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Endocrine Influences on Normal and Neoplastic Ovarian Surface Epithelium (OSE) Cell Growth
and further suggest an active role of InsP3 and its receptor in triggering the Ca21 rise and secondary activation phenomena at fertilization. (Support: Grant awarded by the University of Ghent, Belgium).
Monday, October 23, 2000 4:45 P.M. O-012 Endocrine Influences on Normal and Neoplastic Ovarian Surface Epithelium (OSE) Cell Growth. K.-C. Choi, S. K. Kang, C.-J. Tai, N. Auersperg, P. C. K. Leung. Dept. OBS & GYN, Faculty of Medicine, University of British Columbia, Vancouver, British Columbia, Canada Objectives: Over 85% of human ovarian carcinomas are thought to arise in the OSE, i.e. the mesothelium that covers the ovary. Increasing evidence suggests that endocrine factors play an important role in ovarian tumorigenesis. In the present study, the roles of follicle-stimulating hormone (FSH), estradiol (E2) and gonadotropin-releasing hormone (GnRH) were investigated using normal and neoplastic OSE cell lines directly generated from human OSE. Design: The expression of receptors for and growth modulatory effects of FSH, E2 and GnRH were investigated. Materials and Methods: The presence of receptors of the above-mentioned factors was investigated using quantitative RT-PCR, Southern blot and immunoblot analysis. The cells were treated with various concentrations of FSH, E2, GnRH and TGF-b, and cell proliferative index was measured using a [3H]thymidine incorporation assay. Cell types included normal OSE, IOSE cell lines (SV40 Tag immortalized OSE) and an established ovarian cancer cell line (OVCAR-3). Statistical differences were considered significant at p,0.05. Results: DNA fragments for FSH receptor (FSHR) and GnRH receptor (GnRHR) were obtained from normal and neoplastic OSE cells. The PCR products were confirmed by Southern blot analysis and their nucleotide sequences were determined to be 100% homology to the published sequences. Recombinant FSH stimulated the growth of normal and neoplastic OSE cells. In contrast, a GnRH agonist, (D-Ala6)-GnRH, like transforming growth factor-b (TGF-b), significantly inhibited the growth of normal and neoplastic OSE cells in a dose-dependent manner. The mRNA and protein expressions of estrogen receptors (ER-a and ER-b) were observed in normal OSE, IOSE cell lines, and OVCAR-3 cells. E2 did not affect cell growth in normal OSE or IOSE-29 cells, but significantly stimulated the growth of neoplastic OSE cells, i.e. IOSE-29EC and OVCAR-3 cells. The co-treatment with tamoxifen, an E2 antagonist, blocked the growth stimulatory effect of E2. Conclusion: Presence of receptors of FSH, GnRH and E2 has been demonstrated in normal OSE, IOSE cell lines and OVCAR-3. FSH stimulates, while GnRH and TGF-b inhibit, the growth of normal and neoplastic OSE cells. On the other hand, E2 stimulates the growth of neoplastic OSE but not in normal OSE cells. These results suggest that these factors may play a role in ovarian tumorigenesis. This study was supported by the MRC of Canada.
NURSES IN REPRODUCTIVE MEDICINE PROFESSIONAL GROUP Monday, October 23, 2000 2:00 P.M.
lope were mailed to the 431 members of the Nurses’ Professional Group of the American Society for Reproductive Medicine. Statistical analyses were performed with chi-square analysis and Fisher’s exact test, as appropriate, using the SAS-PC statistical package (SAS Institute, Cary, NC). Results: A total of 228 completed questionnaires (52.9%) were returned from 39 states in the U.S. and 6 from outside the U.S. Respondents indicated that approximately 78% of prescribed gonadotropins were administered SQ. Sixty-nine percent of nurses who administer both urinary (UR) and recombinant (REC) gonadotropins combine them in a single injection; of these, 67% did not specify which diluent to use for reconstitution. SQ injections were administered at a 90° angle by 82.4% of nurses, with a 45° angle used by 15.3%. The preferred SQ injection site was the abdomen (28.6%), anterior thigh (27.7%), and the upper arm (0.4%); rotation of injection sites was recommended by 42.4%. Management of injection site reactions included ice pack to the site (27.8%), expectant management (24.1%), and warm compresses (21.3%). The majority of nurses (51.5%) observed that injection site reactions lessened with continued SQ injections, while only 13.1% noted that the reactions worsened. Among nurses who combined UR and REC gonadotropins, 58% reported resolution of local reactions with continued injections using the diluent supplied with the REC gonadotropins; only 11% of nurses who used the UR gonadotropin diluent observed resolution of injection site reactions (P50.022). There was no association between an improvement in injection site reaction with the injection site (P50.155), angle of administration (P50.852), recommended needle length (P50.889), administration of UR and REC gonadotropins in a single injection, or the length of time spent giving injection instructions. Most commonly, the primary format for injection instructions was one-on-one (73.7%) compared to group classes (23.2%). Interestingly, more than half (53%) the nurses spent $30 minutes per couple for injection instructions, while the remainder (46%) spent ,30 minutes. Conclusions: Injection techniques and instructions vary widely among nurses. Most injection site reactions improve over time; however, resolution seems to occur less frequently when the diluent supplied with the urinary gonadotropin is used for combined urinary/recombinant administration.
Monday, October 23, 2000 2:15 P.M. O-014 Predictive Value of hCG Levels 14 Days After Blastocyst Transfer. B. Guth, E. S. Critser, M. A. Henry, W. L. Gentry. Advanced Fertility Institute, Indianapolis, IN. Objectives: This study was designed to evaluate predictive value of hCG levels 14 days after blastocyst transfer (ET). Design: To compare 14 day post-ET serum quantitative hCG levels to ongoing pregnancies, as well as multiple pregnancies associated with day 5 blastocyst transfer, performed during 1999. Materials and Methods: Patients undergoing in vitro fertilization (IVF) and ET were monitored by serum quantitative hCG levels 14 days after ET (n5179). If positive, serial values of hCG were obtained and transvaginal ultrasound was performed 3 weeks after ET, and weekly, until fetal cardiac activity was seen (IUP with 1FCA). Results: One hundred one (101/179; 56%) patients had positive serum quantitative hCG levels 14 days post-ET; 7/101 (7%) were biochemical and 94/101 (93%) had an intrauterine pregnancy (IUP), with a spontaneous miscarriage rate of 6/94 (6%). There were no ongoing pregnancies if the hCG level was ,100. Fetal Cardiac Activity— 6 weeks
O-013 Injection Techniques for Subcutaneous (SQ) Gonadotropins: Results of a North American Survey. K. R. Hammond, M. P. Steinkampf. Dept. Ob/ Gyn, The University of Alabama at Birmingham, Birmingham, AL. Objectives: Gonadotropins are increasingly administered subcutaneously (SQ) whether recombinant or derived from urinary products. However, injection procedures and patient instructions have not been standardized. The purpose of this study was to survey infertility nurses to ascertain information regarding specific injection techniques. Design: An eighteen question, two page questionnaire. Materials and Methods: The questionnaire and a stamped, return enve-
FERTILITY & STERILITYt
HCG level ,100 ,300 ,600 ,900 ,1500 $1500
Positive BiohCG chemical 4 18 21 23 15 20 101
3 2 2 0 0 0 7 (7%)
IUP
0
1 1 16 1 19 1 23 3 15 0 20 0 94 (93%) 6 (6%)
1
2
14 (88%) 1 (6%) 15 (79%) 2 (10%) 12 (52%) 7 (30%) 9 (60%) 3 (20%) 4 (20%) 11 (55%) 54 (57%) 24 (26%)
3
1 (5%) 1 (4%) 3 (20%) 4 (15%) 9 (10%)
S5
Monday, October 23, 2000 4:45 P.M. O-012 Endocrine Influences on Normal and Neoplastic Ovarian Surface Epithelium (OSE) Cell Growth. K.-C. Choi, S. K. Kang, C.-J. Tai, N. Auersperg, P. C. K. Leung. Dept. OBS & GYN, Faculty of Medicine, University of British Columbia, Vancouver, British Columbia, Canada Objectives: Over 85% of human ovarian carcinomas are thought to arise in the OSE, i.e. the mesothelium that covers the ovary. Increasing evidence suggests that endocrine factors play an important role in ovarian tumorigenesis. In the present study, the roles of follicle-stimulating hormone (FSH), estradiol (E2) and gonadotropin-releasing hormone (GnRH) were investigated using normal and neoplastic OSE cell lines directly generated from human OSE. Design: The expression of receptors for and growth modulatory effects of FSH, E2 and GnRH were investigated. Materials and Methods: The presence of receptors of the above-mentioned factors was investigated using quantitative RT-PCR, Southern blot and immunoblot analysis. The cells were treated with various concentrations of FSH, E2, GnRH and TGF-b, and cell proliferative index was measured using a [3H]thymidine incorporation assay. Cell types included normal OSE, IOSE cell lines (SV40 Tag immortalized OSE) and an established ovarian cancer cell line (OVCAR-3). Statistical differences were considered significant at p,0.05. Results: DNA fragments for FSH receptor (FSHR) and GnRH receptor (GnRHR) were obtained from normal and neoplastic OSE cells. The PCR products were confirmed by Southern blot analysis and their nucleotide sequences were determined to be 100% homology to the published sequences. Recombinant FSH stimulated the growth of normal and neoplastic OSE cells. In contrast, a GnRH agonist, (D-Ala6)-GnRH, like transforming growth factor-b (TGF-b), significantly inhibited the growth of normal and neoplastic OSE cells in a dose-dependent manner. The mRNA and protein expressions of estrogen receptors (ER-a and ER-b) were observed in normal OSE, IOSE cell lines, and OVCAR-3 cells. E2 did not affect cell growth in normal OSE or IOSE-29 cells, but significantly stimulated the growth of neoplastic OSE cells, i.e. IOSE-29EC and OVCAR-3 cells. The co-treatment with tamoxifen, an E2 antagonist, blocked the growth stimulatory effect of E2. Conclusion: Presence of receptors of FSH, GnRH and E2 has been demonstrated in normal OSE, IOSE cell lines and OVCAR-3. FSH stimulates, while GnRH and TGF-b inhibit, the growth of normal and neoplastic OSE cells. On the other hand, E2 stimulates the growth of neoplastic OSE but not in normal OSE cells. These results suggest that these factors may play a role in ovarian tumorigenesis. This study was supported by the MRC of Canada.
NURSES IN REPRODUCTIVE MEDICINE PROFESSIONAL GROUP Monday, October 23, 2000 2:00 P.M.
lope were mailed to the 431 members of the Nurses’ Professional Group of the American Society for Reproductive Medicine. Statistical analyses were performed with chi-square analysis and Fisher’s exact test, as appropriate, using the SAS-PC statistical package (SAS Institute, Cary, NC). Results: A total of 228 completed questionnaires (52.9%) were returned from 39 states in the U.S. and 6 from outside the U.S. Respondents indicated that approximately 78% of prescribed gonadotropins were administered SQ. Sixty-nine percent of nurses who administer both urinary (UR) and recombinant (REC) gonadotropins combine them in a single injection; of these, 67% did not specify which diluent to use for reconstitution. SQ injections were administered at a 90° angle by 82.4% of nurses, with a 45° angle used by 15.3%. The preferred SQ injection site was the abdomen (28.6%), anterior thigh (27.7%), and the upper arm (0.4%); rotation of injection sites was recommended by 42.4%. Management of injection site reactions included ice pack to the site (27.8%), expectant management (24.1%), and warm compresses (21.3%). The majority of nurses (51.5%) observed that injection site reactions lessened with continued SQ injections, while only 13.1% noted that the reactions worsened. Among nurses who combined UR and REC gonadotropins, 58% reported resolution of local reactions with continued injections using the diluent supplied with the REC gonadotropins; only 11% of nurses who used the UR gonadotropin diluent observed resolution of injection site reactions (P50.022). There was no association between an improvement in injection site reaction with the injection site (P50.155), angle of administration (P50.852), recommended needle length (P50.889), administration of UR and REC gonadotropins in a single injection, or the length of time spent giving injection instructions. Most commonly, the primary format for injection instructions was one-on-one (73.7%) compared to group classes (23.2%). Interestingly, more than half (53%) the nurses spent $30 minutes per couple for injection instructions, while the remainder (46%) spent ,30 minutes. Conclusions: Injection techniques and instructions vary widely among nurses. Most injection site reactions improve over time; however, resolution seems to occur less frequently when the diluent supplied with the urinary gonadotropin is used for combined urinary/recombinant administration.
Monday, October 23, 2000 2:15 P.M. O-014 Predictive Value of hCG Levels 14 Days After Blastocyst Transfer. B. Guth, E. S. Critser, M. A. Henry, W. L. Gentry. Advanced Fertility Institute, Indianapolis, IN. Objectives: This study was designed to evaluate predictive value of hCG levels 14 days after blastocyst transfer (ET). Design: To compare 14 day post-ET serum quantitative hCG levels to ongoing pregnancies, as well as multiple pregnancies associated with day 5 blastocyst transfer, performed during 1999. Materials and Methods: Patients undergoing in vitro fertilization (IVF) and ET were monitored by serum quantitative hCG levels 14 days after ET (n5179). If positive, serial values of hCG were obtained and transvaginal ultrasound was performed 3 weeks after ET, and weekly, until fetal cardiac activity was seen (IUP with 1FCA). Results: One hundred one (101/179; 56%) patients had positive serum quantitative hCG levels 14 days post-ET; 7/101 (7%) were biochemical and 94/101 (93%) had an intrauterine pregnancy (IUP), with a spontaneous miscarriage rate of 6/94 (6%). There were no ongoing pregnancies if the hCG level was ,100. Fetal Cardiac Activity— 6 weeks
O-013 Injection Techniques for Subcutaneous (SQ) Gonadotropins: Results of a North American Survey. K. R. Hammond, M. P. Steinkampf. Dept. Ob/ Gyn, The University of Alabama at Birmingham, Birmingham, AL. Objectives: Gonadotropins are increasingly administered subcutaneously (SQ) whether recombinant or derived from urinary products. However, injection procedures and patient instructions have not been standardized. The purpose of this study was to survey infertility nurses to ascertain information regarding specific injection techniques. Design: An eighteen question, two page questionnaire. Materials and Methods: The questionnaire and a stamped, return enve-
FERTILITY & STERILITYt
HCG level ,100 ,300 ,600 ,900 ,1500 $1500
Positive BiohCG chemical 4 18 21 23 15 20 101
3 2 2 0 0 0 7 (7%)
IUP
0
1 1 16 1 19 1 23 3 15 0 20 0 94 (93%) 6 (6%)
1
2
14 (88%) 1 (6%) 15 (79%) 2 (10%) 12 (52%) 7 (30%) 9 (60%) 3 (20%) 4 (20%) 11 (55%) 54 (57%) 24 (26%)
3
1 (5%) 1 (4%) 3 (20%) 4 (15%) 9 (10%)
S5