Michał Kunicki

Use of ovarian reserve parameters for predicting live births in women undergoing in vitro fertilization

OBJECTIVE To examine common clinical determinants, including patient age; levels of anti-Müllerian hormone (AMH), inhibin B, and follicle-stimulating hormone (FSH); antral follicle count (AFC); and number of oocytes retrieved, to predict live births in women undergoing in vitro fertilization. STUDY DESIGN Women undergoing cycles of intracytoplasmic sperm injection (ICSI) for the first time were reviewed retrospectively, and serum levels of AMH, inhibin B, and FSH, as well as AFC (days 1 and 4 of pre-ICSI menstrual period) and patient age were analyzed as determinants of live birth rates. RESULTS Of the patients studied, 35.71% (891/2495) became pregnant, with live births achieved in 32.20% (806/2495) of cycles initiated and in 46.37% (806/1738) of embryo transfers. Clinical pregnancy rate was 35.71% (891/2495) for cycles initiated and 51.26% (891/2318) for embryo transfers. Univariate analysis revealed that the odds of live birth significantly decreased with increasing age, declining AMH or inhibin B concentrations, and fewer oocytes retrieved. At AMH levels greater than 5.7 ng/ml, the odds of live birth were 3.18 times greater than for AMH levels less than 1.9 ng/ml [95% confidence interval (CI), 1.89-5.43]. Using multivariate logistic regression, only AMH (OR = 1.89; 95% CI, 1.00-3.60; p < 0.05) and AFC (OR = 1.86; 95% CI, 1.02-3.40; p < 0.05) showed statistically significant associations with live birth. Area under the curve for ROC (ROC(AUC)) indicated that AMH (AUC = 0.60) surpassed AFC (AUC = 0.59), number of oocytes retrieved (AUC = 0.59), inhibin B (AUC = 0.55), FSH (ROC(AUC) = 0.54) and chronological age (ROC(AUC) = 0.53) in predicting live birth. CONCLUSIONS In this assessment of various indices (i.e., age; levels of AMH, inhibin B, and FSH; AFC; and quantity of oocytes retrieved) for predicting live births for IVF patients, AMH, AFC and the quantity of oocytes retrieved constituted the most reliable determinants.

Evaluation of Granulocyte Colony-Stimulating Factor Effects on Treatment-Resistant Thin Endometrium in Women Undergoing In Vitro Fertilization

The aim of the study was to assess the granulocyte colony-stimulating factor (G-CSF) effects on unresponsive thin (<7 mm) endometrium in women undergoing in vitro fertilization (IVF). We included thirty-seven subjects who had thin unresponsive endometrium on the day of triggering ovulation. These patients also failed to achieve an adequate endometrial thickness in at least one of their previous IVF cycles. In all the subjects at the time of infusion of G-CSF, endometrial thickness was 6,74 ± 1,75 mm, and, after infusion, it increased significantly to 8,42 ± 1,73 mm. When we divided the group into two subgroups according to whether the examined women conceived, we showed that the endometrium expanded significantly from 6,86 ± 1,65 to 8,80 ± 1,14 mm in the first group (who conceived) and from 6,71 ± 1,80 to 8,33 ± 1,85 mm in the second, respectively. There were no significant differences between the two subgroups in respect to the endometrial thickness both before and after G-CSF infusion. The clinical pregnancy rate was 18,9%. We concluded that the infusion of G-CSF leads to the improvement of endometrium thickness after 72 hours.

Probability of live birth in women with extremely low anti-Müllerian hormone concentrations

The aim of the present study was to investigate the clinical pregnancy and live birth rates in women with extremely low (≤ 0.4 ng/ml) anti-Müllerian hormone (AMH) concentrations. The study included 101 women (188 cycles) with extremely low AMH concentrations undergoing IVF cycles and compared the number of live births in women with low AMH. Moreover, the study compared the number of live births in women with or without endometriosis stage III/IV. Fourteen clinical pregnancies and 14 live births (including one pair of twins) were recorded; one woman miscarried. Significantly higher clinical pregnancy (P = 0.046) and live birth rates (P = 0.018) were found in women aged < 35 years compared with older women. AMH concentration did not differ significantly between women with or without endometriosis and there were six live births in women with endometriosis. This was not significantly different from the rate in healthy women. It is concluded that live births are possible in women with extremely low AMH concentrations. The presence of endometriosis stage III/IV did not affect live birth rates in women with extremely low AMH concentrations although an important limitation of the study is the small number of women included who were affected by that disease.

Anti-Müllerian hormone (AMH) is a strong predictor of live birth in women undergoing assisted reproductive technology.

In the present study, we evaluated the clinical value of the following parameters: basal anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), inhibin B and antral follicle count (AFC) in predicting live birth outcomes. The study involved 603 women undergoing in vitro fertilization (IVF) using the long protocol for controlled ovarian hyperstimulation (COH). Serum levels of AMH, FSH and inhibin B as well as AFC were measured on the first three days of the menstrual cycle prior to the beginning of stimulation. AMH was the only independent parameter that correlated with the chance of live birth. We found that live birth rates of 46.2% (patient age <35 years), 44.7% (35-37 years), 32.1% (38-39) and 15.3% (>39) were associated with concentrations of AMH>1.4 ng/ml. For the AMH range 0.6-1.4 ng/ml, the live birth rates were 29.3%, 12.5%, 5.6% and 2.7%, respectively, and for AMH concentrations below 0.6 ng/ml the rates were 7.1%, 8.3%, 0% and 5.8%, respectively. Independently of other parameters affecting the chance of live birth, the success rate was the highest when the AMH level was >2 ng/ml, significantly lower when the AMH concentration was about 1 ng/ml, and 0% when the AMH concentration was ∼0.1 ng/ml. In conclusion, this is the first report to demonstrate that AMH level correlated better than age, FSH or inhibin B concentrations or AFC with live birth outcome. Therefore, the basal serum concentration of AMH may become a new, substantial prognostic factor regarding live birth above and beyond other currently available predictors of IVF outcome.

Decreasing quality of the new generations of anti-Mullerian hormone assays.

Anti-Müllerian hormone (AMH) measurements are widely used to optimize the stimulation protocols. First generation AMH kits correlated well with ovarian reserve and response to stimulation. In the present study we aimed to asses if the new generation kits share the same accurate correlations. Retrospective data were collected from 8323 blood samples. For comparison we used Immunotech I generation kit (ImI 4035 samples), Beckman Coulter II generation kit RUO (BCII RUO 3449, samples) and Beckman Coulter II generation kit with IVD certificate (BCII IVD 839 samples). We compared average AMH concentrations measured with different kits, as well as correlation between kits. We also compared average AMH concentrations in sera collected on different cycle days and samples of different quality of preservation. AMH serum concentrations differed for each kit, ranging 4.4 ± 4.12 (mean ± SD) for the ImI, 2.68 ± 3.15 for the BCII RUO, and 1.64 ± 2.85 for BCII IVD. The mean differences from an adjusted regression model were −48.7%, −40%, and −69.2%, respectively. In conclusion, the changes of the BC AMH kits are unpredictable; however, the improvement of them is still possible. It would be very dangerous to use elaborated stimulation protocol (based on the Ist generation AMH results) with the results from the IInd generation assays.

Granulocyte colony stimulating factor treatment of resistant thin endometrium in women with frozen-thawed blastocyst transfer

ABSTRACT The aim of the study was to assess the granulocyte-colony stimulating factor (G-CSF) effect on unresponsive thin (<7 mm) endometrium in women undergoing frozen-thawed embryo transfer at the blastocyst stage. A total of 62 women with thin unresponsive endometrium were included in the study, of which, 29 received a G-CSF infusion and 33 who opted out of the study served as controls. Patients in both groups had similar endometrial thickness at the time of the initial evaluation: 6.50 mm (5.50-6.80) in the G-CSF and 6.40 mm (5.50-7.0) in the control group. However, after the infusion endometrial thickness increased significantly in the G-CSF group in comparison with the controls (p=0.01), (Δ) 0.5 (0.02-1.2) (p=0.005). In the G-CSF group endometrium expanded to 7.90 mm (6.58-8.70) while in the control group to 6.90 mm (6.0-7.75). Five women in each group conceived. The clinical pregnancy rate was 5/29 (17.24%) in the G-CSF treated group and 5/33 (15.15%) in the control group (p>0.05). The live birth rate was 2/29 (6.89%) in the G-CSF group and 2/33 (6.06%) in the control group (p>0.05). We concluded that G-CSF infusion leads to an improvement in endometrium thickness but not to any improvement in the clinical pregnancy and live birth rates. Until more data is available G-CSF treatment should be considered to be of limited value in increasing pregnancy rate. Abbreviations: G-CSF: granulocyte colony-stimulating factor; M-CSF: macrophagecolony-stimulating factor; GM-CSF: granulocyte-macrophage colony-stimulating factor; FET: frozen embryo transfer; IVF: in vitro fertilization

Serum Dehydroepiandrosterone Sulphate Concentration Is Not a Predictive Factor in IVF Outcomes before the First Cycle of GnRH Agonist Administration in Women with Normal Ovarian Reserve

Objective The aim of our study was to determine whether serum dehydroepiandrosterone sulphate (DHEAS) concentration and the models incorporating it could help clinicians to predict IVF outcomes in women with normal ovarian reserve undergoing their first long protocol. Study Design We performed a retrospective analysis of 459 women undergoing cycles of intracytoplasmic sperm injection (ICSI) for the first time in a long GnRH agonist protocol. Results Embryo transfer was performed in 407 women (88.7%). The fertilisation rate was 78.6%. The clinical pregnancy rate was 44.8% per started cycle and 50.6% per embryo transfer. Our univariate model revealed that the best predictors of clinical pregnancy were the number of mature oocytes, the number of embryos transferred and the number of good quality embryos, account for the clinical parameters that reflect ovarian reserve the best being AMH level and AFC. DHEAS did not predict clinical pregnancy (OR 1.001, 95% CI, 0.999–1.004). After adjusting for the number of embryos transferred and class of embryos in a multivariate model, the best predictors were age (OR 0.918, 95% CI, 0.867–0.972) and AFC (OR 1.022, 95% CI, 0.992–1.053). Serum DHEAS levels were positively correlated with AFC (r = 0.098, P<0.039) and testosterone levels (r = 0.371, P<0.001), as well as the number of mature oocytes (r = 0.109, P<0.019); serum DHEAS levels were negatively correlated with age (r = -0.220, P<0.001), follicle-stimulating hormone (FSH), (r = -0.116, P<0.015) and sex hormone-binding globulin (SHBG), (r = -0.193, P<0.001). Conclusions DHEAS concentration (in addition to the known factors of ovarian reserve) does not predict clinical pregnancy in women with normal ovarian reserve who are undergoing ICSI.

Age-related decline in AMH is assay dependent limiting clinical interpretation of repeat AMH measures across the reproductive lifespan

Abstract Purpose: The aim of the study was to determine whether the assays exhibit an interaction with age and exhibit heterogeneous age related declines in AMH. Apart of chronological age, AMH variation was investigated with relation to menstrual cycle day (MCD). The goal implicates two questions: Are distributions of AMH concentrations homogenous after adjustment for the specific AMH assay? Does age-assay product has an effect on AMH depletion? Methods: The study was conducted by examining results of AMH tests performed for 12,917 women with four types of AMH assays: Immunotech I generation kit (IMI, 4016 samples), Beckman Coulter II generation kit RUO (BCII RUO, 3430 samples), Beckman Coulter II generation kit with IVD certificate (BCII IVD, 830 samples), and Ansh Labs I generation kit (AnshLabs, 4641 samples). Statistical analysis included ACNOVA and least square regression technique. Results: Menstrual cycle day has no effect on AMH measurements. On the other hand, AMH values differed substantially between the four assays, with a marked discordance in the rate of age-related AMH decline for the four assays (ranging from –8.16% (95% CI: –8.79, –7.54) to –11.53% (95% CI –12.20, –10.87), with a significant interaction between age and assay. Conclusions: (1) The distribution of AMH concentration is heterogeneous after controlling the age across assays; (2) the rate of AMH decline as a function of age is different for the four manual AMH ELISA assays.

Estradiol Valerate Pretreatment in Short Protocol GnRH-Agonist Cycles versus Combined Pretreatment with Oral Contraceptive Pills in Long Protocol GnRH-Agonist Cycles: A Randomised Controlled Trial

The strategy of in vitro fertilization (IVF) procedures relies on the increasing pregnancy rate and decreasing the risk of premature ovulation and ovarian hyperstimulation syndrome. They are also designed to avoid weekend oocyte retrievals. Combined oral contraceptive (OC) pills are among the medicines used to accomplish these objectives. Alternatively, estradiol can be used instead of OC to obtain similar results. The aim of our study was to compare the differences in pregnancy rates (PRs), implantation rates, and miscarriage rates between a short agonist protocol with estradiol priming and a long protocol with combined OC. Of the 298 women who participated in this study, 134 achieved clinical pregnancies (45.0%). A higher PR (58.4%, n = 80, compared to 40.3%, n = 54) was achieved in the long protocol after OC pretreatment group. The implantation rate was also higher for this group (37.8% versus 28.0%; P = 0.03). The miscarriage rate was 15.0% (n = 12) for the long protocol after OC pretreatment group and 20.4% (n = 11) for the short agonist group (P = 0.81). The short agonist protocol required a 5.7% lower human menopausal gonadotropin (hMG) dosage than the long protocol but surprisingly the number of oocytes retrieved was also smaller.

Testing ovarian reserve in pre-menopausal women: why, whom and how?

Numerous social and environmental factors (environmental hazards, social factors such as education and career, higher economic status desired before the decision is made to have children) influence a womens decision to postpone pregnancy until late reproductive age. In turn, age is related to a fall in ovarian reserve. The main goal of testing ovarian reserve is the identification of women with so-called diminished ovarian reserve (DOR). Additionally, it provides assistance in the counselling of women who are planning to use assisted reproductive techniques (ART). This review examines current methods of testing ovarian reserve and their application. The most useful methods of assessing ovarian reserve are ultrasonographic count of ovarian antral follicles (AFC) and serum tests of both the anti-Müllerian hormone (AMH) level and the third-day level of follicle stimulating hormone (FSH). However, there are limitations to the currently used methods of testing ovarian reserve, especially in relation to their specificity and sensitivity. It is also difficult to predict egg quality based on these tests. The value of screening programmes of ovarian reserve is yet to be determined.