MATERIALS AND METHODS: Of 1632 ultrasonographic scans of the pelvis, 49 displayed Mullerian duct defects: 17 arcuate, 8 bicornuate, 18 septate, 2 didelphys and 4 unicornuate. The diagnosis of septate uterus was confirmed using the Accuvix 3D ultrasound. Subendometrial blood flow was measured both qualitatively and quantitatively using an Acuson Sequoia with a 4-8MHz transvaginal probe. The quality of subendometrial flow was graded from 1 to 4 according to the amount of visible color flow (grade 1 when visible in the outer 1⁄2 of the myometrium, grade 2 in the inner 1⁄2 of the myometrium, grade 3 in the subendometrial space and grade 4 color extended into the endometrium). Quantitative subendometrial blood flow was measured as peak systolic velocity (PVS). Subendometrial blood flow was compared between septate uteri and uteri with no Mullerian duct defect. RESULTS: All septate uteri had grade 3 (n⫽2) or grade 4 (n⫽16) blood flow. No differences were observed when PSV measured on cycle day 2-3 was compared between septate uteri (mean 8.6 cm/sec, SD 2.8, SEM 0.5 ) and normal uteri (mean 8.9 cm/sec, SD 2.9, SEM 0.5) (F⫽0.4, P⫽0.5). CONCLUSION: Consideration of amending the AFS classification of Mullerian duct defects to include septate uteri with normal subendometrial blood flow and abnormal blood flow should be undertaken before clinical trials to evaluate efficacy of surgical removal of the septum are done. Supported by: None P-520 TRIFUNART (TRIPLE FUNCTION ASSISTED REPRODUCTIVE TECHNOLOGY) CANNULA - HYSTEROMETRY, ENDOMETRIAL ASPIRATION AND BIOPSY - A NEW TOOL FOR ROUTINE OFFICE USE. D. S. Castellotti, A. S. Cambiaghi. Instituto Paulista de Ginecologia, Obstetricia e Medicina da Reprodução, São Paulo - SP, Brazil. OBJECTIVE: Present a new instrument with triple function of great value for patients in treatment for infertility. Hysteroscopy (HSC) is the gold-standard exam for evaluation of the uterine cavity. Before in vitro fertilization (IVF) treatment, HSC is a valuable source of information providing diagnosis of intrauterine pathologies. Endometrial biopsy provides information of luteal function and hysterometry assesses the length of cervical canal and uterine cavity. That information can help a reproductive endocrinologist to decide where one would transfer embryos to increase implantation rates. DESIGN: A cannula was designed to accomplish precise measures of endocervix and uterine cavity, aspiration and biopsy endometrial tissue while performing a routine HSC under local anesthesia (not always necessary). Usually, after placement of the Pozzi, the endometrial biopsy is carried out following hysteroscopy; followed by the biopsy curette (figure). This step could cause some degree of discomfort to the patient. This new cannula is a dual system; biopsy and aspirates the endometrium. It is a cylindrical tube, longer than the diagnostic sleeve and is introduced by the HSC sleeve. It has in its distal portion, a rectangular window with cutting edges that allows punching a larger amount of endometrial tissue than a pipelle, minimizing repetition of the exam for “insufficient tissue”. The distal portion of this instrument is marked allowing accurate point to biopsy as well as to perform a hysterometry. In the distal portion a disposable syringe is connected that guarantees a negative and constant pressure (picture). The cannula will not scrape the cervix canal diminishing the discomfort of the patient (figure). MATERIALS AND METHODS: Fifty patients were evaluated during the period of July of 2004 to January of 2006. They had previously undergone an endometrial biopsy with conventional curette immediately after a diagnostic HSC. In the early luteal phase of a preceding IVF cycle we performed a HSC with endometrium biopsy with the TRIFUNART cannula. Patients answered a score pain questionnaire comparing the conventional biopsy with the one with the new cannula. RESULTS: Forty four of the fifty evaluations had mentioned lesser pain and discomfort with this cannula in relation to the conventional and six patients had not noticed difference. In all cases enough material for endometrial dating showed the advantage of the biopsy with HSC. The data from hysterometric measurements correlated with the measure of the uterine cavity made by transvaginal ultrasound showed excellent agreement. FERTILITY & STERILITY威 CONCLUSION: The TRIFUNART cannula is a new instrument of easy manipulation which can be used in office with a minimum of discomfort and adding valuable information. Supported by: None. P-521 ROLE OF SALINE INFUSION SONOGRAPHY (SIS) IN UTERINE EVALUATION PRIOR TO FROZEN EMBRYO TRANSFER (FET) CYCLE. P. S. Gera, M. C. Allemand, L. L. Tatpati, T. M. Galanits, D. Morbeck, C. C. Coddington III. Mayo Clinic, Rochester, MN. OBJECTIVE: Saline infusion sonography has been used in the evaluation of structural uterine abnormalities in infertile patients undergoing in-vitro fertilization (IVF). It gives excellent anatomic detail of the uterus & is accurate in the diagnosis of mass lesions, uterine septum & various other anomalies. As compared to hysteroscopy it is less invasive, better tolerated & less expensive. No studies have been found that address the value of a repeat SIS in patients undergoing FET cycles. This study aims to determine the role of SIS in uterine evaluation prior to the FET cycle. DESIGN: Institutional review board approved retrospective data analysis conducted at division of Reproductive Endocrinology & Infertility. MATERIALS AND METHODS: Twenty-nine patients undergoing FET cycles at Mayo clinic from September 2005 to February 2006, using their own or donor embryos, were screened for uterine anomalies using SIS. All these patients had a normal uterine evaluation in the past using either SIS or hysterosalpingography before their fresh IVF cycle or a previous FET cycle between 1998 & 2004. Transvaginal sonography was performed followed S327 by SIS in the follicular phase of the menstrual cycle prior to the actual FET cycle. Prophylactic antibiotics were not routinely given. A pelvic exam was done prior to SIS to determine the position & mobility of the uterus. A Tampa catheter (Cooper Surgicals, Connecticut) was used for SIS. After catheter insertion, sterile saline was instilled to distend the uterine cavity slowly under direct sonographic visualization. The uterine cavity & the endocervical canal were evaluated for any evidence of intracavitary polyps, myomas, focal endometrial thickening, uterine septum, cavity distortion & endometrial changes. The medical records of the patients were screened for SIS findings. The main outcome measures of the study were SIS findings, clinical pregnancy rate & ongoing pregnancy rate. RESULTS: The mean age of the patients was 34.3 ⫾ 4.6 years (range 26-47). Seven out of 29 patients (24.1%) had positive findings on SIS, in spite of a previous normal uterine evaluation. Diagnostic ultrasound prior to SIS in all these patients was suggestive of a uterine abnormality. The specific nature of the abnormality was further delineated clearly by SIS. The findings included endometrial polyp (42.8%), myoma (14.2%), uterine septum (14.2%), cervical stenosis at the level of internal os (14.2%) & cystic changes in the endometrium with negative Doppler findings (14.2%). Five patients underwent subsequent operative hysteroscopy & correction of their anomalies. 18/29 patients conceived (62.0%) with an ongoing pregnancy rate of 41.3% (12/29). The clinical pregnancy rate was 48.2% (14/29), 4 patients had biochemical pregnancy (13.7%) & 2 had pregnancy loss (6.8%). One patient with a uterine septum detected on SIS declined hysteroscopic treatment, she had a pregnancy loss at 7 weeks. CONCLUSION: SIS prior to FET cycle can be helpful in detection and further delineation of uterine abnormalities associated with poor implantation and pregnancy failure. If time since uterine evaluation before IVF/ previous FET cycle has been more than 1-2 years we recommend doing SIS as this may improve FET outcome. Supported by: None P-522 ULTRASOUND-GUIDED EMBRYO TRANSFER AND DIRECT PLACEMENT OF EMBRYOS DIMINISHES ECTOPIC PREGNANCY RATES. A. Alladawi, M. Nouriani, B. Acacio. White Memorial Medical Center, Los Angeles, CA; Sher Institute of Reproductive Medicine, Glendale, CA. OBJECTIVE: To analyze the incidence of ectopic pregnancies (EP’s) after ultrasound-guided IVF-ET, using a specific technique. DESIGN: Retrospective study. MATERIALS AND METHODS: A total of 1018 women who underwent IVF procedures from September 2002 through January 2006 were studied with regard to indications for IVF and factors related to the IVF procedure through a retrospective review of medical charts. The incidence of EP and techniques utilized were reviewed. The review consisted of two experienced clinicians, separately performing ultrasound-guided IVF-ETs. During the stimulation phase of the cycle, the distance between the fundus of the uterine cavity and the external cervical os was measured using colpohydrosonography (CHS). At the time of embryo transfer, the catheter was advanced to a distance equivalent to that calculated from the CHS minus 5 mm. Embryo placement was observed via ultrasound. RESULTS: 431 out of 1018 cycles resulted in a positive pregnancy test (pregnancy rate of 42.3%), 4 of which resulted in EP’s (0.4% of all patients and 0.97% of all pregnancies; P⬍ .0001). This data was compared to a review of 9 previous studies, which collectively reveal an EP rate of 4.5% of all pregnancies. Previous studies reviewed 8,604 pregnancies, with dates ranging from 1983 to 2003. Five of the studies (a total of 3,193 pregnancies) attribute tubal factor as the main cause of the increased EP rates. Of the 4 EP’s in our study, 2 of the patients were advanced maternal age (AMA); 1 procedure had an outcome of a heterotopic pregnancy, which subsequently resulted in 1 successful live birth. Tubal factor was not an indication for IVF for any of the 4 patients with ectopic pregnancies. CONCLUSION: Ultrasound-guided IVF-ET with embryos placed 5mm proximal to the uterine fundus, by experienced clinicians using state of the art equipment, yielded a statistically significant decrease in the EP rates, regardless of IVF indications. AMA may be an independent risk factor for EP. Supported by: None S328 Abstracts P-523 COCULTURE WITH GRANULOSE CELLS OF HUMAN OOCYTES DURING IN VITRO MATURATION AND THE INFLUENCE ON THE MATURATION OF OOPLASM. J. Tong Jr., Y. Li, Z. Chen, X. Gao, M. Li, J. Hu. Reproductive Center of Shandong Univ. and Shandong Provincial Hospital, Shandong Province China, Jinan,Shandong Province, China. OBJECTIVE: IVM technique plays an important role in assisted reproductive technologies, especially for PCOS patients. The current in vitro culture medium can achieve nuclear maturation of oocytes rather than cytoplasmic maturation. This study is designed:firstly, to identify whether nuclear and cytoplasmic maturation can reach simultaneously in vitro maturation. Secondly, to observe if in vitro culture time can affect nuclear and cytoplasmic maturity of oocytes . Thirdly, Many animal experiments have identified that cocultured with monolayer granulose cells(GCs) can greatly improve the ooplasmic maturation, which is manifested in high fertilization rate, high embryo quality and high pregnancy rate. Our objectives were to confirm that cocultured with GCs can surely improve ooplasmic maturation of human oocytes. In this experiment, we used the linear distribution of cortical granules(CG) as the marker of cytoplasmic maturity of oocytes, since many reports have defined that. DESIGN: Prospective cohort study. MATERIALS AND METHODS: Immature oocytes were retrieved from PCOS patients who were undergoing treatment from March to June 2005 at the Reproductive Medical Center in the Provincial Hospital of Shandong,all of whom had signed informed consents explicitly for this study. Pick-up of immature oocytes were performed 36 hours after HCG injection in natural menstruation cycles. Granulosa cells for coculture were taken from the cumulus-oocyte complexes (COC) in classical ICSI cycles. The COCs were digested with 80IU/ml hyaluronidase no more than 30 seconds. Then GCs were suspended with M-199 medium, centrifuged twice, 1000rpm for 10 mins. Then the suspension of granulose cells were dropped into 4-well plates and half of the culture medium was changed after 24 h. At that time, monolayer of GCs was formed and prepared for coculture with immature oocytes. Randomly,some of immature oocytes were cocultured with GCs and others were not cocultured in M-199 medium as control. All of them were evaluated 24h and 48h after in vitro maturation. The oocytes reached MII stage were labeled with FITC-LCA and observed under laser-scanning confocal microscope to investigate the distribution of CG. RESULTS: In comparison with the oocytes cultured in M-199 medium without granulose cells, the percentage of nuclear maturity after 24h, 48h in vitro maturation of cocultured oocytes were not significantly different respectively(36/69 vs 37/71,P⬎0.05;43/69 vs 43/71,P⬎0.05).The nuclear maturation rate of oocytes cultured for 48h are significantly higher than that of 24h in both control and experimental group (37/71 vs 43/71,P⬍0.05; 39/69 vs 43/69,P⬍0.05).As for CG distribution pattern, the rate of peripheral distribution pattern is significantly higher in cocultured oocytes compared with control, either after cultured for 24h (27/39 vs 10/37,P⬍0.01) or cultured for 48h(31/43 vs 11/43,P⬍0.05),but for the comparison of maturality after in vitro maturation for 24hs between for 48hs in either control group or in co-culture group,no significant difference(10/37vs11/ 43,P⬎0.05;27/39vs31/43,P⬎0.05). CONCLUSION: It surely occurs that nuclear maturation and cytoplasmic maturation can not reach simutaneously during in vitro maturation. Cocultured with granulose cells monolayer can prompt greatly the cytoplasmic maturation of oocytes.Prolonged culture time can increase nuclear maturate rate but seem no helpful in cytoplasmic maturation. Supported by: None P-524 PREGNANCY OUTCOME IN PATIENTS UNDERGOING UTERINE EVALUATION BY SALINE INFUSION SONOGRAPHY (SIS) BEFORE A FROZEN EMBRYO TRANSFER (FET) CYCLE. P. S. Gera, M. C. Allemand, L. L. Tatpati, D. Morbeck, C. C. Coddington III. Mayo Clinic, Rochester, MN. OBJECTIVE: 1.3 % of the patients undergoing IVF-FET have structural abnormalities of the uterus & the live birth rate in this group is 27.3%. Uterine anomalies may be associated with implantation failure & early pregnancy loss (PL) & their accurate identification followed by surgical Vol. 86, Suppl 2, September 2006