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Factors affecting early pregnancy loss following assisted conception
TABLE 1.
Endomedtrium thickness (mm) No. of patients Ages Days of stimulation E2 levels (pmol/l) No. of oocytes recovered No. of MII oocytes No. of 2 PN zygotes No. of 8-16 cells No. of blastocyts No. of embryos transferred % of HCG positives % of gestational pregnancies % of implantation
7.5 (3.2-8.9) 28 32.4 (28-41) 9.9 12959 13.7 (383/28) 12 (336/28) 9.5 (266/28) 7.4 (207/28) 5 (140/28) 2.1 (59/28)
10.4 (9-11.8)
13 (12-17)
P value
60 35 31.4 (24-42) 34 (25-42.2) 9.6 9.3 12451 13581 14.5 (870/60) 14 (490/35) 12 (720/60) 9.3 (558/60) 6.7 (402/60) 5.2 (312/60) 2.1 (126/60)
39.3 (11/28)* 25 (7/28)*
12.3 (430/35) 9.9 (346/35) 6.6 (231/35) 4.6 (161/35) 2.2 (77/35)
75 (45/60) 68.6 (24/35) 60 (36/60) 57.1 (20/35)
0.002 0.003
18.7 (11/59)* 42.9 (54/126) 42.9 (33/77)
0.001
Data with * are statistically different (p<0.05).
OBJECTIVE: To perform assisted oocyte activation (AOA) by calcium ionophore treatment in five patients with fertilization failure or low fertilization rate after intracytoplasmic sperm injection (ICSI) to evaluate fertilization, implantation, pregnancy, miscarriage and ongoing pregnancy rates. DESIGN: Prospective study. MATERIALS AND METHODS: Case 1:32 year-old female, polycystic ovarian syndrome, 3 years of infertility, globozoospermia. 15% fertilization rate in a previous attempt with ICSI.Case 2: 38 year-old patient, primary sterility of 2 years, premature ovarian failure, globozoospermia without any ICSI attempt. Case 3: 36 year-old patient, secondary sterility of 2 years, normal female etiology, severe asthenozoospermia, recurrent implantation failure and low fertilization rate (48,8%). Case 4: 39 yearold patient, primary sterility of 10 years with low fertilization (33,3%) in 2 previous ICSI attempts. Case 5: 35 year-old patient, endometriosis, primary sterility of 10 years due to criptozoospermia. No fertilization was obtained by routine ICSI. After ovarian stimulation, oocyte retrieval and ICSI were undergone in all cases. AOA with calcium ionophore at a concentration of 1 mmol/l was done on half of the oocytes in the two first cases and on all oocytes in the remaining ones. Fertilization, implantation, pregnancy, miscarriage and ongoing pregnancy rates were assessed. Embryo transfer was carried out on day 3 in all cases except in case 4 we transferred blastocysts. RESULTS: The results are:
Supported by: Private fertility center.
P-364 FACTORS AFFECTING EARLY PREGNANCY LOSS FOLLOWING ASSISTED CONCEPTION. H. S. Clarke, R. C. Cutting, T. C. Li. Assisted Conception Unit, Centre for Reproductive Medicine and Fertility, Sheffield, United Kingdom; Centre for Reproductive Medicine and Fertility, Sheffield, United Kingdom.
TABLE 1. Results Patient 1
Patient 2
Patient 3
No. oocytes
27
14
10
14
No. MII
23
11
10
14
7
9
5
10
14
7
No. oocytes þCaI No. fertilized oocytes(%)
5 (55.5)
3 (60)
% Pregnancies/transfer
100
100
Implantation rate (%)
50
50
% Miscarriage Baby at birth
OBJECTIVE: To be able to better advise our patients of their likelihood of a live birth outcome when they have tested positive for beta human chorionic gonadotrophin (bhCG) at 14 days post-egg collection (þ14d). DESIGN: Retrospective analysis of all patients, who had a positive pregnancy test following IVF, was used to determine which factors were most predictive of early pregnancy loss (EPL). MATERIALS AND METHODS: The effect of age, BMI, smoking, previous IVF attempts, previous miscarriages, embryo quality, þ14 level of bhCG and increase in bhCG levels between þ14 and þ21d on pregnancy loss were analysed. Logistic regression analysis of 427 IVF/ICSI cycles was used to determine which of these variables were most predictive of EPL (significance level <0.05). Multiple ongoing pregnancies and live births were excluded. RESULTS: 65% of pregnancies resulted in a live birth/ongoing clinical pregnancy (LB) compared with 35% miscarriage (MC), including biochemical pregnancy. The most important factor in predicting pregnancy loss was the level [meanSD] of bhCG at þ14d (LB 11670; MC 5043), followed by the rise in bhCG between þ14d and þ21d (LB 22.310.6; MC 158.9), the number of previous IVF attempts (LB 0.71.1; MC 1.01.2), age (LB 33.24.2; MC 34.64.7) then BMI (LB 24.33.7;MC 25.14.1). No correlation was observed between smoking nor embryo quality and EPL. CONCLUSIONS: Measurement of bhCG and the rise in bhCG between þ14 and þ21d were the most predictive variables. However, the number if previous IVF attempts, BMI and age were also significantly related to EPL in our patient population. Patients achieving a positive pregnancy test following IVF have heightened anxiety, and the potential risk of early pregnancy loss adds to the patients’ emotional burden. Data from the current study may now be used to lessen this anxiety, as further information can be given to the patient to predict their likelihood of EPL. Supported by: None.
P-365 SUCCESSFUL PREGNANACIES ACHIEVED AFTER CALCIUM IONOPHORE OOCYTE ACTIVATION. M. Molla´ Silva, M. Ojeda Varela, L. Muriel Rios, S. Portela Perez, A. Pellicer, E. Mu~ noz Mu~ noz. IVI Vigo, Vigo, Spain.
FERTILITY & STERILITYÒ
0 Female without malformations
0 Male without malformations
Patient 4 Patient 5 8
7 (70)
11 (78.5) 3 (42.9)
100
100
33.3 0 Female without
on course
50 0 on course
malformations
CONCLUSIONS: Assisted oocyte activation improved fertilization rates in all cases and four pregnancies were achieved up to date. AOA may become a reasonable and efficient treatment in cases of sperm or oocyte- related fertilization failure. The clinical use of ionophores in assisted reproduction is limited by insufficient knowledge about their potential toxic effect on oocytes and embryos. However, healthy children seemed no to be affected by ionophore treatment in our study. More prospective controlled studies should be performed to confirm these results. Supported by: None.
P-366 L- CARNITINE PREVENTS DNA DAMAGE OF OOCYTES INCUBATED IN PERITONEAL FLUID OF ENDOMETRIOSIS. G. Mansour, G. Lotfy, T. Falcone, R. Sharma, A. Agarwal. Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH; Suez Canal University Hospital, Ismailia, Egypt; Cleveland Clinic, Cleveland, OH. OBJECTIVE: Endometriosis affects approximately 14% of all women 30 50% of infertile women, and 84% of women with infertility and pelvic pain. Previous studies by our group demonstrated damage to oocyte cytoskeleton and embryos after exogenous exposure to peritoneal fluid from patients with endometriosis. L-Carnitine (LC) is able to stabilize mitochondrial membranes, increase the supply of energy to the organelle and protect the cell from apoptotic death. The objective was to investigate the protective effect of LC on the oocyte DNA against the toxic effects of peritoneal fluid of patients with endometriosis. DESIGN: Experimental study at Reproductive Research laboratory at an academic hospital. MATERIALS AND METHODS: 480 oocytes were divided into 4 groups Group I: 180 mature mouse oocytes (metaphase II) were incubated in peritoneal fluid (PF) of patients with endometriosis (29 mild endometriosis and 31 severe endometriosis). Group II: 180 mature oocytes incubated with LC 0.6 mg/mL þ PF (1:1 dilution) and group III (control): 30 oocytes added to human tubal fluid (HTF media) and 90 oocytes added to PF of tubal ligation patients. Oocytes in each group were incubated for 30 min, 1.5 h and 4 h. TUNEL staining was done for measuring apoptosis and oocytes were examined under
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Endomedtrium thickness (mm) No. of patients Ages Days of stimulation E2 levels (pmol/l) No. of oocytes recovered No. of MII oocytes No. of 2 PN zygotes No. of 8-16 cells No. of blastocyts No. of embryos transferred % of HCG positives % of gestational pregnancies % of implantation
7.5 (3.2-8.9) 28 32.4 (28-41) 9.9 12959 13.7 (383/28) 12 (336/28) 9.5 (266/28) 7.4 (207/28) 5 (140/28) 2.1 (59/28)
10.4 (9-11.8)
13 (12-17)
P value
60 35 31.4 (24-42) 34 (25-42.2) 9.6 9.3 12451 13581 14.5 (870/60) 14 (490/35) 12 (720/60) 9.3 (558/60) 6.7 (402/60) 5.2 (312/60) 2.1 (126/60)
39.3 (11/28)* 25 (7/28)*
12.3 (430/35) 9.9 (346/35) 6.6 (231/35) 4.6 (161/35) 2.2 (77/35)
75 (45/60) 68.6 (24/35) 60 (36/60) 57.1 (20/35)
0.002 0.003
18.7 (11/59)* 42.9 (54/126) 42.9 (33/77)
0.001
Data with * are statistically different (p<0.05).
OBJECTIVE: To perform assisted oocyte activation (AOA) by calcium ionophore treatment in five patients with fertilization failure or low fertilization rate after intracytoplasmic sperm injection (ICSI) to evaluate fertilization, implantation, pregnancy, miscarriage and ongoing pregnancy rates. DESIGN: Prospective study. MATERIALS AND METHODS: Case 1:32 year-old female, polycystic ovarian syndrome, 3 years of infertility, globozoospermia. 15% fertilization rate in a previous attempt with ICSI.Case 2: 38 year-old patient, primary sterility of 2 years, premature ovarian failure, globozoospermia without any ICSI attempt. Case 3: 36 year-old patient, secondary sterility of 2 years, normal female etiology, severe asthenozoospermia, recurrent implantation failure and low fertilization rate (48,8%). Case 4: 39 yearold patient, primary sterility of 10 years with low fertilization (33,3%) in 2 previous ICSI attempts. Case 5: 35 year-old patient, endometriosis, primary sterility of 10 years due to criptozoospermia. No fertilization was obtained by routine ICSI. After ovarian stimulation, oocyte retrieval and ICSI were undergone in all cases. AOA with calcium ionophore at a concentration of 1 mmol/l was done on half of the oocytes in the two first cases and on all oocytes in the remaining ones. Fertilization, implantation, pregnancy, miscarriage and ongoing pregnancy rates were assessed. Embryo transfer was carried out on day 3 in all cases except in case 4 we transferred blastocysts. RESULTS: The results are:
Supported by: Private fertility center.
P-364 FACTORS AFFECTING EARLY PREGNANCY LOSS FOLLOWING ASSISTED CONCEPTION. H. S. Clarke, R. C. Cutting, T. C. Li. Assisted Conception Unit, Centre for Reproductive Medicine and Fertility, Sheffield, United Kingdom; Centre for Reproductive Medicine and Fertility, Sheffield, United Kingdom.
TABLE 1. Results Patient 1
Patient 2
Patient 3
No. oocytes
27
14
10
14
No. MII
23
11
10
14
7
9
5
10
14
7
No. oocytes þCaI No. fertilized oocytes(%)
5 (55.5)
3 (60)
% Pregnancies/transfer
100
100
Implantation rate (%)
50
50
% Miscarriage Baby at birth
OBJECTIVE: To be able to better advise our patients of their likelihood of a live birth outcome when they have tested positive for beta human chorionic gonadotrophin (bhCG) at 14 days post-egg collection (þ14d). DESIGN: Retrospective analysis of all patients, who had a positive pregnancy test following IVF, was used to determine which factors were most predictive of early pregnancy loss (EPL). MATERIALS AND METHODS: The effect of age, BMI, smoking, previous IVF attempts, previous miscarriages, embryo quality, þ14 level of bhCG and increase in bhCG levels between þ14 and þ21d on pregnancy loss were analysed. Logistic regression analysis of 427 IVF/ICSI cycles was used to determine which of these variables were most predictive of EPL (significance level <0.05). Multiple ongoing pregnancies and live births were excluded. RESULTS: 65% of pregnancies resulted in a live birth/ongoing clinical pregnancy (LB) compared with 35% miscarriage (MC), including biochemical pregnancy. The most important factor in predicting pregnancy loss was the level [meanSD] of bhCG at þ14d (LB 11670; MC 5043), followed by the rise in bhCG between þ14d and þ21d (LB 22.310.6; MC 158.9), the number of previous IVF attempts (LB 0.71.1; MC 1.01.2), age (LB 33.24.2; MC 34.64.7) then BMI (LB 24.33.7;MC 25.14.1). No correlation was observed between smoking nor embryo quality and EPL. CONCLUSIONS: Measurement of bhCG and the rise in bhCG between þ14 and þ21d were the most predictive variables. However, the number if previous IVF attempts, BMI and age were also significantly related to EPL in our patient population. Patients achieving a positive pregnancy test following IVF have heightened anxiety, and the potential risk of early pregnancy loss adds to the patients’ emotional burden. Data from the current study may now be used to lessen this anxiety, as further information can be given to the patient to predict their likelihood of EPL. Supported by: None.
P-365 SUCCESSFUL PREGNANACIES ACHIEVED AFTER CALCIUM IONOPHORE OOCYTE ACTIVATION. M. Molla´ Silva, M. Ojeda Varela, L. Muriel Rios, S. Portela Perez, A. Pellicer, E. Mu~ noz Mu~ noz. IVI Vigo, Vigo, Spain.
FERTILITY & STERILITYÒ
0 Female without malformations
0 Male without malformations
Patient 4 Patient 5 8
7 (70)
11 (78.5) 3 (42.9)
100
100
33.3 0 Female without
on course
50 0 on course
malformations
CONCLUSIONS: Assisted oocyte activation improved fertilization rates in all cases and four pregnancies were achieved up to date. AOA may become a reasonable and efficient treatment in cases of sperm or oocyte- related fertilization failure. The clinical use of ionophores in assisted reproduction is limited by insufficient knowledge about their potential toxic effect on oocytes and embryos. However, healthy children seemed no to be affected by ionophore treatment in our study. More prospective controlled studies should be performed to confirm these results. Supported by: None.
P-366 L- CARNITINE PREVENTS DNA DAMAGE OF OOCYTES INCUBATED IN PERITONEAL FLUID OF ENDOMETRIOSIS. G. Mansour, G. Lotfy, T. Falcone, R. Sharma, A. Agarwal. Center for Reproductive Medicine, Cleveland Clinic, Cleveland, OH; Suez Canal University Hospital, Ismailia, Egypt; Cleveland Clinic, Cleveland, OH. OBJECTIVE: Endometriosis affects approximately 14% of all women 30 50% of infertile women, and 84% of women with infertility and pelvic pain. Previous studies by our group demonstrated damage to oocyte cytoskeleton and embryos after exogenous exposure to peritoneal fluid from patients with endometriosis. L-Carnitine (LC) is able to stabilize mitochondrial membranes, increase the supply of energy to the organelle and protect the cell from apoptotic death. The objective was to investigate the protective effect of LC on the oocyte DNA against the toxic effects of peritoneal fluid of patients with endometriosis. DESIGN: Experimental study at Reproductive Research laboratory at an academic hospital. MATERIALS AND METHODS: 480 oocytes were divided into 4 groups Group I: 180 mature mouse oocytes (metaphase II) were incubated in peritoneal fluid (PF) of patients with endometriosis (29 mild endometriosis and 31 severe endometriosis). Group II: 180 mature oocytes incubated with LC 0.6 mg/mL þ PF (1:1 dilution) and group III (control): 30 oocytes added to human tubal fluid (HTF media) and 90 oocytes added to PF of tubal ligation patients. Oocytes in each group were incubated for 30 min, 1.5 h and 4 h. TUNEL staining was done for measuring apoptosis and oocytes were examined under
S229