onic villi sections yielded amplifiable DNA in 5 of 8 cases. These 5 cases had clear evidence of trisomy 16 by PCR when analyzed with a panel of 9 polymorphic chromosome 16-specific markers. Unamplifiable samples were not spe- cific for chromosome 16 primers, and likely reflected gener- alized DNA damage. PCR products for X, Y, and 16 accu- rately correlated for the 69,XXY case.

Conclusions: Common chromosomal aneuploides can be diagnosed retrospectively from archival pregnancy loss tissues and DNA, utilizing PCR-based strategies involving informative polymorphic chromosome-specific markers. REACH TM has several potential applications, offering: 1) recurrent loss patients more comprehensive diagnostic in- formation; 2) an adjunct to confirm the accuracy of cytoge- netic evaluation; 3) a back-up when cytogenetic evaluation fails; and 4) a methodology to screen for uniparental dis- omy. Supported by NIH HD 29729.

0 - 0 8 9

A C o m p a r i s o n o f P r e g n a n c y R a t e s in C l o m i p h e n e Ci trate (CC) O v a r i a n S t i m u l a t i o n Cyc les C o m b i n e d With I n t r a u t e r i n e I n s e m i n a t i o n (IUI) in Which Ovu- l a t i o n O c c u r r e d S p o n t a n e o u s l y or Was I n d u c e d With H u m a n C h o r i o n i c G o n a d o t r o p i n (hCG). P.Y. Ling, D. Maier, K. Thornton, J. C. Nulsen. Division of Re- productive Endocrinology and Infertility, Univ. of Con- necticut Health Center, Farmington, CT.

Objective: To determine the effect ofhCG induced ovula- tion vs spontaneous ovulation on clinical pregnancy and live birth rates in patients undergoing CC ovarian stimu- lation combined with IUI.

Design: A retrospective analysis of 711 CC/IUI cycles performed in 204 patients between Feb. 1992 and May 1996.

Materials and Methods: All patients had infertility of greater than one year duration and had undergone an infertility evaluation including: 1) assessment of hormonal and ovulatory status, 2) HSG _+ laparoscopy and 3) semen analysis. Diagnostic categories included unexplained in- fertility, endometriosis, anovulation, male factor and do- nor insemination. CC was administered for five consecu- tive days starting on cycle day 2, 3 or 5 at a dose of 50 to 150 mg daily. Sperm samples were prepared utilizing a discontinuous Percoll gradient. IUI was performed: 1) 36 hours after the IM administration of hCG, 10,000 units, given when at least one follicle had an average diameter -> 20 mm or 2) 18-22 hours following a spontaneous LH surge detected utilizing a home urinary assay kit. A clini- cal pregnancy was defined as the presence of a fetal pole/ fetal heartbeat seen on ultrasound. The data was analyzed using proportion analysis, chi square test and regression analysis.

Results: The overall clinical pregnancy rate per cycle was 9.6% (28/291) after spontaneous ovulation and 6% (25/ 420) after HCG-induced ovulation (P=0.07). The overall live birth rate per cycle was 8.2% (24/291) after spontane- ous ovulation and 6% (25/420) after HCG-induced ovula- tion (P=0.23). Given the sample size of the current investi-

gation and a power level of 0.8, a two-tailed test at the 0.05 level would be able to detect a minimum difference of 4%. These findings were independent of patient age, diagnosis, CC dosage or initial day of CC administration.

Conclusion: The findings of this investigation suggest that for patients undergoing ovulation induction with CC combined with IUI, there is no clinically significant differ- ence in live birth rates between cycles with spontaneous ovulation and hCG induced ovulation regardless of the diagnostic category. If a statistical difference exists, it is likely to be less than 4%. The routine utilization of ultra- sound monitoring and hCG administration therefore does not appear to be justified as the utilization of a home uri- nary assay results in similar live birth rates at a signifi- cantly lower cost to the patient.

O-090

Effect o f C o n s e c u t i v e E j a c u l a t i o n o n the Tota l Num- ber o f Mot i le S p e r m I n s e m i n a t e d . S. E. Brown, V. C. Montgomery Rice, D. L. Stewart, M. A. Harris, S. Jordan, T. K. Clinton. University of Kansas Medical Center, Kan- sas City, KS.

Objective: It has been suggested that a significant de- crease in sperm count occurs with consecutive ejaculation, thereby decreasing the total number of sperm inseminated (TMI), and limiting the effectiveness of two-day insemina- tion protocols. The objective of this study was to determine the changes in pre count/ml, % motility, and TMI in nor- mal and abnormal semen samples after two consecutive ejaculations.

Design: This was a retrospective case control study of semen samples collected from patients undergoing intra- uterine insemination (IUI).

Materials and Methods: Couples were asked to abstain from intercourse for 3 - 4 days prior to the initial IUI. Each patient produced two samples: the first on the day of the LH surge, the second a day later. Male factor patients were classified as follows: oligozoospermic (OZ; count <20 mill/ml), asthenozoospermic (AZ; motility <40%), and oli- goasthenozoospermic (OAZ), as determined by the Day 1 sample. ANOVA or Student's t-test were used for statisti- cal comparisons (indicated by an asterisk, p<0.05).

Results: The sample population was comprised of 85.8% and 14.2% normal and abnormal samples, respectively. A comparison of Day 1 values indicated a significant differ- ence between normal and abnormal samples. Although count/ml and TMI values for normal samples on Day 2 were statistically different from Day 1, the changes not clinically significant. Consecutive ejaculation had no effect on OZ samples. The motility of AZ and OAZ samples in- creased on Day 2, while count/ml and TMI were un- changed. The TMI of OZ and AZ samples were similar on both days. In comparison, the TMI of OAZ samples on Day i was significantly reduced. However, the increase in motility observed in OAZ samples on Day 2 equalized the TMI on this day. Of those patients with normal values on Day 1, a total of 34 (9.4%) became abnormal on Day 2.

Abstracts S45

Comparison of Semen Samples: Classified by Day 1 Diagnosis

Diagnosis n Count/ml % Motility TMI

Normal: Day 1 339 80.3 _+ 3.0 60.1 _+ 0.49 55.4 ± 2.3 Day 2 339 69.9 ± 2.8* 59.5 -+ 0.58 43.3 ± 1.6"

Abnormal: Day 1 56 27.3 ± 5.4* 40.0 _+ 2.3* 12.9 ± 1.9" Day2 56 27.2 -- 3.9* 44.6 -- 2.3* 13.8 ± 1.9"

Oligozoospermic: Day 1 28 13.3 ± 1.0 54.6 _+ 1.4 14.6 ± 2.6 Day2 28 16.4 ± 1.9 50.0 ± 2.8 13.4 ± 2.1

Asthenozoospermic: Day 1 17 63.0 ± 15.3 27.0 ± 2.4 15.7 ± 4.4 Day2 17 48.2 + 9.5 41.1 ± 4.9* 12.9 ± 2.9

Oligoasthenozoospermic: Day 1 11 9.5 -+ 1.8 22.5 ± 3.9 4.3 ± 2.1 Day2 11 17.7 ± 8.8 35.3 ± 5.9* 8.8 ± 4.4*

* P < 0.05.

Conclusion: Consecutive ejaculation does not adversely affect the count/ml, motility, or TMI of either normal or abnormal semen samples. Less than 10% of normal pa- tients convert to abnormal after consecutive ejaculation. In the case of AZ and OAZ samples, a significant improve- ment in motility was observed on Day 2. For OAZ samples, this increase was sufficient to raise the TMI.

O-091

Effectiveness of High Dose Progesterone in the Treatment of Ovarian Hyperstimulation Syndrome in Women Undergoing a Superovulation Program

1 2 for Infertility. F. Scarpellini, R. Boccadoro, 1M. Sbracia, 3j. A. Grasso. 1FARM, 2Dept. Ob/Gyn Ospedale Civile di Colleferro, Italy. 8Yale University School of Medicine, New Haven, CT.

Objective: The ovarian hyperstimu]ation syndrome (OHSS) is becoming a common complication of pharmaco- logical ovulation induction in IVF programs and in super- ovulation protocols for the t reatment of infertility. Several therapies have been suggested but at the moment the best t reatment is the prevention of OHSS by biophysical and chemical monitoring while refraining from the infusion of hCG. In order to study the effectiveness of progesterone administration in the t reatment of OHSS we compared this therapy with infusion of albumin in women with OHSS.

Design: We have t reated 24 women undergoing a super- ovulation program which developped OHSS randomly with progesterone or in t ra venous infusion of albumin, after the clinical manifestation of OHSS.

Materials and Methods: Twenthy four consecutive women which developped moderate or severe OHSS after superovulation treatment protocol for IVF were included in the study. The women were randomly assigned to one of the two groups of patients, 12 women each, which were treated one group with progesterone, 200 mg daily, and the other with intra venous infusion of albumin, 100 rag daily. Dimension of ovary were recorded at beginning of treat- ment, after 5 days and after 10 days of t reatment in the two groups. Hematocrite estradiol plasma levels and creatinine were checked every two days during the treatment time.

Results: The women treated with progesterone showed

a dramatic improvment of both subjective and objective symptoms. The reduction in ovarian size and estradiol lev- els was faster in progesterone t reated patients with re- spect to the albumin t reated patients (P<0.01).

Conclusion: Progesterone t reatment prevent the clinical manifestation of OHSS more than albumine infusion. If these data are confirmed by larger controlled studies it would be the solution to one of the most important atro- genic problems of the t reatment of infertility.

Tuesday, October 21, 1997 2:00 P.M. to 5:00 P.M.

D. Clinical ART

0-092

Expression of Apoptosis Related Genes in Human Oocytes, Embryos, and Reproductive Cells. H. C. Liu, Z. Y. He, C. Mele, O. Davis, Z. Rosenwaks. The Center for Reproductive Medicine and Infertility, Department of OB/ GYN, The New York Hospital-Cornell Medical Center, New York, NY.

Objectives: To study whether apoptosis occurs in human embryogenesis.

Design: The balance of Bax, Fas, Fas-ligand, P53 and BCL-2 expression has been shown to determine cell fate: survival vs. apoptosis. Increasing BCL-2 promotes sur- vival whereas increasing Bax, Fas, Fas-ligand, and P-53 favors apoptosis. In our study, the expression of these apoptosis related genes were monitored in human oocytes/ embryos, granulosa-luteal and late secretory endometrial stromal cells to evaluate the presence of apoptosis in these biological materials.

Materials and Methods: Human non-viable and degen- erated embryos, immature and non-fertilized oocytes, and granulosa-luteal and endometrial stromal cells donated by our IVF-ET patients were used to detect transcripts of apoptosis related genes by reverse-transcriptase-polymer- ase chain reaction (RT-PCR).

Results: The frequencies of gene expression (+/n) are shown in the following table:

Bax (+/n) Fas (+/n) P53 (+/n)

Degenerated Embryos 5/5 5/5 0/2 Non-viable Embryos 5/6 7/7 0/3 Immature Oocytes/Eggs 0/6 0/4 0/6 Non-fertilized Eggs 0/3 0/5 0/2 Granulosa Cells 3/3 3/3 3/3 Stromal Cells 3/3 3/3 3/3

BCL-2 Fas-ligand Actin (+/n) (+/n) (+/n)

Degenerated Embryos 0/2 1/3 17/17 Non-viable Embryos 0/3 0/2 21/21 Immature Oocytes/Eggs 0/5 0/3 24/24 Non-fertilized Eggs 0/3 0/2 15/15 Granulosa Cells 3/3 0/3 3/3 Stromal Cells 0/3 0/3 3/3

846 Abstracts