- Email: [email protected]
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Monday, October 23, 2006 4:15 pm O-76 TAURINE TRANSPORTER (TAUT), A NOVEL TARGET OF SODIUM BUTYRATE IN SUPRESSION OF AROMATASE EXPRESSION IN STROMAL CELLS DERIVED FROM ENDOMETRIOSIS. E. Attar, V. Fenkci, M. B. Yilmaz, M. Demura, H. Utsunomiya, S. E. Bulun. Northwestern Univ, Feinberg Medical School, Chicago, IL. OBJECTIVE: Endometriosis is a common estrogen-dependent disorder characterized by the presence of uterine endometrial tissue outside of the normal location. Aberrant activation of aromatase promoter II region is responsible for the excessive production of aromatase enzyme activity and estrogen synthesis in endometriosis. We have previously shown that NaBu has a promoter-specific inhibitory effect on aromatase expression in endomeriotic cells. This effect is mediated, at least in part, via binding of transcription factors. Now, we propose a novel molecular mechanism that taurine transpoter (TauT) mediates NaBu induced aromatase inhibition in endometriotic cells. DESIGN: Prospective experimental study. MATERIALS AND METHODS: Human endometriotic tissue samples were obtained at the time of surgery from women undergoing laparascopy. Primary human endometriotic sromal cell cultures were generated from endometrioma cyst walls. Experiments were conducted when cells reached 80% confluency. At confluence, the cells were placed in serum-free medium for 24 h to wash out the effects of serum and then maintained in serum-free DMEM/F-12 containing NaBu and taurine for varying intervals and doses. Five g of DNase I-treated total RNA were used for reverse transcription (RT) reaction. Five l of the RT mix, 5⬘-end sense primer and 3⬘-end antisense primer complementary to coding exons were used to amplify the coding region of the genes studied. The aromatase activity was measured by [3H]-water release assay directly on endometriotic cell cultures. RESULTS: NaBu decreased dibutyryl (Bt2) cAMP induction of aromatase mRNA expression and enzyme activity in endometriotic cells. Instead, taurine increased aromatase mRNA expression and enzyme activity at a concentration of 0.5 mM/mL in these cells. Moreover, NaBu strikingly decreased the TauT mRNA expression in a concentration dependent manner in human endometriotic cells. Maximal effect of NaBu on TauT expression was observed at the concentration of 10 mM/mL. This effect was reversible at higher concentrations. CONCLUSION: Our results indicate that TauT is a target of NaBu and estrogen synthesis may be regulated by decreasing taurine concentrations in endometriotic cells. Our findings are especially important in that they do not only elucidate the mechanism of NaBu action, but also demonstrate a new pathway in aromatase gene activation and estrogen synthesis. We predict that these findings will make an important impact on understanding and the future treatment strategies of endometriosis. Moreover, relationship between the taurine and estrogen synthesis in endometriosis may also be important in defining new environmental factors in endometriosis development. Supported by: NIH grant HD 36891 and a grant from Friends of Prentice.
Monday, October 23, 2006 4:30 pm O-77 INHIBITION OF CD44 N-LINKED GLYCOSYLATION INHIBITS ENDOMETRIAL CELL BINDING TO PERITONEAL MESOTHELIAL CELLS. A. S. Nair, C. A. Witz, H. B. Nair, R. R. Tekmal, R. S. Schenken. Univ of Texas Health Science Center at San Antonio, San Antonio, TX. OBJECTIVE: CD44 is a transmembrane glycoprotein that is the major ligand for hyaluronan. We have previously demonstrated that abrogation of peritoneal mesothelial cell-associated hyaluronan inhibits endometrial cell (EC) binding to peritoneal mesothelial cells (PMCs), suggesting that CD44/ hyaluronan is involved in the early development of the endometriotic lesion. Other studies using ovarian cancer cell lines have demonstrated that CD44 binding to hyaluronan is regulated in part by the degree of CD44 glycosyl-
FERTILITY & STERILITY威
ation. Here, we assessed the effect of EC CD44 glycosylation inhibition on EC binding to PMCs. DESIGN: In vitro study. MATERIALS AND METHODS: EM- 42 cells, an endometrial epithelioid cell line, were grown to near confluence and treated with the N-linked glycosylation inhibitor, tunicamycin (TUNICA) (5 mcg/ml) for 24 hours. Cell viability was assessed in TUNICA-treated and -untreated EM42 cells using propidium iodide (PI) staining and flow cytometry. The binding of the fluorescence-conjugated lectin, Ricinus communis agglutinin-1 (RCA), to TUNICA-treated and -untreated EM42 cells was used to assess the degree of glycosylation inhibition. Western blots analysis of TUNICA-treated and -untreated EM42 cell lysates were carried out using the anti-CD44 antibody 5F12. The effect of TUNICA on the density of cell surface CD44 was also determined using flow cytometry with the anti-CD44 antibody 5F12. Binding of TUNICA-treated and -untreated EM42 cells to the PMC line LP9 was compared at one hour of co-culture using an established in vitro assay [1]. RESULTS: Tunicamycin did not affect cell viability as assessed by PI labeling. TUNICA-treated EM-42 cells demonstrated a 19% decreased rate of RCA lectin binding, suggesting significant glycosylation inhibition. Western blot analysis and flow cytometry using the 5F12 antibody confirmed that TUNICA treatment did not decrease the expression of cell surface CD44. TUNICA treatment of EM42 cells lead to a 20% decreased rate of binding to PMCs (37% vs. 46%, p ⬍ .004). CONCLUSION: The present study demonstrates that inhibition of EC N-linked glycosylation decreases EC binding to PMCs. The degree of EM-42 cell CD44 glycosylation inhibition, as determined by RCA lectin binding following TUNICA treatment, was highly correlated with the decrease in EM-42 cell binding to PMCs. The observation that EC surface CD44 density was not decreased by TUNICA suggests that EC CD44 glycosylation patterns are involved in the pathogenesis of the early endometriotic lesion. 1. Fertil Steril 84:16-21, 2005 Supported by: National Institutes of Health 1R01 HD044135-01.
Monday, October 23, 2006 4:45 pm O-78 EXPERIMENTAL ENDOMETRIOSIS IN IMMUNE COMPROMISED MICE FOLLOWING ADOPTIVE TRANSFER OF HUMAN LEUKOCYTES. K. L. Bruner-Tran, S. Gladson, J. N. Higginbotham, G. R. Yeaman, E. Eisenberg, K. G. Osteen. Vanderbilt Univ School of Medicine, Nashville, TN. OBJECTIVE: Endometriosis, the growth of endometrial glands and stroma outside the uterus, may result, in part, from a compromised immune surveillance resulting in a failure to clear displaced endometrium. Certainly, previous studies indicate that women with endometriosis have an altered immune status, but it is currently not known whether these alterations are a causal factor or are simply a consequence of the disease. In order to better understand the mechanisms of ectopic tissue growth, we have utilized an experimental model of endometriosis in which human endometrium is injected into nude mice. However, nude mice are not useful for examining the role of the immune system in the development of experimental endometriosis. In the current study, we present a model system in which the role of human immune cells can be studied during establishment and progression of endometriosis. Specifically, we have established experimental endometriosis in Rag2 ␥(c) (Recombinant Activating Gene 2/common cytokine receptor ␥ chain (␥c) double null) mice in which human immune cells have been transferred. DESIGN: Laboratory-based study. MATERIALS AND METHODS: Human peripheral blood mononuclear cells and neutrophils (collectively, PBMCs) were obtained at the time of endometrial biopsy during the late proliferative phase from normally cycling women. PBMCs were labeled with pKH-26 and immediately injected intraperitoneally into ovariectomized Rag2␥(c) mice implanted with a slowrelease estradiol capsule. Human endometrial tissues were maintained as organ cultures in the presence of 1 nM estradiol for 24 hrs prior to intraperitoneal injection into mice. Mice were sacrificed and examined for the presence of experimental endometriosis 17-20 days after receiving human PBMCs and endometrial tissue. Spleen, lymph nodes and experimental lesions were removed from mice at the time of necropsy and analyzed by flow cytometry for the presence of pKH-labeled human PB-
S33
MCs. A timecourse analysis was also conducted in order to examine the presence of human immune cells in mice over time. Control Rag2 ␥ (c) mice were also injected with human endometrium, but did not receive immune cells. RESULTS: Rag2 ␥ (c) mice readily accept human immune cells and endometrium, exhibiting experimental peritoneal endometriosis. Interestingly, mice receiving endometrium in the absence of immune cells demonstrated larger, more numerous lesions than mice receiving human immune cells. Compared to the lymph nodes, flow cytometry revealed a slightly greater concentration of cells within the spleen, which peaked (5.2%) 4 hrs after injection of cells. Human PBMCs within the spleen measured 2.7% at 5 days and averaged 1.2% at the last timepoint examined. Interestingly, the concentration of immune cells within endometrial lesions was maintained over time. CONCLUSION: The Rag2 ␥ (c) “human:mouse double chimera” may prove to be a useful model system in which to examine the role of immune cells in the establishment and progression of endometriosis. Importantly, our data suggest that the presence of normal human immune cells may impede the growth of ectopic endometrial tissue. Our preliminary data indicates that the immune cells track to the ectopic lesions, supporting a possible role of these cells in the natural course of the disease. Whether immune cells from women with endometriosis behave in a similar fashion remains to be determined. Supported by: K12 HD043483-04 (KBT) and The Endometriosis Association (KGO).
Monday, October 23, 2006 5:00 pm O-79 EFFECTS OF PROTEIN KINASE C INHIBITOR ON INITIAL DEVELOPMENT OF ECTOPIC IMPLANTS IN A MOUSE MODEL OF ENDOMETRIOSIS. S. Matsuzaki, M. Canis, C. Darcha, N. Bourdel, J. Pouly, G. Mage. Centre d⬘Endoscopie et des Nouvelles Techniques Interventionnelles, Universite´ d’Auvergne, Clermont-Ferrand, France; CHU Clermont-Ferrand, Clermont-Ferrand, France. OBJECTIVE: Our recent cDNA microarray study demonstrated that protein kinase C (PKC) beta is up-regulated in endometriosis. In the present study, we further investigated whether PKC beta could be functionally involved in payhophysiology of endometriosis. We evaluated the effects of PKC inhibitor use on initial development of ectopic implants in a mouse model of endometriosis. Our prior real-time RT-PCR analysis had revealed PKC beta is expressed in uterus, ectopic implants and peritoneum in a mouse model of endometriosis. Thus, we concluded that our mouse model was useful in the present study to evaluate the effects of PKC inhibitor use. DESIGN: Prospective, randomized study. MATERIALS AND METHODS: Donor female C57BJ mice were injected with estrogen depot on day -7 (Donor Group A: n⫽10) or -14 and -7 (Donor group B: n⫽10). On day -7, oral gavage of PKC inhibitor (100 mg/kg/day, once/day) was started for 1 week in Donor Group 2. On day 0, donor mice were sacrificed and uterine horns were removed. Uterine fragments from Donor Group A were implanted into a total of 20 recipient mice (three uterine fragments to each mouse) and estrogen depot was injected. Then, recipient mice were divided randomly into two groups of 10 animals each: Group 1 (vehicle) and Group 2 (PKC inhibitor). Similarly, uterine fragments from Donor Group B were implanted into a total of 20 recipient mice (three uterine fragments to each mouse) and estrogen depot was injected. Then, recipient mice were divided randomly into two groups of 10 animals each: Group 3 (vehicle) and Group 4 (PKC inhibitor). Oral gavage of PKC inhibitor (100 mg/kg/day, once/day) or vehicle was started for 1 week. On day 7, a laparotomy was performed. RESULTS: In Group 1 (controls), ectopic implants were detected in all mice. In Groups 2, 3 and 4, ectopic implants were detected in seven, four and three mice, respectively. The number of mice that developed ectopic implants was significantly lower in Groups 3 and 4 than Group 1 (P ⬍0.02, P ⬍0.004, respectively). The number of ectopic implants was significantly lower in Groups 2, 3 and 4 than Group 1 (P ⬍0.004, P ⬍0.0001, P ⬍0.0001, respectively). In addition, the number of ectopic implants was significantly lower in Groups 3 and 4 than Group 2 (P ⬍0.04, P ⬍0.02, respectively). CONCLUSION: PKC inhibitor use, particularly, PKC inhibitor pretreatment could prevent the development of ectopic implants in a mouse
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Abstracts
model of endometriosis. The present findings suggested that PKC beta might be functionally involved in initial development of endometriosis. Supported by: None.
Monday, October 23, 2006 5:15 pm O-80 MENTAL DISORDERS IN WOMEN WITH ENDOMETRIOSIS. S. Ferrero, M. Giordano, L. H. Abbamonte, N. Ragni, V. Remorgida. Dept of Obstetrics and Gynecology, San Martino Hospital and Univ of Genoa, Genoa, Italy. OBJECTIVE: To compare the prevalence of mental disorders in women with and without endometriosis. In addition, pain symptoms and mental disorders were re-evaluated in women with endometriosis at 1 year from surgery. DESIGN: Prospective study. MATERIALS AND METHODS: This study included women of reproductive age who underwent laparoscopy because of pain symptoms (dysmenorrhea, dyspareunia, chronic pelvic pain), infertility, uterine myomas, and ovarian cysts. No patient included in the study had used GnRH analogs in the 6 months prior to surgery. Before surgery, mental disorders were diagnosed by using the Primary Care Evaluation of Mental Disorders Patient Health Questionnaire (PRIME-MD PHQ); in case of depression, its severity was scored using the Beck Depression Inventory (BDI). The presence and intensity (measured on a 100-mm visual analogue scale, VAS) of pain symptoms were investigated. All visible endometriotic lesions were excised at surgery; the diagnosis of endometriosis was histologically confirmed. A follow-up of BDI scores and pain symptoms was performed at 1 year from surgery. Statistical analysis was performed by using the chisquare test, Student t-test, and Mann-Whitney U test. RESULTS: Out of 260 consecutive women approached for the study, 222 accepted to participate yielding a response rate of 85.4%. 114 patients had histologically confirmed endometriosis while 108 did not have endometriosis. No significant difference was observed in the mean age and marital status between women with and without endometriosis. 45.6% (52/114) of the women with endometriosis were classified as depressed compared to 24.1% (26/108) of women without endometriosis (p ⬍ 0.001); this difference was confirmed in the subgroup of patients with pain symptoms (p ⫽ 0.003) but not in the infertile subjects (p ⫽ 0.151). According to the BDI, 1 woman with endometriosis and no patient without endometriosis had extreme depression, 6 women with endometriosis and one patient without endometriosis had severe depression. 42 (36.8%) women with endometriosis and 24 (22.2%) subjects without endometriosis screened positive for a somatoform disorder (p ⫽ 0.017). Panic syndrome was significantly more frequent among women with endometriosis (20/114) than in those without endometriosis (9/108; p ⫽ 0.042); no significant difference between the two groups was observed in other anxiety disorders, bulimia nervosa, binge eating disorders, and alcohol abuse. At 1 year follow-up, absence (VAS ⬍ 30-mm) or improvement (⬎ 40-mm in the VAS scale) of pain symptoms was observed in 78.1% of women with endometriosis (77/105). Depressed women with endometriosis were significantly less likely to report absence or improvement in pain symptoms than non-depressed women (p ⬍ 0.001). Among women with endometriosis, no significant change in the BDI score was observed in subjects with severe and extreme depression (p ⫽ 0.212); the BDI was significantly decreased in subjects with borderline and moderate depression (p ⫽ 0.012). CONCLUSION: This study demonstrates that women with endometriosis have an increased risk of depression, somatoform disorder, and panic syndrome. Depressed patients often do not report a significant improvement in pain symptoms at one year from the surgical excision of endometriosis. Supported by: None.
OUTCOME PREDICTORS-CLINICAL: ART Monday, October 23, 2006 3:30 pm O-81 ADJUNCTIVE PGD DOES NOT ENHANCE EMBRYO SELECTION BEYOND ANY IMPROVEMENT PROVIDED BY BLASTOCYT
Vol. 86, Suppl 2, September 2006
Monday, October 23, 2006 4:30 pm O-77 INHIBITION OF CD44 N-LINKED GLYCOSYLATION INHIBITS ENDOMETRIAL CELL BINDING TO PERITONEAL MESOTHELIAL CELLS. A. S. Nair, C. A. Witz, H. B. Nair, R. R. Tekmal, R. S. Schenken. Univ of Texas Health Science Center at San Antonio, San Antonio, TX. OBJECTIVE: CD44 is a transmembrane glycoprotein that is the major ligand for hyaluronan. We have previously demonstrated that abrogation of peritoneal mesothelial cell-associated hyaluronan inhibits endometrial cell (EC) binding to peritoneal mesothelial cells (PMCs), suggesting that CD44/ hyaluronan is involved in the early development of the endometriotic lesion. Other studies using ovarian cancer cell lines have demonstrated that CD44 binding to hyaluronan is regulated in part by the degree of CD44 glycosyl-
FERTILITY & STERILITY威
ation. Here, we assessed the effect of EC CD44 glycosylation inhibition on EC binding to PMCs. DESIGN: In vitro study. MATERIALS AND METHODS: EM- 42 cells, an endometrial epithelioid cell line, were grown to near confluence and treated with the N-linked glycosylation inhibitor, tunicamycin (TUNICA) (5 mcg/ml) for 24 hours. Cell viability was assessed in TUNICA-treated and -untreated EM42 cells using propidium iodide (PI) staining and flow cytometry. The binding of the fluorescence-conjugated lectin, Ricinus communis agglutinin-1 (RCA), to TUNICA-treated and -untreated EM42 cells was used to assess the degree of glycosylation inhibition. Western blots analysis of TUNICA-treated and -untreated EM42 cell lysates were carried out using the anti-CD44 antibody 5F12. The effect of TUNICA on the density of cell surface CD44 was also determined using flow cytometry with the anti-CD44 antibody 5F12. Binding of TUNICA-treated and -untreated EM42 cells to the PMC line LP9 was compared at one hour of co-culture using an established in vitro assay [1]. RESULTS: Tunicamycin did not affect cell viability as assessed by PI labeling. TUNICA-treated EM-42 cells demonstrated a 19% decreased rate of RCA lectin binding, suggesting significant glycosylation inhibition. Western blot analysis and flow cytometry using the 5F12 antibody confirmed that TUNICA treatment did not decrease the expression of cell surface CD44. TUNICA treatment of EM42 cells lead to a 20% decreased rate of binding to PMCs (37% vs. 46%, p ⬍ .004). CONCLUSION: The present study demonstrates that inhibition of EC N-linked glycosylation decreases EC binding to PMCs. The degree of EM-42 cell CD44 glycosylation inhibition, as determined by RCA lectin binding following TUNICA treatment, was highly correlated with the decrease in EM-42 cell binding to PMCs. The observation that EC surface CD44 density was not decreased by TUNICA suggests that EC CD44 glycosylation patterns are involved in the pathogenesis of the early endometriotic lesion. 1. Fertil Steril 84:16-21, 2005 Supported by: National Institutes of Health 1R01 HD044135-01.
Monday, October 23, 2006 4:45 pm O-78 EXPERIMENTAL ENDOMETRIOSIS IN IMMUNE COMPROMISED MICE FOLLOWING ADOPTIVE TRANSFER OF HUMAN LEUKOCYTES. K. L. Bruner-Tran, S. Gladson, J. N. Higginbotham, G. R. Yeaman, E. Eisenberg, K. G. Osteen. Vanderbilt Univ School of Medicine, Nashville, TN. OBJECTIVE: Endometriosis, the growth of endometrial glands and stroma outside the uterus, may result, in part, from a compromised immune surveillance resulting in a failure to clear displaced endometrium. Certainly, previous studies indicate that women with endometriosis have an altered immune status, but it is currently not known whether these alterations are a causal factor or are simply a consequence of the disease. In order to better understand the mechanisms of ectopic tissue growth, we have utilized an experimental model of endometriosis in which human endometrium is injected into nude mice. However, nude mice are not useful for examining the role of the immune system in the development of experimental endometriosis. In the current study, we present a model system in which the role of human immune cells can be studied during establishment and progression of endometriosis. Specifically, we have established experimental endometriosis in Rag2 ␥(c) (Recombinant Activating Gene 2/common cytokine receptor ␥ chain (␥c) double null) mice in which human immune cells have been transferred. DESIGN: Laboratory-based study. MATERIALS AND METHODS: Human peripheral blood mononuclear cells and neutrophils (collectively, PBMCs) were obtained at the time of endometrial biopsy during the late proliferative phase from normally cycling women. PBMCs were labeled with pKH-26 and immediately injected intraperitoneally into ovariectomized Rag2␥(c) mice implanted with a slowrelease estradiol capsule. Human endometrial tissues were maintained as organ cultures in the presence of 1 nM estradiol for 24 hrs prior to intraperitoneal injection into mice. Mice were sacrificed and examined for the presence of experimental endometriosis 17-20 days after receiving human PBMCs and endometrial tissue. Spleen, lymph nodes and experimental lesions were removed from mice at the time of necropsy and analyzed by flow cytometry for the presence of pKH-labeled human PB-
S33
MCs. A timecourse analysis was also conducted in order to examine the presence of human immune cells in mice over time. Control Rag2 ␥ (c) mice were also injected with human endometrium, but did not receive immune cells. RESULTS: Rag2 ␥ (c) mice readily accept human immune cells and endometrium, exhibiting experimental peritoneal endometriosis. Interestingly, mice receiving endometrium in the absence of immune cells demonstrated larger, more numerous lesions than mice receiving human immune cells. Compared to the lymph nodes, flow cytometry revealed a slightly greater concentration of cells within the spleen, which peaked (5.2%) 4 hrs after injection of cells. Human PBMCs within the spleen measured 2.7% at 5 days and averaged 1.2% at the last timepoint examined. Interestingly, the concentration of immune cells within endometrial lesions was maintained over time. CONCLUSION: The Rag2 ␥ (c) “human:mouse double chimera” may prove to be a useful model system in which to examine the role of immune cells in the establishment and progression of endometriosis. Importantly, our data suggest that the presence of normal human immune cells may impede the growth of ectopic endometrial tissue. Our preliminary data indicates that the immune cells track to the ectopic lesions, supporting a possible role of these cells in the natural course of the disease. Whether immune cells from women with endometriosis behave in a similar fashion remains to be determined. Supported by: K12 HD043483-04 (KBT) and The Endometriosis Association (KGO).
Monday, October 23, 2006 5:00 pm O-79 EFFECTS OF PROTEIN KINASE C INHIBITOR ON INITIAL DEVELOPMENT OF ECTOPIC IMPLANTS IN A MOUSE MODEL OF ENDOMETRIOSIS. S. Matsuzaki, M. Canis, C. Darcha, N. Bourdel, J. Pouly, G. Mage. Centre d⬘Endoscopie et des Nouvelles Techniques Interventionnelles, Universite´ d’Auvergne, Clermont-Ferrand, France; CHU Clermont-Ferrand, Clermont-Ferrand, France. OBJECTIVE: Our recent cDNA microarray study demonstrated that protein kinase C (PKC) beta is up-regulated in endometriosis. In the present study, we further investigated whether PKC beta could be functionally involved in payhophysiology of endometriosis. We evaluated the effects of PKC inhibitor use on initial development of ectopic implants in a mouse model of endometriosis. Our prior real-time RT-PCR analysis had revealed PKC beta is expressed in uterus, ectopic implants and peritoneum in a mouse model of endometriosis. Thus, we concluded that our mouse model was useful in the present study to evaluate the effects of PKC inhibitor use. DESIGN: Prospective, randomized study. MATERIALS AND METHODS: Donor female C57BJ mice were injected with estrogen depot on day -7 (Donor Group A: n⫽10) or -14 and -7 (Donor group B: n⫽10). On day -7, oral gavage of PKC inhibitor (100 mg/kg/day, once/day) was started for 1 week in Donor Group 2. On day 0, donor mice were sacrificed and uterine horns were removed. Uterine fragments from Donor Group A were implanted into a total of 20 recipient mice (three uterine fragments to each mouse) and estrogen depot was injected. Then, recipient mice were divided randomly into two groups of 10 animals each: Group 1 (vehicle) and Group 2 (PKC inhibitor). Similarly, uterine fragments from Donor Group B were implanted into a total of 20 recipient mice (three uterine fragments to each mouse) and estrogen depot was injected. Then, recipient mice were divided randomly into two groups of 10 animals each: Group 3 (vehicle) and Group 4 (PKC inhibitor). Oral gavage of PKC inhibitor (100 mg/kg/day, once/day) or vehicle was started for 1 week. On day 7, a laparotomy was performed. RESULTS: In Group 1 (controls), ectopic implants were detected in all mice. In Groups 2, 3 and 4, ectopic implants were detected in seven, four and three mice, respectively. The number of mice that developed ectopic implants was significantly lower in Groups 3 and 4 than Group 1 (P ⬍0.02, P ⬍0.004, respectively). The number of ectopic implants was significantly lower in Groups 2, 3 and 4 than Group 1 (P ⬍0.004, P ⬍0.0001, P ⬍0.0001, respectively). In addition, the number of ectopic implants was significantly lower in Groups 3 and 4 than Group 2 (P ⬍0.04, P ⬍0.02, respectively). CONCLUSION: PKC inhibitor use, particularly, PKC inhibitor pretreatment could prevent the development of ectopic implants in a mouse
S34
Abstracts
model of endometriosis. The present findings suggested that PKC beta might be functionally involved in initial development of endometriosis. Supported by: None.
Monday, October 23, 2006 5:15 pm O-80 MENTAL DISORDERS IN WOMEN WITH ENDOMETRIOSIS. S. Ferrero, M. Giordano, L. H. Abbamonte, N. Ragni, V. Remorgida. Dept of Obstetrics and Gynecology, San Martino Hospital and Univ of Genoa, Genoa, Italy. OBJECTIVE: To compare the prevalence of mental disorders in women with and without endometriosis. In addition, pain symptoms and mental disorders were re-evaluated in women with endometriosis at 1 year from surgery. DESIGN: Prospective study. MATERIALS AND METHODS: This study included women of reproductive age who underwent laparoscopy because of pain symptoms (dysmenorrhea, dyspareunia, chronic pelvic pain), infertility, uterine myomas, and ovarian cysts. No patient included in the study had used GnRH analogs in the 6 months prior to surgery. Before surgery, mental disorders were diagnosed by using the Primary Care Evaluation of Mental Disorders Patient Health Questionnaire (PRIME-MD PHQ); in case of depression, its severity was scored using the Beck Depression Inventory (BDI). The presence and intensity (measured on a 100-mm visual analogue scale, VAS) of pain symptoms were investigated. All visible endometriotic lesions were excised at surgery; the diagnosis of endometriosis was histologically confirmed. A follow-up of BDI scores and pain symptoms was performed at 1 year from surgery. Statistical analysis was performed by using the chisquare test, Student t-test, and Mann-Whitney U test. RESULTS: Out of 260 consecutive women approached for the study, 222 accepted to participate yielding a response rate of 85.4%. 114 patients had histologically confirmed endometriosis while 108 did not have endometriosis. No significant difference was observed in the mean age and marital status between women with and without endometriosis. 45.6% (52/114) of the women with endometriosis were classified as depressed compared to 24.1% (26/108) of women without endometriosis (p ⬍ 0.001); this difference was confirmed in the subgroup of patients with pain symptoms (p ⫽ 0.003) but not in the infertile subjects (p ⫽ 0.151). According to the BDI, 1 woman with endometriosis and no patient without endometriosis had extreme depression, 6 women with endometriosis and one patient without endometriosis had severe depression. 42 (36.8%) women with endometriosis and 24 (22.2%) subjects without endometriosis screened positive for a somatoform disorder (p ⫽ 0.017). Panic syndrome was significantly more frequent among women with endometriosis (20/114) than in those without endometriosis (9/108; p ⫽ 0.042); no significant difference between the two groups was observed in other anxiety disorders, bulimia nervosa, binge eating disorders, and alcohol abuse. At 1 year follow-up, absence (VAS ⬍ 30-mm) or improvement (⬎ 40-mm in the VAS scale) of pain symptoms was observed in 78.1% of women with endometriosis (77/105). Depressed women with endometriosis were significantly less likely to report absence or improvement in pain symptoms than non-depressed women (p ⬍ 0.001). Among women with endometriosis, no significant change in the BDI score was observed in subjects with severe and extreme depression (p ⫽ 0.212); the BDI was significantly decreased in subjects with borderline and moderate depression (p ⫽ 0.012). CONCLUSION: This study demonstrates that women with endometriosis have an increased risk of depression, somatoform disorder, and panic syndrome. Depressed patients often do not report a significant improvement in pain symptoms at one year from the surgical excision of endometriosis. Supported by: None.
OUTCOME PREDICTORS-CLINICAL: ART Monday, October 23, 2006 3:30 pm O-81 ADJUNCTIVE PGD DOES NOT ENHANCE EMBRYO SELECTION BEYOND ANY IMPROVEMENT PROVIDED BY BLASTOCYT
Vol. 86, Suppl 2, September 2006