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Thirty Years of Advances in
Reproductive Endocrinology and ART: A Celebratory Congress
Centre Mont-Royal, Montreal, Canada, May 27-31, 2011
 
  Poster Presentations

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OVULATION INDUCTION IN POLYCYCTIC OVARIAN DISEASE THROUGH TRANSVAGINAL ULTRASOUND-GUIDED FOLLICULAR ASPIRATION
M.Y.K. Amin
, M.R Gaber,  M.H Mahmoud, E.A Khalifa                  
Alexandria University, School of Medicine, OB/GYN

Objective: To evaluate the feasibility and the effect of Transvaginal Ultrasound (TV/US) guided follicular aspiration on inducing ovulation, hormonal profile. And subsequent pregnancy rate in patients with PCOD. Methods: Fifteen patients had anovulatory infertility due to PCOD and not. Responding to Clomiphen Citrate (CC) for at least 3 months. After informed Consent, All were subjected to ovarian stimulation using 100mg CC daily for 5 days. Three days later, aspiration of all follicles more than one cm in diameter were done, using aspiration needle attached to TV probe under general anesthesia .Serum level of basal LH, FSH, and LH/FSH ratio were measured, then, one week and one month after aspiration. Evidence of ovulation and pregnancy were evaluated by US for the following 3 months. Results : Nine cases menstruated regularly after aspiration, while 6 demonstrated oligomenrroea.There was a significant decrease in LH, increase FSH and subsequently a significant decrease in the ratio.Ovulation detected in 9 cases (60%), 2 cases after one month, 5 in the second month, and 2 cases became pregnant(13%) at the third month of follow up. It is convenient to conclude that, the use of US-guided aspiration can be advised as a second step after failure of CC alone in PCOD, before proceeding to more expensive therapy. The subject is still open for further assessment, on large scale.
   
PREIMPLANTATION GENETIC DIAGNOSIS (PGD): THE USERS’ PERSPECTIVE ON ETHICAL ISSUES
C. Bouffard 
                      
Division of Genetics, Department of Paediatrics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada
   
Around the world, the medical and social uses of PGD are in process of diversification. As well, with little heed for legislative contexts, reproductive tourism is making these uses increasingly accessible. With users currently exercising more power than the experts over the development, marketing, and institutionalization of this reprogenetics technique, the positions adopted by the experts are no longer sufficient to ensure adequate ethical governance of this and other reprogenetic procedures. In this context, it appears timely to inquire into users’ perspective on what is ethical and unethical in connection with PGD. Methods: Multi-site ethnographic study in medical anthropology conducted in the three centres accredited to perform PGD in France. 990 hours of participant observation at PGD clinics and laboratories (clinical consultations, diagnosis laboratories, FIV, embryo-cell puncture, and embryo-transfer procedures). 40 semi-structured interviews were conducted with 20 couples undergoing PGD. Results: For these couples, PGD is an act of love toward one’s partner and unborn child, an expiatory rite, a means of assuming responsibility, and a way to compensate for one’s biological inability to prevent a child from being afflicted with a serious disease. Participating couples’ representations about what is ethical coherently integrate individual, familial, cultural, social, moral, economic, and medical aspects of the issue. Conclusion: These French PGD users had developed substantially different representations about the procedure than those held within the bioethics milieu. Ignoring users’ perspective in the debate on ethical governance of PGD and other reprogenetics practices is likely to make efforts at governance irrelevant.
    
ETHICAL CONSIDERATIONS ASSOCIATED WITH THE CROSS-BORDER MOVEMENT OF GAMETES: A CANADIAN PERSPECTIVE
V. Couture
1, C. Bouffard 1, J.M. Moutquin 2, R. Drouin 1                  
1 Division of Genetics, Department of Paediatrics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada
2 Department of Obstetrics and Gynecology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada

Introduction: The Canadian Assisted Human Reproduction Act (2004) has the particularity to ban the sale of human biological material. The objective behind this measure is to avoid all forms of objectification of the person. This position had the effect of reducing the number of donors and led to a shortage of gametes for Canadian banks, thus limiting infertile couples to altruistic donations. In 2011, almost all of the donations that are not directed towards relatives are coming from abroad. Thus, the Act had included, despite themselves, the practitioners and the users in an international movement of cross-border reproductive transactions. Methods: Review and analysis of the literature on cross-border transactions associated with gamete donation from various databases (OVID, EBSCO, SCOPUS, CAIRN and FRANCIS). Results: There is a limited number of studies on the Canadian situation regarding the cross-border flow of gametes. From our legal analysis of gamete donation in the West, we can affirm that the situation in Canada is similar to several other Western countries. From an ethical standpoint, this poses several problems: (1) it encourages the purchase of gametes from abroad; (2) it promotes the use of gametes from countries where paying donors is allowed; (3) it leaves children from such donations in total ignorance of their biological origins; (4) it increases the weight of this trade on vulnerable populations in poorer countries. Conclusion: Beyond the concerns about the availability, Canadian law raises several ethical questions as regards the fundamental principles of justice and respect for human dignity.

MATERNAL CAFFEINE CONSUMPTION DURING LACTATION AFFECTS POSTNATAL DEVELOPMENT OF TESTIS IN OFFSPRING WISTAR RATS
M. Dorostghoal
1, A.A. Moazedi 2, P. Nooraei 3                    
1 Assistant Professor of Embryology, Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz, Ahwaz, Iran
2 Professor of Physiology, Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz, Ahwaz, Iran
3 Msc of Embryology, Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz, Ahwaz, Iran
 
During recent decades, there are reports about the decrease of sperm production ability in men affected by different factors. Caffeine is one of the popular nutrients that used extensive in human populations that its reproductive effects have been reported. So, present study was aimed to determine the effects of Caffeine consumption during lactation on postnatal development of testis in offspring Wistar rat. Female Wistar rats were exposed to low and high doses (25 and 35 mg/kg) of Caffeine via drinking water during lactation. The testes of pups were removed at 21, 28, 60 and 90 days of postnatal development, their weights recorded and were fixed in Bouin’s solution. Then, the volumes of testes were estimated by Cavellieri method. Results: Mean body weight decreased significantly (p<0.05) in high dose group at all ages of postnatal development. Mean testis relative weight and total volume were decreased significantly (p<0.05) at 28, 60 and 90 days of age in 35 mg/kg dose group. Significant (p<0.05) decreases were seen in means of seminiferous tubules diameter and germinal epithelium at 28, 60 and 90 days of age in high dose treatment group. Also, serum testosterone levels decreased significantly (p<0.05) at puberty in high dose treatment group.Present study indicates that Caffeine consumption during lactation can reduce efficiency of spermatogenesis and fertility in offspring Wistar rats at puberty.
 
DEVELOPMENTAL EFFECTS OF MATERNAL CAFFEINE CONSUMPTION ON OVARIAN FOLLICLES IN OFFSPRING WISTAR RATS
M. Dorostghoal 1, A. Moazedi 2, S. Adham 3                   
1 Assistant Professor of Embryology, Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz, Ahwaz, Iran
2 Professor of Physiology, Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz, Ahwaz, Iran
3 MSc of Embryology, Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz, Ahwaz, Iran
  
In recent years concerns have been raised about incidence of human reproductive disorders. So, present study was designed to assess the effects of maternal caffeine consumption on ovarian follicles development in offspring rats.60 Pregnant female rats randomly divided in control group and two treated groups received caffeine via drinking water during gestation (26 and 45 mg/kg). The ovaries of offspring were removed at 28, 60 and 90 days after birth, their weights recorded and fixed in Bouin’s solution. Followed tissue processing, 5µm serial sections were prepared, then, structural changes of ovarian follicles and corpora lutea were studied in offspring during postnatal development. Ovary weight decreased significantly (p<0.05) in high dose caffeine-treated group at all stages of postnatal development. Significant (p<0.05) decreases were seen in number of primordial follicles from at 28, 60 and 90 days after birth in high dose caffeine-treated group. Primary and secondary follicles numbers and secondary and antral follicles diameters were not shown any significant differences between groups. Also, no statistically significant differences were seen in number of corpora lutea between different groups.Present study showed that caffeine consumption during gestation affects early stages of ovarian follicle development so that reduces reproductive efficiency in offspring Wistar rats.

ULTRA SHORT, HIGHLY ECONOMIC AND EFFECTIVE PROTOCOL FOR ICSI PATIENTS
M.W. El Deeb
1, S.S. Said 1, A.M. Sallam 2, E.A. Ismail 1, E.M. Mohsen 1, Y.E. Darwish 3, A.M. El Sharaki 1               
1 Intergrated Fertility Center, Alexandria, Egypt
2 Elsalama Hospital, Alexandria, Egypt
3 Alexandria School of Medicine, Egypt

Keeping things simple without altering success rate of ICSI is the idea behind our novel protocol. This simple protocol aims to reduce the burden of ICSI procedures and its related complications, thereby giving a couple the chances to conceive using procedures less costly in terms of physical, emotional, social and financial costs. We had 65 cases underwent ICSI, their age ranges from (23-47) years. Clomiphene citrate (CC ),3 tablets daily started from the second day of the cycle for 5 days, In the last day of CC administration HMG was given in the form of 2 amps of Menegon 75 (Ferring) or Merional 75 (Ibsa) + 2 amps of Gonal F 75 ( Merck Serono) , till we reach one or more follicles > 16 mm. E2 is measured. Next day early morning one amp. Of Cetroide 0.25 mg. and the full HMG dose was received… HCG 10,000 was administrated when one or more follicles > 18 m.m as usual. Average number of oocytes retrieved ranges from (2-25) oocytes, number of embryos transferred (ET) ranges from (1-5) embryos, only one case had no ET. We had 25 pregnant cases 38.4 % five of them had blastocyst transfer. These encouraging results of this highly economic protocol should be put in consideration as a simple and successful ICSI protocol.
   
THE RELATIONSHIP OF THE FSH-R POLYMORPHISM TO MODERATE AND SEVERE TYPES
P. Feldmar
1, M. Macek Sr 2, H. Kluckova 2, M. Hrehorcak 2, J. Diblik 2, P. Paulasova 2, M. Turnovec 2, M. Brandejska 1, A. Langerova 1, M. Macek Jr 1          
1 Gennet Ltd, Prague, Czech Republic
2 University Hospital Motol, Prague, Czech Republic
    
Aim: Our study aimed to compare the relationship between the genotype of the FSH receptor (FSHR) and ovarian hyperstimulation syndrome (OHSS) class III-V of Golans classification in Czech females. We based our study on the fact that polymorphism of the FSHR influences the reaction of the FSH receptor and thus could determine the risk of OHSS development. The exact prediction of severe OHSS development might help prevent this life threatening syndrome. Method: The FSH-R polymorphism genotypes were ascertained in 317 randomly selected fertile controls. These results were compared to 24 females with type III a 17 with type IV/V OHSS, according to Golan´s classification. Females with OHSS had median: BMI 23.12; age 31 years; estradiol 7993.50 pmol/L (3533-41335); number of aspirated oocytes 28. FSH-R polymorphism was examined by real-time PCR on ABI Prism 7000. Results: Controls were characterized by the prevalence of the Asn/Asn (37.2%), Asn/Ser (47.3%) and Ser/Ser (15.5%). In OHSS type III the prevalence of Asn/Ser genotype increased from 47.3 % to 62.5%. In type IV/V Asn/Asn increased from 37.2% to 58.8% and Asn/Ser decreased from 47,3% to 29,4%. Due to the small numbers of tested cases compared to 317 controls, one cannot offer statistics with any certainty. Conclusion: The results of our pilot study indicate that the risk of less severe type III of OHSS is associated with Asn/Ser heterozygosity, whereas the most severe types IV/V are associated with the “highest sensitivity” genotype Asn/Asn. These results support the hypothesis that the FSH-R genotype might predict the clinical severity of OHSS in Czech females. These results support pharmacogenetic approach in controlled ovarian hyperstimulation.
   
OVARIAN HYPESTIMULATION SYNDROME - OHSS – STRATEGY TREATMENT – BASED ON THE MANAGEMENT OF OHSS PATIENTS IN THE 2ND DEPARTMENT OF OBSTETRISCS, GYNAECOLOGY AND NEONATOLOGY MEDICALU UNIVERSITY WROCLAW, POLAND
T. Fuchs
, M. Zimmer, M. Pomorski, M. Tomialowicz, A. Wiarowski                  
2nd Department of Obstetrics, Gynaecology and Neonatology Medicalu University Wroclaw, Poland
    
OHSS is a complications of ovarian stimulation. It’s mild form is quite common, especially betwen women undervent gonadotroipin stimulation and presesce of PCOS ovaries . Patients with mild and moderte form can be managed as out-patients , while severe forms need multidisciplinary approach , intensive observation and treatment . There are some ciritical poinst how to manage these cases. Mild and moderate form, Oral fluid intake, Bed rest to avoid ovarian torsion, Daily weight recording, Monitoring biochemical parameters, Severe form, Admision to hospital and bed rest are obligatory. Monitoring vital signs. Physical examination is obligatory while bimanual examination is contraindcatated – risk of ovarian rupture. Replacement intravascular fluid violume is crucial to prevent thrombosis and multiorgan disfunction. Saline, Albumin and Hydroxyethyl stach solution ( HAES) are used . Role of intravenous albumin infusion and it‘s doubtfull benefits are still discussed. Monitoring fluid intake and urine output. Thorough monitoring of complete blood counts, electrolytes and biochemical parameters is essential to properly reacts on patients‘ conditions. Parameters describing liver and kidneys function are crucial. ECG monitoring to prevent cardiac rate dysfunction due to hyperkaleia Thromboprophylaxis with low molecular weight hepatin is nesessary to prevent thrombo-embolic disease Admision to Intensive Care Unit and mechanical ventilation can be necessary in cases of cases of respiratory distress syndrome. Paracentesis and/or aditionally thoracocentesis can be helpful to improve ventilation parameters and decrease lecel of ascites Further study are needed to answer the question if new methods ie dopamine agonist and doxycycline have positve impact on ovaries during OHSS treatment.
  
COMBINING LUTEAL ESTRADIOL PATCH AND GNRH ANTAGONIST SUPPRESSION WITH CO-FLARE AND MICRODOSE FLARE STIMULATION FOR IVF IN POOR RESPONDERS
J. Huang
, T. Singer, S. Spandorfer, O. Davis, Z. Rosenwaks
The Center for Reproductive Medicine and Infertility, Weill Cornell Medical College
   
OBJECTIVE: Luteal administration of estradiol (E2) patch and GnRH antagonist (Ant) is effective in preventing follicular recruitment in the preceding luteal phase. The objective is to compare the efficacy of combining E2 patch/Ant pretreatment with 3 commonly applied poor responder IVF protocols: Co-flare (CF), microdose lupron (MDL), and Ant. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Patients who underwent the 3 protocols from 1/1/2004-12/01/2009 were included.  All patients applied 0.1mg E2 patch every alternate day, starting 10 days post-LH surge. On second day of E2 patch, patients started Ant for 3 days. CF protocol entailed administering 1mg leuprolide acetate (LA) from cycle day (CD) 2 to 4 then decreasing LA dose to 0.5mg on CD5. High dose gonadotropins (FSH/HMG) started on CD3. MDL protocol involved starting 40ug MDL twice daily on CD2 followed by FSH/HMG administration  on CD4. Ant protocol involved starting FSH/HMG on CD2. Ant was started when leading follicle reached 13mm or E2 measured 300pg/ml. HCG was given when at least 2 follicles attained 17 mm and retrieval occurred 35 hours later. The main outcome measures were: no-start rate,  days of stimulation, and cancellation and clinical pregnancy rates. Statistical analyses included X2 test and Mann-Whitney test.  P<0.05 was deemed statistically significant. RESULTS: 89 CF, 56 MDL, and 2125 Ant cycles were identified. Both groups had comparable baseline characteristics and antral follicle count. The no-start rates were very low in all 3 groups.  Compared to E2/Ant/Ant group, the E2/Ant/MDL group had a significantly higher cancellation rate (36.4% vs. 18.3%), required more days of stimulation (12.6±2.1 vs. 10.8±2.2) and higher dose of FSH/HMG (6534.1±1517.6IU vs. 5754.9±1459.8IU). The clinical pregnancy rates per cycle started were comparable in 3 groups (16.3% vs. 20.0% vs. 22.7% respectively). CONCLUSIONS: Combinations of luteal E2 patch/Ant pretreatment with CF and MDL protocols yielded similar clinical pregnancy rates compared to E2 patch/Ant/Ant protocol and represent viable treatment options for poor responders. SUPPORT: Institutional
  
WHAT IS THE OPTIMAL FOLLICULAR SIZE IN INTRA-UTERINE INSEMINATION CYCLES WITH OVULATION INDUCTION WITH CLOMIPHENE CITRATE OR LETROZOLE?
A. Livshits
, P. Robb, E. Jr. Strawn                       
Medical College of Wisconsin, Milwaukee, Wisconsin
   
Objective: To determine the optimal size of the leading follicle that needs to be reached prior to the administration of human chorionic gonadotropin (hCG), in intra-uterine insemination (IUI) with Clomiphene Citrate(CC) or Letrozole. Secondary objective was to look at the effect of endometrial lining thickness on pregnancy rates. Methods: Retrospective chart review of 988 women who underwent ovulation induction with CC or Letrozole. All had mid-cycle ultrasound, when the mean diameter of the follicle was calculated.All received hCG and had IUI. Pregnancy was recorded as clinical pregnancy with fetal heart motion seen at a 6-7 week transvaginal ultrasound scan. Results: Cycles with Letrozole achieved higher pregnancy rates than CC (23% vs 16%). There was no difference in endometrial lining between the two cycle types. There was no difference in the optimal follicular size was between cycles with Letrozole and CC, but for each endometrial lining thickness, the optimal follicular size of the leading follicle was different. In our study, for the mean endometrial thickness of 9 mm, the optimal size of the leading follicle was 23.8mm for CC and 24.7mm for Letrozole. For each additional millimeter increase in endometrial thickness, the optimal follicular diameter increased by 0.4mm. For each cycle type, thicker endometrium yielded higher pregnancy rates (p=0.03). Conclusions: There is no difference in the optimal size of the leading follicles between cycles with CC and Letrozole. The size of the optimal follicle depends on the endometrial thickness. Thicker endometrium was associated with higher pregnancy rates.
     
ENDOMETRIAL STROMAL CELLS SUBJECTED TO HIGH GLUCOSE SHOW INSULIN-RESISTANCE IN VITRO
P. Ormazabal
1,2, C. Romero 1,3, A. Quest 2, M. Vega 1,3                    
1 Endocrinology and Biology of Reproduction Laboratory, University of Chile Clinical Hospital
2 Cellular Communication Laboratory, Center for Molecular Studies of the Cell (CEMC), Faculty of Medicine, University of Chile
3 Obstetrics and Gynecology Department, University of Chile Clinical Hospital
  
Endometrium requires an adequate glucose supply for appropriate function. We reported that endometria from patients with Polycystic Ovary Syndrome and insulin-resistance express lower levels of IRS-1, GLUT4 and phospho-T642AS160 compared with control. Binding of insulin to insulin receptors (IRs) activates the PI3K/Akt pathway, thereby favoring GLUT4 translocation to plasma membrane and glucose uptake. Interaction between IR and Caveolin-1 protects the receptor against proteasomal degradation and IR activation leads to Y14caveolin-1 phosphorylation. Aim: To study the effect of excess glucose (condition that induces insulin-resistance in other cell types) on expression, protein phosphorylation and deoxiglucose (DOG) uptake in endometrial stromal cells. Methods: T-HESCs cells were cultured in the presence or absence of 25mM glucose (24h). Samples from control and experimental conditions were evaluated for Caveolin-1 gene expression by RT-PCR and Caveolin-1, IR, IRS-1, Akt, GLUT4 and GLUT1 protein expression by western blot analysis. After acute insulin stimulation (100nM, 20min) phospho-S473Akt, phospho-Y14Caveolin-1 and DOG uptake were also evaluated.  Results:  Treatment with 25mM glucose reduced IR and Caveolin-1 protein levels to 34% and 55% of controls, respectively (p<0.05). Increased phospho-S473Akt, phospho-Y14Caveolin-1 and DOG uptake were observed in control cells after insulin stimulation (p<0.05), while for cells pre-treated with glucose neither increase in phosphorylations nor DOG uptake were detected. Conclusion: High glucose impairs insulin induced DOG uptake probably due to the lower content of Caveolin-1 and IR, together with the inability of insulin to increase phospho-S473Akt. Therefore, these results suggest that exposure to glucose of T-HESCs induces resistance to insulin in-vitro. Fondecyt1095127, Fondap15010006, ConicytATD24100029.
 
PREMATURE ELEVATION IN SERUM LH DESPITE UTILIZING AN ANTAGONIST IN YOUNG POOR RESPONDERS (AGE<40) : NEGATIVE IMPACT ON OUTCOME
T. Singer, J.Y. Huang, S. Lin, S. Spandorfer, Z. Rosenwaks
The Center for Reproductive Medicine and Infertility, Weill Cornell Medical College

Objective: To evaluate the impact of premature elevation of serum LH levels on IVF outcomes in young patients (<40) utilizing an antagonist protocol. Design: Retrospective cohort study of 146 consecutive patients using a single protocol over 12 months. Materials and Methods: The stimulation protocol consisted of luteal phase(10 days post LH surge) E2 and a GnRH-Antagonist prior to the administration of gonadotropins on cycle day 2. Patients older than 40 were excluded. Only the first cycle was included. After ROC analysis, peak LH level was subdivided into LH<4.0 and LH 4.0 for the purpose of this analysis. LH > 4 was considered premature elevation of LH. Statistical analysis was performed using the student t test, Mann-Whitney U test and the 2 test when appropriate. Results: A total of 146 patients were studied (129 LH≤4.0 and 17 LH>4.0). Those with LH 4.0 had significantly more oocytes retrieved (8.9± 4.9 vs. 5.6± 4.3 p=0.001) and more mature oocytes (7.1 ±4.2 vs. 4.9±4.0 p<.05). There was no difference in terms of age, peak E2, E2 to oocyte ratio, fertilization rate, # of embryos transferred, implantation rate or pregnancy outcome. Conclusions: Our study suggests that in younger patients with diminished ovarian reserve undergoing an E2 Patch/GnRH antagonist protocol for IVF stimulation a premature rise in LH levels is associated with a lower yield of oocytes. Support: Institutional.

THE EFFECT OF INTRACYTOPLASMIC SPERM INJECTION ON GENDER IN IVF CYCLES IN PATIENTS UNDERGOING PREIMPLANTATION GENETIC SCREENING (PGS)
T. Singer
, J.Y. Huang, M. Noel, A. Melnick, S.D. Spandorfer, Z. Rosenwaks
The Center for Reproductive Medicine and Infertility, Weill Cornell Medical Center

Objective: In the past 2 decades, studies have suggested differences in sex ratio following IVF treatment. Recently, analysis showed a trend towards male embryos when transferring fresh blastocysts (JARG 2009). It has also been suggested that ICSI might be associated with the delivery of more females (F&S 2009). Our study's objective was to determine whether the use of ICSI in IVF cycles for PGS is associated with gender. We studied all the fertilized embryos that were biopsies in patient undergoing PGS. Design: Retrospective analysis. Materials and Methods: 87 patients who underwent IVF-PGS cycles between 1/1/2007-12/31/2009 and total of 334 embryos were included in the analysis. The main outcome measures included the use of ICSI or insemination and embryo gender as determined by PGS. Statistical analyses was performed using Fisher's exact test. P<0.05 was deemed statistically significant. Results: 140 IVF-PGS embryos (42%) were fertilized using insemination whereas 194 embryos (58%) were created via the use of ICSI. Though not statistically significant, a trend was noted towards more female embryos when ICSI (55.15%) was used for fertilization as compared to insemination (47.14%). Conclusions: Our study suggests that with the use of ICSI over insemination in the IVF-PGS cycles, there is a trend toward female embryos over male (P=0.15). A larger prospective randomized study is needed to further investigate this trend. Support: Institutional.

SIGNIFICANT VARIATIONS OF ANTI-MÜLLERIAN HORMONE SERUM LEVELS DURING PREGNANCY AND DURING ORAL CONTRACEPTION
S. Tsepelidis
1,2, S. Barant 1, R. Imbert 2, A. Flahaut 3, C. Gervy 3, Y. Englert 1,2              
1 Research Laboratory on Human Reproduction, Faculty of Medicine, Free University of Brussels (ULB), Belgium
2 Fertility Clinic, Erasme Hospital, Brussels, Belgium
3 Biochemistry Department, Erasme Hospital, Brussels, Belgium

Introduction: Anti-Müllerian hormone (AMH), secreted by the granulosa cells of preantral and small antral follicle, is now worldwide used as a marker of the ovarian reserve. It has now been demonstrated that AMH serum levels are stable during the menstrual cycle but the data during the pregnancy and hormonal contraception are still controversial. The aim of these two clinical studies was to investigate the potential stability of AMH during the pregnancy and after hormonal exposition. Material & Methods: To study the effect of oral contraception on AMH serum levels, 18 healthy volunteers aged from 18 to 35 years old using oral contraception were recruited. AMH serum levels dosages were realized on day 7 of the hormone-free interval and after 21 days of hormonal exposition. To assess the stability of AMH during pregnancy, AMH serum levels measurements were realized on frozen sera of 22 pregnant patients at 5-10, 11-15, 16-20, 21-25, 26-30, 31-35, 36-40 weeks of pregnancy and in the post partum. Results : A significant difference in AMH serum levels was observed after 21 days of hormonal treatment (p<0.0001) both in 20 and 30 µg of ethinyl estradiol groups (p<0.05 and p<0.0005). During pregnancy, significant variations of AMH serum levels were detected (p<0.005) with a maximum at 16-20 weeks of gestation and a minimum at the end of the pregnancy. Conclusions: These results show that AMH is not stable during pregnancy and after hormonal treatment. Clinical studies and fundamental researches are still essential to understand the physiology of this hormone and its relationship with other hormones.

EFFECT OF LAPAROSCOPIC TUBECTOMY ON OVARIAN RESPONSE TO CONTROLLED OVARIAN STIMULATION FOR IVF-ET
I. Vagman
, R. Kapustiansky, J.B, Lessing, A. Amit, Y. Raz, G. Bibi, I. Eldar, B. Almog            
VF Unit, Tel-Aviv Sourasky Medical Center affiliated to the Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel

Objective: To evaluate the influence of salpingectomy on ovarian response in controlled ovarian hyperstimulation protocol (COH). Setting: University-based tertiary medical center. Study design: Retrospective paired matched case control study. Patients: 36 women who underwent two controlled ovarian stimulation cycles for IVF-ET. Interventions: Laparoscopic salpingectomy
Main outcome measures: The number of dominant follicles and oocyte aspirated before and after laparoscopic tubectomy. Additionally the effects of tubectomy on maximal estradiol levels, duration of stimulation and average daily dose of gonadotrophins were assessed. Results: Overall, the number of dominant follicles and the number of oocyte aspirated before and after tubectomy were comparable (8.2±3.8vs. 8.3±3.7, p=0.7, and 10.2±6.6 vs., 10.3±7.4, p=0.3). Maximal E2 levels before (1899±185pg/ml) and after the surgery (1997±231pg/ml) were not different (p=0.9), as well as the average daily dose of gonadotrophins used (217.8±10.0IU vs. 239±16.3IU, p=0.1). The same results were observed when number of dominant follicles in the operated side of the same patient were compared before and after surgery (4.8±2.2 vs. 4.7±2.0, p=0.8 respectively). Regression analysis done to assess the effect of unilateral vs. bilateral tubectomy showed no effect on main outcome measured. Conclusions: In contrast to previous reports we found that tubectomy weather unilateral or bilateral had no effects on ovarian response in controlled ovarian hyperstimulation protocol for IVF-ET.
   
PREGNANCY OUTCOMES IN WOMEN OF ADVANCED MATERNAL AGE
M. Valadan
, A. Sepahi, M. Bandegi                    
Tehran University of Medical Sciences

The aim of this study was to investigate perinatal and obstetric outcome in aged 40 years or older. A prospective comparative study was conducted for the women aged 40 years and over who delivered at 20 week’s gestation or beyond from January 2004 to December 2005 at four Hospitals of Tehran University of Medical Sciences. For comparison, a control group of patient who were 20-29 years of age was obtained. There were statistically significant increases in the rates of gestational diabetes, preeclampsia, caesarean section, breech presentation and stillbirth in women 40 years of age or older. There is a need to offer older women special counseling both before and after conception so that they become informed of the increased risks involved.

THE RELATIONSHIP BETWEEN FETAL MACROSOMIA AND MATERNAL HBA1C CONCENTRATION IN MATERNAL TYPE 1 AND 2 DIABETES MELLITUS
M. Valadan
, S. Rostamzade, A.B. Heidari, M. Khahani, M. Bandegi              
Tehran University of Medical Sciences, Tehran, Iran

To examine the relationships between maternal HbA1c concentration at the first and third trimester and birth weight in pregnancies complicated by pre-existing type 1 and 2 diabetes, a comprehensive dataset was collected prospectively for 2 years on all deliveries in 2 hospitals for women with pre-existing type 1 and 2 diabetes. Data included HbA1c concentrations in first and third trimesters of pregnancy and birth weight. Relationships between standardized birth weight and HbA1c concentrations at each of the 2 time points were examined using correlation analysis. Standardized birth weight (Z scores) could be calculated for 196 of 201 singleton live-born infants. HbA1c concentrations were standardized to correct for assay differences among different labs. There was a significant negative correlation between first trimester HbA1c and birth weight (Spearman’s R, _0.217; P _ 0.019). There were no statistically significant correlations for third trimester HbA1c. Maternal weigh gain during pregnancy had significant positive relationship with birth weight. In conclusion, there is no simple relationship between maternal glycemic status in third trimester and birth weight, but there seems to be an inverse relationship between first trimester glycemic control and standardized birth weight.
 
CUMULATIVE SUCCESS RATES AFTER IVF TREATMENT: LONGITUDINAL STUDY FROM A SINGLE CENTRE
V. Viardot-Foucault
1, B.C. Tai 2, S.Y. Oo 1, G.H. Lim 3, J.K.Y. Chan 1,2, S.F. Loh 1            
1 KK Women’s and Children’s Hospital, Singapore
2 National University of Singapore, Singapore
3 National University Health System, Singapore

Introduction. For most couple undergoing IVF/ICSI (Intra-cytoplasmic sperm injection), multiple treatment cycles are required to achieve a live-birth (LB). Therefore, a longitudinal cohort study where couples are followed-up over time provides a more realistic cumulative live-birth rate (CLBR) and informative counseling. Here, we present the CLBR of a cohort of patients undergoing IVF/ICSI treatment in KKIVF Centre (Singapore). Methods. We retrospectively analysed 5006 IVF/ ICSI cycles from 2779 patients who started their first cycle at KK Hospital, Singapore from 2000 to 2008. Frozen embryo transfer (ET) cycles were analysed within the stimulated cycle from which they were formed. The CLBR was calculated based on a maximum of 3 cycles where the primary outcome was a live birth. Results. Due to a high drop-out rate (69-76%), the CLBR calculated based on the initial cohort was biased as it has likely underestimated the true rates (LB rate was 33.7%, 5.0%, 0.7% after cycle 1, 2 and 3 respectively, with a CLBR of 39.4%). However, by assuming that the drop-out population had the same probability of LB than the one who continued their treatment, the CLBR reaches 59.3% (LB was 33.7%, 16.3%, 9.3% respectively). Conclusion. The CLBR determined in a longitudinal cohort study can provide meaningful data for patients requiring multiple cycles of IVF treatment. However, this data can be skewed by the high drop out rates experienced in our series, a likely result of a privately-funded treatment model. It is likely that the true CLBR falls between 39.4 to 59.3%.

DOSE-DEPENDENT EFFECT OF KETOROLAC ADMINISTRATION AROUND OVARIAN STIMULATION ON IN VIVO AND IN VITRO FERTILIZATION AND SUBSEQUENT EMBRYO DEVELOPMENT
H.W. Youm
1, B.C. Jee 1,2,3, J.H. Lee 4, C.S. Suh 1,2,3, S.H. Kim 2,3,4
1 Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Korea
2 Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, Korea
3 Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea.
4 Department of Obstetrics and Gynecology, Seoul National University Hospital, Seoul, Korea

Aim: To investigate the effect of ketorolac administration around ovarian stimulation on in vivo and in vitro fertilization and subsequent embryonic development. Methods: Female mice were injected with ketorolac (7.5, 15 and 30 µg/d) for 3 days starting from the day of eCG treatment. Control group was treated by saline. Two-cell embryos were obtained by hCG triggering and mating from 41 mouse and in vitro blastocyst-forming rate was assessed. Mature oocytes were collected after hCG triggering from 23 mouse and in vitro fertilization and blastocyst-forming rate were assessed. Results: The number of in vivo fertilized 2-cell embryos per mouse was similar across four groups, however, the blastocyst-forming rates showed an inverse relationship with dosage of ketorolac (64.2%, 35.4%, and 25.9%; P<0.001 when compared with control [97.6%]). Degenerated oocytes were frequently observed in three ketorolac-treating groups (22.9%, 22.4%, and 75.0%) and the rate was significantly higher in 30-µg-treating group when compared with control (4.3%). Lower fertilization rates were noted in three ketorolac-treating groups (63.2%, 62.1%, and 40.0%) but not significant when compared with control (78.8%). Blastocyst-forming rates were similar in 7.5- and 15-µg-treating group (75% and 88.9%) when compared with control (88.5%). No blastocyst was obtained in 30-µg-treating group. Conclusion: Administration of ketorolac around ovarian stimulation does not affect in vivo fertilization but significantly affects their in vitro developmental competence in a dose-dependent manner. In in vitro fertilization process, high-dose ketorolac could result in poor oocyte quality and decreased embryo developmental competence.

CD9 IS EXPRESSED ON PUTATIVE HUMAN SPERMATOGONIAL STEM CELLS AND IS AN EFFECTIVE MARKER FOR STEM CELL ENRICHMENT
K. Zohni
1, X. Zhang 1, S.L. Tan 1, P. Chan 2, M. Nagano 1
1 Department of obstetrics and gynecology, McGill University
2 Department of surgery, McGill University

BACKGROUND: Human spermatogonial stem cells (SSCs) play critical roles in life-long maintenance of male fertility and regeneration of spermatogenesis. These cells are expected to provide an important resource for male fertility preservation and restoration. For instance, testis biopsy specimens can be harvested from a cancer patient prior to cancer therapies, and SSCs included in the specimens can be transplanted back to the patient at a later time. To achieve this strategy in clinical settings, it is important to isolate live human SSCs. In this study, we investigated if CD9, a known rodent SSC marker, is expressed on human SSCs and can be used as an effective marker to enrich human testis cells for SSCs. METHODS AND RESULTS: Testicular tissues were obtained from men with obstructive azoospermia. Using immunohistochemistry, we found that CD9 was expressed in human spermatogonia, as confirmed by co-staining with antibodies against MAGEA4, an antigen specific to human spermatogonia. Following immunomagnetic cell sorting, CD9-positive cells were significantly enriched for spermatogonia expressing MAGEA4, compared to unsorted cells. When these cells were transplanted into the testes of immunocompromised mice, we detected ~3 to 4-fold enrichment of putative human SSCs at two and four months after transplantation, compared to unsorted cells. CONCLUSIONS: Out results demonstrate that CD9 is a marker of putative human SSCs and can be used effectively to enrich human testis cells for SSCs.


 
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