The Prediction of IVF Outcomes with Autologous Oocytes and the Optimal MII Oocyte/Embryo Number for Live Birth at Advanced Maternal Age.

Background and Objectives: Delayed childbearing in advanced age might be associated with a low prognosis for achieving pregnancy. Therefore, it is important to establish a predictive tool that will optimize the likelihood of a live birth at advanced age. Material and Methods: The retrospective study was conducted at the Ferona Fertility Clinic in Novi Sad (Republic of Serbia), between January 2020 and May 2021. The survey included 491 women aged ≥35 who met the inclusion criteria and who were subjected to an IVF (in vitro fertilization) treatment cycle. Results: The average number of retrieved oocytes, MII (metaphase II) oocytes, and developed embryos significantly decreased in advanced age. Age was also found to have a significant adverse effect on pregnancy and live birth rates. In women aged ≥35, 10/12 MII oocytes or 10/11 embryos are required for reaching an optimal live birth rate/cumulative live birth rate. Optimal CLBR (cumulative live birth rate) per one oocyte was achieved when 9 MII oocyte were retrieved. Conclusions: The study indicates that the cut-off for increased risk is ≥42 year. However, despite low live birth rates, autologous IVF for these women is not futile. An increase in the number of retrieved mature oocytes and a generation of surplus cryopreserved embryos could reinforce LBR (live birth rate) and CLBR. Clinicians should be very cautious in counseling, as autologous IVF may only be applicable to women with good ovarian reserve.

DEHP Decreases Steroidogenesis through the cAMP and ERK1/2 Signaling Pathways in FSH-Stimulated Human Granulosa Cells.

DEHP is an endocrine disruptor that interferes with the function of the female reproductive system. Several studies suggested that DEHP affects steroidogenesis in human and rodent granulosa cells (GC). Some studies have shown that DEHP can also affect the FSH-stimulated steroidogenesis in GC; however, the mechanism by which DEHP affects hormone-challenged steroidogenesis in human GC is not understood. Here, we analyzed the mechanism by which DEHP affects steroidogenesis in the primary culture of human cumulus granulosa cells (hCGC) stimulated with FSH. Cells were exposed to DEHP and FSH for 48 h, and steroidogenesis and the activation of cAMP and ERK1/2 were analyzed. The results show that DEHP decreases FSH-stimulated STAR and CYP19A1 expression, which is accompanied by a decrease in progesterone and estradiol production. DEHP lowers cAMP production and CREB phosphorylation in FSH but not cholera toxin- and forskolin-challenged hCGC. DEHP was not able to decrease steroidogenesis in cholera toxin- and forskolin-stimulated hCGC. Furthermore, DEHP decreases FSH-induced ERK1/2 phosphorylation. The addition of EGF rescued ERK1/2 phosphorylation in FSH- and DEHP-treated hCGC and prevented a decrease in steroidogenesis in the FSH- and DEHP-treated hCGC. These results suggest that DEHP inhibits the cAMP and ERK1/2 signaling pathways, leading to the inhibition of steroidogenesis in the FSH-stimulated hCGC.

T-2 toxin downregulates LHCGR expression, steroidogenesis, and cAMP level in human cumulus granulosa cells.

Our goals were to investigate whether environmentally relevant doses of T-2 toxin can affect human ovarian granulosa cells' function and to reveal the potential mechanism of T-2 toxin's action. Results showed that T-2 toxin strongly attenuated luteinizing hormone/choriogonadotropin receptor (LHCGR) mRNA expression in follicle-stimulating hormone (FSH)-stimulated human cumulus granulosa cells. Addition of human chorionic gonadotropin was not able to elicit maximal response of ovulatory genes amphiregulin, epiregulin, and progesterone receptor. T-2 toxin reduced mRNA levels of CYP19A1 and steroidogenic acute regulatory protein (STAR) and lowered FSH-stimulated estradiol and progesterone production. Mechanistic experiments demonstrated that T-2 toxin decreased FSH-stimulated cyclic adenosine monophosphate (cAMP) production. Addition of total PDE inhibitor 3-isobutyl-1-methylxanthine prevented T-2 toxin's action on LHCGR, STAR, and CYP19A1 mRNA expression in FSH-stimulated human cumulus granulosa cells. Furthermore, T-2 toxin partially decreased 8-bromoadenosine 3'5'-cyclic monophosphate (8-Br-cAMP)-stimulated LHCGR and STAR, but did not affect 8-Br-cAMP-stimulated CYP19A1 mRNA expression in human cumulus granulosa cells. Overall, our data indicate that environmentally relevant dose of T-2 toxin decreases steroidogenesis and ovulatory potency in human cumulus granulosa cells probably through activation of PDE, thus posing a significant risk for female fertility.

The effect of serum vitamin D levels on ovarian reserve markers: a prospective cross-sectional study.

STUDY QUESTION: Is there any association between serum 25-OH vitamin D levels and ovarian reserve markers in infertile women? SUMMARY ANSWER: Vitamin D is not associated with the ovarian reserve markers, anti-mullerian hormone (AMH) and antral follicle count (AFC), in infertile women. WHAT IS KNOWN ALREADY: The mechanism underlying the relationship between vitamin D deficiency and reproduction is still unclear; however, evidence indicates a potential direct negative impact on ovarian function. This is mainly due to the fact that gonadal function may be altered by vitamin D deficiency, as observed by the expression of vitamin D receptor mRNA in human ovaries, mixed ovarian cell cultures and granulosa cell cultures. On the other hand, results from clinical studies are conflicting, with some suggesting that vitamin D status is associated with ovarian reserve, whereas other cross-sectional studies have not found any significant correlation between vitamin D and AMH levels. STUDY DESIGN, SIZE, DURATION: This study was a prospective cross-sectional study from the Centre for Reproductive Medicine at the University Hospital of Brussels. The duration of the study was one year. PARTICIPANTS/MATERIALS, SETTING, METHODS: Overall, the study included 283 consecutive infertile women younger than 42 years old and undergoing their first treatment cycle in our institution. All patients were recruited within a time interval of 12 months from the initiation of the study, before undergoing infertility treatment. Women consuming vitamin D supplements or taking medication for systematic disease or women who had undergone ovarian surgery were excluded from the study. All infertile women had serum AMH and vitamin D sampled on the same day. AFC was measured on the second or third day of the first cycle following the blood sampling for the determination of AMH and 25-OH vitamin D levels. MAIN RESULTS AND THE ROLE OF CHANCE: Among all patients, 30.7% (n = 87) were vitamin D deficient (<20 ng/mL) whereas 69.3% (n = 196) had normal vitamin D levels (≥20 ng/mL). The mean AMH and AFC levels did not differ significantly between the two groups: AMH 3.9 µ/L (±3.8) versus 4.3 µ/L (±4.8), (P value = 0.5) and AFC 13.9 (±13.3) versus 12.7 (±11.4), (P = 0.7), respectively. No correlation was observed between 25-O H vitamin D and AMH (spearman's r = 0.02, P value = 0.7) or AFC (spearman's r = -0.02, P value = 0.7). In multiple linear regression analysis, after adjusting for potential confounders (age, BMI, smoking status, infertility cause and season of blood sampling), the regression slope in all participants for total 25OH-D predicting log10 AMH was 0.006 [standard error (SE) = 0.07, P value = 0.9]. Similarly, no significant association was observed between AFC and vitamin D levels, even after controlling for relevant co-variants (regression coefficient -0.09. SE 0.08, P value = 0.2). LIMITATIONS, REASONS FOR CAUTION: Although this is the first prospective study to evaluate the relationship between vitamin D and the most important ovarian reserve markers (AMH and AFC), we need to acknowledge that the data used to generate the study findings are cross-sectional in nature. In this regard, we cannot generate or exclude any causal effect hypothesis. Nevertheless, our data support that an association between vitamin D and ovarian reserve markers is highly unlikely to exist. WIDER IMPLICATIONS OF THE FINDINGS: Although data from basic research indicate that vitamin D deficiency may have an effect on steroidogenesis and follicular development, our study, by prospectively recruiting a large number of infertile women, clearly demonstrates that vitamin D deficiency is highly unlikely to have a detrimental effect on ovarian reserve. Ongoing prospective and translational research projects are currently being conducted in order to evaluate the potential effect of vitamin D deficiency on reproductive outcome mediated through either an effect on the oocyte quality or on endometrial receptivity and embryo implantation. STUDY FUNDING/COMPETING INTERESTS: No external funding was used for this study. No conflicts of interest are declared. TRIAL REGISTRATION NUMBER: N/A.

[Prediction of labor induction outcome using different clinical parameters].

INTRODUCTION: Induction of labor is one of the most common obstetric interventions in contemporary obstetrics. OBJECTIVE: The aim of the study was to evaluate the clinical and sonographic parameters in prediction of success of labor induction. METHODS: The prospective study included 422 women in whom induction of labor was carried out at the Department of Obstetrics and Gynecology of Clinical Centre of Vojvodina. The role of body mass index and age of women, parity Bishop score, cervical length measured by transvaginal ultrasound was evaluated in regard of the success of induction, which was considered successful if a vaginal delivery occurred within 24 hours after the onset of induction. Data were statistically analyzed by univariate statistical analysis and Pearson's chi2 test. RESULTS: Out of 422 women, induction of labor was successful in 356 (84.4%), and it failed in 66 (15.6%) cases. The values of Bishop score and cervical length had positive correlation with the success of induction. CONCLUSION: Bishop score and transvaginal cervical length were both reliable predictors in determining the success of labor induction, as well as parity and BMI. These parameters are mostly complementary, not competitive in prediction of labor induction success.

[The role and contribution of transvaginal endoscopy to diagnostics and treatment of infertility].

Transvaginal endoscopy (TVE) presents minimal invasive endoscopic method that involves hysteroscopy, transvaginal laparoscopy and salpingoscopy. It gives a new approach to both basic evaluation of marital infertility and its treatment. The role of TVE is a subject of controversies regarding its justification as standard infertility treatment. Another aspect is a role of TVE prior to methods of assisted reproductive technologies (ART). The aim of this paper was to try, through the analysis of the available literature, to clarify the role of TVE in reproductive medicine, as well as to show our experience. The concept of one-day diagnostics, which is so-called one stop fertility clinic, is performed in the Clinical Center of Vojvodina, Department of Gynecology and Obstetrics on a daily basis. It consists of history, gynecology and ultrasound exam, spermiogram, hormone tests, and TVE. Patients are informed about results on the very same day and advised on the proper infertility treatment. By forming the infertility diagnostics protocols, which use the methods of TVE, we consider it possible to evaluate adequately and accurately the fertility within the shortest possible time. It replaced standard laparoscopy in certain indication fields; it eventually confirmed the necessity of its use in recurrent IVF implantation failure, raising the question of its routine use prior to the first IVF cycle that is a topic requiring further randomized trials.

[Results of in vitro fertilization cycles at the Clinic for Gynecology and Obstetrics, Clinical Center of Nis].

INTRODUCTION: Infertility affects 15-17% of reproductive age couples in our country. In vitro fertilization brought revolution in treatment of this problem, bringing hope to many couples around the world for more than 3 decades. The aim of this paper was to present results and experiences of implementation of this method of treatment at the Clinic of Gynecology and Obstetrics in Nis. MATERIAL AND METHODS: The study included the first 402 women who had undergone in vitro fertilization program at the Clinical Center of Nis. The data were statistically analyzed by basic descriptive methods. The main outcome measures were demographic features, cause of infertility, duration of stimulation, average gonadothropine consumption, number of oocytes per aspiration and embryos transferred, mode of conception as well as clinical pregnancy and aspiration rate. RESULTS: The two main factors were the male infertility and tubal factor inferitily, being 51.61% and 48.39%, respectively The classical method of in vitro fertilization constituted 72.40% of all cycles, while intra cytoplasmic sperm injection method was used in 27.60% of all cycles. The average number of embryos transferred was 2.75. The cycle cancellation rate was 15.05%. The clinical pregnancy rate per ermbryo transfer was 35.44%; while the live birth rate per embryo transfer was 26.53%. DISCUSSION AND CONCLUSION: Our success rates are comparable with those in other European countries, where for in vitro fertilization the clinical pregnancy rates per aspiration and per transfer were 29.0 and 32.4%, respectively in the observed period. For intra cytoplasmic sperm injection, the corresponding rates were 29.9 and 33.0%. The main difference from the European average was the average number of transferred embryos and lower percentage rate of intra cytoplasmic sperm injection as a method of conception. Results

Validity of clinical and ultrasound variables to predict the risk of cesarean delivery after induction of labor.

OBJECTIVE: To evaluate a previously described score to predict the risk of cesarean delivery after induction of labor. METHODS: We conducted a multicenter prospective study among women at 36 weeks of gestation or more undergoing induction of labor in the maternity units of Geneva, Switzerland, and Novi Sad, Serbia. Before induction, we calculated the risk score for cesarean delivery including data on maternal height, body mass index, parity, and transvaginal ultrasonographic cervical length. We calculated the sensitivity and specificity of the score using different cutoffs of calculated risk. RESULTS: Of the 537 women included in the analysis, 92 (17%) had a cesarean delivery. Among the variables tested, only the transvaginal ultrasonographic cervical length was associated with the risk of cesarean delivery (P<.001). Using the different cutoffs of calculated risk of cesarean delivery (20%, 30%, and 40%), we calculated the sensitivity (69.6%, 54.3%, and 45.7%, respectively), specificity (42.0%, 58.2%, and 69.2%, respectively), and positive predictive value (19.9%, 21.0%, and 23.5%, respectively) of the risk score. The area under the receiver operating characteristic curve was 0.59. There was a poor association between the outcome of labor induction (vaginal delivery or cesarean delivery) and the predicted risk. CONCLUSION: The evaluated score was not useful to predict the outcome of women undergoing labor induction. Our results show the necessity of validating existing scores in different settings and patient populations before widespread implementation in clinical care.

Results and experiences after 2000 performed hysteroscopies.

INTRODUCTION: Hysteroscopy is one of the oldest endoscopic procedures which uses the cervix for introducing a telescope to place a camera into the uterine cavity. OBJECTIVE: The aim of the study was to present our experiences with this procedure during the long-term work starting from the time when hysteroscopic method of treatment was first introduced at this Clinic until today. METHODS: This prospective study involved 2000 female patients referred to the Clinic for Gynecology and Obstetrics in Novi Sad from January 2005 till January 2011 for diagnostic and operative hysteroscopy. The following parameters were analyzed: the presence of minor and major pathology of the endometrium, type of anesthesia, technique of operative work, instruments and energy used during hysteroscopy and complications. RESULTS: Seventy-eight percent of all procedures were done under intravenous anesthesia. The most common operative procedure was polypectomy and the most complicated one was myomectomy. By histopathological examination of hysteroscopic biopsy specimens four endometrial carcinomas were revealed. The combination of mechanical instrument and bipolar energy were used in most of the cases, while the percentage of complications was extremely low. CONCLUSION: Hysteroscopy is a safe, highly sensitive, precise diagnostic and operative endoscopic procedure. Our experiences and dilemmas open a field for discussion and offer salutations to everyday problems. The introduction of this procedure into out-patients conditions has contributed to the efficiency of the treatment of vaginal pathological processes, thus enabling that the method has become available to all gynecologists. This fact requires further study and new results.

[Results and experiences after 1000 IVF cycles at the Clinic for Gynecology and Obstetrics in Novi Sad].

INTRODUCTION: Infertility affects 15-17% of reproductive age couples in our country, and 10-15% of couples worldwide. The aim of this paper was to present results and experience gained after the first 1000 cycles of the national In Vitro Fertilization (IVF) program, to offer professional standard of work in our country and to compare it with results obtained in Europe and worldwide. MATERIAL AND METHODS: The study prospectively included 1000 women who had undergone national In Vitro Fertilization program from October 2006 until November 2009 at the Department of Gynecology and Obstetrics, Clinical Center of Vojvodina, Novi Sad. RESULTS: The analysis included 1000 in vitro fertilization cycles. Male factor infertility was the leading cause (56.9%) followed by tubal factor (45.9%). The classic method of in vitro fertilization constituted 72.3% of all cycles, while intracytoplasmic sperm injection (ICSI) method was used in 27.7% of all cycles. The average number of embryos transferred was 2.67. The cycle cancellation rate was 14.10% and the aspiration rate was 94.40%. The clinical pregnancy and live birth rate were 33.41% and 26.78% per embryo transfer respectively. DISCUSSION AND CONCLUSION: The results in our study showed that in our setting there are far less intracytoplasmic sperm injection cycles compared with the European average of 66.5% of all fresh cycles, and that we transferred more embryos on average. Our success rates are comparable with those in other European countries where the clinical pregnancy rates per aspiration and per transfer for in vitro fertilization were 29 and 32.4%, respectively in the period of observation. For intracytoplasmic sperm injection, the corresponding rates were 29.9 and 33%.