819 ANDROGEN RECEPTOR IS NEGATIVELY REGULATED BY FEMINIZATION FACTOR 1B (FEM1B) IN MAMMALIAN TESTIS

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THE JOURNAL OF UROLOGY姞

patients were asked to complete the IIEF. The physical examination included BMI. Blood samples were obtained in the same day: a routine serum profile was performed, comprehensive of plasma level of U-II. RESULTS: The results obtained from clinical study are reported as mean ⫾ SD. The first result showing the difference between the UII plasma levels measured both in controls and ED patients,indicated that the mean values were 1662,06 pg/ml and 3513,21pg/ml respectively (ratio 2,11). A correlation test was performed to evaluate association between UII plasma values in ED patients and IIEF score. It was found a Pearson product-moment correlation coefficient (PPMCC) of ⫺0,82 suggesting a strong, statistically significant, negative correlation between the IIEF score and The UII plasma levels. Correlation tests were made also between UII plasma levels and BMI, blood total cholesterol and glycemia. Results obtained showed a moderate, statistically significant, positive correlation with BMI (PPMCC⫽0,48); a small statistically significant positive correlation with Blood total cholesterol (PPMCC⫽ 0,34)and no statistically significant correlation was founded with glycemia (PPMCC⫽ 0,09). CONCLUSIONS: UII plasma levels are doubled in ED patients compared to healthy controls and inversely correlated to IIEF score. Further studies are warranted to evaluate a possible role of urotensin II as serum marker of ED. Source of Funding: None

818 MURINE EXPERIMENTAL MODEL OF PEYRONIE’S DISEASE: A PILOT STUDY Carolina D’Elia*, Maria Angela Cerruto, Alberto Molinari, Francesca Maria Cavicchioli, Antonio D’Amico, Walter Artibani, Verona, Italy INTRODUCTION AND OBJECTIVES: The Peyronie’s disease (PD) is an idiopathic disorder of connective tissue ot the penis, that involves the tunica albuginea of the corpora cavernosa and the adjacent areolar space. Many studies have tried to identify risk factors associated with PD and the role of inflammation and free radicals in the pathogenesis of plaque seems to be important. It is a growing clinical evidence to support the therapeutic potential of mesenchymal stem cells for the revascularization of ischemic tissues and the recovery of their function and histological findings has assumed a possible application of lipofilling technique in patients with PD. The objective of this experimental study is the creation of a murine experimental model of PD, evaluating with MRI the penis of the rats (feasibility study), in order to plane the application of lipofilling technique in an animal model. METHODS: Four male Wistar rats Before were anesthetized, fixed in prone position and subjected to MRI; image acquisition was performed using a scanner Biospec (Bruker, Karlsruhe, Germany) equipped with a horizontal magnet operating at 4.7 Tesla with opening of 33 cm (Oxford Ltd, Oxford, UK). The animals underwent, subsequently, an injection of thrombin in the tunica albuginea, using a 1 mL syringe with needle 30 G, opening the dartos. MRI images were acquired in the same manner as described above at 7 and 21 days after injection with incision of the Dartos. RESULTS: The MRI acquisitions, both in coronal and axial projection, showed an adequate visibility of the anatomical structures. At 7 days after thrombin injection with the Dartos incision it was evident an oedematous portion, visible as a hyperintense area, located at the injection area. At 21 days after injection, oedema was partially resolved: the injection part of the hyperintense area remains unchanged, while the remaining area appears to be part of a re-absorption and re-organization process. CONCLUSIONS: Since none of the various treatment modalities currently available for the management of PD is able to bring healing, the researchers’ attention is increasingly directed towards innovative treatment programs, such as the use of stem cells of mesenchymal origin. Since there is no data in the literature concerning the role of stem cells in management of PD, we planned to develop a murine model of stable IPP, monitored with imaging (MRI). At the

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present time, the research in PD is hampered by the lack of universally accepted animal model and this is likely attributed to the limited insight into PD mechanisms and the difficulties faced by current animal models to truly represent the complexity. Source of Funding: None

819 ANDROGEN RECEPTOR IS NEGATIVELY REGULATED BY FEMINIZATION FACTOR 1B (FEM1B) IN MAMMALIAN TESTIS Shoichiro Iwatsuki*, Shoichi Sasaki, Yasue Kubota, Hiroki Kubota, Yukihiro Umemoto, Yutaro Hayashi, Kenjiro Kohri, Nagoya, Japan INTRODUCTION AND OBJECTIVES: Mammalian Fem1b (Feminization Factor 1b) is a homolog of nematode fem-1, which is a sex-determining factor of gametocytes. Although it is suggested that mammalian Fem1b plays roles in maintenance of testicular function, its mechanism remains to be solved. In this study, we evaluated expression patterns of Fem1b in rat testis and its contribution to androgen regulation in Leydig and Sertoli cell. METHODS: Gene expression of Fem1b in normal SpragueDawley rat testes according to the age in weeks (4 to 12 weeks) was assessed by real-time polymerase chain reaction (RT-PCR) and the loci of expressions were assessed by immunohistochemistry. To assess androgen regulation in Leydig and Sertoli cells, we knocked down Fem1b gene expression by siRNA against Fem1b using I-10 cell (Leydig cell line) and TM4 cell (Sertoli cell line). After 48 hours from administration of siRNA, sex steroidogenesis enzymes (Cyp11a1 and Hsd3␤ mRNA levels in the I-10 cells and androgen receptor mRNA level in the TM4 cells were evaluated. RESULTS: Fem1b mRNA expression remarkably increased up to 2.5-fold at 4 to 6 weeks of age, when spermatogenesis began. By immunohistochemistry, Fem1b was detected in the cytoplasm of spermatocytes, round spermatids and elongate spermatids. It was also detected in interstitial cells, especially in Leydig cells. Cyp11a1 and Hsd3␤ mRNA levels in the I-10 cells were not affected by gene knocking down of Fem1b. However, androgen receptor mRNA levels in the TM4 cells increased by Fem1b knockdown. CONCLUSIONS: Fem1b mRNA expression remarkably increased up to 2.5-fold at 4 to 6 weeks of age, when spermatogenesis began. By immunohistochemistry, Fem1b was detected in the cytoplasm of spermatocytes, round spermatids and elongate spermatids. It was also detected in interstitial cells, especially in Leydig cells. ` mRNA levels in the I-10 cells were not affected Cyp11a1 and Hsd3aˆO by gene knocking down of Fem1b. However, androgen receptor mRNA levels in the TM4 cells increased by Fem1b knockdown. CONCLUSIONS: In this study, we showed that Fem1b expression is localized in spermatocytes, spermatids and Leydig cells in rat testis. We also showed that knocking down Fem1b expression resulted in an increase of androgen receptor expression, suggesting that Fem1b does not contribute to steroidogenesis, which is stimulated by the hypothalamus-pituitary-testis axis, but contributes to the regulation of androgen receptor expression. It is suggested that Fem1b may be a negative-regulatory factor of androgen sensitivity in the mammalian testis. Source of Funding: None

820 TEMPORAL CHANGES IN NEUROTROPHIC FACTORS AND NEURITE GROWTH FOLLOWING CAVERNOUS NERVE INJURY Johanna L. Hannan*, Bernard L. Stopak, Baltimore, MD; Maarten Albersen, Leuven, Belgium; Xiaopu Liu, Arthur L. Burnett, Baltimore, MD; Frank van der Aa, Leuven, Belgium; Petter Hedlund, Lund, Sweden; Trinity J. Bivalacqua, Baltimore, MD INTRODUCTION AND OBJECTIVES: Surgical treatments for prostate, bladder and colorectal cancers often lead to neuronal damage and erectile dysfunction (ED). Despite nerve-sparing techniques and