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P-431
PR between the BMI groups were determined statistically by Chi Square analysis. p⬍0.05 was considered statistically significant. RESULTS: Significant differences in PR based on BMI were not seen in women undergoing IUI cycles except in the obese group (30-34.9 kg/m2) vs. the morbidly obese (⬎35 kg/m2), p⬍0.05.
37%, 66% and 53% respectively. This inhibition was reversible with the addition of RU486. CONCLUSION: Our results demonstrate that TIMP-2 is down-regulated by progestogens at different levels, and this effect can be reversed by RU486. Thus, not all estrogen/progestins in HRT have the same effect on breast cancer cells. Additional studies are needed to provide best options for HRT use. Supported by: No current extramural financial support for this work. P-432
CONCLUSION: Significant differences in PR were not observed for women in the different BMI groups. However, there was a statistically significant difference between the obese vs. morbidly obese patients which may potentially be related to fertility diagnosis. The morbidly obese group seems to have a higher incidence of anovulation. Therefore, this patient population may be more easily treated with medicated IUI in comparison to the other groups which have a more varied diagnosis. Supported by: None
P-431 PROGESTOGEN INHIBITION OF TISSUE INHIBITOR OF METALLOPROTEINASE (TIMP)-2 IN BREAST CANCER CELLS. M. A. Abdallah, D. D. Taylor, S. T. Nakajima, C. Gercel-Taylor. Pacific Reproductive Center, Torrance, CA; Univ. of Louisville School of Medicine, Louisville, KY. OBJECTIVE: The potential benefit of hormone replacement therapy (HRT) has been superceded by the concern of increased risk in breast cancer incidence. Analysis of the world’s literature concluded that ever users of HRT had an overall risk of breast cancer of 1.14. Current users for 5 or more years had a relative risk of 1.35 and the risk increased with increasing duration of use. The Women’s Health Initiative (WHI) concluded that conjugated equine estrogen combined with progestogen (medroxyprogesterone acetate, MPA) therapy has a small but significant effect on breast cancer risk. Clinical and in vitro studies are needed to understand this association. End-points involving matrix metalloproteinase and inhibitors tissue inhibitors of matrix metalloproteinases (TIMPs) may be used to investigate the molecular mechanisms involved in this process. Our objective in this study was to study effects of the components of hormone replacement therapy on TIMP-2 in breast cancer cells. DESIGN: Breast cancer cells were treated with estradiol (E2), equilin (Eq), norethinderone acetate (NET), medroxyprogesterone acetate (MPA), progesterone (P) alone and in combination. MATERIALS AND METHODS: MCF-7 and T47D cells were used in these studies however TIMP-2 was below detectable levels in MCF-7 cells. These cells are estrogen and progesterone receptor positive. Cells were treated for 48 hours with estrogen (0.1-10.0 nM) and progestogens (0.0110,000 nM). RU486 (100nM) was included as progesterone receptor antagonist as indicated. The Biotrak ELISA assay kits were used to determine the effect of estrogen and progestogens on TIMP-2. Samples were prepared by removing the supernatants from various treated cells, centrifuging to remove any cells or debris, and frozen at -70C. Determinations were performed in triplicate. Specificity of TIMP-2 ELISA is such that free TIMP-2 and that complexed with the active form of MMPs are recognized, but not TIMP-2 complexed with proMMP-2. The sensitivity of the detection system used in our experiments was 3 ng/ml, and allows the detection of free TIMP-2 and those associated with active MMP-2 and not with pro-MMP-2. Data were analyzed by calculating the mean and standard error of the mean (SEM), comparing the treated sample values in all groups to controls using one way ANOVA and Tukey-Kramer post test. RESULTS: In T47-D breast cancer cell lines NET and progesterone alone significantly decreased TIMP-2. This effect was partially reversible with RU486 in the case of NET, and returned to control levels in progesterone treated cells. We demonstrated that equilin induces TIMP-2 in T47D cells (1.13 fold compared to untreated controls). When progestogens were combined with equilin, Eq-NET resulted in decreased TIMP-2 37% which was partially reversed by the addition of RU486. Estradiol/progestogen combinations E2MPA, E2NET, E2P resulted in significant reduction of TIMP-2,
S296
Abstracts
HIGH DOSE VERSUS LOW DOSE PREDNISOLONE ADMINISTRATION IN ROUTINE IVF PATIENTS DOES NOT IMPROVE IMPLANTATION AND PREGNANCY RATES. C. Tempfer, K. Sator, M. Sator, E. Bentz, J. Huber. Univ. of Vienna School of Medicine, Vienna, Austria. OBJECTIVE: A wide variety of factors are thought to influence the success rates of reproductive techniques, among them immunologic factors. The aim of this study was to evaluate wether administration of a high dose corticosteroid regimen (20mg prednisolone daily) during ovarian stimulation and embryo transfer produces a higher implantation and pregnancy rate compared to a low dose corticosteroid regimen (7,5mg prednisolone daily) during ovarian stimulation and embryo transfer. DESIGN: Prospective randomized study. MATERIALS AND METHODS: A total of 132 patients were randomly assigned by a computer generated list to receive either prednisolone 20mg/d (group A) or prednisolone 7,5mg/d (group B) starting on the first day of ovarian stimulation for 4 weeks. RESULTS: The mean age, body mass index, and proportion of women with autoantibodies (anticardiolipin, anti double-strand DNA, anti nuclear antibodies) were comparable between groups. In an intention-to-treat (ITT) analysis comprising all 132 women randomized, the biochemical pregnancy rates were not statistically different between group A and group B (13/64 [64%] versus 19/68 [27,9];p⫽0,4). The clinical pregnancy rates (embryo with recognisable heartbeat) were also not different between group A and group B (9/64 [14,0%] vs 14/68 [20,5%];p⫽0,4). In a secondary analysis of the 99 women who completed the study, the biochemical pregnancy rates were also not statistically different between group A and group B (13/44 [29,5%] versus 19/55 [35,1%];p⫽0,7). The clinical pregnancy rates (embryo with recognisable heartbeat) were also not statistically significant different between group A and group B (9/44 [20,4%] versus 14/54[25,9]; p⫽ 0,6). Secondary outcome parameters (side effects, proportion of lowresponders, endometrial thickness, number of oocytes, number of embryos) were also not different between groups. CONCLUSION: High-dose compared to low-dose prednisolone administration during ovarian stimulation and embryo transfer in women undergoing in vitro fertilisation-embryo transfer dies not improve implantation and pregnancy rates. Supported by: None P-433 DOES IMPAIRED THYROID FUNCTION OR ANTITHYROID ANTIBODIES INFLUENCE SUCCESS RATES IN ASSISTED REPRODUCTIVE TECHNOLOGIES? A. Akdogan, E. G. Tavmergen, M. Ulukus, C. Barut, R. Levi, E. Tavmergen. Ege Univ. Family Planning and Infertility Research and Treatment Center, Izmir, Turkey; Ege Univ. Medical Faculty, Dept. of Obstetrics and Gynecology, Izmir, Turkey. OBJECTIVE: The aim of this study was to evaluate thyroid function and antithyroid antibodies in women undergoing assisted reproductive technology and compare pregnancy outcomes. DESIGN: Retrospective study in a university hospital. MATERIALS AND METHODS: One hundred and fifty-one women treated by ART cycles were included in the study. Previous medical history, age, infertility duration and infertility etiology, smoking , basal FSH, basal LH, basal E2, basal prolactine levels were evaluated. TSH, free T3, free T4. Antithyroglobulin antibodies (antiTG) and antithyroid peroxidase antibodies (antiTPO) were analysied in 90 patients. The number of total retrivied oocytes, MII oocytes and number of transfered embrios, endometrial thickness and pregnancy outcomes were evaluated. Data were analysied by spss 10.0.
Vol. 86, Suppl 2, September 2006
37%, 66% and 53% respectively. This inhibition was reversible with the addition of RU486. CONCLUSION: Our results demonstrate that TIMP-2 is down-regulated by progestogens at different levels, and this effect can be reversed by RU486. Thus, not all estrogen/progestins in HRT have the same effect on breast cancer cells. Additional studies are needed to provide best options for HRT use. Supported by: No current extramural financial support for this work. P-432
CONCLUSION: Significant differences in PR were not observed for women in the different BMI groups. However, there was a statistically significant difference between the obese vs. morbidly obese patients which may potentially be related to fertility diagnosis. The morbidly obese group seems to have a higher incidence of anovulation. Therefore, this patient population may be more easily treated with medicated IUI in comparison to the other groups which have a more varied diagnosis. Supported by: None
P-431 PROGESTOGEN INHIBITION OF TISSUE INHIBITOR OF METALLOPROTEINASE (TIMP)-2 IN BREAST CANCER CELLS. M. A. Abdallah, D. D. Taylor, S. T. Nakajima, C. Gercel-Taylor. Pacific Reproductive Center, Torrance, CA; Univ. of Louisville School of Medicine, Louisville, KY. OBJECTIVE: The potential benefit of hormone replacement therapy (HRT) has been superceded by the concern of increased risk in breast cancer incidence. Analysis of the world’s literature concluded that ever users of HRT had an overall risk of breast cancer of 1.14. Current users for 5 or more years had a relative risk of 1.35 and the risk increased with increasing duration of use. The Women’s Health Initiative (WHI) concluded that conjugated equine estrogen combined with progestogen (medroxyprogesterone acetate, MPA) therapy has a small but significant effect on breast cancer risk. Clinical and in vitro studies are needed to understand this association. End-points involving matrix metalloproteinase and inhibitors tissue inhibitors of matrix metalloproteinases (TIMPs) may be used to investigate the molecular mechanisms involved in this process. Our objective in this study was to study effects of the components of hormone replacement therapy on TIMP-2 in breast cancer cells. DESIGN: Breast cancer cells were treated with estradiol (E2), equilin (Eq), norethinderone acetate (NET), medroxyprogesterone acetate (MPA), progesterone (P) alone and in combination. MATERIALS AND METHODS: MCF-7 and T47D cells were used in these studies however TIMP-2 was below detectable levels in MCF-7 cells. These cells are estrogen and progesterone receptor positive. Cells were treated for 48 hours with estrogen (0.1-10.0 nM) and progestogens (0.0110,000 nM). RU486 (100nM) was included as progesterone receptor antagonist as indicated. The Biotrak ELISA assay kits were used to determine the effect of estrogen and progestogens on TIMP-2. Samples were prepared by removing the supernatants from various treated cells, centrifuging to remove any cells or debris, and frozen at -70C. Determinations were performed in triplicate. Specificity of TIMP-2 ELISA is such that free TIMP-2 and that complexed with the active form of MMPs are recognized, but not TIMP-2 complexed with proMMP-2. The sensitivity of the detection system used in our experiments was 3 ng/ml, and allows the detection of free TIMP-2 and those associated with active MMP-2 and not with pro-MMP-2. Data were analyzed by calculating the mean and standard error of the mean (SEM), comparing the treated sample values in all groups to controls using one way ANOVA and Tukey-Kramer post test. RESULTS: In T47-D breast cancer cell lines NET and progesterone alone significantly decreased TIMP-2. This effect was partially reversible with RU486 in the case of NET, and returned to control levels in progesterone treated cells. We demonstrated that equilin induces TIMP-2 in T47D cells (1.13 fold compared to untreated controls). When progestogens were combined with equilin, Eq-NET resulted in decreased TIMP-2 37% which was partially reversed by the addition of RU486. Estradiol/progestogen combinations E2MPA, E2NET, E2P resulted in significant reduction of TIMP-2,
S296
Abstracts
HIGH DOSE VERSUS LOW DOSE PREDNISOLONE ADMINISTRATION IN ROUTINE IVF PATIENTS DOES NOT IMPROVE IMPLANTATION AND PREGNANCY RATES. C. Tempfer, K. Sator, M. Sator, E. Bentz, J. Huber. Univ. of Vienna School of Medicine, Vienna, Austria. OBJECTIVE: A wide variety of factors are thought to influence the success rates of reproductive techniques, among them immunologic factors. The aim of this study was to evaluate wether administration of a high dose corticosteroid regimen (20mg prednisolone daily) during ovarian stimulation and embryo transfer produces a higher implantation and pregnancy rate compared to a low dose corticosteroid regimen (7,5mg prednisolone daily) during ovarian stimulation and embryo transfer. DESIGN: Prospective randomized study. MATERIALS AND METHODS: A total of 132 patients were randomly assigned by a computer generated list to receive either prednisolone 20mg/d (group A) or prednisolone 7,5mg/d (group B) starting on the first day of ovarian stimulation for 4 weeks. RESULTS: The mean age, body mass index, and proportion of women with autoantibodies (anticardiolipin, anti double-strand DNA, anti nuclear antibodies) were comparable between groups. In an intention-to-treat (ITT) analysis comprising all 132 women randomized, the biochemical pregnancy rates were not statistically different between group A and group B (13/64 [64%] versus 19/68 [27,9];p⫽0,4). The clinical pregnancy rates (embryo with recognisable heartbeat) were also not different between group A and group B (9/64 [14,0%] vs 14/68 [20,5%];p⫽0,4). In a secondary analysis of the 99 women who completed the study, the biochemical pregnancy rates were also not statistically different between group A and group B (13/44 [29,5%] versus 19/55 [35,1%];p⫽0,7). The clinical pregnancy rates (embryo with recognisable heartbeat) were also not statistically significant different between group A and group B (9/44 [20,4%] versus 14/54[25,9]; p⫽ 0,6). Secondary outcome parameters (side effects, proportion of lowresponders, endometrial thickness, number of oocytes, number of embryos) were also not different between groups. CONCLUSION: High-dose compared to low-dose prednisolone administration during ovarian stimulation and embryo transfer in women undergoing in vitro fertilisation-embryo transfer dies not improve implantation and pregnancy rates. Supported by: None P-433 DOES IMPAIRED THYROID FUNCTION OR ANTITHYROID ANTIBODIES INFLUENCE SUCCESS RATES IN ASSISTED REPRODUCTIVE TECHNOLOGIES? A. Akdogan, E. G. Tavmergen, M. Ulukus, C. Barut, R. Levi, E. Tavmergen. Ege Univ. Family Planning and Infertility Research and Treatment Center, Izmir, Turkey; Ege Univ. Medical Faculty, Dept. of Obstetrics and Gynecology, Izmir, Turkey. OBJECTIVE: The aim of this study was to evaluate thyroid function and antithyroid antibodies in women undergoing assisted reproductive technology and compare pregnancy outcomes. DESIGN: Retrospective study in a university hospital. MATERIALS AND METHODS: One hundred and fifty-one women treated by ART cycles were included in the study. Previous medical history, age, infertility duration and infertility etiology, smoking , basal FSH, basal LH, basal E2, basal prolactine levels were evaluated. TSH, free T3, free T4. Antithyroglobulin antibodies (antiTG) and antithyroid peroxidase antibodies (antiTPO) were analysied in 90 patients. The number of total retrivied oocytes, MII oocytes and number of transfered embrios, endometrial thickness and pregnancy outcomes were evaluated. Data were analysied by spss 10.0.
Vol. 86, Suppl 2, September 2006