- Email: [email protected]
Set regulates androgen production by PP2A in the cultured preantral follicles
Mural GCs were collected from the follicular fluid (FF). RESULTS: The fertilization rate was higher in the GnRH agonist group (70% vs. 54% p¼0.07). The mRNA expression of aromatase (0.50 vs 1, arbitrary unit, p<0.01), CYP 11 (0.6 vs. 1, P¼0.02) and 3 beta hydroxysteroiddehydrogenase (0.39 vs 1, p¼0.03) was significantly lower in the GnRH group. The expression of VEGF (0.74 vs. 1, P¼0.05) and inhibin beta B (0.38 vs 1 p¼0.01) was significantly lower in the GnRH analog triggered group. CONCLUSION: : Expression of genes related to steroidogenesis is already impaired at the time of oocyte retrieval in patients triggered with GnRH agonist. The decreased expression of VEGF and inhibin beta B in the GnRH agonist group can explain the mechanism of early OHSS prevention.
P-587 Wednesday, October 16, 2013 ART OUTCOME IN YOUNG WOMEN WITH PREMATURE OVARIAN AGEING. S. Malik, M. Dua. Reproductive Medicine, Southend Fertility & IVF Centre, Delhi, India. OBJECTIVE: Young women with signs of ovarian ageing are a matter of concern as far as their reproductive performance is concerned. With more women approaching infertility centers with this problem it becomes necessary to understand what reproductive outcomes are possible in such cases. Female age and basal FSH level, both are strong independent predictors of IVF outcome. Objective: To correlate age related basal FSH with IVF outcome in women with premature ovarian ageing in gonadotropins induced cycles. DESIGN: Retrospective observational study. MATERIALS AND METHODS: Between January 2011 and October 2012, a total of 135 women undergoing IVF and ICSI cycles with antagonist protocol were included in this retrospective cohort study. Basal FSH concentrations were measured and the women’s ages were calculated before they were undergoing pituitary desensitization and its correlation with ART outcome was evaluated. RESULTS: Increasing FSH was associated significantly with reduced number of oocytes retrieved, and embryos obtained. Young women with high FSH up to 20 produced less but good quality embryo’s resulting in sound pregnancy rate. CONCLUSION: FSH is a quantitative & age is a qualitative measure of ovarian reserve. Both are equally important in predicting IVF outcome. Basal FSH concentration should be restricted to counseling of patients on probability of achieving pregnancy, but should not be used to exclude them from fertility treatment. P-588 Wednesday, October 16, 2013 AN INEXPENSIVE METHOD FOR GENDER PRESELECTION. S. Cheung, Q. V. Neri, P. J. Husserl, Z. Rosenwaks, G. D. Palermo. Reproductive Medicine - CRM, Weill Cornell Medical College, New York, NY. OBJECTIVE: To test a simple, reliable, and inexpensive method to attempt selection of gender specific spermatozoa. The only available technique today is flow cytometry, however, it is expensive and requires DNA staining with a fluorochrome coadjuvated by laser beam excitation. DESIGN: We assessed multilayer discontinuous density gradient (DG) to measure which layer is capable of enriching X- or Y-bearing spermatozoa. MATERIALS AND METHODS: Semen specimens were processed by various discontinuous DG: 1) 60-90% (double), 2) 40-60-90% (triple), 3) 20-40-60-90% (quad). Putatively X-bearing spermatozoa were searched in 90% fraction while Y-bearing spermatozoa were identified from the lightest DG fraction. Sperm suspensions were smeared on slides for FISH analysis using centromeric probes for chromosomes 18, X, and Y. The ratio of Xto Y- chromosome bearing spermatozoa was blindly assessed as a percentage on at least 200 cells per slide. Aneuploid cells and those without signals were omitted. Unselected fractions of each sample served as controls. RESULTS: A total of 6 samples with a concentration of 48.3 17 x 106/ ml, motility of 50.9 6%, and normal morphology of 2.4 1% were included. The overall proportion of gender specific spermatozoa in unselected samples was 50.1% for X and 49.9% for Y. When selecting for X, a two-layer gave 63.2%, a triple 75.5%, and the quad 80.3%. This provided a direct correlation with the increasing number of layers (P ¼ 0.0001). When assessing for Y, a double layer yielded 62.7%, triple 75.5%, and
S318
ASRM Abstracts
quad 78.5%. As the density gradient became lighter, the proportion of Ybearing spermatozoa increased (P ¼ 0.0001). CONCLUSION: Selection of spermatozoa is an ethically accepted method for gender pre-selection. The only commercially available method to gender select spermatozoa is MicrosortÒ. We were able, with an inexpensive, safe, and reproducible way, to enrich gender specific spermatozoa at a comparable level with MicrosortÒ. Supported by: Institutional. FEMALE REPRODUCTIVE ENDOCRINOLOGY P-589 Wednesday, October 16, 2013 SET REGULATES ANDROGEN PRODUCTION BY PP2A IN THE CULTURED PREANTRAL FOLLICLES. L. Gao,a X. Liu,a B. Xu,b Y. Cui,a J. Liu.a aThe State Key Laboratory of Reproductive Medicine, Clinical Center of Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, China; bDepartment of Obstetrics and Gynecology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China. OBJECTIVE: By cDNA microarray it is found that SET in PCOS ovarias was twice higher than that in normal ones. In ovaries SET is predominantly expressed in theca cells and oocytes.So we investigate whether SET participates in ovarian androgen biosynthesis and the pathogenesis of PCOS. DESIGN: Basic research. MATERIALS AND METHODS: The in vitro culture model of mouse preantral follicle was applied to investigate the role of SET in ovarian androgen biosynthesis. The recombinant adenoviral vectors, AdCMV-SET and AdH1SiRNA/SET, were constructed to overexpress and knockdown SET expression respectively. The culture media of infected follicleswere collected to evaluate the testosterone production. RESULTS: SET overexpression in theca cells stimulated testosterone production whereas SET knockdown decreased testosterone production. Moreover, we found that PP2A activity was inhibited by SET overexpression, but stimulated by SET knockdown.Treatment with PP2A inhibitor okadaic acid (OA) led to the increased testosterone synthesis, while treatment with PP2A activators 1, 9-dideoxyforskolin and FTY720 resulted in the decreased testosterone synthesis. Furthermore, PP2A knockdown in theca cells confirmed the key role of PP2A in the testosterone synthesis, and OA was able to block the AdH1-SiRNA/SET-mediated inhibition on testosterone production. The central role of PP2A in SET-mediated regulation of testosterone production was confirmed by the finding that SET promoted the lyase activity of P450c17 and that PP2A inhibited its lyase activity. CONCLUSION: SET-mediated activation of testosterone biosynthesis by inhibition of PP2A activity and further activation of the 17,20-lyase activity. Supported by: Since our previous studies demonstrated the overexpression of SET in ovaries of polycystic ovary syndrome (PCOS) patients, this regulatory pathway may contribute to the hyperandrogenemia associated with PCOS. P-590 Wednesday, October 16, 2013 KNOWLEDGE OF REPRODUCTIVE MEDICINE AMONG GRADUATING OBSTETRICS AND GYNECOLOGY RESIDENTS: ARE THEY READY? T. Gipps, C. E. Sueldo. Obstetrics and Gynecology, University of California San Francisco-Fresno, Clovis, CA. OBJECTIVE: The purpose of this study was to assess the REI knowledge among our graduating Ob-Gyn residents and correlate it with the degree of comfort in managing various areas of this sub-specialty. DESIGN: Prospective analysis of parameters related to Ob-Gyn residency education in REI. MATERIALS AND METHODS: An analysis of the CREOG REI scores for 2012 and 2013 among our four graduating residents was performed; in addition, a self-assessment questionnaire was individually completed on the degree of comfort in managing clinical areas of REI, which was scored from 1 to 10 (10 being highest), in endoscopy, reproductive endocrinology, ART and fertility preservation; finally a written test comprised of 29 questions and an individual oral discussion of 7 clinical cases in the same REI areas (10 points maximum score for each case), was carried out and analyzed. RESULTS: The four graduating residents had a REI score in 2012 & 2013 CREOG, with average correct answers of 62.2% and 71 % respectively (ranked >50% nationwide). The answers in the comfort questionnaire
Vol. 100, No. 3, Supplement, September 2013
P-587 Wednesday, October 16, 2013 ART OUTCOME IN YOUNG WOMEN WITH PREMATURE OVARIAN AGEING. S. Malik, M. Dua. Reproductive Medicine, Southend Fertility & IVF Centre, Delhi, India. OBJECTIVE: Young women with signs of ovarian ageing are a matter of concern as far as their reproductive performance is concerned. With more women approaching infertility centers with this problem it becomes necessary to understand what reproductive outcomes are possible in such cases. Female age and basal FSH level, both are strong independent predictors of IVF outcome. Objective: To correlate age related basal FSH with IVF outcome in women with premature ovarian ageing in gonadotropins induced cycles. DESIGN: Retrospective observational study. MATERIALS AND METHODS: Between January 2011 and October 2012, a total of 135 women undergoing IVF and ICSI cycles with antagonist protocol were included in this retrospective cohort study. Basal FSH concentrations were measured and the women’s ages were calculated before they were undergoing pituitary desensitization and its correlation with ART outcome was evaluated. RESULTS: Increasing FSH was associated significantly with reduced number of oocytes retrieved, and embryos obtained. Young women with high FSH up to 20 produced less but good quality embryo’s resulting in sound pregnancy rate. CONCLUSION: FSH is a quantitative & age is a qualitative measure of ovarian reserve. Both are equally important in predicting IVF outcome. Basal FSH concentration should be restricted to counseling of patients on probability of achieving pregnancy, but should not be used to exclude them from fertility treatment. P-588 Wednesday, October 16, 2013 AN INEXPENSIVE METHOD FOR GENDER PRESELECTION. S. Cheung, Q. V. Neri, P. J. Husserl, Z. Rosenwaks, G. D. Palermo. Reproductive Medicine - CRM, Weill Cornell Medical College, New York, NY. OBJECTIVE: To test a simple, reliable, and inexpensive method to attempt selection of gender specific spermatozoa. The only available technique today is flow cytometry, however, it is expensive and requires DNA staining with a fluorochrome coadjuvated by laser beam excitation. DESIGN: We assessed multilayer discontinuous density gradient (DG) to measure which layer is capable of enriching X- or Y-bearing spermatozoa. MATERIALS AND METHODS: Semen specimens were processed by various discontinuous DG: 1) 60-90% (double), 2) 40-60-90% (triple), 3) 20-40-60-90% (quad). Putatively X-bearing spermatozoa were searched in 90% fraction while Y-bearing spermatozoa were identified from the lightest DG fraction. Sperm suspensions were smeared on slides for FISH analysis using centromeric probes for chromosomes 18, X, and Y. The ratio of Xto Y- chromosome bearing spermatozoa was blindly assessed as a percentage on at least 200 cells per slide. Aneuploid cells and those without signals were omitted. Unselected fractions of each sample served as controls. RESULTS: A total of 6 samples with a concentration of 48.3 17 x 106/ ml, motility of 50.9 6%, and normal morphology of 2.4 1% were included. The overall proportion of gender specific spermatozoa in unselected samples was 50.1% for X and 49.9% for Y. When selecting for X, a two-layer gave 63.2%, a triple 75.5%, and the quad 80.3%. This provided a direct correlation with the increasing number of layers (P ¼ 0.0001). When assessing for Y, a double layer yielded 62.7%, triple 75.5%, and
S318
ASRM Abstracts
quad 78.5%. As the density gradient became lighter, the proportion of Ybearing spermatozoa increased (P ¼ 0.0001). CONCLUSION: Selection of spermatozoa is an ethically accepted method for gender pre-selection. The only commercially available method to gender select spermatozoa is MicrosortÒ. We were able, with an inexpensive, safe, and reproducible way, to enrich gender specific spermatozoa at a comparable level with MicrosortÒ. Supported by: Institutional. FEMALE REPRODUCTIVE ENDOCRINOLOGY P-589 Wednesday, October 16, 2013 SET REGULATES ANDROGEN PRODUCTION BY PP2A IN THE CULTURED PREANTRAL FOLLICLES. L. Gao,a X. Liu,a B. Xu,b Y. Cui,a J. Liu.a aThe State Key Laboratory of Reproductive Medicine, Clinical Center of Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu, China; bDepartment of Obstetrics and Gynecology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, China. OBJECTIVE: By cDNA microarray it is found that SET in PCOS ovarias was twice higher than that in normal ones. In ovaries SET is predominantly expressed in theca cells and oocytes.So we investigate whether SET participates in ovarian androgen biosynthesis and the pathogenesis of PCOS. DESIGN: Basic research. MATERIALS AND METHODS: The in vitro culture model of mouse preantral follicle was applied to investigate the role of SET in ovarian androgen biosynthesis. The recombinant adenoviral vectors, AdCMV-SET and AdH1SiRNA/SET, were constructed to overexpress and knockdown SET expression respectively. The culture media of infected follicleswere collected to evaluate the testosterone production. RESULTS: SET overexpression in theca cells stimulated testosterone production whereas SET knockdown decreased testosterone production. Moreover, we found that PP2A activity was inhibited by SET overexpression, but stimulated by SET knockdown.Treatment with PP2A inhibitor okadaic acid (OA) led to the increased testosterone synthesis, while treatment with PP2A activators 1, 9-dideoxyforskolin and FTY720 resulted in the decreased testosterone synthesis. Furthermore, PP2A knockdown in theca cells confirmed the key role of PP2A in the testosterone synthesis, and OA was able to block the AdH1-SiRNA/SET-mediated inhibition on testosterone production. The central role of PP2A in SET-mediated regulation of testosterone production was confirmed by the finding that SET promoted the lyase activity of P450c17 and that PP2A inhibited its lyase activity. CONCLUSION: SET-mediated activation of testosterone biosynthesis by inhibition of PP2A activity and further activation of the 17,20-lyase activity. Supported by: Since our previous studies demonstrated the overexpression of SET in ovaries of polycystic ovary syndrome (PCOS) patients, this regulatory pathway may contribute to the hyperandrogenemia associated with PCOS. P-590 Wednesday, October 16, 2013 KNOWLEDGE OF REPRODUCTIVE MEDICINE AMONG GRADUATING OBSTETRICS AND GYNECOLOGY RESIDENTS: ARE THEY READY? T. Gipps, C. E. Sueldo. Obstetrics and Gynecology, University of California San Francisco-Fresno, Clovis, CA. OBJECTIVE: The purpose of this study was to assess the REI knowledge among our graduating Ob-Gyn residents and correlate it with the degree of comfort in managing various areas of this sub-specialty. DESIGN: Prospective analysis of parameters related to Ob-Gyn residency education in REI. MATERIALS AND METHODS: An analysis of the CREOG REI scores for 2012 and 2013 among our four graduating residents was performed; in addition, a self-assessment questionnaire was individually completed on the degree of comfort in managing clinical areas of REI, which was scored from 1 to 10 (10 being highest), in endoscopy, reproductive endocrinology, ART and fertility preservation; finally a written test comprised of 29 questions and an individual oral discussion of 7 clinical cases in the same REI areas (10 points maximum score for each case), was carried out and analyzed. RESULTS: The four graduating residents had a REI score in 2012 & 2013 CREOG, with average correct answers of 62.2% and 71 % respectively (ranked >50% nationwide). The answers in the comfort questionnaire
Vol. 100, No. 3, Supplement, September 2013