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thawed embryo treatment following cryopreservation of all embryos in cycles with excessive ovarian response
(ET). Multivariable discrete-time survival analyses were performed to assess the association between luteal supplementation and clinical pregnancy (CP), defined by the presence of at least one viable fetus (FH) at 8 weeks. RESULTS: 283 IVF-AH cycles were evaluated. Cycle characteristics were similar among patients receiving AH alone (n ¼ 85) vs. AH with either E2 (n ¼ 107), or E2 þ ASA (n ¼ 91) (Table 1). The CP rates were 24.7% for AH only, 28.0% for AH þ E2, and 31.9% for AH þ E2 þ ASA. The multivariable model (Table 1), revealed that CP was more likely in those patients having supplementation either with E2 (odds ratio (OR) ¼ 1.6, 95% confidence interval (CI) ¼ 0.6–4.6, P¼0.39) or with E2 þ ASA (OR ¼ 2.2, 95% CI 0.8–6.2, P¼0.15), compared to women without luteal support. E2 patients were 60% more likely, while E2þASA patients were twice as likely, to achieve CP than AH only patients, although these results were not statistically significant. TABLE 1. Parameter Age (years) Day 3 FSH (mIU/ml) Oocytes retrieved Peak estradiol Embryos transferred Endometrial Thickness (mm) @HCG Days of Stimulation Total FSH (amps) Total HMG (amps)
AH (N ¼ 85)
AHþE2 (N ¼ 107)
AHþE2þASA (N ¼ 91)
P-value
34.6 2.7 8.5 3.6 13.1 7.4 2025 1042 2.8 1.0 11.5 2.73
34.7 2.7 8.1 3.1 14.4 8.2 2112 92 2.3 0.73 11.5 2.4
35.1 2.5 8.2 3.1 13.9 7.6 2004 962 2.5 0.86 11.1 2.6
0.5 0.9 0.7 0.4 0.4 0.4
11.3 1.9 31.9 17.8 21.3 18.2
11.0 2.0 30.0 14.1 25.9 18.0
11.6 2.0 34.6 18.0 24.4 19.4
0.1 0.2 0.3
CONCLUSIONS: The findings suggest that among women <38 years undergoing IVF-AH, luteal supplementation with E2 appears to be associated with an increased CP rate, which is further enhanced by additional supplementation with ASA. Although these associations failed to reach statistical significance, the wide confidence intervals suggest that our ongoing analyses among a larger population may demonstrate a definitive association. Supported by: None.
P-44 PREGNANCY OUTCOME AFTER FROZEN/THAWED EMBRYO TREATMENT FOLLOWING CRYOPRESERVATION OF ALL EMBRYOS IN CYCLES WITH EXCESSIVE OVARIAN RESPONSE. G. R. Verwoerd, K. Marikinti, T. Mathews, M. C. Macnamee. Bourn Hall Clinic, Bourn Hall LCG, Cambridge, Cambridgeshire, United Kingdom. OBJECTIVE: Cryopreservation of all embryos (freeze all embryos, FAE) is widely performed when the risk of ovarian hyperstimulation syndrome (OHSS) is considered too high to go ahead with fresh embryo transfer. The aim of this study was to compare the clinical pregnancy rate after the first frozen/thawed embryo treatment cycle (FET) following FAE in the context of excessive ovarian response, with our unit’s background pregnancy rates for all fresh and frozen/thawed embryo treatment cycles. DESIGN: Retrospective case-control comparative study in a tertiary Fertility Centre. MATERIALS AND METHODS: Between January 2003 and December 2006, 4725 IVF, ICSI and FET cycles were performed, and FAE was carried out for 110 patients in one hundred and eleven (111) cycles (2.3%). Excessive ovarian response was the indication for FAE in 88 cycles. Criteria for FAE were as follows: R30 oocytes, estradiol >4000 pg/ml on the day of HCG administration, or significant symptoms of OHSS. Subsequently 72 patients underwent at least one FET. RESULTS: The clinical pregnancy rate per treatment cycle for the first FET following FAE, was significantly higher than for all FET cycles performed over this period (33.3%, 24/72 vs. 18.8%, 252/1340, P<0.01), but not significantly different from all fresh treatment cycles over the same period (33.3%, 24/72 vs. 27.9%, 944/3385, P¼0.38). The female age at the time of oocyte retrieval was lower, and the proportion of blastocyst culture treatment cycles was higher in FET cycles after FAE, compared with all FET (Table 1). When cases with <20 oocytes retrieved during the FAE cycle were compared with R20, there was a trend towards a lower pregnancy rate for the first FET following FAE, but this did not reach statistical significance (9.1% vs. 37.7%, P¼0.12).
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Abstracts
TABLE 1. Factors that may affect pregnancy rate among all frozen embryo treatment cycles (FET) compared with FET following freeze all embryos (FAE)
All FET FET following FAE Mean age (years) at time of oocyte retrieval Proportion of blastocyst culture treatment cycles Mean number of embryos transferred
P-value
34.6
32.2
<0.0001
18.0%
37.5%
<0.001
1.82
1.84
Not significant
CONCLUSIONS: When FAE is advised to patients who have evidence of excessive ovarian response, they may expect a similar clinical pregnancy rate after one cycle of FET, as after fresh embryo treatment. However, the retrieval of fewer than 20 oocytes is associated with a lower pregnancy rate. Supported by: None.
P-45 EFFICACY OF L-CARNITINE IN REVERSING THE ANTIPROLIFERATIVE EFFECTS OF TNF-a ON MOUSE EMBRYOS IN VITRO. H. Abdelrazik, A. Agarwal, G. Mansour, S. Gupta, E. Sabanegh, R. Sharma. Reproductive Research Center, Glickman Urological Institute and Department of Obstetrics & Gynecology, Cleveland Clinic, Cleveland, OH. OBJECTIVE: TNF-a levels are elevated in the serum, follicular fluid and peritoneal fluid of infertile women with endometriosis and PCOS. Elevated levels of TNF-a have been reported to restrict inner cell mass proliferation in the mouse blastocyst. This leads to retardation of embryo development, reduced embryo viability and embryonic death. L-Carnitine (LC) has been reported to decrease the elevated serum level of TNF-a in certain tumors and inflammatory diseases. Reports on the role of LC as an anti-TNF-a agent in in vitro culture of embryos are lacking. Our objective was to investigate the role of LC in antagonizing the inhibition of cell proliferation effects of TNF-a on the mouse embryo development. DESIGN: Prospective in vitro experiment. MATERIALS AND METHODS: A total of 100 two cell mouse embryos were divided into 4 groups: group 1: control (HTF media only); groups 2: (TNF-a 500 ng); and group 3–4: TNF-a 500 ng þ LC (0.3 and 0.6 mg/ mL). Results of our pilot study have shown that LC is not embryotoxic at concentrations of 0.3 and 0.6 mg/mL. Embryos were incubated at 37 C in 5% CO2. Assessment of embryo development was done after 72 hours by examining the percentage blastocyst development rate (%BDR). Detection of apoptosis was done after TUNEL staining and by measuring the percentage of damaged blastomeres using confocal microscopy. RESULTS: Comparison of %BDR and apoptosis between control, TNF-a alone, and TNF-a þ Carnitine are shown in the Table. TABLE 1.
a P<0.05 considered significant using Fisher’s exact test for %BDR differences; b P<0.05 considered significant for differences in apoptosis using Kruskal-Wallis or Wilcoxon test; c P differences between TNF-a group and TNF-a þ LC groups.
CONCLUSIONS: TNF-a reduces the blastocyst development rate in mouse embryos. L-Carnitine could reverse the TNF-a induced inhibition in BDR. The lack of any significant effect on DNA damage by TNF-a suggests its antiproliferative effect on developing mouse embryos. Inclusion of L-carnitine in culture media may help in antagonizing the antiproliferative action of TNF-a thereby improving embryo development. Supported by: None.
Vol. 88, Suppl 1, September 2007
AH (N ¼ 85)
AHþE2 (N ¼ 107)
AHþE2þASA (N ¼ 91)
P-value
34.6 2.7 8.5 3.6 13.1 7.4 2025 1042 2.8 1.0 11.5 2.73
34.7 2.7 8.1 3.1 14.4 8.2 2112 92 2.3 0.73 11.5 2.4
35.1 2.5 8.2 3.1 13.9 7.6 2004 962 2.5 0.86 11.1 2.6
0.5 0.9 0.7 0.4 0.4 0.4
11.3 1.9 31.9 17.8 21.3 18.2
11.0 2.0 30.0 14.1 25.9 18.0
11.6 2.0 34.6 18.0 24.4 19.4
0.1 0.2 0.3
CONCLUSIONS: The findings suggest that among women <38 years undergoing IVF-AH, luteal supplementation with E2 appears to be associated with an increased CP rate, which is further enhanced by additional supplementation with ASA. Although these associations failed to reach statistical significance, the wide confidence intervals suggest that our ongoing analyses among a larger population may demonstrate a definitive association. Supported by: None.
P-44 PREGNANCY OUTCOME AFTER FROZEN/THAWED EMBRYO TREATMENT FOLLOWING CRYOPRESERVATION OF ALL EMBRYOS IN CYCLES WITH EXCESSIVE OVARIAN RESPONSE. G. R. Verwoerd, K. Marikinti, T. Mathews, M. C. Macnamee. Bourn Hall Clinic, Bourn Hall LCG, Cambridge, Cambridgeshire, United Kingdom. OBJECTIVE: Cryopreservation of all embryos (freeze all embryos, FAE) is widely performed when the risk of ovarian hyperstimulation syndrome (OHSS) is considered too high to go ahead with fresh embryo transfer. The aim of this study was to compare the clinical pregnancy rate after the first frozen/thawed embryo treatment cycle (FET) following FAE in the context of excessive ovarian response, with our unit’s background pregnancy rates for all fresh and frozen/thawed embryo treatment cycles. DESIGN: Retrospective case-control comparative study in a tertiary Fertility Centre. MATERIALS AND METHODS: Between January 2003 and December 2006, 4725 IVF, ICSI and FET cycles were performed, and FAE was carried out for 110 patients in one hundred and eleven (111) cycles (2.3%). Excessive ovarian response was the indication for FAE in 88 cycles. Criteria for FAE were as follows: R30 oocytes, estradiol >4000 pg/ml on the day of HCG administration, or significant symptoms of OHSS. Subsequently 72 patients underwent at least one FET. RESULTS: The clinical pregnancy rate per treatment cycle for the first FET following FAE, was significantly higher than for all FET cycles performed over this period (33.3%, 24/72 vs. 18.8%, 252/1340, P<0.01), but not significantly different from all fresh treatment cycles over the same period (33.3%, 24/72 vs. 27.9%, 944/3385, P¼0.38). The female age at the time of oocyte retrieval was lower, and the proportion of blastocyst culture treatment cycles was higher in FET cycles after FAE, compared with all FET (Table 1). When cases with <20 oocytes retrieved during the FAE cycle were compared with R20, there was a trend towards a lower pregnancy rate for the first FET following FAE, but this did not reach statistical significance (9.1% vs. 37.7%, P¼0.12).
S124
Abstracts
TABLE 1. Factors that may affect pregnancy rate among all frozen embryo treatment cycles (FET) compared with FET following freeze all embryos (FAE)
All FET FET following FAE Mean age (years) at time of oocyte retrieval Proportion of blastocyst culture treatment cycles Mean number of embryos transferred
P-value
34.6
32.2
<0.0001
18.0%
37.5%
<0.001
1.82
1.84
Not significant
CONCLUSIONS: When FAE is advised to patients who have evidence of excessive ovarian response, they may expect a similar clinical pregnancy rate after one cycle of FET, as after fresh embryo treatment. However, the retrieval of fewer than 20 oocytes is associated with a lower pregnancy rate. Supported by: None.
P-45 EFFICACY OF L-CARNITINE IN REVERSING THE ANTIPROLIFERATIVE EFFECTS OF TNF-a ON MOUSE EMBRYOS IN VITRO. H. Abdelrazik, A. Agarwal, G. Mansour, S. Gupta, E. Sabanegh, R. Sharma. Reproductive Research Center, Glickman Urological Institute and Department of Obstetrics & Gynecology, Cleveland Clinic, Cleveland, OH. OBJECTIVE: TNF-a levels are elevated in the serum, follicular fluid and peritoneal fluid of infertile women with endometriosis and PCOS. Elevated levels of TNF-a have been reported to restrict inner cell mass proliferation in the mouse blastocyst. This leads to retardation of embryo development, reduced embryo viability and embryonic death. L-Carnitine (LC) has been reported to decrease the elevated serum level of TNF-a in certain tumors and inflammatory diseases. Reports on the role of LC as an anti-TNF-a agent in in vitro culture of embryos are lacking. Our objective was to investigate the role of LC in antagonizing the inhibition of cell proliferation effects of TNF-a on the mouse embryo development. DESIGN: Prospective in vitro experiment. MATERIALS AND METHODS: A total of 100 two cell mouse embryos were divided into 4 groups: group 1: control (HTF media only); groups 2: (TNF-a 500 ng); and group 3–4: TNF-a 500 ng þ LC (0.3 and 0.6 mg/ mL). Results of our pilot study have shown that LC is not embryotoxic at concentrations of 0.3 and 0.6 mg/mL. Embryos were incubated at 37 C in 5% CO2. Assessment of embryo development was done after 72 hours by examining the percentage blastocyst development rate (%BDR). Detection of apoptosis was done after TUNEL staining and by measuring the percentage of damaged blastomeres using confocal microscopy. RESULTS: Comparison of %BDR and apoptosis between control, TNF-a alone, and TNF-a þ Carnitine are shown in the Table. TABLE 1.
a P<0.05 considered significant using Fisher’s exact test for %BDR differences; b P<0.05 considered significant for differences in apoptosis using Kruskal-Wallis or Wilcoxon test; c P differences between TNF-a group and TNF-a þ LC groups.
CONCLUSIONS: TNF-a reduces the blastocyst development rate in mouse embryos. L-Carnitine could reverse the TNF-a induced inhibition in BDR. The lack of any significant effect on DNA damage by TNF-a suggests its antiproliferative effect on developing mouse embryos. Inclusion of L-carnitine in culture media may help in antagonizing the antiproliferative action of TNF-a thereby improving embryo development. Supported by: None.
Vol. 88, Suppl 1, September 2007