lumen oocyte aspiration needle was introduced into the endometrioma.Normal saline solution was sequentially injected and aspirated into the cystuntil all the contents had been evacuated. Tetracycline 5% (5-15cc) was theninjected into the empty cyst cavity and the needle was partially withdrawn.After a waiting period of 2-3 minutes to allow any leakage of the tetracy-cline into the pelvic cavity to cease, the residual saline solution wasaspirated from the pelvic cavity. The patients were discharged within 1 hourof the procedure, free of pain, in all cases.

Results: Of the 22 women who underwent sclerotherapy, all had fol-low-up ultrasound examinations 6 weeks following the procedure. Two hada recurrence necessitating a second procedure. There were no significantcomplications. Of this group, 17 patients subsequently underwent IVF.None had any sign of recurrence of the endometrioma at egg retrieval. Theclinical pregnancy and implantation rates were 59% (10/17,) and 35% (15sacs/ 43 embryos), respectively.

Conclusion: Sclerotherapy is a safe, effective and relatively inexpensivealternative to surgery in women with ovarian endometriomas who arepreparing for treatment with IVF.

P-313

Effects of peritoneal fluid from endometriosis patients on pre-implan-tation mouse embryo development and apoptosis. Navid Esfandiari,Tommaso Falcone, Ashok Agarwal, Marjan Attaran, Matthew Karafa,Rakesh K. Sharma. Cleveland Clin Fdn, Cleveland, OH.

Objective: The cause of infertility in women with endometriosis remainscontroversial. The aim of our study was to assess the effect of peritonealfluid (PF) from endometriosis patients on pre-implantation mouse embryodevelopment and apoptosis.

Design: Prospective experimental study in an embryology research lab-oratory.

Materials and Methods: Peritoneal fluid from patients with endometriosis(n � 7) was obtained during laparoscopy. Two-cell mouse embryos werecultured for 72h in HTF medium supplemented with 10% serum substitutesupplement (SSS) � 5%PF (group I), 10% SSS � 10% PF (group II) and0% PF as control (group III). Grading of embryos from 2-cell to hatchingblastocyst was recorded at 24, 48, and 72 hrs. Blastocyst development rate(BDR), total cell number (TCN), and allocation of inner cell mass (ICM)and trophectoderm (TE) in blastocyst were recorded. ICM and TE wererecorded based on images obtained by confocal microscopy. Total blas-tomere count per embryo was determined by staining with bisbenzimide(Hoechst 33258). A terminal transferase-mediated dUPT end labeling(TUNEL) kit was used to determine apoptosis. To distinguish apoptosisfrom necrosis, embryos were stained with propidium iodide (PI) before theTUNEL assay.

Results: Embryo development to blastocyst, as well as TCN and ICM/TEratio was significantly different between the groups (Table I). However,exposure to PF did not change the incidence of apoptosis or the number ofapoptotic cells in cultured embryos. However, the number of apoptotic cellsin TE was significantly higher in group I and II (P � 0.001).

Conclusion: The developmental alterations observed following culture ofmouse embryos in presence of different concentrations of peritoneal fluidfrom patients with endometriosis provides evidence for the negative role ofendometriosis on pre-implantation embryo development; however, this roleis independent from the apoptosis process.

P-314

Differential contribution of reactive oxygen species and tumor necrosisfactor-� to the peritoneal fluid-induced embryotoxicity in endometrio-sis patients. Javier Noriega, Mohamed A. Bedaiwy, Sarah Worley, Rakesh

K. Sharma, Ashok Agarwal, Tommaso Falcone. Cleveland Clin Fdn,Cleveland, OH.

Objectives: Alterations in the production of peritoneal fluid cytokines andoxidative stress parameters contribute to the evolution of endometriosis.Peritoneal fluid (PF) TNF-� and reactive oxygen species (ROS) are indirectmeasures of both pathogenic mechanisms. Peritoneal fluid embryotoxicity isa presumptive mechanism of endometriosis-associated infertility. The ob-jectives of this study were to study the embryotoxic effects of PF in womenwith endometriosis, and to relate any embryotoxicity to the PF levels ofROS and TNF-�.

Design: Experimental study in a research laboratory in a tertiary carefacility.

Materials and methods: Reactive oxygen species levels were measured inlaparoscopically obtained PF of 12 endometriosis patients by enhancedchemiluminescence assay using luminol as the probe. Concentrations ofTNF-� in PF of those patients were measured using quantitative ELISAkits. Two hundered eighty four thawed mouse embryos were pooledandrandomly distributed between 5 groups: (A) composed of plain HTF withoutPF as a control group, (B) HTF supplemented with 10% PF, (C) HTFsupplemented with 15% PF, (D) HTF supplemented with 25% PF, and (E)HTF supplemented with 50% PF for each patient. The numbers of embryoswere from 62 to 132 embryos per concentration. Blastocyst developmentrates (BDR) were checked after 72 hours of incubation. Logistic regressionwith generalized estimating equations (GEE) to assess the effect of ROS,TNF-� and PF concentration on blastocyst development rate while adjustingfor correlation between embryos grown together and embryos grown in PFfrom the same patient was performed.

Results: Higher PF concentration was significantly associated with lowerBDR (P�0.001), both with and without adjusting for ROS and TNF-�(Figure). The median and interquartile ranges of PF ROS and TNF-� were1.21 (0.66-2.32) x106 cpm and 1.69 (0.21-6.33) pg/mL respectively. Thereis no evidence that either ROS or TNF-� was associated with BDR afteradjusting for PF concentration.

Conclusions: Peritoneal fluid embryotoxicity may be a contributing factorto endometriosis associated infertility. Peritoneal fluid ROS or TNF-� wasnot the culprit. The embryotoxic effect may be due to dilutional effects.Given the fact that this is an interim analysis, the results may change whenwe analyze data from the planned sample size of 25 endometriosis patients.

P-315

Polymorphisms of Glutathione S-Transferase mu-1 (GSTM1) and ofCatechol-O-Methyltransferase (COMT) and risk of endometriosis. Jen-nifer L. Marino, Chu Chen, Delia Scholes, Victoria L. Holt. Fred Hutchin-son Cancer Research Ctr, Seattle, WA; Group Health Cooperative Ctr forHealth Study, Seattle, WA.

S226 Abstracts Vol. 80, Suppl. 3, September 2003