- Email: [email protected]
Cross Talk Between Inducible Nitric Oxide Syntheses (iNOS) and Myeloperoxidase (MPO) in Fibroblasts Isolated From Normal Peritoneal and Adhesion Tissues
achieved more than one positive test and of the patients using more than one donor, similar numbers of pregnancies occurred before and after switching donors. Overall, biochemical pregnancy rate was 13.7% per cycle and the live birth rate was 9.4% per cycle. CONCLUSION: The results of the study indicate that switching donors does not increase the likelihood of pregnancy for TDI. Supported by: None
REPRODUCTIVE SURGERY P-846 Can Hypoxia Impact Lymphocyte-Mediated Elimination of Peritoneal Fibroblasts Leading to Development of Adhesion Phenotype? Z. Alpay, M. S. Ozgonenel, S. Savasan, S. Buck, G. M. Saed, M. P. Diamond. Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, Wayne State University, Detroit, MI; Children’s Hospital of Michigan, Division of Hematology/Oncology, Wayne State University, Detroit, MI. OBJECTIVE: Several characteristics of fibroblasts obtained from postsurgical intraperitoneal adhesion tissue (AT) could be induced by hypoxia treatment in normal peritoneal fibroblasts suggesting a central role for local hypoxic environment in the adhesion development. The natural immune response participates in the elimination of altered cells maintaining body homeostasis. The rates of this response and tissue regeneration ultimately determine the outcome of tissue healing. We have shown that lymphokineactivated killer (LAK) cells kill AT fibroblasts more efficiently than normal peritoneal (NP) fibroblasts. In this study, we investigated the effect of hypoxia on the expression of immune response-related surface molecules and LAK cell-mediated fibroblast elimination of NP and AT fibroblasts in vitro. DESIGN: Primary cell cultures of both NP and AT fibroblasts obtained from patients with post-surgical adhesions were expanded and used in the experiments. Immune response-related surface marker expression of untreated and hypoxia-treated NP and AT fibroblasts was studied at least three times. The effect of allogeneic LAK cells on those cells was simultaneously investigated following a 4-hour co-incubation of LAK cells. MATERIALS AND METHODS: Hypoxia treatment was achieved by incubating fibroblasts in an airtight Plexiglas chamber, which was deoxygenated by positive infusion of 2% CO2 /nitrogen gas mixture. Cultures in hypoxic chamber were then placed in a standard humidified tissue incubator for 24 hours prior to experimentation. The expression of intercellular adhesion molecule-1 (CD54), co-stimulatory molecule (CD40), tumor necrosis factor-alpha receptor type II (CD120b) and transferrin receptor (CD71) was studied by flow cytometry in hypoxia-treated and untreated NP and AT primary culture fibroblasts obtained from four different individuals. Allogeneic LAK cells were generated by incubating peripheral blood mononuclear cells from healthy donors with interleukin 2 and interleukin 15 for five days. The LAK cell-mediated fibroblast elimination was studied by our established flow cytometric cell mediated cytotoxicity assay. Paired t test of the mean values of individual cases was used for statistical evaluation. RESULTS: Hypoxia treatment of fibroblasts resulted in increased average LAK cell-mediated fibroblast killing compared to untreated cells. This effect was more prominent in NP than in AT fibroblasts (1.5 ⫾ 0.4 fold vs. 1.1 ⫾ 0.2 fold). The surface expression of CD54, CD40, CD120b and CD71 were not affected significantly by short term hypoxia exposure, however it resulted in a measurable decrease in CD71 expression in AT fibroblasts (0.95 ⫾ 0.08 in NP vs. 0.88 ⫾ 0.03 in AT). CONCLUSION: The finding of a more prominent increase in LAK cell-mediated fibroblast elimination following hypoxia exposure in NP than AT fibroblasts was supportive of a role of hypoxia in the pathogenesis of adhesion development. In other words, hypoxia treatment of NP could induce a phenotype similar to AT, but could not further affect AT fibroblasts. However, this increase in lymphocyte response was not correlated with enhanced immune response associated marker expressions following short term hypoxia exposure. The decrease in transferrin receptor expression might reflect an adaptive response to hypoxia resulting in decreased cellular iron uptake and this might be a transient phenomenon. Supported by: None.
FERTILITY & STERILITY威
P-847 If This Is No Pollution the Solution Is Not Dilution: The Consequences of Irrigation on Peritoneal Fluid and Cell Homeostasis. D. E. Ott. Mercer University, Macon, GA. OBJECTIVE: Since peritoneal fluid and its contents are intimately involved with peritoneal homeostasis and repair it is necessary under normal circumstances to maintain as normal a physiologic condition and environment as possible. Removal of peritoneal fluid cells may effect tissue repair and immunologic response to peritoneal damage and have significant local and peritoneal effects. Thus the objective of this work is to evaluate the effect of peritoneal irrigation on peritoneal cell count. DESIGN: Prospective analysis of forty-eight (48), four groups of twelve (12) patients, having sequential pelvic irrigation sampling to determine the effect of irrigation on peritoneal cell count done in University research laboratory and a private research facility. MATERIALS AND METHODS: Forty-eight female patients undergoing benign, non-endometriosis, non-infectious laparoscopy had all visible peritoneal fluid evacuated from the cul-de-sac after establishing a pneumoperitoneum. Irrigation was done using lactated ringer’s solution as 500 milliliter (cc) aliquots for three (3) liters by using ten (10) 500 cc lavages, or 250 cc aliquots (12 lavages) or 100 cc aliquots (thirty (30) lavages). Samples were measured for cell type and count after each lavage. Data compared the number of cells retrieved in each lavage cycle and was analyzed using a Mann-Whitney U test and variance between groups was analyzed using analysis of variance. Probability values of ⬍ 0.05 were deemed significant. RESULTS: Initial cell type and number of groups did not significantly differ. A similar proportion of cells were removed from each group in each successive irrigation suction cycle. A significantly greater number of cells were removed from the first to third lavage irrigations than from subsequent irrigations. A distinctive washout curve was found showing progressive cell count reduction with the greatest change occurring within the first lavage irrigation retrieval progressing to 99.9% removal by the third cycle and no significant changes after further irrigations. Peritoneal cell depletion as a result of lavage was not statistically different regardless of the volume of irrigation used. CONCLUSION: Cells within the peritoneal fluid, adherent mesothelial cells and free floating cells are rapidly removed from the peritoneal cavity by three lavage irrigation suction cycles. Three irrigations of any volume or irrigation from 100 to 500 cc at a time removes virtually all peritoneal cells. Three or more peritoneal lavages regardless of volume used significantly reduces peritoneal resident cell population. Elimination of peritoneal fluid and peritoneal resident cell population by irrigation severely affects at least the immediate homeostatic balance of the peritoneal cavity. Supported by: Georgia BioMedical, Inc. P-848 Cross Talk Between Inducible Nitric Oxide Syntheses (iNOS) and Myeloperoxidase (MPO) in Fibroblasts Isolated From Normal Peritoneal and Adhesion Tissues. G. M. Saed, H. Lu, Z. Jiang, S. Abuolba, H. Abu-Soud, M. P. Diamond. Wayne State University/Detroit Medical Center, Detroit, MI. OBJECTIVE: Fibroblasts beneath the mesothelium in the human peritoneum play an important role in the healing process of the peritoneum. Adhesion fibroblasts are characterized by lower levels of nitric oxide (NO) although there were no differences in iNOS, the enzyme which generates NO, between normal and adhesion fibroblasts. Additionally, adhesion fibroblasts exhibited significantly lower levels of MPO. We have previously shown that there is a cross talk between MPO and iNOS utilizing purified enzymes. MPO upregulate the catalytic activity of iNOS by preventing the feed back inhibition attributed to the formation of iNOS nitrosyl complex. Our objective is to determine whether there is a cross talk between MPO and iNOS in vivo utilizing fibroblasts isolated from normal peritoneum and from adhesions. DESIGN: Laboratory Study. MATERIALS AND METHODS: We have obtained fibroblast primary cultures from normal peritoneum and adhesion tissues of the same patients. We have utilized the small interfering RNA (SiRNA) technology to specifically knock-out the expression of MPO alone, iNOS alone, and combination of both and then quantify the expression levels of MPO and iNOS using the real-time RTR/PCR developed in our laboratory.
S463
RESULTS: There was no significant difference in iNOS mRNA levels between normal (5.0 X105 iNOS mRNA copies and adhesion fibroblasts (4.8 X105). When iNOS gene expression is knocked-out, the iNOS mRNA levels were reduced by 98% in both normal and adhesion fibroblasts. Similar finding were observed for MPO SiRNA, confirming the efficiency of SiRNA to completely destroy iNOS and MPO mRNAs in both cell lines. Use of MPO mRNA resulted in a 62 % decrease in iNOS mRNA levels in normal fibroblasts and completely destroyed iNOS mRNA in adhesion fibroblasts. Destruction of mRNA for iNOS and MPO together resulted in a complete destruction of iNOS mRNA in both cell lines. CONCLUSION: Our data suggests that there is intricate cross talk between iNOS and MPO in normal and adhesion fibroblasts. This may be the cause for the disturbance of iNOS/NO/protein nitration system observed in adhesions, as compared to, normal fibroblasts. Supported by: None
P-849 Differential Expression of Estrogen, Progesterone, Androgen, and Prolactin Receptors in Human Fibroblasts Isolated From Normal Peritoneum and Adhesions. G. M. Saed, L. Detti, H. Lu, Z. Jiang, S. Aboulba, M. P. Diamond. Wayne State University/Detroit Medical Center, Detroit, MI. OBJECTIVE: Adhesions are believed to contribute to infertility in 40% of infertile couples and are the most common cause of small bowel obstruction. Also abdominal-pelvic pain is attributed to adhesions in a little less of 30% of cases. Our objective is to characterize fibroblasts isolated from normal peritoneum and adhesion tissues of the same patient in their ability to express hormone receptors. DESIGN: Prospective experimental study. MATERIALS AND METHODS: We have utilized the real time RT-PCR to determine mRNA levels for estrogen (ER-␣, ER-), progesterone A and B isoforms (P4), androgen (A), and prolactin (PRL) receptors in primary cultures of fibroblasts isolated from normal peritoneum and adhesion tissues of the same patient. Our main outcome measure was the quantification of mRNA levels (mRNA copies/g RNA) of ER-␣, ER-, P4, A, PRL receptors in both cell lines. RESULTS: Our results have shown a differential expression in all hormone receptors studied normal peritoneal and adhesion fibroblasts. There were no detectable levels of ER-␣ in adhesion fibroblast as compared to 1,487 copies in normal fibroblasts. Normal fibroblasts (713 x 105) expressed significantly higher levels of ER- receptor (713 x 105) as compared to adhesion fibroblasts (24.8 x 105). There were no detectable levels of progesterone in normal fibroblasts as compared to (194 mRNA copies/g RNA) of progesterone in adhesion fibroblasts. The levels of androgen receptors were also markedly different in normal peritoneal (245 x 105) and in adhesion fibroblasts (180 x 105). There were higher prolactin levels in normal fibroblasts (280 copies) as compared to adhesion fibroblasts (72 copies). CONCLUSION: Expression of hormone receptors in adhesion fibroblasts is different from that in normal peritoneum fibroblasts. This may be used for future design of intervention therapies for postoperative adhesions. Supported by: None
P-850 The Role of Robotic-Assisted Laparoscopy in Gynecological Surgeries. N. Salamat Saberi, B. Shahmohamady, C. H. Nezhat, F. Nezhat, C. Nezhat. Stanford University Hospital, Palo Alto, CA; Mount Sinai Medical, New York, NY. OBJECTIVE: To evaluate the role of robotic-assisted laparoscopy in gynecological surgeries DESIGN: Retrospective chart review study MATERIALS AND METHODS: In a tertiary referral center for laparoscopic gynecologic surgery, a total of 15 patients who were undergoing various gynecologic surgeries were consented for combined laparoscopy and roboticassisted laparoscopic surgery. The da Vinci robot was used in each case. Four ports were inserted: umbilicus, suprapubic, and two lateral ports. These surgeries were performed both using laparoscopy and robotic-assisted laparoscopy surgery.
S464
Abstracts
RESULTS: Fifteen patients underwent variety of gynecologic surgeries such as myomectomies, treatment of endometriosis, total and supracervical hysterectomy, ovarian cystectomy, sacral colpopexy and Moskowitz procedure. The assembly time to switch from laparoscopy to robotic assisted surgery was 18.9 minutes (14-27 minutes), and the disassembly time was 2.1 minutes (1-3 minutes). Robotic assisted surgery was used for suturing after myomectomy, tissue manipulation, suturing of cervical stump after supracervical hysterectomy, vaginal cuff closure and suspension after total laparoscopic hysterectomy, ovarian suturing after cystectomy and performing Moskowitz procedure. CONCLUSION: Robotic-assisted laparoscopic surgeries have advantages in providing a 3 dimensional visualization of operative field, decreasing fatigue and tension tremor of surgeon, and added wrist motion for improved dexterity and greater surgical precision. The disadvantages include cost and added operating time for assembly and disassembly. The greatest practical use lies in the ability to perform easier suturing in laparoscopic surgery. Supported by: None
P-851 The Value of 3D Ultrasound in the Diagnosis and Treatment of Asherman’s Syndrome. J. Cohen, A. B. Copperman. Mount Sinai Medical Center, New York, NY. OBJECTIVE: To assess the value of 3D ultrasound in the management of patients with suspected Asherman’s syndrome DESIGN: A prospective comparative study of infertile patients with suspected Asherman’s undergoing both HSG and 3D ultrasound prior to hysteroscopy, between the years of 1998-2004 MATERIALS AND METHODS: 54 women, ranging in age from 23-49, presented to a tertiary care center with secondary infertility between the years of 1998-2004 and were given a diagnosis of Asherman’s. 49 presented with the diagnosis and 5 were newly diagnosed. The results of both imaging modalities were compared with those obtained by hysteroscopy. Sensitivity, specificity, positive predictive and negative predictive values were calculated. In addition, the ability to re-establish a functional uterine cavity and attain fertility post-operatively were evaluated. RESULTS: Intra-uterine adhesions were demonstrated on 3D ultrasound and HSG in all 54 patients and confirmed at hysteroscopy. 3D ultrasound had a sensitivity of 100% and PPV of 100%, when compared with the hysteroscopic findings, for correctly grading the extent of intra-uterine adhesions in terms of percent cavity of obstructed, while HSG showed a sensitivity of 66.7% and a PPV of 66.7% for accurately identifying the extent of intrauterine adhesions. In 61.1% of these cases lower segment outflow obstruction was present and the HSG was erroneously read as severe Asherman’s with complete cavity obstruction. Following targeted hysteroscopy, 45/50 (90%) of patients conceived. CONCLUSION: 3D ultrasound, a non-invasive office based exam, has a better prognostic value with a higher sensitivity and PPV than HSG when classifying the grade of cavity damage in patients with suspected Asherman’s syndrome. 3D ultrasound is particularly helpful in differentiating severe IUA from lower uterine segment outflow obstruction. In addition, the superior quality of the diagnostic images produced by the 3D ultrasound allows the hysteroscopist to perform a more precise procedure and this, combined with good surgical technique and post-operative care, may ultimately improve reproductive outcome. Our study is one of the first and largest to demonstrate the benefit of 3D ultrasound in the diagnosis and treatment of Asherman’s Syndrome. We propose re-evaluating current grading systems that classify severity according to amenorrhea and/or HSG findings, as they clearly mis-classify patients with lower outflow tract obstruction. In fact, these patients actually have an excellent prognosis if there is normal cavity beyond the obstruction. Supported by: None
P-852 Outcome of Pregnancies Achieved Following Laparoscopic Surgery for Correction of Tubo-Peritoneal Factor Infertility Associated With Endometriosis or Pelvic Inflammatory Disease (PID). M. F. Mitwally, H. Albuarki, Z. Al-Wahab, M. P. Diamond, M. Abuzeid. Wayne State University, Detroit, MI; IVF-Michigan and Michigan State University, Rochester Hills & East Lansing, MI.
Vol. 84, Suppl 1, September 2005
REPRODUCTIVE SURGERY P-846 Can Hypoxia Impact Lymphocyte-Mediated Elimination of Peritoneal Fibroblasts Leading to Development of Adhesion Phenotype? Z. Alpay, M. S. Ozgonenel, S. Savasan, S. Buck, G. M. Saed, M. P. Diamond. Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, Wayne State University, Detroit, MI; Children’s Hospital of Michigan, Division of Hematology/Oncology, Wayne State University, Detroit, MI. OBJECTIVE: Several characteristics of fibroblasts obtained from postsurgical intraperitoneal adhesion tissue (AT) could be induced by hypoxia treatment in normal peritoneal fibroblasts suggesting a central role for local hypoxic environment in the adhesion development. The natural immune response participates in the elimination of altered cells maintaining body homeostasis. The rates of this response and tissue regeneration ultimately determine the outcome of tissue healing. We have shown that lymphokineactivated killer (LAK) cells kill AT fibroblasts more efficiently than normal peritoneal (NP) fibroblasts. In this study, we investigated the effect of hypoxia on the expression of immune response-related surface molecules and LAK cell-mediated fibroblast elimination of NP and AT fibroblasts in vitro. DESIGN: Primary cell cultures of both NP and AT fibroblasts obtained from patients with post-surgical adhesions were expanded and used in the experiments. Immune response-related surface marker expression of untreated and hypoxia-treated NP and AT fibroblasts was studied at least three times. The effect of allogeneic LAK cells on those cells was simultaneously investigated following a 4-hour co-incubation of LAK cells. MATERIALS AND METHODS: Hypoxia treatment was achieved by incubating fibroblasts in an airtight Plexiglas chamber, which was deoxygenated by positive infusion of 2% CO2 /nitrogen gas mixture. Cultures in hypoxic chamber were then placed in a standard humidified tissue incubator for 24 hours prior to experimentation. The expression of intercellular adhesion molecule-1 (CD54), co-stimulatory molecule (CD40), tumor necrosis factor-alpha receptor type II (CD120b) and transferrin receptor (CD71) was studied by flow cytometry in hypoxia-treated and untreated NP and AT primary culture fibroblasts obtained from four different individuals. Allogeneic LAK cells were generated by incubating peripheral blood mononuclear cells from healthy donors with interleukin 2 and interleukin 15 for five days. The LAK cell-mediated fibroblast elimination was studied by our established flow cytometric cell mediated cytotoxicity assay. Paired t test of the mean values of individual cases was used for statistical evaluation. RESULTS: Hypoxia treatment of fibroblasts resulted in increased average LAK cell-mediated fibroblast killing compared to untreated cells. This effect was more prominent in NP than in AT fibroblasts (1.5 ⫾ 0.4 fold vs. 1.1 ⫾ 0.2 fold). The surface expression of CD54, CD40, CD120b and CD71 were not affected significantly by short term hypoxia exposure, however it resulted in a measurable decrease in CD71 expression in AT fibroblasts (0.95 ⫾ 0.08 in NP vs. 0.88 ⫾ 0.03 in AT). CONCLUSION: The finding of a more prominent increase in LAK cell-mediated fibroblast elimination following hypoxia exposure in NP than AT fibroblasts was supportive of a role of hypoxia in the pathogenesis of adhesion development. In other words, hypoxia treatment of NP could induce a phenotype similar to AT, but could not further affect AT fibroblasts. However, this increase in lymphocyte response was not correlated with enhanced immune response associated marker expressions following short term hypoxia exposure. The decrease in transferrin receptor expression might reflect an adaptive response to hypoxia resulting in decreased cellular iron uptake and this might be a transient phenomenon. Supported by: None.
FERTILITY & STERILITY威
P-847 If This Is No Pollution the Solution Is Not Dilution: The Consequences of Irrigation on Peritoneal Fluid and Cell Homeostasis. D. E. Ott. Mercer University, Macon, GA. OBJECTIVE: Since peritoneal fluid and its contents are intimately involved with peritoneal homeostasis and repair it is necessary under normal circumstances to maintain as normal a physiologic condition and environment as possible. Removal of peritoneal fluid cells may effect tissue repair and immunologic response to peritoneal damage and have significant local and peritoneal effects. Thus the objective of this work is to evaluate the effect of peritoneal irrigation on peritoneal cell count. DESIGN: Prospective analysis of forty-eight (48), four groups of twelve (12) patients, having sequential pelvic irrigation sampling to determine the effect of irrigation on peritoneal cell count done in University research laboratory and a private research facility. MATERIALS AND METHODS: Forty-eight female patients undergoing benign, non-endometriosis, non-infectious laparoscopy had all visible peritoneal fluid evacuated from the cul-de-sac after establishing a pneumoperitoneum. Irrigation was done using lactated ringer’s solution as 500 milliliter (cc) aliquots for three (3) liters by using ten (10) 500 cc lavages, or 250 cc aliquots (12 lavages) or 100 cc aliquots (thirty (30) lavages). Samples were measured for cell type and count after each lavage. Data compared the number of cells retrieved in each lavage cycle and was analyzed using a Mann-Whitney U test and variance between groups was analyzed using analysis of variance. Probability values of ⬍ 0.05 were deemed significant. RESULTS: Initial cell type and number of groups did not significantly differ. A similar proportion of cells were removed from each group in each successive irrigation suction cycle. A significantly greater number of cells were removed from the first to third lavage irrigations than from subsequent irrigations. A distinctive washout curve was found showing progressive cell count reduction with the greatest change occurring within the first lavage irrigation retrieval progressing to 99.9% removal by the third cycle and no significant changes after further irrigations. Peritoneal cell depletion as a result of lavage was not statistically different regardless of the volume of irrigation used. CONCLUSION: Cells within the peritoneal fluid, adherent mesothelial cells and free floating cells are rapidly removed from the peritoneal cavity by three lavage irrigation suction cycles. Three irrigations of any volume or irrigation from 100 to 500 cc at a time removes virtually all peritoneal cells. Three or more peritoneal lavages regardless of volume used significantly reduces peritoneal resident cell population. Elimination of peritoneal fluid and peritoneal resident cell population by irrigation severely affects at least the immediate homeostatic balance of the peritoneal cavity. Supported by: Georgia BioMedical, Inc. P-848 Cross Talk Between Inducible Nitric Oxide Syntheses (iNOS) and Myeloperoxidase (MPO) in Fibroblasts Isolated From Normal Peritoneal and Adhesion Tissues. G. M. Saed, H. Lu, Z. Jiang, S. Abuolba, H. Abu-Soud, M. P. Diamond. Wayne State University/Detroit Medical Center, Detroit, MI. OBJECTIVE: Fibroblasts beneath the mesothelium in the human peritoneum play an important role in the healing process of the peritoneum. Adhesion fibroblasts are characterized by lower levels of nitric oxide (NO) although there were no differences in iNOS, the enzyme which generates NO, between normal and adhesion fibroblasts. Additionally, adhesion fibroblasts exhibited significantly lower levels of MPO. We have previously shown that there is a cross talk between MPO and iNOS utilizing purified enzymes. MPO upregulate the catalytic activity of iNOS by preventing the feed back inhibition attributed to the formation of iNOS nitrosyl complex. Our objective is to determine whether there is a cross talk between MPO and iNOS in vivo utilizing fibroblasts isolated from normal peritoneum and from adhesions. DESIGN: Laboratory Study. MATERIALS AND METHODS: We have obtained fibroblast primary cultures from normal peritoneum and adhesion tissues of the same patients. We have utilized the small interfering RNA (SiRNA) technology to specifically knock-out the expression of MPO alone, iNOS alone, and combination of both and then quantify the expression levels of MPO and iNOS using the real-time RTR/PCR developed in our laboratory.
S463
RESULTS: There was no significant difference in iNOS mRNA levels between normal (5.0 X105 iNOS mRNA copies and adhesion fibroblasts (4.8 X105). When iNOS gene expression is knocked-out, the iNOS mRNA levels were reduced by 98% in both normal and adhesion fibroblasts. Similar finding were observed for MPO SiRNA, confirming the efficiency of SiRNA to completely destroy iNOS and MPO mRNAs in both cell lines. Use of MPO mRNA resulted in a 62 % decrease in iNOS mRNA levels in normal fibroblasts and completely destroyed iNOS mRNA in adhesion fibroblasts. Destruction of mRNA for iNOS and MPO together resulted in a complete destruction of iNOS mRNA in both cell lines. CONCLUSION: Our data suggests that there is intricate cross talk between iNOS and MPO in normal and adhesion fibroblasts. This may be the cause for the disturbance of iNOS/NO/protein nitration system observed in adhesions, as compared to, normal fibroblasts. Supported by: None
P-849 Differential Expression of Estrogen, Progesterone, Androgen, and Prolactin Receptors in Human Fibroblasts Isolated From Normal Peritoneum and Adhesions. G. M. Saed, L. Detti, H. Lu, Z. Jiang, S. Aboulba, M. P. Diamond. Wayne State University/Detroit Medical Center, Detroit, MI. OBJECTIVE: Adhesions are believed to contribute to infertility in 40% of infertile couples and are the most common cause of small bowel obstruction. Also abdominal-pelvic pain is attributed to adhesions in a little less of 30% of cases. Our objective is to characterize fibroblasts isolated from normal peritoneum and adhesion tissues of the same patient in their ability to express hormone receptors. DESIGN: Prospective experimental study. MATERIALS AND METHODS: We have utilized the real time RT-PCR to determine mRNA levels for estrogen (ER-␣, ER-), progesterone A and B isoforms (P4), androgen (A), and prolactin (PRL) receptors in primary cultures of fibroblasts isolated from normal peritoneum and adhesion tissues of the same patient. Our main outcome measure was the quantification of mRNA levels (mRNA copies/g RNA) of ER-␣, ER-, P4, A, PRL receptors in both cell lines. RESULTS: Our results have shown a differential expression in all hormone receptors studied normal peritoneal and adhesion fibroblasts. There were no detectable levels of ER-␣ in adhesion fibroblast as compared to 1,487 copies in normal fibroblasts. Normal fibroblasts (713 x 105) expressed significantly higher levels of ER- receptor (713 x 105) as compared to adhesion fibroblasts (24.8 x 105). There were no detectable levels of progesterone in normal fibroblasts as compared to (194 mRNA copies/g RNA) of progesterone in adhesion fibroblasts. The levels of androgen receptors were also markedly different in normal peritoneal (245 x 105) and in adhesion fibroblasts (180 x 105). There were higher prolactin levels in normal fibroblasts (280 copies) as compared to adhesion fibroblasts (72 copies). CONCLUSION: Expression of hormone receptors in adhesion fibroblasts is different from that in normal peritoneum fibroblasts. This may be used for future design of intervention therapies for postoperative adhesions. Supported by: None
P-850 The Role of Robotic-Assisted Laparoscopy in Gynecological Surgeries. N. Salamat Saberi, B. Shahmohamady, C. H. Nezhat, F. Nezhat, C. Nezhat. Stanford University Hospital, Palo Alto, CA; Mount Sinai Medical, New York, NY. OBJECTIVE: To evaluate the role of robotic-assisted laparoscopy in gynecological surgeries DESIGN: Retrospective chart review study MATERIALS AND METHODS: In a tertiary referral center for laparoscopic gynecologic surgery, a total of 15 patients who were undergoing various gynecologic surgeries were consented for combined laparoscopy and roboticassisted laparoscopic surgery. The da Vinci robot was used in each case. Four ports were inserted: umbilicus, suprapubic, and two lateral ports. These surgeries were performed both using laparoscopy and robotic-assisted laparoscopy surgery.
S464
Abstracts
RESULTS: Fifteen patients underwent variety of gynecologic surgeries such as myomectomies, treatment of endometriosis, total and supracervical hysterectomy, ovarian cystectomy, sacral colpopexy and Moskowitz procedure. The assembly time to switch from laparoscopy to robotic assisted surgery was 18.9 minutes (14-27 minutes), and the disassembly time was 2.1 minutes (1-3 minutes). Robotic assisted surgery was used for suturing after myomectomy, tissue manipulation, suturing of cervical stump after supracervical hysterectomy, vaginal cuff closure and suspension after total laparoscopic hysterectomy, ovarian suturing after cystectomy and performing Moskowitz procedure. CONCLUSION: Robotic-assisted laparoscopic surgeries have advantages in providing a 3 dimensional visualization of operative field, decreasing fatigue and tension tremor of surgeon, and added wrist motion for improved dexterity and greater surgical precision. The disadvantages include cost and added operating time for assembly and disassembly. The greatest practical use lies in the ability to perform easier suturing in laparoscopic surgery. Supported by: None
P-851 The Value of 3D Ultrasound in the Diagnosis and Treatment of Asherman’s Syndrome. J. Cohen, A. B. Copperman. Mount Sinai Medical Center, New York, NY. OBJECTIVE: To assess the value of 3D ultrasound in the management of patients with suspected Asherman’s syndrome DESIGN: A prospective comparative study of infertile patients with suspected Asherman’s undergoing both HSG and 3D ultrasound prior to hysteroscopy, between the years of 1998-2004 MATERIALS AND METHODS: 54 women, ranging in age from 23-49, presented to a tertiary care center with secondary infertility between the years of 1998-2004 and were given a diagnosis of Asherman’s. 49 presented with the diagnosis and 5 were newly diagnosed. The results of both imaging modalities were compared with those obtained by hysteroscopy. Sensitivity, specificity, positive predictive and negative predictive values were calculated. In addition, the ability to re-establish a functional uterine cavity and attain fertility post-operatively were evaluated. RESULTS: Intra-uterine adhesions were demonstrated on 3D ultrasound and HSG in all 54 patients and confirmed at hysteroscopy. 3D ultrasound had a sensitivity of 100% and PPV of 100%, when compared with the hysteroscopic findings, for correctly grading the extent of intra-uterine adhesions in terms of percent cavity of obstructed, while HSG showed a sensitivity of 66.7% and a PPV of 66.7% for accurately identifying the extent of intrauterine adhesions. In 61.1% of these cases lower segment outflow obstruction was present and the HSG was erroneously read as severe Asherman’s with complete cavity obstruction. Following targeted hysteroscopy, 45/50 (90%) of patients conceived. CONCLUSION: 3D ultrasound, a non-invasive office based exam, has a better prognostic value with a higher sensitivity and PPV than HSG when classifying the grade of cavity damage in patients with suspected Asherman’s syndrome. 3D ultrasound is particularly helpful in differentiating severe IUA from lower uterine segment outflow obstruction. In addition, the superior quality of the diagnostic images produced by the 3D ultrasound allows the hysteroscopist to perform a more precise procedure and this, combined with good surgical technique and post-operative care, may ultimately improve reproductive outcome. Our study is one of the first and largest to demonstrate the benefit of 3D ultrasound in the diagnosis and treatment of Asherman’s Syndrome. We propose re-evaluating current grading systems that classify severity according to amenorrhea and/or HSG findings, as they clearly mis-classify patients with lower outflow tract obstruction. In fact, these patients actually have an excellent prognosis if there is normal cavity beyond the obstruction. Supported by: None
P-852 Outcome of Pregnancies Achieved Following Laparoscopic Surgery for Correction of Tubo-Peritoneal Factor Infertility Associated With Endometriosis or Pelvic Inflammatory Disease (PID). M. F. Mitwally, H. Albuarki, Z. Al-Wahab, M. P. Diamond, M. Abuzeid. Wayne State University, Detroit, MI; IVF-Michigan and Michigan State University, Rochester Hills & East Lansing, MI.
Vol. 84, Suppl 1, September 2005