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intra-lesional treatment of vehicle, and the PD⫹ADSC group received 1 million human labeled ADSC in 50 ␮L vehicle. Five weeks after treatment, 6 rats per group underwent erectile function measurement. Following euthanasia, penisses were harvested for histology and western blot. outcomes were measured using intracavernous pressure/ mean arterial pressure ratio (ICP/MAP) upon cavernous nerve stimulation, Elastin and Collagen III protein expression (western blot), and histomorphometric analysis of cross-sections of the penis. Statistical analysis was performed by ANOVA followed by Tukey Kramer test for post-hoc comparisons, or Mann-Whitney test when applicable. RESULTS: Erectile function significantly improved after ADSC treatment (ICP/MAP 0,37 in PD vs. 0,59 in PD⫹ADSC at 5 V stimulation, P⫽0,03). PD animals developed areas of fibrosis and elastosis with significant upregulation of Collagen III and Elastin protein expression as displayed by western blot and immunohistochemistry. These fibrotic changes were prevented by ADSC treatment. CONCLUSIONS: This study is the first to test stem cell therapy in an animal model of PD. Injection of ADSC into the TA during the active phase of PD prevents formation of fibrosis and elastosis in the TA and corpus cavernosum. Source of Funding: ESSM grant for medical research & departmental funding.

807 THE EFFECT OF HYDROXYFASUDIL ON PENILE DYSFUNCTION IN THE MALE SPONTANEOUSLY HYPERTENSIVE RAT Motoaki Saito*, Shogo Shimizu, Shuhei Tomita, Yonago, Japan INTRODUCTION AND OBJECTIVES: Hypertension represents a major risk factor for erectile dysfunction. Although the etiology of hypertension-induced erectile dysfunction is multifactorial and still unknown, Rho-Rho kinase pathway is one of the key factors. We investigated whether administration of hydroxyfasudil, a Rho kinase inhibitor could alternate dysfunction of NO-induced relaxation in corpus cavernosum smooth muscle in the SHR (spontaneously hypertensive rat). METHODS: Twelve-week-old male SHRs were treated with hydroxyfasudil (3 or 10 mg/kg, i.p.) once a day for 6 weeks. Wistar rats and SHRs treated with vehicle were used as age-matched controls. Penile cGMP concentrations and Rho kinase activities were determined, and penile function was estimated by organ bath studies with norepinephrine-induced contractions and acetylcholine-induced relaxations. The participation mRNA levels of eNOS and participation protein levels of eNOS and phosphorylated eNOS were investigated by quantitative real-time PCR methods and immunoblot analysis, respectively. RESULTS: The SHR showed significantly decreased cGMP concentrations, increased Rho kinase activities, norepinephrine-induced hyper-contractions, and acetylcholine-induced hypo-relaxations in the penile tissue. Treatment with hydroxyfasudil significantly improved the decreased penile cGMP concentrations, the increased Rho kinase activities, the increased norepinephrine-induced contractions, and the decreased acetylcholine-induced relaxation in a dose-dependent manner. Although there were no significant differences in expression protein levels of eNOS among any of the groups, down-regulation of eNOS mRNAs as well as phosphorylated eNOS were significantly ameliorated after treatment with hydroxyfasudil. CONCLUSIONS: Our data suggest that hydroxyfasudil ameliorates hypertension-associated dysfunction of NO-induced relaxation in corpus cavernosum smooth muscle possibly via inhibition of the RhoRho kinase pathway and activation of NO-eNOS pathway in the SHR.

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Source of Funding: None

808 PHOSPHODIESTERASE TYPE 5 (PDE5) IS CO-LOCALIZED WITH KEY MEDIATORS OF THE CYCLIC GMP SIGNALING IN THE HUMAN CLITORIS ¨ ckert*, Matthias Oelke, Knut Albrecht, Markus Kuczyk, Stefan U Hannover, Germany; Petter Hedlund, Lund, Sweden INTRODUCTION AND OBJECTIVES: The clitoris contributes mainly to the normal female sexual response cycle. The expression of the phosphodiesterase type 5 (PDE5, cyclic GMP PDE) in clitoral erectile tissue has been shown and a significance of cyclic GMP assumed in the control of clitoral vascular and non-vascular smooth muscle. However, up until today, only little is known on the mechanisms controlling this female genital organ. The aim of the present study was to evaluate in the human clitoris the expression and distribution of key mediators/proteins of the cyclic GMP pathway, such as the endothelial nitric oxide synthase (eNOS), cyclic GMP-dependent protein kinase G type I (cGKI) and calcitonin gene-related peptide (CGRP), in relation to the PDE5. METHODS: Specimens of the human clitoris (mid to proximal portion) were obtained within 4 hours - 6 hours postmortem from four female cadavers (age at death: 18 years - 42 years). Clitoral sections were exposed to antibodies directed against eNOS, PDE5, cGKI and CGRP. Then, fluorochrome-labelled secondary antibodies were applied. Visualization was commenced by means of laser fluorescence microscopy. RESULTS: Immunohistochemistry revealed immunosignals specific for the PDE5 and cGKI in the smooth musculature of small arteries transversing the supepithelial and stromal space. In addition, these blood vessels were characterized by the expression of NOS in the vascular endothelium. Immunosignals specific for PDE5 were also identified in slender varicose nerve fibers interspersing the tissue sections. Here, PDE5 was found co-localized with CGRP. CONCLUSIONS: The results are in favour of a role of the cyclic GMP signaling in the control of clitoral function. It seems likely that PDE5/cGKI/eNOS are involved in the maintenance of local blood perfusion whereas PDE5/CGRP may contribute to the mechanism of neuronal transmission. Source of Funding: Institutional (Excellence in Research Support Program of the Hannover Medical School).

809 TISSUE SEALING SHEET ATTENUATES ERECTILE DYSFUNCTION IN A RAT MODEL OF NERVE-SPARING RADICAL PROSTATECTOMY Shinichi Yamashita*, Yuko Iki, Hideaki Izumi, Yohei Satake, Yasuhiro Kaiho, Haruo Nakagawa, Yoichi Arai, Sendai, Japan INTRODUCTION AND OBJECTIVES: Recovery rates for erectile dysfunction (ED) following radical prostatectomy (RP) remain unsatisfactory, even with cavernous nerve (CN)-sparing surgery. We established an experimental rat model similar to the actual nervesparing RP in patients. In this model, erectile function was significantly

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decreased at 4 weeks after surgery, not in the earlier phase, indicating that ED following CN dissection occurred due to not only technical nerve injury, but also other factors such as inflammatory changes or surgical stresses. We found intraoperative bleeding, i.e., one of the parameters of the surgical stresses, influenced the recovery of erectile function after nerve-sparing RP, suggesting that reducing the amount of intraoperative bleeding would be beneficial for recovery of erectile function. A tissue sealing sheet, TachoSil®, has recently been used for intraoperative bleeding in RP. However, the efficacy of tissue sealing sheet for erectile function following nerve-sparing RP remains unclear. We evaluated the effect of the tissue sealing sheet on nerve injuryrelated ED. METHODS: Male Sprague-Dawley rats were randomly divided into 3 groups and subjected to sham operation or bilateral CN dissection with or without TachoSil®. In the group with TachoSil®, immediately after CNs were dissected bilaterally from the major pelvic ganglion to the apex of the prostate without crushing or cutting, CNs were sealed with TachoSil®. One urologist performed these operations. Another researcher assessed erectile function by measuring intracavernous pressure along with mean arterial pressure (ICP/MAP) during electrical pelvic nerve stimulation. Erectile function was evaluated 4 weeks postoperatively. RESULTS: Four weeks after CN dissection, the TachoSil® group showed significantly greater ICP/MAP than the group without TachoSil®. ICP/MAP in the TachoSil® group was similar to that in the sham group. CONCLUSIONS: The TachoSil® tissue sealing sheet attenuated ED following CN dissection in a rat model of nerve-sparing RP. Tissue sealing sheet appears effective in preventing intraoperative bleeding and postoperative adhesion, suggesting that the tissue sealing sheet may reduce surgical stress. TachoSil® tissue sealing sheet may thus represent a useful therapeutic approach to improve ED occurring after nerve-sparing RP, and is available for robotic-assisted laparoscopic prostatectomy using the da Vinci surgical system. Source of Funding: This study was supported by a Young Researcher Promotion Grant from the Japanese Urological Association (S.Y.) and Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (24791632).

810 OCCLUDED SEMINAL VESICLES INCREASE SEXUAL ACTIVITY IN MALE MICE: SYSTEMATIC LITERATURE REVIEW AND RESULTS OF A NOVEL MOUSE EXPERIMENT Frederic D. Birkhäuser*, Celine Schumacher, Roland Seiler, Louis de Meuron, Pascal Zehnder, Beat Roth, Antoinette Wetterwald, George N. Thalmann, Marco G. Cecchini, Urs E. Studer, Bern, Switzerland INTRODUCTION AND OBJECTIVES: The physiological role of seminal vesicles in fertility is well known. However, a relation between seminal vesicles and sexual activity has never been found although repeatedly investigated in various animal studies. We performed a systematic literature review and created a novel mouse experiment in order to assess whether occluded seminal vesicles affect sexual activity. METHODS: Based on a systematic review of published animal experiments on the relation between seminal vesicles and sexual activity and of contemporary literature about animal experiments, we created a novel mouse experiment with adult CD1 mice. Seventyseven male mice were assigned randomly to three experimental groups: mice with seminal vesicle occlusion (SVO, n⫽24), mice with seminal vesicle resection (SVR, n⫽23), and mice undergoing sham operation (SO, n⫽30). Females were brought into estrus by the Whitten effect, i.e. by exposure to male pheromones. Ten to 14 days postoperatively, three observers blinded to the experimental conditions separately observed male sexual activity during the dark phase of the

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day/night cycle during sessions of 3 h each. Primary endpoint was the percentage of sessions with intromission, secondary endpoints were number of intromissions and latency until first intromission. RESULTS: One hundred forty-one sessions were analyzed for a total of 423 h. In the SVO group intromissions were scored in 20 of 42 (48%) sessions. This rate was significantly higher than the 8 of 39 (21%) sessions in the SVR group (P⫽0.001) and 18 of 60 (30%) sessions in the SO group (P⫽0.004). Number of intromissions and latency until first intromission were statistically not different in all three groups (P⫽0.303; P⫽0.450). CONCLUSIONS: Male mice with occluded seminal vesicles were significantly more often sexually active than were mice after resection of seminal vesicles or after sham operation. This suggests that occluded and thus engorged seminal vesicles increase sex drive in male mice. Source of Funding: None

811 LONG-TERM CONTINUOUS TREATMENT WITH AVANAFIL PROMOTES RECOVERY OF ERECTILE FUNCTION IN A RAT MODEL OF POSTPROSTATECTOMY INDUCED ERECTILE DYSFUNCTION Ahmet Gokce*, George Lasker, Sree Harsha Mandava, Zakaria Abd Elmageed, Landon Trost, Philip Kadowitz, Asim Abdel-Mageed, Suresh Sikka, Wayne Hellstrom, New Orleans, LA INTRODUCTION AND OBJECTIVES: Surgical management of localized adenocarcinoma of the prostate is frequently associated with post-operative erectile dysfunction (ED). Several management strategies for penile rehabilitation have been evaluated with varying results. The objective of this study was to evaluate whether avanafil is successful in preventing ED following bilateral cavernosal nerve crush injury (BCNI), a model for human ED after bilateral nerve sparing radical prostatectomy (BNSRP). METHODS: Seventeen Sprague-Dawley rats (300-350 g) were divided three groups: Sham group (n⫽5), BCNI exposed to no avanafil treatment (BCNI, n⫽6) or avanafil (BCNI ⫹ treatment, n⫽6). In the treatment group, avanafil was administered in drinking water at a dose of 20 mg/day to each rat for four weeks beginning from the first day of BCNI. Rats in the sham and BCNI groups received drinking water alone. Four weeks following BCNI, animals underwent electrophysiologic assessment of erectile function by measuring intracavernosal pressure/mean arterial blood pressure (ICP/MAP) ratios. RESULTS: Daily oral avanafil treatment resulted in significant improvements in ICP/MAP ratios compared to BCNI alone (34⫾5 vs. 19⫾1% for 2.5 V (p ⫽ 0.01); 49⫾4 vs. 27⫾3% for 5.0 V (p ⫽ 0.002); 59⫾3 vs. 40⫾7% for 7.5 V (p ⫽ 0.02)). Sham animals had an ICP/MAP ratio of 29⫾9, 50⫾5 and 71⫾4% at stimulation voltages of 2.5, 5.0 and 7.5 V, respectively. BCNI rats showed significant decreases in ICP/ MAP compared to sham animals (p ⬍ 0.05), while avanafil treated rats were statistically similar to sham rates at all stimulation voltages except for 7.5 V (p ⫽ 0.02). CONCLUSIONS: Continuous long-term avanafil treatment following BCNI resulted in restoration of erectile function in a rat model of postprostatectomy ED. Further validation of this study is required to assess it’s potential role in human penile rehabilitation therapies following BNSRP. Source of Funding: None