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Effects of ketoconazole on ovarian response in patients with resistant polycystic ovarian syndrome undergoing ovulation induction
and pregnancy. We would therefore expect a low level of apoptosis in egg donors. The present study was designed to test the effect of different ovulation induction protocols (antagonist/hCG, antagonist/GnRH agonist, downregulation/hCG) on in vitro GL cells apoptosis in young donors. We were particularly interested in whether administration of a GnRH agonist to trigger final oocyte maturation instead of hCG in the antagonist group would increase the rate of in vitro apoptosis of gonadotropin-exposed GL cells. DESIGN: In vitro assays. MATERIALS AND METHODS: 19 egg donors with a mean age of 22.42 2.87 underwent ovulation induction with recombinant FSH combined with recombinant LH or HMG using a downregulation or an antagonist protocol. The ovarian response was monitored by transvaginal ultrasound and blood estradiol levels as needed. Final oocyte maturation was triggered by either hCG or leuprolide acetate and the egg retrieval was performed 35.5 hours later. GL cells were collected from pooled follicular fluid after oocyte recovery. Cells were washed and further purified using a 50% percoll gradient and anti-CD45 coated magnetic beads. After 48 hour culture without serum the percentage of apoptosis was determined under a fluorescent microscope after staining with 10 mM caspACE FITC-VAD-FMK (a fluorescent marker for activated caspases) and 0.05 mg/ml propidium iodide. Statistical analysis was carried out using the Student’s t-test. RESULTS: Overall the apoptosis rate was low, as expected for a group of young donors (19.06% 4.01, n ¼ 19). There were no significant differences in the percentage of apoptosis among the three groups of egg donors analyzed (antagonist/hCG: 17.35 4.64, n ¼ 5; antagonist/GnRH agonist 19.55 2.61, n ¼ 5; downregulation/hCG: 19.73 4.42, n ¼ 9; P>0.05). Moreover, the level of apoptosis observed in the donors was significantly lower than that observed in a group of infertility patients with a mean age of 32.47 3.22 undergoing IVF during the same period of time (19.06% 4.01, n ¼ 19 vs. 24.92 4.25, n ¼ 19; P<0.05). CONCLUSIONS: Triggering of ovulation with a GnRH agonist does not increase apoptosis of gonadotrophin exposed granulosa-lutein cells. Furthermore, there is no difference between downregulation and antagonist protocols. Supported by: Grant PI060772 from Fondo Investigaciones Sanitarias, Spain; Cajacanarias, Spain ; Fundacion Salud 2000, Serono, Spain.
were likely to differ by less than twenty percent of values from immediately frozen samples. Supported by: DSL and Immunotech systems donated by Beckman Coulter, Inc. Experiments and statistical analysis supported by Repromedix.
P-199 EFFECTS OF KETOCONAZOLE ON OVARIAN RESPONSE IN PATIENTS WITH RESISTANT POLYCYSTIC OVARIAN SYNDROME UNDERGOING OVULATION INDUCTION. M. Sadeghi, B. Hossein Rashidi, E. Sahrokh Tehraninejad, N. Memarpour. Obstetrics and Gynecology, Vali-e-Asr Hospital of Medicine, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran. OBJECTIVE: To assess the efficacy of low dose ketoconazole in addition to CC (clomiphen citrate) and hMG on ovulation induction. DESIGN: A double blind, randomized, clinical trial. MATERIALS AND METHODS: Fifty infertile patients with PCOS who had failed to respond to a daily dose of 150 mg of CC were randomly divided into two equal groups; group A received 50 mg ketoconazole every other day while group B received placebo. All the patients were induced by CC and hMG for controlled ovarian hyperstimulation. Ovarian response to induction, numbers of mature follicles, estradiol and progesterone levels, endometrial thickness, OHSS, pregnancy outcome, abortion, multiple pregnancy were assessed by the use of Chi square test, fisher’s exact test, t-test and Mann Whitney test were used to compare the two groups and to analyze the statistical findings. RESULTS: No significant statistical differences exist in the number of mature follicle, estradiol and progesterone levels prior to hCG administration and endometrial thickness between the two groups. Estradiol level was lower among those receiving ketoconazole. Ovarian hyperstimulation were present in only three patients receiving placebo (P¼0.1). Three patients in group A and two in group B became pregnant (P¼0.99). No abortion and multiple pregnancy were found. CONCLUSIONS: Ketoconazole may suppress steroid production in resistant PCOS patients undergoing CC and hMG induction. But it has no effects on follicular maturation and OHSS prevention. Supported by: None.
P-198 ¨ LLERIAN HORMONE (AMH) MEASUREMENT: SERUM ANTI-MU INTER-ASSAY AGREEMENT AND TEMPERATURE STABILITY. J. Zhao, S. Y. Ng, B. Rivnay, B. S. Leader. Repromedix Corporation, Woburn, MA; Immunology, Harvard Medical School, Boston, MA. OBJECTIVE: Two different immunoassay systems are currently in use to measure serum AMH, one produced by Diagnostic Systems Laboratories (DSL) and one produced by Immunotech (both Beckman Coulter, Inc companies). Previously, the Immunotech system was more widely used in publications and is currently in use outside the United States. However, the DSL system is currently the only commercially available system in the United States. Limited published data is available which compares the two systems. There were two objectives to this study: 1) better characterize the relationship between the Immunotech and DSL systems; 2) test the stability of serum specimens over 48 hours at different temperatures. DESIGN: ELISA analysis of serum samples. MATERIALS AND METHODS: Objective 1) AMH measurement was performed on samples from serum from 38 different donors using both the Immunotech and DSL AMH assays. Objective 2) Multiple AMH measurements were conducted using the DSL assay and serum from seven different donors. Specimens were separated into different aliquots, the first immediately frozen at 20 C and the subsequent samples frozen after 4, 8, 24, and 48 hours at 4 C and room temperature (22 C). RESULTS: Data from the 38 serum samples comparing the two AMH assays revealed a relationship defined by the line Immunotech ¼ (1.5) DSL þ 0.7 (ng/ml) with good agreement (R2 ¼ 0.98). The mean AMH value ratio of samples incubated for 48 hours at 4 C to samples immediately frozen, was 1.01 with a 95% CI of (0.94–1.08). After 48 hours at 22 C, the mean AMH value ratio to immediately frozen samples was 1.09 with a 95% CI of (0.98–1.20). CONCLUSIONS: Values obtained using the DSL AMH immunoassay can be linearly converted to Immunotech AMH values with some degree of variation. DSL AMH measurement is stable for at least 48 hours at 4 C, with values likely to differ by less than eight percent of values from immediately frozen samples. With samples at room temperature for 48 hours, AMH values
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Abstracts
P-200 ASSESSMENT OF DAY-3 FOLLICLE STIMULATING HORMONE (FSH) LEVELS ON THE BECKMAN COULTER ACCESS2 FOR OVARIAN RESERVE ASSESSMENT IMPROVES IN VITRO FERTILIZATION OUTCOME. B. Combs, C. A. Jacobs, A. L. Davis, T. H. Taylor, Z. P. Nagy, R. J. Straub. IVF Laboratory, Reproductive Biology Associates, Atlanta, GA. OBJECTIVE: Day-3 follicle stimulating hormone (FSH) levels are commonly used to help determine ovarian reserve, stimulation regimen and potential pregnancy outcome in infertile patients seeking treatment via in vitro fertilization (IVF). However, there is a controversy as to what the day-3 FSH cut-off level should be. Therefore, the study objective was to identify the day-3 FSH cut-off value on the Beckman Coulter Access2 for improving the outcome prediction in patients undergoing IVF. DESIGN: Retrospective data analysis of 1298 infertile women undergoing IVF. MATERIALS AND METHODS: FSH was measured on the Access2 (Beckman Coulter, Inc., Chaska, MN) platform. The main endpoints were Day-3 FSH levels, eggs recovered per retrieval and pregnancy rates. RESULTS: Patient age, number of eggs harvested at IVF retrieval and pregnancy rates are presented in the figure below. As expected, day-3 FSH levels reflected the patients’ age, i.e. as a woman reaches and ages above 36 years her respective day-3 FSH level will rise. There was a significant difference in day-3 FSH levels between patients <36 (mean, 5.03; SEM þ0.16 mIU/mL) and those >36 (mean, 6.11; SEM þ0.15 mIU/mL) years. Additionally, as day-3 FSH levels rose, the number of eggs collected at retrieval decreased as did the corresponding pregnancy rate. Regression analysis for day-3 FSH on collecting 10 eggs calculated that the suggested cut-off value for FSH is 12.7 mIU/mL. There was a significant difference (P<0.05) in pregnancy rates between patients with FSH values <12.7 mIU/L (47.1%) and >12.7 mIU/mL (35.0%).
Vol. 88, Suppl 1, September 2007
were likely to differ by less than twenty percent of values from immediately frozen samples. Supported by: DSL and Immunotech systems donated by Beckman Coulter, Inc. Experiments and statistical analysis supported by Repromedix.
P-199 EFFECTS OF KETOCONAZOLE ON OVARIAN RESPONSE IN PATIENTS WITH RESISTANT POLYCYSTIC OVARIAN SYNDROME UNDERGOING OVULATION INDUCTION. M. Sadeghi, B. Hossein Rashidi, E. Sahrokh Tehraninejad, N. Memarpour. Obstetrics and Gynecology, Vali-e-Asr Hospital of Medicine, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran. OBJECTIVE: To assess the efficacy of low dose ketoconazole in addition to CC (clomiphen citrate) and hMG on ovulation induction. DESIGN: A double blind, randomized, clinical trial. MATERIALS AND METHODS: Fifty infertile patients with PCOS who had failed to respond to a daily dose of 150 mg of CC were randomly divided into two equal groups; group A received 50 mg ketoconazole every other day while group B received placebo. All the patients were induced by CC and hMG for controlled ovarian hyperstimulation. Ovarian response to induction, numbers of mature follicles, estradiol and progesterone levels, endometrial thickness, OHSS, pregnancy outcome, abortion, multiple pregnancy were assessed by the use of Chi square test, fisher’s exact test, t-test and Mann Whitney test were used to compare the two groups and to analyze the statistical findings. RESULTS: No significant statistical differences exist in the number of mature follicle, estradiol and progesterone levels prior to hCG administration and endometrial thickness between the two groups. Estradiol level was lower among those receiving ketoconazole. Ovarian hyperstimulation were present in only three patients receiving placebo (P¼0.1). Three patients in group A and two in group B became pregnant (P¼0.99). No abortion and multiple pregnancy were found. CONCLUSIONS: Ketoconazole may suppress steroid production in resistant PCOS patients undergoing CC and hMG induction. But it has no effects on follicular maturation and OHSS prevention. Supported by: None.
P-198 ¨ LLERIAN HORMONE (AMH) MEASUREMENT: SERUM ANTI-MU INTER-ASSAY AGREEMENT AND TEMPERATURE STABILITY. J. Zhao, S. Y. Ng, B. Rivnay, B. S. Leader. Repromedix Corporation, Woburn, MA; Immunology, Harvard Medical School, Boston, MA. OBJECTIVE: Two different immunoassay systems are currently in use to measure serum AMH, one produced by Diagnostic Systems Laboratories (DSL) and one produced by Immunotech (both Beckman Coulter, Inc companies). Previously, the Immunotech system was more widely used in publications and is currently in use outside the United States. However, the DSL system is currently the only commercially available system in the United States. Limited published data is available which compares the two systems. There were two objectives to this study: 1) better characterize the relationship between the Immunotech and DSL systems; 2) test the stability of serum specimens over 48 hours at different temperatures. DESIGN: ELISA analysis of serum samples. MATERIALS AND METHODS: Objective 1) AMH measurement was performed on samples from serum from 38 different donors using both the Immunotech and DSL AMH assays. Objective 2) Multiple AMH measurements were conducted using the DSL assay and serum from seven different donors. Specimens were separated into different aliquots, the first immediately frozen at 20 C and the subsequent samples frozen after 4, 8, 24, and 48 hours at 4 C and room temperature (22 C). RESULTS: Data from the 38 serum samples comparing the two AMH assays revealed a relationship defined by the line Immunotech ¼ (1.5) DSL þ 0.7 (ng/ml) with good agreement (R2 ¼ 0.98). The mean AMH value ratio of samples incubated for 48 hours at 4 C to samples immediately frozen, was 1.01 with a 95% CI of (0.94–1.08). After 48 hours at 22 C, the mean AMH value ratio to immediately frozen samples was 1.09 with a 95% CI of (0.98–1.20). CONCLUSIONS: Values obtained using the DSL AMH immunoassay can be linearly converted to Immunotech AMH values with some degree of variation. DSL AMH measurement is stable for at least 48 hours at 4 C, with values likely to differ by less than eight percent of values from immediately frozen samples. With samples at room temperature for 48 hours, AMH values
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Abstracts
P-200 ASSESSMENT OF DAY-3 FOLLICLE STIMULATING HORMONE (FSH) LEVELS ON THE BECKMAN COULTER ACCESS2 FOR OVARIAN RESERVE ASSESSMENT IMPROVES IN VITRO FERTILIZATION OUTCOME. B. Combs, C. A. Jacobs, A. L. Davis, T. H. Taylor, Z. P. Nagy, R. J. Straub. IVF Laboratory, Reproductive Biology Associates, Atlanta, GA. OBJECTIVE: Day-3 follicle stimulating hormone (FSH) levels are commonly used to help determine ovarian reserve, stimulation regimen and potential pregnancy outcome in infertile patients seeking treatment via in vitro fertilization (IVF). However, there is a controversy as to what the day-3 FSH cut-off level should be. Therefore, the study objective was to identify the day-3 FSH cut-off value on the Beckman Coulter Access2 for improving the outcome prediction in patients undergoing IVF. DESIGN: Retrospective data analysis of 1298 infertile women undergoing IVF. MATERIALS AND METHODS: FSH was measured on the Access2 (Beckman Coulter, Inc., Chaska, MN) platform. The main endpoints were Day-3 FSH levels, eggs recovered per retrieval and pregnancy rates. RESULTS: Patient age, number of eggs harvested at IVF retrieval and pregnancy rates are presented in the figure below. As expected, day-3 FSH levels reflected the patients’ age, i.e. as a woman reaches and ages above 36 years her respective day-3 FSH level will rise. There was a significant difference in day-3 FSH levels between patients <36 (mean, 5.03; SEM þ0.16 mIU/mL) and those >36 (mean, 6.11; SEM þ0.15 mIU/mL) years. Additionally, as day-3 FSH levels rose, the number of eggs collected at retrieval decreased as did the corresponding pregnancy rate. Regression analysis for day-3 FSH on collecting 10 eggs calculated that the suggested cut-off value for FSH is 12.7 mIU/mL. There was a significant difference (P<0.05) in pregnancy rates between patients with FSH values <12.7 mIU/L (47.1%) and >12.7 mIU/mL (35.0%).
Vol. 88, Suppl 1, September 2007