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Decreased expression of angiogenin in the eutopic endometrium from infertile women with advanced stage endometriosis
(P⬍0.05). In group V basal FSH, and resistance increased (P⬍0.05) and number of oocyte and good quality oocytes ratio decreased (P⬍0.05). CONCLUSION: Sclerotherapy is one of the proper therapeutic modalities performing before IVF-ET cycle in the patients with ovarian endometriomas. Further study on a large scale is necessary to identify effective clinical outcome of sclerotherapy. Supported by: None
P-62 Association between the human alpha 2-Heremans Schmidt glycoprotein (AHSG) polymorphism and endometriosis. J. G. Kim, H. Kim, S.-Y. Ku, S. H. Kim, Y. M. Choi, S. Y. Moon. Seoul National University College of Medicine, Seoul, Republic of Korea. OBJECTIVES: Many studies have been undertaken to identify the candidate genes for endometriosis with the conflicting data in various populations. To date, the genes responsible for development of endometriosis have not been defined. Recently, it has been demonstrated that the the alpha 2-Heremans Schmidt glycoprotein (AHSG) gene is expressed in the endometrium of endometriosis patients, and that endometriosis patients have high levels of AHSG in serum and peritoneal fluid. Genetic polymorphism of AHSG with two common alleles, AHSG 1 and AHSG 2 has been described on isoelectric focusing. The objective was to evaluate the relationship between the AHSG polymorphism and endometriosis. DESIGN: Case-control study. MATERIALS AND METHODS: Genetic polymorphism in AHSG gene was analyzed in seventy-nine women with endometriosis, and one hundred and five women without endometriosis by polymerase chain reactionrestriction fragment length polymorphism at polymorphic NlaIII and SacI sites inside the exon 6 and exon 7 of the AHSG gene. The absence of both restriction sites was designated as the AHSG 1 allele, and the presence of these sites was designated as the AHSG 2 allele. According to the revised American Fertility Society classification, 46 women had early stage endometriosis (stage I and stage II), 33 had late stage endometriosis (stage III, and stage IV). RESULTS: The allele frequencies of AHSG 1 and AHSG 2 were found to be 0.69 and 0.31 respectively. The proportion of noncarriers of the AHSG 2 allele was significantly higher in women with endometriosis than in women without endometriosis (55.7% versus 39.0%). Women not carrying the AHSG 2 allele were found to have 2 times the risk of endometriosis than those carrying at least one copy of this allele. No significant differences were noted in the distribution of the AHSG allele or AHSG genotypes between early stage endometriosis and late stage endometriosis. CONCLUSION: Endometriosis is associated with the AHSG gene polymorphism. Supported by: This work was supported by Korea Research Foundation Grant (KRF-2002– 015-EP0090).
P-63 Endometriosis has no effect on ART results after ICSI procedure. K. Sakhel, M. Khedr, F. N. Shamma, J. Ayers, M. H. Fakih, M. Abuzeid. IVF Michigan and Michigan State University, Rochester Hills, MI. OBJECTIVE: The effects of endometriosis on ART results remain controversial. Many studies suggested reduced fertilization rate after traditional IVF in endometriosis patients. The purpose of this study was to compare ART results after ICSI procedure in patients with endometriosis to those with tubal factors. DESIGN: Retrospective study MATERIALS AND METHODS: Charts of all patients with single diagnosis of endometriosis or tubal factor infertility who underwent ART in the period between 05/2000 –12/2003 were reviewed. Patients were divided into 3 groups: group 1(n⫽81) had stages 1 or 2 endometriosis, group 2 (n⫽40) had stages 3 or 4 endometriosis and group 3 (n⫽52) had tubal factors. Patients underwent similar controlled ovarian stimulation protocols with ICSI and uterine embryo transfer performed in all cycles. RESULTS: The mean age was 34.3 ⫾ 4.5, 32.2 ⫾ 4.3 and 34.9 ⫾ 4.2 years for Group 1, Group 2 and Group 3 respectively. The mean duration of infertility for the 3 groups was 3.4 ⫾ 2.6, 3.5 ⫾ 2.3 and 2.75 ⫾ 2.5 years respectively. No significant difference was found among the 3 groups with
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respect to type of infertility, day3 FSH levels, number of ampules of gonadotropins used and number of follicles ⱖ 16mm on hCG day. There was no significant difference in the number of mature oocytes, fertilization rate, number of zygotes, number of cleaved embryos, cleavage rate, or the number of embryos transferred among the 3 groups. In addition there was no significant difference in the number of grade 1, grade 2 and grade 3 embryos among the 3 groups. Furthermore, there was no significant difference in the pregnancy rate per cycle among the 3 groups (42% for Group 1, 45% for Group 2 and 34.6% for Group 3). CONCLUSION: Our data suggest that endometriosis, irrespective of its stage, does not affect embryo quality or ART results when ICSI procedure is performed. Supported by: None
P-64 Decreased expression of angiogenin in the eutopic endometrium from infertile women with advanced stage endometriosis. S. H. Kim Sr., Y. M. Choi Sr., I.-A. Park Sr., H.-D. Chae Sr., C.-H. Kim Sr., B.-M. Kang Sr. Asan medical center, Seoul, Republic of Korea; Seoul National University Hospital, Seoul, Republic of Korea. OBJECTIVE: Angiogenin, a potent inducer of angiogenesis, is expressed in human endometrium. This study was performed to compare the expression of angiogenin mRNA level in the eutopic endometrium from women with and without endometriosis. DESIGN: Molecular studies in human tissues MATERIALS AND METHODS: Thirty-two women with advanced stage endometriosis and 29 control women were recruited. Following isolation of total RNA from endometrial tissue and reverse transcription, cDNA samples were amplified by real time PCR to quantify the expression of angiogenin and GAPDH genes. In selected patients, immunohistochemistry with an anti-angiogenin monoclonal antibody was utilized to localize the area of angiogenin expression. RESULTS: Angiogenin mRNA level was significantly lower in the endometriosis group than in the control group during the secretory phase (0.72 ⫾ 0.14 vs. 2.21 ⫾ 0.92, mean ⫾ SEM, P⬍0.05), especially the mid-secretory phase (0.55 ⫾ 0.16 vs. 1.69 ⫾ 0.45, P⬍0.05), and the decline was observed only in the women who presented with infertility (0.40 ⫾ 0.15 vs. 1.69 ⫾ 0.45, P⬍0.05). Within the endometriosis group, angiogenin mRNA levels did not differ between the proliferative and secretory phases, but, in the control group, the level in the secretory phase was higher than that during the proliferative phase (0.96 ⫾ 0.28 vs. 2.21 ⫾ 0.92, P⬍0.05). Immunohistochemistry showed that the glandular epithelial cell layer was decorated positively in both groups. CONCLUSION: These findings suggest that the relative deficiency of angiogenin expression in the secretory endometrium could impair implantation in women with advanced stage endometriosis. Supported by: This study was supported by a grant of the Korea Health 21 R&D Project, Ministry of Health & Welfare, Republic of Korea. (01PJ10-PG6 – 01GN13– 0002)
P-65 Toxicity of tumor necrosis factor (TNF)-␣ on human spermatozoa ⴚ Possible role in endometriosis associated infertility. T. M. Said, R. K. Sharma, M. A. Bedaiwy, A. Agarwal, T. Falcone. Cleveland Clinic Foundation, Cleveland, OH; Assuit University, Assuit, Egypt. OBJECTIVE: TNF-␣ is a pleiotropic cytokine with numerous biological functions that include modulation of spermatogenesis and apoptosis in addition to acting as a pro-inflammatory cytokine. Spermatozoa may be exposed to abnormal levels of TNF-␣ in the male reproductive tract or along the passage in the female reproductive tract in cases of endometriosis. The objective of our study was to examine the toxic effects of TNF-␣ on human spermatozoa and to establish the minimum concentration that can affect both the sperm function and genomic integrity. DESIGN: Prospective-controlled study. MATERIALS AND METHODS: Semen samples were collected from 25
Vol. 82, Suppl. 2, September 2004
P-62 Association between the human alpha 2-Heremans Schmidt glycoprotein (AHSG) polymorphism and endometriosis. J. G. Kim, H. Kim, S.-Y. Ku, S. H. Kim, Y. M. Choi, S. Y. Moon. Seoul National University College of Medicine, Seoul, Republic of Korea. OBJECTIVES: Many studies have been undertaken to identify the candidate genes for endometriosis with the conflicting data in various populations. To date, the genes responsible for development of endometriosis have not been defined. Recently, it has been demonstrated that the the alpha 2-Heremans Schmidt glycoprotein (AHSG) gene is expressed in the endometrium of endometriosis patients, and that endometriosis patients have high levels of AHSG in serum and peritoneal fluid. Genetic polymorphism of AHSG with two common alleles, AHSG 1 and AHSG 2 has been described on isoelectric focusing. The objective was to evaluate the relationship between the AHSG polymorphism and endometriosis. DESIGN: Case-control study. MATERIALS AND METHODS: Genetic polymorphism in AHSG gene was analyzed in seventy-nine women with endometriosis, and one hundred and five women without endometriosis by polymerase chain reactionrestriction fragment length polymorphism at polymorphic NlaIII and SacI sites inside the exon 6 and exon 7 of the AHSG gene. The absence of both restriction sites was designated as the AHSG 1 allele, and the presence of these sites was designated as the AHSG 2 allele. According to the revised American Fertility Society classification, 46 women had early stage endometriosis (stage I and stage II), 33 had late stage endometriosis (stage III, and stage IV). RESULTS: The allele frequencies of AHSG 1 and AHSG 2 were found to be 0.69 and 0.31 respectively. The proportion of noncarriers of the AHSG 2 allele was significantly higher in women with endometriosis than in women without endometriosis (55.7% versus 39.0%). Women not carrying the AHSG 2 allele were found to have 2 times the risk of endometriosis than those carrying at least one copy of this allele. No significant differences were noted in the distribution of the AHSG allele or AHSG genotypes between early stage endometriosis and late stage endometriosis. CONCLUSION: Endometriosis is associated with the AHSG gene polymorphism. Supported by: This work was supported by Korea Research Foundation Grant (KRF-2002– 015-EP0090).
P-63 Endometriosis has no effect on ART results after ICSI procedure. K. Sakhel, M. Khedr, F. N. Shamma, J. Ayers, M. H. Fakih, M. Abuzeid. IVF Michigan and Michigan State University, Rochester Hills, MI. OBJECTIVE: The effects of endometriosis on ART results remain controversial. Many studies suggested reduced fertilization rate after traditional IVF in endometriosis patients. The purpose of this study was to compare ART results after ICSI procedure in patients with endometriosis to those with tubal factors. DESIGN: Retrospective study MATERIALS AND METHODS: Charts of all patients with single diagnosis of endometriosis or tubal factor infertility who underwent ART in the period between 05/2000 –12/2003 were reviewed. Patients were divided into 3 groups: group 1(n⫽81) had stages 1 or 2 endometriosis, group 2 (n⫽40) had stages 3 or 4 endometriosis and group 3 (n⫽52) had tubal factors. Patients underwent similar controlled ovarian stimulation protocols with ICSI and uterine embryo transfer performed in all cycles. RESULTS: The mean age was 34.3 ⫾ 4.5, 32.2 ⫾ 4.3 and 34.9 ⫾ 4.2 years for Group 1, Group 2 and Group 3 respectively. The mean duration of infertility for the 3 groups was 3.4 ⫾ 2.6, 3.5 ⫾ 2.3 and 2.75 ⫾ 2.5 years respectively. No significant difference was found among the 3 groups with
S158
respect to type of infertility, day3 FSH levels, number of ampules of gonadotropins used and number of follicles ⱖ 16mm on hCG day. There was no significant difference in the number of mature oocytes, fertilization rate, number of zygotes, number of cleaved embryos, cleavage rate, or the number of embryos transferred among the 3 groups. In addition there was no significant difference in the number of grade 1, grade 2 and grade 3 embryos among the 3 groups. Furthermore, there was no significant difference in the pregnancy rate per cycle among the 3 groups (42% for Group 1, 45% for Group 2 and 34.6% for Group 3). CONCLUSION: Our data suggest that endometriosis, irrespective of its stage, does not affect embryo quality or ART results when ICSI procedure is performed. Supported by: None
P-64 Decreased expression of angiogenin in the eutopic endometrium from infertile women with advanced stage endometriosis. S. H. Kim Sr., Y. M. Choi Sr., I.-A. Park Sr., H.-D. Chae Sr., C.-H. Kim Sr., B.-M. Kang Sr. Asan medical center, Seoul, Republic of Korea; Seoul National University Hospital, Seoul, Republic of Korea. OBJECTIVE: Angiogenin, a potent inducer of angiogenesis, is expressed in human endometrium. This study was performed to compare the expression of angiogenin mRNA level in the eutopic endometrium from women with and without endometriosis. DESIGN: Molecular studies in human tissues MATERIALS AND METHODS: Thirty-two women with advanced stage endometriosis and 29 control women were recruited. Following isolation of total RNA from endometrial tissue and reverse transcription, cDNA samples were amplified by real time PCR to quantify the expression of angiogenin and GAPDH genes. In selected patients, immunohistochemistry with an anti-angiogenin monoclonal antibody was utilized to localize the area of angiogenin expression. RESULTS: Angiogenin mRNA level was significantly lower in the endometriosis group than in the control group during the secretory phase (0.72 ⫾ 0.14 vs. 2.21 ⫾ 0.92, mean ⫾ SEM, P⬍0.05), especially the mid-secretory phase (0.55 ⫾ 0.16 vs. 1.69 ⫾ 0.45, P⬍0.05), and the decline was observed only in the women who presented with infertility (0.40 ⫾ 0.15 vs. 1.69 ⫾ 0.45, P⬍0.05). Within the endometriosis group, angiogenin mRNA levels did not differ between the proliferative and secretory phases, but, in the control group, the level in the secretory phase was higher than that during the proliferative phase (0.96 ⫾ 0.28 vs. 2.21 ⫾ 0.92, P⬍0.05). Immunohistochemistry showed that the glandular epithelial cell layer was decorated positively in both groups. CONCLUSION: These findings suggest that the relative deficiency of angiogenin expression in the secretory endometrium could impair implantation in women with advanced stage endometriosis. Supported by: This study was supported by a grant of the Korea Health 21 R&D Project, Ministry of Health & Welfare, Republic of Korea. (01PJ10-PG6 – 01GN13– 0002)
P-65 Toxicity of tumor necrosis factor (TNF)-␣ on human spermatozoa ⴚ Possible role in endometriosis associated infertility. T. M. Said, R. K. Sharma, M. A. Bedaiwy, A. Agarwal, T. Falcone. Cleveland Clinic Foundation, Cleveland, OH; Assuit University, Assuit, Egypt. OBJECTIVE: TNF-␣ is a pleiotropic cytokine with numerous biological functions that include modulation of spermatogenesis and apoptosis in addition to acting as a pro-inflammatory cytokine. Spermatozoa may be exposed to abnormal levels of TNF-␣ in the male reproductive tract or along the passage in the female reproductive tract in cases of endometriosis. The objective of our study was to examine the toxic effects of TNF-␣ on human spermatozoa and to establish the minimum concentration that can affect both the sperm function and genomic integrity. DESIGN: Prospective-controlled study. MATERIALS AND METHODS: Semen samples were collected from 25
Vol. 82, Suppl. 2, September 2004