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Effects of Laparoscopic Ovarian Drilling on Serum Insulin and Adrenal Steroids in Women with Polycystic Ovary Syndrome
oocyte-degeneration rates and abnormal fertilization rates especially when the zona pellucida is particularly thick and/or hard to be penetrated and when the oolemma is more fragile. This laser assisted ICSI procedure could be the method of choice in case of fragile oocytes or when only very few oocytes are obtained.
Wednesday, October 25, 2000 4:45 P.M. O-223 Pronuclear Formation of Human Oocytes Excluded from IVF-ET Program in an Oocyte Microsurgery Program.* 1S. H. Jun, 1,2J. M. Lim, 1 S. E. Park, 1,2H. M. Chung, 1,2H. Shim, 1,2K. Y. Cha. 1Infertility Medical Center of CHA General Hospital, 2College of Medicine, Pochon CHA University, Seoul, Korea. Objective: We have attempted to develop a pronuclear transfer system for rescuing unfertilized oocytes (UFOs) and developmentally incompetent oocytes (IOs) that excluded from IVF-ET program. This study was undertaken to establish an effective method for pronucleus (PN) formation of UFOs and IOs in the system. Design: Randomized, prospective study using human oocytes retrieved from stimulated cycle. Materials and Methods: Oocytes were retrieved from consenting patients stimulated with a long protocol using GnRHa and gonadotropins. In Experiment 1, mature oocytes with a first polar body (PB) were fertilized in vitro by our conventional IVF program and UFOs collected at 24 h after IVF were activated by one of following methods: 1) no treatment (control), 2) 10% ethanol, 5 min (ET), 3) 5 mg/ml calcium ionophore, 5 min (CI) and 4) a single electrical DC pulse of 1.5 kV/cm, 30 msec (EP). Treated UFOs were subsequently cultured in 100 ml droplets of TCM-199 medium at 37°C, 5% CO2 in air atmosphere and PN formation was evaluated by staining with Hoechst 33342 at 24 h after the treatments. In Experiment 2, IOs either arrested at the germinal vesicle (GV), GV breakdown (GVBD) to metaphase-I (MI) stage or classified as a mature oocyte with abnormal morphology at the time of retrieval were further cultured for 24 –28, 8 –15 and 0 h, respectively. IOs were then activated by the optimal protocol of Experiment 1 and PN formation was observed at 24 h after the treatment. Data from these experiments were analyzed by ANOVA and the least square method in PLOC-GLM of SAS. Results: In Experiment 1, there was a significant (P50.0001) treatment effect on PN formation in UFOs. No spontaneous activation was occurred in the control, but activation treatments induced PN formation with various efficacy (table 1). More UFOs formed PN after ET or EP treatment than after CI treatment and a highest proportion (64.3%) was obtained by ET. EP was as effective (63.5%) as ET, but fragmentation was observed in 20% of UFOs activated by EP. Proportion of UFOs that formed presumptive haploid PN (2 PNs 1 1 PB or 1 PN 1 2 PBs) was 33.3, 0 and 28.6% after ET, CI and EP treatments, respectively. In Experiment 2, a great (P50.0362) effect of IOs’ status on PN formation was found. IOs at the GVBD-MI oocytes had higher potential to form PN than those at the GV stage or with abnormal morphology (25 vs. 77.8%). Conclusion: The results of this study clearly demonstrated that the treatment of 10% ethanol for 5 min effectively induced PN formation of UFOs. IOs could form pronucleus with high efficacy by this ethanol treatment, as long as they grew beyond the GVBD stage and maintained morphological normality under the hormonal stimulation before oocyte retrieval. * Supported by a grant from the Interdisciplinary Research Program of the KOSEF (1999-2-205-002-5).
and Infertility, Department of Obstetrics and Gynecology, McGill University, Montreal, Quebec, Canada. Objectives: To evaluate insulin response to oral glucose tolerance test (OGTT) and adrenal steroid responses to ACTH stimulation in clomipheneresistant anovulatory women with polycystic ovary syndrome (PCOS) before and after laparoscopic ovarian drilling. Design: Prospective study. Materials and Methods: 20 clomiphene-resistant anovulatory women with PCOS underwent OGTT and ACTH tests before and after ovarian drilling. Results: Of a total 20 patients, 15 patients completed the study. 10 patients with the body mass index of .24 kg/m2 revealed evidence of insulin resistance. Positive correlation was found between serum insulin and body weight (r:0.6, P5.04). Insulin responses to OGTT before ovarian drilling at 215, 0, 30, 60 and 120 minutes were 192 6 108, 182 6 102, 863 6 534, 1074 6 713 and 1200 6 901 pmol/L and after drilling were 162 6 99, 158 6 98, 742 6 470, 963 6 660 and 1085 6 917 pmol/L respectively (P:NS). 17 a-OH progesterone responses to ACTH before and after ovarian drilling at 215, 0, 30 and 60 minutes were 2.9 6 1.3, 2.7 6 1.3, 8.9 6 9.4 and 10.4 6 4.4 nmol/L and 3.0 6 2.2, 2.7 6 2.9, 6.3 6 2.8 and 6.6 6 3.0 nmol/L respectively (P:NS). Conclusions: laparoscopic ovarian drilling does not influence insulin and adrenal steroids dynamics.
Wednesday, October 25, 2000 2:15 P.M. O-225 The Effect of Glucose on the Expression of Type I Collagen and Transforming Growth Factor-Beta 1 (TGF-b1) in Human Peritoneal Fibroblast Cells in Culture. G. Saed, W. Zhang, M. P. Diamond. Departments of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, MI. Objective: Since high glucose levels contribute to the pathogenesis of fibrosis in organs in diabetics, we hypothesized that high glucose may also stimulate extracellular matrix accumulation following peritoneal injury, thereby leading to adhesion development. A potential mediator for these effects is TGF-b1, a major profibrogenic factor produced by peritoneal tissues. Design: To test this hypothesis, we utilized the multiplex RT/PCR technique to determine the effect of increasing glucose concentrations on the mRNA levels of type I collagen and TGF-b1 in human peritoneal fibroblasts in culture (HPF). Materials and Methods: Primary cultures of HPF were incubated with varying amounts of glucose (0 –5 mg/L) for 24 hours in DMEM. Total RNA was extracted from HPF and converted to cDNA by reverse transcriptase. Multiplex RT/PCR simultaneously amplifying b-actin with TGF-b1 or type I collagen mRNAs in the same tube was developed in our laboratory to quantitate type I collagen and TGF-b1 mRNA levels in response to increasing glucose concentrations. PCR products were analyzed by agarose gel electrophoresis and density of each ethidium bromide stained band was measured by a scanning densitometer. Results: Multiplex RT/PCR showed that there was a significant increase in the mRNA for type I collagen and TGF-b1 in response to increasing glucose concentration. Conclusion: Increasing glucose concentration stimulated type I collagen expression in peritoneal fibroblasts in culture. A potential mediator for this effect may be TGF-b1. Our results suggest that individuals with diabetes mellitus may be at risk for greater levels of postoperative adhesions.
Wednesday, October 25, 2000 2:30 P.M.
THE SOCIETY OF REPRODUCTIVE SURGEONS Wednesday, October 25, 2000 2:00 P.M. O-224 Effects of Laparoscopic Ovarian Drilling on Serum Insulin and Adrenal Steroids in Women with Polycystic Ovary Syndrome. A. Saleh, D. Morris, S. Lin Tan, T. Tulandi. Division of Reproductive Endocrinology
S84
Abstracts
O-226 A Randomized Trial on the Effects of Local Installation of Bupivacaine on Postoperative Pain after Operative Laparoscopy. A. Saleh, G. Fox, A. Felemban, C. Guerra, T. Tulandi. Department of Obstetrics and Gynecology, and Department of Anesthesia. McGill University, Royal Victoria Hospital, Montreal, Quebec, Canada.
Vol. 74, No. 3, Suppl. 1, September 2000
Wednesday, October 25, 2000 4:45 P.M. O-223 Pronuclear Formation of Human Oocytes Excluded from IVF-ET Program in an Oocyte Microsurgery Program.* 1S. H. Jun, 1,2J. M. Lim, 1 S. E. Park, 1,2H. M. Chung, 1,2H. Shim, 1,2K. Y. Cha. 1Infertility Medical Center of CHA General Hospital, 2College of Medicine, Pochon CHA University, Seoul, Korea. Objective: We have attempted to develop a pronuclear transfer system for rescuing unfertilized oocytes (UFOs) and developmentally incompetent oocytes (IOs) that excluded from IVF-ET program. This study was undertaken to establish an effective method for pronucleus (PN) formation of UFOs and IOs in the system. Design: Randomized, prospective study using human oocytes retrieved from stimulated cycle. Materials and Methods: Oocytes were retrieved from consenting patients stimulated with a long protocol using GnRHa and gonadotropins. In Experiment 1, mature oocytes with a first polar body (PB) were fertilized in vitro by our conventional IVF program and UFOs collected at 24 h after IVF were activated by one of following methods: 1) no treatment (control), 2) 10% ethanol, 5 min (ET), 3) 5 mg/ml calcium ionophore, 5 min (CI) and 4) a single electrical DC pulse of 1.5 kV/cm, 30 msec (EP). Treated UFOs were subsequently cultured in 100 ml droplets of TCM-199 medium at 37°C, 5% CO2 in air atmosphere and PN formation was evaluated by staining with Hoechst 33342 at 24 h after the treatments. In Experiment 2, IOs either arrested at the germinal vesicle (GV), GV breakdown (GVBD) to metaphase-I (MI) stage or classified as a mature oocyte with abnormal morphology at the time of retrieval were further cultured for 24 –28, 8 –15 and 0 h, respectively. IOs were then activated by the optimal protocol of Experiment 1 and PN formation was observed at 24 h after the treatment. Data from these experiments were analyzed by ANOVA and the least square method in PLOC-GLM of SAS. Results: In Experiment 1, there was a significant (P50.0001) treatment effect on PN formation in UFOs. No spontaneous activation was occurred in the control, but activation treatments induced PN formation with various efficacy (table 1). More UFOs formed PN after ET or EP treatment than after CI treatment and a highest proportion (64.3%) was obtained by ET. EP was as effective (63.5%) as ET, but fragmentation was observed in 20% of UFOs activated by EP. Proportion of UFOs that formed presumptive haploid PN (2 PNs 1 1 PB or 1 PN 1 2 PBs) was 33.3, 0 and 28.6% after ET, CI and EP treatments, respectively. In Experiment 2, a great (P50.0362) effect of IOs’ status on PN formation was found. IOs at the GVBD-MI oocytes had higher potential to form PN than those at the GV stage or with abnormal morphology (25 vs. 77.8%). Conclusion: The results of this study clearly demonstrated that the treatment of 10% ethanol for 5 min effectively induced PN formation of UFOs. IOs could form pronucleus with high efficacy by this ethanol treatment, as long as they grew beyond the GVBD stage and maintained morphological normality under the hormonal stimulation before oocyte retrieval. * Supported by a grant from the Interdisciplinary Research Program of the KOSEF (1999-2-205-002-5).
and Infertility, Department of Obstetrics and Gynecology, McGill University, Montreal, Quebec, Canada. Objectives: To evaluate insulin response to oral glucose tolerance test (OGTT) and adrenal steroid responses to ACTH stimulation in clomipheneresistant anovulatory women with polycystic ovary syndrome (PCOS) before and after laparoscopic ovarian drilling. Design: Prospective study. Materials and Methods: 20 clomiphene-resistant anovulatory women with PCOS underwent OGTT and ACTH tests before and after ovarian drilling. Results: Of a total 20 patients, 15 patients completed the study. 10 patients with the body mass index of .24 kg/m2 revealed evidence of insulin resistance. Positive correlation was found between serum insulin and body weight (r:0.6, P5.04). Insulin responses to OGTT before ovarian drilling at 215, 0, 30, 60 and 120 minutes were 192 6 108, 182 6 102, 863 6 534, 1074 6 713 and 1200 6 901 pmol/L and after drilling were 162 6 99, 158 6 98, 742 6 470, 963 6 660 and 1085 6 917 pmol/L respectively (P:NS). 17 a-OH progesterone responses to ACTH before and after ovarian drilling at 215, 0, 30 and 60 minutes were 2.9 6 1.3, 2.7 6 1.3, 8.9 6 9.4 and 10.4 6 4.4 nmol/L and 3.0 6 2.2, 2.7 6 2.9, 6.3 6 2.8 and 6.6 6 3.0 nmol/L respectively (P:NS). Conclusions: laparoscopic ovarian drilling does not influence insulin and adrenal steroids dynamics.
Wednesday, October 25, 2000 2:15 P.M. O-225 The Effect of Glucose on the Expression of Type I Collagen and Transforming Growth Factor-Beta 1 (TGF-b1) in Human Peritoneal Fibroblast Cells in Culture. G. Saed, W. Zhang, M. P. Diamond. Departments of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, MI. Objective: Since high glucose levels contribute to the pathogenesis of fibrosis in organs in diabetics, we hypothesized that high glucose may also stimulate extracellular matrix accumulation following peritoneal injury, thereby leading to adhesion development. A potential mediator for these effects is TGF-b1, a major profibrogenic factor produced by peritoneal tissues. Design: To test this hypothesis, we utilized the multiplex RT/PCR technique to determine the effect of increasing glucose concentrations on the mRNA levels of type I collagen and TGF-b1 in human peritoneal fibroblasts in culture (HPF). Materials and Methods: Primary cultures of HPF were incubated with varying amounts of glucose (0 –5 mg/L) for 24 hours in DMEM. Total RNA was extracted from HPF and converted to cDNA by reverse transcriptase. Multiplex RT/PCR simultaneously amplifying b-actin with TGF-b1 or type I collagen mRNAs in the same tube was developed in our laboratory to quantitate type I collagen and TGF-b1 mRNA levels in response to increasing glucose concentrations. PCR products were analyzed by agarose gel electrophoresis and density of each ethidium bromide stained band was measured by a scanning densitometer. Results: Multiplex RT/PCR showed that there was a significant increase in the mRNA for type I collagen and TGF-b1 in response to increasing glucose concentration. Conclusion: Increasing glucose concentration stimulated type I collagen expression in peritoneal fibroblasts in culture. A potential mediator for this effect may be TGF-b1. Our results suggest that individuals with diabetes mellitus may be at risk for greater levels of postoperative adhesions.
Wednesday, October 25, 2000 2:30 P.M.
THE SOCIETY OF REPRODUCTIVE SURGEONS Wednesday, October 25, 2000 2:00 P.M. O-224 Effects of Laparoscopic Ovarian Drilling on Serum Insulin and Adrenal Steroids in Women with Polycystic Ovary Syndrome. A. Saleh, D. Morris, S. Lin Tan, T. Tulandi. Division of Reproductive Endocrinology
S84
Abstracts
O-226 A Randomized Trial on the Effects of Local Installation of Bupivacaine on Postoperative Pain after Operative Laparoscopy. A. Saleh, G. Fox, A. Felemban, C. Guerra, T. Tulandi. Department of Obstetrics and Gynecology, and Department of Anesthesia. McGill University, Royal Victoria Hospital, Montreal, Quebec, Canada.
Vol. 74, No. 3, Suppl. 1, September 2000