistant bacteria began to appear in 2010 (quinolone-resistance era). In the univariate analysis, core number> or =12 (p=0.024), body mass index (BMI)>25 kg/m2 (p=0.004), and biopsy done in the quinolone-resistance era (p=0.014) were significant factors. However, in the multivariate analysis adjusted for core number, the results were not significant, with the exception of BMI>25 kg/m2 (p=0.011) and biopsy during the quinolone-resistance era (p=0.035), which were significantly associated with febrile UTI. CONCLUSIONS: Quinolone resistance is the main cause of postbiopsy infections in our center. We suggest that further evaluation is required to validate similar trends. Novel strategies to find alternative prophylactic agents are also necessary.

Beta-Defensin 124 Is Required for Efficient Innate Immune Responses in Prostate Epithelial RWPE-1 Cells

PURPOSE: The present study aimed to determine the role played by beta-defensin 124 (DEFB124) in the innate immunity of prostate epithelial RWPE-1 cells during bacterial infection. MATERIALS AND METHODS: The expression of DEFB124 was examined by quantitative real-time polymerase chain reaction (PCR), Western blotting, and immunocytochemistry. Enzyme-linked immunosorbent assays and quantitative real-time PCR were performed to determine the production of cytokines and chemokines. Western blotting and chromatin immunoprecipitation studies were performed to assess the interaction between DEFB124 and nuclear factor-kappa B (NF-kappaB) in peptidoglycan (PGN)-stimulated RWPE-1 cells. By chemotaxis assay, we assessed the effect of DEFB124 on the migration of monocytes. RESULTS: Exposure to PGN induced DEFB124 upregulation and NF-kappaB activation through IkappaBalpha phosphorylation and IkappaBalpha degradation. Bay11-7082, an NF-kappaB inhibitor, blocked PGN-induced DEFB124 production. Also, NF-kappaB was shown to be a direct regulator and to directly bind to the -3.14 kb site of the DEFB124 promoter in PGN-treated human prostate epithelial RWPE-1 cells. When DEFB124 was overexpressed in RWPE-1 cells, interestingly, the production of cytokines (interleukin [IL] 6 and IL-12) and chemokines (CCL5, CCL22, and CXCL8) was significantly increased. These DEFB124-upregulated RWPE-1 cells markedly induced chemotactic activity for THP-1 monocytes. CONCLUSIONS: Taken together, these results provide strong evidence for the first time that increased DEFB124 expression via NF-kappaB activation in PGN-exposed RWPE-1 cells enhances the production of cytokines and chemokines, which may contribute to an efficient innate immune defense.

Pilot Study of Low-Dose Nonenhanced Computed Tomography With Iterative Reconstruction for Diagnosis of Urinary Stones

PURPOSE: To evaluate the efficacy of low-dose computed tomography (LDCT) for detecting urinary stones with the use of an iterative reconstruction technique for reducing radiation dose and image noise. MATERIALS AND METHODS: A total of 101 stones from 69 patients who underwent both conventional nonenhanced computed tomography (CCT) and LDCT were analyzed. Interpretations were made of the two scans according to stone characteristics (size, volume, location, Hounsfield unit [HU], and skin-to-stone distance [SSD]) and radiation dose by dose-length product (DLP), effective dose (ED), and image noise. Diagnostic performance for detecting urinary stones was assessed by statistical evaluation. RESULTS: No statistical differences were found in stone characteristics between the two scans. The average DLP and ED were 384.60+/-132.15 mGy and 5.77+/-1.98 mSv in CCT and 90.08+/-31.80 mGy and 1.34+/-0.48 mSv in LDCT, respectively. The dose reduction rate of LDCT was nearly 77% for both DLP and ED (p<0.01). The mean objective noise (standard deviation) from three different areas was 23.0+/-2.5 in CCT and 29.2+/-3.1 in LDCT with a significant difference (p<0.05); the slight increase was 21.2%. For stones located throughout the kidney and ureter, the sensitivity and specificity of LDCT remained 96.0% and 100%, with positive and negative predictive values of 100% and 96.2%, respectively. CONCLUSIONS: LDCT showed significant radiation reduction while maintaining high image quality. It is an attractive option in the diagnosis of urinary stones.

Pilot Study of Low-Dose Nonenhanced Computed Tomography With Iterative Reconstruction for Diagnosis of Urinary Stones

PURPOSE: To evaluate the efficacy of low-dose computed tomography (LDCT) for detecting urinary stones with the use of an iterative reconstruction technique for reducing radiation dose and image noise. MATERIALS AND METHODS: A total of 101 stones from 69 patients who underwent both conventional nonenhanced computed tomography (CCT) and LDCT were analyzed. Interpretations were made of the two scans according to stone characteristics (size, volume, location, Hounsfield unit [HU], and skin-to-stone distance [SSD]) and radiation dose by dose-length product (DLP), effective dose (ED), and image noise. Diagnostic performance for detecting urinary stones was assessed by statistical evaluation. RESULTS: No statistical differences were found in stone characteristics between the two scans. The average DLP and ED were 384.60+/-132.15 mGy and 5.77+/-1.98 mSv in CCT and 90.08+/-31.80 mGy and 1.34+/-0.48 mSv in LDCT, respectively. The dose reduction rate of LDCT was nearly 77% for both DLP and ED (p<0.01). The mean objective noise (standard deviation) from three different areas was 23.0+/-2.5 in CCT and 29.2+/-3.1 in LDCT with a significant difference (p<0.05); the slight increase was 21.2%. For stones located throughout the kidney and ureter, the sensitivity and specificity of LDCT remained 96.0% and 100%, with positive and negative predictive values of 100% and 96.2%, respectively. CONCLUSIONS: LDCT showed significant radiation reduction while maintaining high image quality. It is an attractive option in the diagnosis of urinary stones.

Effectiveness of Flexible Ureteroscopic Stone Removal for Treating Ureteral and Ipsilateral Renal Stones: A Single-Center Experience

PURPOSE: The aim of this study was to evaluate the effectiveness of simultaneous flexible ureteroscopic removal of stones (URS) for ureteral and ipsilateral renal stones and to analyze the predictive factors for renal stone-free status. MATERIALS AND METHODS: We retrospectively reviewed the records of patients who underwent simultaneous flexible URS of ureteral and ipsilateral renal stones from January 2010 to May 2012. All operations used a flexible ureteroscope. We identified 74 cases of retrograde intrarenal surgery and 74 ureteral stones (74 patients). Stone-free status was respectively defined as no visible stones and clinically insignificant residual stones <3 mm on a postoperative image study. Predictive factors for stone-free status were evaluated. RESULTS: The immediate postoperative renal stone-free rate was 70%, which increased to 83% at 1 month after surgery. The immediate postoperative ureteral stone-free rate was 100%. Among all renal stones, 15 (20.3%) were separately located in the renal pelvis, 11 (14.8%) in the upper calyx, 15 (20.3%) in the mid calyx, and 33 (44.6%) in the lower calyx. The mean cumulative stone burden was 92.22+/-105.75 mm2. In a multivariate analysis, cumulative stone burden <100 mm2 was a significant predictive factor for postoperative renal stone-free status after 1 month (p<0.01). CONCLUSIONS: Flexible URS can be considered simultaneously for both ureteral and renal stones in selected patients. Flexible URS is a favorable option that promises high stone-free status without significant complications for patients with a stone burden <100 mm2.

Relaxing Effect of Acetylcholine on Phenylephrine-Induced Contraction of Isolated Rabbit Prostate Strips Is Mediated by Neuronal Nitric Oxide Synthase

PURPOSE: The location of acetylcholinesterase-containing nerve fibers suggests a role for acetylcholine in both contractility and secretion in the prostate gland. The colocalization of nitrergic nerves with cholinergic nerves, and the cotransmission of nitric oxide with acetylcholine in cholinergic nerves, has been demonstrated in the prostate glands of various species. Thus, we investigated the effects of acetylcholine on phenylephrine-induced contraction and the correlation between cholinergic transmission and nitric oxide synthase by using isolated prostate strips of rabbits. MATERIALS AND METHODS: Isolated prostate strips were contracted with phenylephrine and then treated with cumulative concentrations of acetylcholine. Changes in acetylcholine-induced relaxation after preincubation with NG-nitroarginine methyl ester, 7-nitroindazole, and aminoguanidine were measured. The effects of selective muscarinic receptor antagonists were also evaluated. RESULTS: In the longitudinal phenylephrine-contracted strip, the cumulative application of acetylcholine (10(-9) to 10(-4) M) elicited a concentration-dependent relaxation effect. Acetylcholine-induced relaxation was inhibited not only by nitric oxide synthase inhibitors (10 microM L-NAME or 10 microM 7-nitroindazole) but also by 10 microM atropine and some selective muscarinic receptor antagonists (10(-6) M 11-([2-[(diethylamino)methyl]-1-piperdinyl]acetyl)-5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepine-6-one and 10(-6) M 4-diphenylacetoxy-N-methyl-piperidine). In contrast, relaxation was significantly increased by pretreatment of the strips with 10 mM L-arginine. CONCLUSIONS: Acetylcholine relaxed phenylephrine-induced contractions of isolated rabbit prostate strips. This relaxation may be mediated via both cholinergic and constitutive nitric oxide synthase with both the M2 and M3 receptors possibly playing key roles.

Improvement of andropause symptoms by dandelion and rooibos extract complex CRS-10 in aging male

Many aging male suffer various andropause symptoms including loss of physical and mental activities. This study evaluated the putative alleviative effects of CRS-10 dandelion and rooibos extract complex (CRS-10) on the symptoms of andropause. The survival rate of TM3 Leydig cells (TM3 cells) treated with CRS-10 was measured based on typical physiological stress. After daily intake of CRS-10 for 4 weeks, the level of testosterone, physical activity and both the number and activity of sperm in older rats (18 weeks) were measured. Furthermore, thirty males were surveyed with AMS (Aging Males' Symptoms) questionnaire after intake of 400 mg of CRS-10. Overall, CRS-10 protected TM3 cells from serum restriction and oxidative stress via activation of ERK and Akt pathways. The level of testosterone and activation of spermatogenesis in rats were significantly enhanced. In addition, physical locomotion was markedly improved. Daily intake of 400 mg of CRS-10 improved the quality of life among agingmale respondents, according to a clinical survey using the AMS. The results indicate the potential of CRS-10 as a safe and efficacious natural substance for reducing or alleviating andropause symptoms.

Delayed Presentation of Intravesical Bone Penetration after Pelvic Ring Fracture

Retrograde cystography and computed tomography (CT) are considered the gold standard for investigating bladder and pelvic bone injury. However, these methods can miss extraperitoneal bladder rupture caused by a penetrating bone fragment from a pelvic bone fracture. We experienced a routine conventional cystography and CT scan that failed to identify penetration of the bladder by a bone fragment, which thus delayed optimal treatment. Therefore, different diagnostic methods such as CT cystography or cystoscopy should be considered to rule out penetrating injury by a bony fragment in patients with extraperitoneal bladder rupture.

Influence of Overweight on 24-Hour Urine Chemistry Studies and Recurrent Urolithiasis in Children

PURPOSE: We investigated the influence of overweight on 24-hour urine chemistry studies and recurrent urolithiasis (UL) in children. MATERIALS AND METHODS: A retrospective cohort study was designed to assess children who presented with UL at a pediatric institution between 1985 and 2010. We calculated body mass index percentile (BMIp) adjusted for gender and age according to the 2007 Korean Children and Adolescents Growth Chart and stratified the children into 3 BMI categories: lower body weight (LBW, BMIp or =85). Twenty-four hour urine chemistry studies (urine volume, creatinine, calcium, oxalate, citrate, and pH) were compared between the 3 BMIp groups. Univariate and multivariate analyses were performed to assess independent risk factors for stone recurrence. RESULTS: A total of 125 patients were included. The age of the patients in the NBW group was older than that of patients in the LBW group, but 24-hour urine chemistry studies did not differ significantly between the three groups. Mean urine citrate levels were lower (0.273+/-0.218 mg/mg/d vs. 0.429+/-0.299 mg/mg/d, p<0.05) and the incidence of hypocitraturia was higher (81.5% vs. 45.7%, p<0.05)) in the recurrent stone former group. In the univariate analysis, hypocitraturia and acidic urinary pH were risk factors, but in the multivariate analysis, only hypocitraturia was a risk factor for stone recurrence (hazard ratio, 3.647; 95% confidence interval, 1.047 to 12.703). In the Kaplan-Meier curve, the hypocitraturia group showed higher recurrence than did the normocitraturia group (p<0.05). CONCLUSIONS: Unlike in adults, in children, overweight adjusted for gender and age was not associated with 24-hour urine chemistry studies and was not a risk factor for recurrent UL. Hypocitraturia was the only risk factor for UL in children.

Impact of Changing Trends in Medical Therapy on Surgery for Benign Prostatic Hyperplasia Over Two Decades

PURPOSE: Following the introduction of medical therapy for benign prostatic hyperplasia (BPH), we determined the effect of the change in trends in medical therapy on the indication and outcome of surgical intervention for BPH. MATERIALS AND METHODS: We compared the basic characteristics of, weight of resected tissue of, transfusions in, and postoperative complications of patients who underwent surgery between 1985 and 1989 (before the advent of medical therapy for BPH), between 1995 and 1999 (when medical therapy was developed and became widely used as alternative treatment), and between 2005 and 2009 (when medical therapy superseded surgical intervention to become first-line treatment and when combination therapy became widely adopted). RESULTS: At our institution, the mean age and BMI of patients increased over the past two decades (p<0.001). Hypertension, operation history, and other comorbidities also increased significantly (p<0.001, p=0.005, and p<0.001, respectively). The indications for surgery in 1985 to 1989, 1995 to 1999, and 2005 to 2009 were as follows: acute urinary retention in 34.7%, 20.2%, and 15.1% of patients and symptomatic deterioration in 61.1%, 72.3%, and 73.0% of patients, respectively. Prostate volume and the weight of resected tissue increased from 34.4+/-14.5 ml to 61.3+/-32.4 ml and from 7.2+/-6.4 g to 10.8+/-7.6 g, respectively, over two decades. Patients who underwent surgery in 2005 to 2009 had their catheters removed earlier (p<0.001). Secondary hemorrhage within four postoperative weeks and repeat transurethral resection of the prostate within 1 year decreased significantly (p=0.03 and p=0.003, respectively). No statistically significant change in impaired detrusor contractility was found (p=0.523). CONCLUSIONS: Although patients who underwent surgery were older after widespread use of medical therapy for BPH, advancements in surgical techniques have benefitted these patients.

A Novel Pathway Underlying the Inhibitory Effects of Melatonin on Isolated Rat Urinary Bladder Contraction

The aim of the present study was to elucidate the direct effects of melatonin on bladder activity and to determine the mechanisms responsible for the detrusor activity of melatonin in the isolated rat bladder. We evaluated the effects of melatonin on the contractions induced by phenylephrine (PE), acetylcholine (ACh), bethanechol (BCh), KCl, and electrical field stimulation (EFS) in 20 detrusor smooth muscle samples from Sprague-Dawley rats. To determine the mechanisms underlying the inhibitory responses to melatonin, melatonin-pretreated muscle strips were exposed to a calcium channel antagonist (verapamil), three potassium channel blockers [tetraethyl ammonium (TEA), 4-aminopyridine (4-AP), and glibenclamide], a direct voltage-dependent calcium channel opener (Bay K 8644), and a specific calcium/calmodulin-dependent kinase II (CaMKII) inhibitor (KN-93). Melatonin pretreatment (10(-8)~10(-6) M) decreased the contractile responses induced by PE (10(-9)~10(-4) M) and Ach (10(-9)~10(-4) M) in a dose-dependent manner. Melatonin (10(-7) M) also blocked contraction induced by high KCl ([KCl]ECF; 35 mM, 70 mM, 105 mM, and 140 mM) and EFS. Melatonin (10(-7) M) potentiated the relaxation response of the strips by verapamil, but other potassium channel blockers did not change melatonin activity. Melatonin pretreatment significantly decreased contractile responses induced by Bay K 8644 (10(-11)~10(-7) M). KN-93 enhanced melatonin-induced relaxation. The present results suggest that melatonin can inhibit bladder smooth muscle contraction through a voltage-dependent, calcium-antagonistic mechanism and through the inhibition of the calmodulin/CaMKII system.

Leptin Enhances Nitric Oxide-Dependent Relaxation of the Clitoral Corpus Cavernosum

PURPOSE: The effects of leptin on female sexual behaviors are controversial, and studies on this topic are limited. The objectives of this study were to evaluate the direct effects of leptin on clitoral vasoreactivity in vitro and to determine the mechanism of action. MATERIALS AND METHODS: Isometric tension studies were conducted to determine the effects of pretreatment with leptin (10(-8) M) on the contractile responses of rabbit clitoral corpus cavernosal smooth muscle strips. The effects of leptin were assessed on precontraction induced by phenylephrine (PE; 10(-9)-10(-4) M) and KCl (35-140 mM). We also examined the effect of leptin on relaxation induced by acetylcholine (ACh; 10(-9)-10(-4) M), verapamil (10(-10)-10(-6) M), and sodium nitroprusside (10(-9)-10(-4) M) in PE-precontracted (10(-5) M) strips. RESULTS: Leptin enhanced ACh-induced relaxation in PE-precontracted strips. L-NAME pretreatment significantly reduced the effect of leptin on ACh-induced relaxation, whereas L-arginine potentiated the effect of leptin. Leptin decreased the KCl-induced contractile responses. Leptin increased verapamil-induced relaxation responses. The relaxation effects of leptin on KCl-induced contraction were inhibited by 10(-5) M methylene blue and L-NAME pretreatment. CONCLUSIONS: A high concentration of leptin enhances ACh-dependent relaxation in clitoral cavernosal smooth muscles. These relaxation effects of leptin may occur through an NO-dependent mechanism and voltage-dependent calcium channel blockade.

Detection of Nanobacteria in Patients with Chronic Prostatitis and Vaginitis by Reverse Transcriptase Polymerase Chain Reaction

PURPOSE: We aimed to investigate the detection of nanobacteria (NB) from expressed prostatic secretions (EPS) in patients with category III chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and from vaginal swabs in patients with vaginitis by reverse transcriptase polymerase chain reaction (RT-PCR) and to evaluate the association between NB and Neisseria gonorrhea, Chlamydia trachomatis, Ureaplasma urealyticum (U. urealyticum), Mycoplasma hominis, Trichomonas vaginalis, and Mycoplasma genitalium. MATERIALS AND METHODS: A group of 11 men attending a specialized CP/CPPS clinic and a group of 157 women who reported symptoms of lower genital tract infection were enrolled in this study. NB were detected by RT-PCR. A Seeplex Sexually Transmitted Disease Detection assay (Seegene Inc., Seoul, Korea) was used that could detect DNA for 6 types of sexually transmitted pathogens. RESULTS: In EPS samples, the detection rate of NB in patients with CP/CPPS was 9.1%, and 9 (5.7%) of 157 vaginitis patients showed positive results in RT-PCR for NB in vaginal swabs. Associations observed among the 7 microorganisms included 6 (54.5%) patients who tested positive on EPS and 75 (47.8%) patients who tested positive on vaginal swabs. Five patients with vaginitis were found to have monoinfection of NB (6.7%). CONCLUSIONS: We found that conventional RT-PCR for NB was rapid, simple, low in cost, and easily available for the detection of NB, and that NB may be a possible etiological factor for vaginitis and CP/CPPS. The prevalence of U. urealyticum among the four patients with NB coinfection was 75%; the presence of U. urealyticum might therefore raise suspicion for nanobacterial infection.

The Effects of 5-alpha Reductase Inhibitor on Contr action of Erectile Tissue and Secretion of Seminal Fluid in Male Rabbits: Preliminary Study / 대한남성과학회지

PURPOSE: Adverse sexual experiences such as erectile dysfunction (ED), loss of libido, and ejaculation disorders have been consistent side effects of 5-alpha reductase inhibitors (5ARI). The aim of this study was to evaluate the effects of 5ARI (finasteride) treatment on semen parameters and contraction of the corpus cavernosum and seminal vesicles in male rabbits. MATERIALS AND METHODS: Adult male New Zealand White rabbits (n=10) were randomized into 2 groups: finasteride-treatment (5ARI) group and vehicle-treatment (control) group. The 5ARI group received daily oral finasteride (10 mg/day) by gavage for 4~6 weeks, and the control group received the same concentration of the vehicle. The semen volume and semen parameters between the 2 groups were compared; thereafter, contraction or relaxation responses of smooth muscle strips of the corpus cavernosum and seminal vesicles were observed in an organ bath. RESULTS: Semen magnesium (14.2 vs 5.1 mg/dl) and protein (2.2 vs 1.6 g/dl) concentrations were significantly lower in the 5ARI group than in the control group. The concentrations of other parameters such as electrolytes (Na/K/Cl), fructose, and citrate did not differ between the 2 groups. The contractile responses to norepinephrine (NE) significantly increased in the 5ARI group compared to the control group and the relaxation responses to acetylcholine (ACh) or sodium nitroprusside (SNP) also increased in the 5ARI group. The contractile responses of the seminal vesicular strips to NE significantly decreased in the 5ARI group compared with the control group. CONCLUSIONS: The results suggest that finasteride may decrease contraction of seminal vesicle tissue and alter semen parameters. The effect of finasteride on erectile tissue was double-faced; enhancing both contraction and relaxation. Further study is needed in this respect.

Effect of Prostate Biopsy Hemorrhage on MRDW and MRS Imaging

PURPOSE: To retrospectively evaluate the effect of post-prostate-biopsy hemorrhage on the interpretation of magnetic resonance diffusion-weighted (MRDW) and magnetic resonance spectroscopic (MRS) imaging in the detection of prostate cancer. We also investigated the optimal timing for magnetic resonance examination after prostate biopsy. MATERIALS AND METHODS: We reviewed the records of 135 men. All patients underwent prostate magnetic resonance imaging (MRI). The prostate was divided into eight regions according to the biopsy site. Subsequently, we measured hemorrhage on apparent diffusion coefficient (ADC) values and (choline+creatinine)/citrate ([Cho+Cr]/Cit) ratios in the same regions on the MRI. We investigated the effect of hemorrhage at ADC values and (Cho+Cr)/Cit ratios on MRI and the relationship between prostate biopsy results and MRI findings. RESULTS: The mean patient age was 68.7 years and the mean time between biopsy and MRI was 23.5 days. The total hemorrhagic score demonstrated no significant associations with intervals from biopsy to MRI. Higher hemorrhagic scores were associated with higher ADC values, prostate cancer, and noncancer groups, respectively (p<0.001). ADC values were lower in tumors than in normal tissue (p<0.001), and ADC values were inversely correlated with tumor Gleason score in biopsy cores (p<0.001). However, (Cho+Cr)/Cit ratios did not exhibit any association with prostate biopsy results and hemorrhage. CONCLUSIONS: Hemorrhage had no significant associations with the interval from biopsy to MRI. ADC values may help to detect prostate cancer and predict the aggressiveness of cancer; however, it is important to consider the bias effect of hemorrhage on the interpretation of MRDW imaging given that hemorrhage affects ADC values.