anti-Müllerian Hormone Concentrations Research Articles

P–328 Dienogest significantly decreases the size of the cyst and alters Anti-Müllerian Hormone concentration in patients with endometrioma

Abstract Study question Does utilization of dienogest make any impact on the size of cyst and Anti-Müllerian Hormone (AMH) concentration in patients with endometrioma throughout 12-months? Summary answer Although dienogest makes a gradual reduction in the size of endometrioma cyst throughout 12-months, a significant drop in AMH serum concentration was also noticed. What is known already According to recent studies, pre-operative serum AMH levels might be illusively increased with parallel to the size of endometrioma which will be a misleading factor while deciding to operate the patient via cystectomy. Although dienogest is one of the medical options that might be commenced in patients with endometrioma cyst, there is limited data about its effect on the size of the endometrioma and hence serum AMH concentration throughout 12 months of follow up. Study design, size, duration The current observational cohort study was conducted among patients with endometrioma those treated with dienogest from January 2017 to January 2020. The primary outcome was alteration in diameter of endometrioma cyst at 6th and 12th months of treatment. Secondary outcome was alteration in serum AMH concentration in the same period. Of 104 patients treated with dienogest, 44 patients were excluded due to being treated with any type of surgical intervention during follow up period. Participants/materials, setting, methods A total of 60 patients were recruited for the final analysis. Of them, primary symptom was dysmenorrhea, chronic pelvic pain and menstrual irregularity in 16 (26.7%), 25 (41.7%) and 8 (13.3%) patients, respectively. Eighteen patients (30%) were asymptomatic. As 21 patients had bi-lateral endometrioma, size of the leading cyst was considered to be analyzed for the primary outcome measure. Paired-t test was used for comparison of numerical values and p ≤ 0.05 was taken as statistical significance. Main results and the role of chance The mean age was 31.5±8.0 years. In the time point when dienogest was started, the mean size of the endometrioma was 46.3±17.4 mm. The mean serum AMH concentration was 3.6±2.4 ng/ml. After 6 months of treatment, the mean size of the endometrioma decreased to 38.6±14.0 mm which corresponds to a mean difference of 7.8 mm (95% CI: 3.0 to 12.6; p: 0.003). The respective figure for AMH was 3.3±2.7 ng/ml which corresponds to a mean difference of 0.3 ng/ml (95% CI: –0.2 to 0.8; p: 0.23) at 6 months. After 12 months of treatment, the mean size of the endometrioma was 37.5±15.7 mm which corresponds to a mean difference of 8.9 mm (95% CI: 2.9 to 14.9; p: 0.005) at the end of 12 months. The respective figure for AMH was 2.7±1.9 ng/ml which corresponds to a mean difference of 0.9 ng/ml (95% CI: 0.1 to 1.7; p: 0.045) at the end of 12 months. The mean diameter of endometrioma and AMH concentration did not differ throughout the time period between 6th and 12th months of the treatment. Limitations, reasons for caution Although herein we present the largest data that depicts the alteration of endometrioma cyst and AMH concentration with the application of dienogest, the lack of control group is a limitation that avoids to perform any comparison. Wider implications of the findings: A shrinkage after commencement of treatment suggest that dienogest might present improvement in patients with endometrioma with respect to radiological findings, but further studies are required whether a decline in AMH concentration after 12 months refers to a genuine decrease in ovarian reserve or resolution of misleading high pre-treatment levels. Trial registration number not available

P–659 Artificial intelligence (AI) as an assisting tool in generating patient-friendly corifollitropin alfa ovarian stimulation protocol during in vitro fertilization

Abstract Study question How machine learning assisted in generating patient-friendly corifollitropin alfa protocol in normal responders? Summary answer In retrospective experiments, our machine learning model integrated physiological measurements of patients and clinical experience to generate a patient-friendly corifollitropin alfa protocol. What is known already Long-acting corifollitropin alfa can simplify the regimen, minimizing injections during the whole cycle. The previous study has described the patient-friendly protocol using corifollitropin alfa without routine pituitary suppression in normal responder can result in non-compromised clinical outcomes. Some studies showed machine learning can help with making clinical decisions and have the ability to learn from physiological measurements. Those methods effectuate certain points throughout short-acting menotropin protocols, however, there are still no robust AI tools for long-acting corifollitropin alfa protocols. Study design, size, duration 1,309 cycles were collected at Stork Fertility Center from November 2016 to October 2019, and 1,221 cycles were available after data cleaning and applying exclusion criteria, which Anti-Mullerian Hormone (AMH) is lower than 2. The data from electronic medical records (EMRs) consisted of age, AMH, body weight, luteinizing hormone (LH), and estradiol (E2) concentrations measured on revisit. Evaluation is performed by one physician who has more than 20 years of experience in IVF. Participants/materials, setting, methods: The protocol generator consisted of 5 parts: doses of Elonva, trigger type, doses of recombinant follicle-stimulating hormone (rFSH), doses of recombinant luteinizing hormone (rLH), and day of oocyte retrieval. The protocol was predicted by age, AMH, and weight firstly, then fine-tuned by LH and E2 after the first revisit. We used the gradient boosting decision tree algorithm to learn the protocol. The dataset was randomly split into 80% for training and 20% for testing. Main results and the role of chance In classification, the model predicted the dose of Elonva achieved an accuracy of 0.913 and an AUC of 0.946, and trigger type got an accuracy of 0.901 and AUC of 0.852 only using features on stimulation day (SD) 1 and gained 0.012 and 0.056 in accuracy and AUC correspondingly after adding features on the first revisit day. In regression, the mean absolute error (MAE) of rFSH dose, rLH dose, and oocytes retrieved day was 156.30 IU, 232.75 IU, and 0.80 days respectively, and after refining, the MAE dropped to 92.37 IU, 100.07 IU, and 0.46 days. The error of predictions in rFSH and rLH was almost equal to half increments of rFSH (150 IU) and one increment rLH (75 IU). This indicated that our model could provide a better prediction of these clinical decisions with one revisit only. Limitations, reasons for caution The present study was a single-center retrospective, and only analyzed the data from normal responders, whose AMH was equal or greater than 2. Though, the recommendations of our system act as references, the physician will make the final decision. Wider implications of the findings: Our result showed the potential of machine learning in generating protocols is promising. Recommendations generated by our model can provide the junior clinical teams to optimize the clinical plans and learn from the experience of experts. We look forward to applying our machine learning model to different protocols. Trial registration number Not applicable

P–679 Magnetic resonance imaging (MRI), an alternative method to evaluate the ovarian reserve

Abstract Study question Is magnetic resonance imaging (MRI) a valid method to access antral follicles count (AFC) compared to two-dimensional (2D) transvaginal ultrasonography (USG) and Anti-Mullerian Hormone (AMH) to evaluate the ovarian reserve? Summary answer AFC has a high agreement between MRI and USG methods, as well as with AMH. MRI can be an alternative method to evaluate ovarian reserve. What is known already In fact, two methods are the most used today to assess ovarian reserve: USG and AMH. The USG is considered the gold standard method for AFC, and contributes to predict and tailor treatment strategies, such as in-vitro fertilization. The major limitations of USG are its user dependency and equipment. Also, there are limitations in displaying a global view of the pelvis and large ovarian lesions. Magnetic resonance imaging (MRI), with its excellent soft-tissue contrast resolution and characteristics, is a useful non-invasive alternative modality to USG. Just one study evaluated MRI and revealed more small size antral follicles compared to 3D-USG. Study design, size, duration A prospective cross-sectional observational study was performed in an assisted reproduction techniques (ART) post-graduation program setting from an assisted reproductive center in Sao Paulo, Brazil, which ran throughout 2019–2020, with a total number of 59 patients that were in fertility treatment and needed to undergo to a MRI procedure to evaluate uterine or pelvic diseases as, Myoma, Adenomyosis, endometriosis, and adnexal cysts. Participants/materials, setting, methods Patients were evaluated to access the AFC by the MRI method and 2D transvaginal USG, and Anti-Mullerian Hormone (AMH) concentration to evaluate the ovarian reserve. Comparison between methods was done through Wilcoxon signed ranks test and Bland-Altman analysis. Ovarian reserve was classified as follows: very low (<4 follicles/AMH<0.5); low (5–7 follicles/AMH=0.5–1.1); normal (8–15 follicles/AMH=1.1–3.5); normal-high (>15 follicles/AMH>3.5). Weighted Cohen’s kappa was used to verify agreement between MRI, USG and AMH classifications of ovarian reserve. Main results and the role of chance Average AFC for USG were 5,55±4.01 for left ovary and 5.55±3.8 for right ovary. Average follicle count for both ovaries was 10±7.07. Regarding MRI, mean counts were 6.44±4.81 for left ovary, 5.65±3.85 for right ovary, and a 11.89±7.89 follicle sum mean count. Average concentration of AMH was 1.79±1.44. The Wilcoxon test and Bland-Altman analyses found differences and systematic biases for comparison between USG and MRI for both ovaries (–2.58; limits of agreement=–14.56 to 9.40, Wilcoxon p = 0,008) and for the right ovary (–1.48; limits of agreement=–8.32 to 5.35, Wilcoxon p = 0,031). There was no difference between methods for the left ovary. Weighted Cohen Kappa coefficients showed substantial agreement between ovarian reserve classifications based on AMH levels, USG, and MRI. The conducted paired comparisons were USG with MRI (k = 0.676), AMH with MRI (k = 0.760) and USG (k = 0.609). Limitations, reasons for caution The systematic biases found when comparing USG to MRI methods may suggest a consistent detection of more follicles with MRI procedures in comparison to the USG method. This bias found warrants caution as it must be confirmed, in future studies. Wider implications of the findings: The MRI method reveals similar ovarian reserve to USG when used the same classification, and a higher agreement to AMH. This suggests that MRI is a reliable method of quantifying antral follicles and can also be adopted when the patient will need to evaluate pelvic pathologies. Trial registration number Not applicable

O-173 Decline in anti-Müllerian hormone (AMH) concentrations following radioactive iodine (RAI) treatment in women with differentiated thyroid cancer (DTC): a systematic review and meta-analysis

Abstract Study question Does radioactive iodine (RAI) treatment in premenopausal women with differentiated thyroid cancer (DTC) affects ovarian reserve, as evaluated by anti-Müllerian hormone (AMH), antral follicle count (AFC) and follicle-stimulating hormone (FSH)? Summary answer AMH concentrations decreased at three, six and 12 months following RAI treatment in women with DTC, whereas no difference was observed regarding FSH. What is known already Thyroid cancer is the third most common type of female malignancy and DTC is the most common histopathological type. Thyroidectomy constitutes the mainstay of treatment for DTC, followed by administration of RAI, which acts as an adjuvant therapy to destroy residual cancerous thyroid tissue. However, the effect of RAI on ovarian reserve of these women, as assessed by AMH, AFC and FSH, remains controversial. Study design, size, duration A systematic review and meta-analysis was performed aiming to identify studies evaluating the effect of RAI treatment on ovarian reserve in women with DTC. For this purpose, a literature search in the electronic databases PubMed, Scopus and CENTRAL was carried out until 06/12/2020. The primary outcome measure was the effect of RAI on ovarian reserve, as evaluated by AMH, AFC and FSH. Participants/materials, setting, methods Studies were eligible if they included premenopausal women with DTC, treated with a single RAI dose and assessed for at least one marker of ovarian reserve repeatedly within 12 months post-RAI.Meta-analysis of weighted data was performed using random effects model. Results were reported as weighted mean difference (WMD) with 95% confidence interval (CI). Main results and the role of chance Four prospective eligible studies, published between 2005 and 2020, were eligible for the meta-analysis, evaluating a total of 154 women. The number of participants ranged from 24 to 50. The single dose of RAI used to treat DTC ranged from 50 to 150 mCi. AMH concentrations decreased at three (WMD -1.66 ng/ml, 95% CI -2.42 to -0.91, p < 0.0001; I2 0%), six (WMD -1.58 ng/ml, 95% CI -2.63 to -0.52, p = 0.003; I2 54.7%) and 12 months (WMD -1.62 ng/ml, 95% CI -2.02 to -1.22, p < 0.0001; I2 15.5%) following a single RAI dose compared with baseline (three studies; n = 104). With respect to FSH concentrations, no difference was observed at six (WMD +3.29 IU/l, 95% CI -1.12 to + 7.70, p = 0.14; I2 96.8%) and 12 months (WMD +0.13 IU/l, 95% CI -1.06 to + 1.32, p = 0.83; I2 55.2%) post-RAI compared with baseline (two studies; n = 83). No data on AFC was available. Limitations, reasons for caution The small number of studies and patients included, as well as the lack of data on AFC may have compromised the validity of the conclusions drawn. Moreover, subgroup analysis according to female age was not feasible, due to the lack of relevant data. Wider implications of the findings The negative effect of RAI on ovarian reserve in premenopausal women with DTC, as indicated by the decreased AMH, should be confirmed by data on AFC, which are currently not available. These findings necessitate close monitoring of ovarian reserve in such women, counselling them regarding the need for fertility preservation. Trial registration number N/A

Serum anti-Müllerian hormone concentration as a diagnostic tool to identify testicular tissue in canine disorders of sexual development

Disorders of sexual development (DSD) may have their origin in alterations of the chromosomal, gonadal or phenotypic sex. Affected animals are usually presented because of ambiguous external genitalia, seldom because of reproductive disorders. Anti-Müllerian hormone (AMH) is secreted in the gonads with higher amounts in males than in females and can be used to identify gonadal tissue in sexually normally developed dogs. The aim of this study was to examine the diagnostic potential of serum AMH to identify testicular tissue in 11 dogs with DSD. The diagnostic procedures applied were: determination of the phenotypic sex (n = 11), genital ultrasound (n = 9), determination of the SRY gene (n = 11), karyogram (n = 6), gonadectomy (n = 11), pathohistology of the gonads (n = 10), serum AMH measurement (n = 11). 39 female dogs described in a previous study and 19 male dogs with a normal spermiogram served as controls for the AMH serum concentrations in sexually intact dogs. The 11 dogs with DSD were classified as 7 XY DSD and 4 XX DSD. Presumptive testes were obtained in 10 dogs and 1 dog had an ovotestis combined with a testis. Mean serum AMH values of the dogs with DSD were significantly higher (P < 0.001) than in male and female controls. The upper limit of the AMH test (≥ 23ng/ml) was reached in 6 dogs. High AMH concentrations have been described previously in cryptorchid dogs. 1 dog with a male phenotype and 2 with a female phenotype had AMH values within the range of the male controls, although all of them had cryptorchid testes. A Poodle, in which epididymis were identified but no definitive gonads, had an AMH concentration of the lower limit of the test (≤ 0.01 ng/ml), comparable to previously described castrated dogs. This study indicates that serum AMH levels are a useful diagnostic tool to identify testicular tissue in dogs with DSD and suggests the possible use of AMH to diagnose testicular dysgenesis.

Body Composition and Serum Anti-Müllerian Hormone Levels in Euthyroid Caucasian Women With Hashimoto Thyroiditis.

ObjectiveWomen with Hashimoto thyroiditis (HT) are characterized by increased incidence of infertility and disturbances in body composition. Serum anti-Müllerian hormone (AMH), which reflects functional ovarian reserve, is decreased in women with HT and it be related to body mass. The aim of the present study was to investigate the relation between serum levels of AMH and body composition in HT compared to control group.Patients and MethodsWe examined 85 euthyroid women: 39 subjects with HT and 46 control women. Body composition was analysed by dual-energy X-ray absorptiometry and with bioimpedance method. Serum concentrations of AMH, leptin, TSH, thyroid hormones were assessed.ResultsWe observed lower serum concentration of AMH in women with HT in comparison to the control group (p=0.01), but without differences in serum concentration of leptin between studied groups (p=0.28). Women with HT were characterized by higher %body fat (p=0.01) estimated with bioimpedance method without differences in BMI, android and gynoid fat mass and visceral adipose tissue (VAT) mass estimated with DXA method when compared to the control group (all p>0.05). We found a negative relationship between serum concentration of AMH and %body fat (r=-0.38,p=0.03) in women with HT. Additionally, in HT group, the relationship between serum levels of AMH and leptin was not statistically significant (r=0.01,p=0.96). We observed a relationship between serum concentration of leptin and BMI, %body fat mass, android, gynoid and VAT mass in HT and in the control group (all p<0.01).ConclusionsWomen with HT are characterized by lower levels of AMH and it is associated with higher fat mass, independently of serum levels of leptin.

Ovarian Reserve after Chemotherapy in Breast Cancer: A Systematic Review and Meta-Analysis.

Background: Worldwide, breast cancer (BC) is the most common malignancy in the female population. In recent years, its diagnosis in young women has increased, together with a growing desire to become pregnant later in life. Although there is evidence about the detrimental effect of chemotherapy (CT) on the menses cycle, a practical tool to measure ovarian reserve is still missing. Recently, anti-Mullerian hormone (AMH) has been considered a good surrogate for ovarian reserve. The main objective of this paper is to evaluate the effect of CT on AMH value. Methods: A systematic review and meta-analysis were conducted on the PubMed and Scopus electronic databases on articles retrieved from inception until February 2021. Trials evaluating ovarian reserves before and after CT in BC were included. We excluded case reports, case-series with fewer than ten patients, reviews (narrative or systematic), communications and perspectives. Studies in languages other than English or with polycystic ovarian syndrome (PCOS) patients were also excluded. AMH reduction was the main endpoint. Egger’s and Begg’s tests were used to assess the risk of publication bias. Results: Eighteen trials were included from the 833 examined. A statistically significant decline in serum AMH concentration was found after CT, persisting even after years, with an overall reduction of −1.97 (95% CI: −3.12, −0.82). No significant differences in ovarian reserve loss were found in the BRCA1/2 mutation carriers compared to wild-type patients. Conclusions: Although this study has some limitations, including publication bias, failure to stratify the results by some important factors and low to medium quality of the studies included, this metanalysis demonstrates that the level of AMH markedly falls after CT in BC patients, corresponding to a reduction in ovarian reserve. These findings should be routinely discussed during oncofertility counseling and used to guide fertility preservation choices in young women before starting treatment.

Association of antimullerian hormone with the size of the appendix testis, the androgen and estrogen receptors and their expression in the appendix testis, in congenital cryptorchidism.

Antimullerian hormone (AMH) causes regression of the mullerian ducts in the male fetus. The appendix testis (AT) is a vestigial remnant of mullerian duct origin, containing both androgen (AR) and estrogen (ER) receptors. The role of both AMH and AT in testicular descent is yet to be studied. We investigated the possible association of AMH with AT size, the AR and ER, and their expression in the AT, in congenital cryptorchidism. A total of 26 patients with congenital unilateral cryptorchidism and 26 controls with orthotopic testes were investigated, and 21 ATs were identified in each group. AMH and insulin-like three hormone (INSL3) concentrations were measured with spectrophotometry. AR and ER receptor expression was assessed with immunohistochemistry using monoclonal antibodies R441 for AR and MAB463 for ER. For the estimation of receptor expression, the Allred Score method was used. AMH concentrations did not present significant differences between patients with congenital cryptorchidism and the controls. Also, no correlation was found between AMH, INSL3, and AT length. Allred scores did not present significant differences. However, expression percentiles and intensity for both receptors presented significant differences. Three children with cryptorchidism and the highest AMH levels also had the highest estrogen receptor scores in the AT. No association was found between AMH and the studied major parameters. However, higher AMH concentrations, in combination with higher estrogen receptor scores in the AT, may play a role in cryptorchidism in some children. Larger population samples are needed to verify this observation.

INTERRELATIONSHIP OF AMH WITH CLINICAL, HORMONAL AND SONOGRAPHIC PARAMETERS IN PATIENTS WITH POLYCYSTIC OVARIAN DISEASE

INTRODUCTION: Polycystic ovary syndrome (PCOS) is a frequently encountered problem in reproductive endocrinology, affecting approximately 6% of women of reproductive age. anti-mullerian hormone (AMH) also known as Mullerian inhibiting, substance is produced by granulosa cells of the preantral and small antral ovarian follicles in women and reect the ovarian reserve. Women with PCOS will often have a high number of antral follicles and, as a result, an equally high level of AMH in their blood. it has an inhibitory inuence on the actions of FSH and positively correlated with LH. Various studies demonstrated that oligo/anovulatory women with PCOS have signicantly higher serum concentrations of AMH. MATERIAL AND METHOD: This is a retro respective study of 54 PCOS patients from rst January 2019 to 13 April 2021 at a private gynaecology clinic in district Shivpuri. Data were collected from the medical records of the patients including age, height, weight, waist circumference, BP and modied Ferriman gallway score for hirsutism, Biochemical and hormonal values like LH, FSH, LH/FSH ratio, TSH, Prolactin and AMH, and lipid prole values were also obtained from the records. Abdominal or Vaginal ultrasound was used to assess the ovarian volume antral follicular count. RESULTS: In our study, the age of patients ranges from 20-39 years and a majority of the patients were in the age group of 21-30 years 75.28%) The Mean age of PCOS patients was 23.89. In our study, the mean cycle length of patients was 54.08 and it ranged from 28-190 days. The mean 2 BMI 25.8kg/m2 and modied FG score were 8.1. On ultrasound mean antral follicle count was 26.2 and the mean ovarian volume was 9.4 cm . In hormonal studies mean LH was 11.5, mean FSH was 3.4 and AMH was 4.34 and Testosterone was 2.1. In bio-chemical parameters increased Cholesterol, triglycerides, LDL, VLDL level and decreased HDL level were found in 31.48% of patients and with increased AMH. CONCLUSION: In our study, we found increased AMH concentration in correlation with increased cycle length in cases of oligomenorrhea, clinical hyperandrogenism, increased LH, increased AFC and increased ovarian volume. Our study concluded that AMH levels as an adjunct to existing Rotterdam criteria for diagnosis of PCOS had good diagnostic potential.

Breast Cancer Risk Factors and Circulating Anti-Müllerian Hormone Concentration in Healthy Premenopausal Women.

We previously reported that anti-Müllerian hormone (AMH), a marker of ovarian reserve, is positively associated with breast cancer risk, consistent with other studies. This study assessed whether risk factors for breast cancer are correlates of AMH concentration. This cross-sectional study included 3831 healthy premenopausal women (aged 21-57, 87% aged 35-49) from 10 cohort studies among the general population. Adjusting for age and cohort, AMH positively associated with age at menarche (P < 0.0001) and parity (P = 0.0008) and inversely associated with hysterectomy/partial oophorectomy (P = 0.0008). Compared with women of normal weight, AMH was lower (relative geometric mean difference 27%, P < 0.0001) among women who were obese. Current oral contraceptive (OC) use and current/former smoking were associated with lower AMH concentration than never use (40% and 12% lower, respectively, P < 0.0001). We observed higher AMH concentrations among women who had had a benign breast biopsy (15% higher, P = 0.03), a surrogate for benign breast disease, an association that has not been reported. In analyses stratified by age (<40 vs ≥40), associations of AMH with body mass index and OCs were similar in younger and older women, while associations with the other factors (menarche, parity, hysterectomy/partial oophorectomy, smoking, and benign breast biopsy) were limited to women ≥40 (P-interaction < 0.05). This is the largest study of AMH and breast cancer risk factors among women from the general population (not presenting with infertility), and it suggests that most associations are limited to women over 40, who are approaching menopause and whose AMH concentration is declining.

Predictive markers for superovulation response and embryo production in beef cattle managed in northwest Mexico are influenced by climate

Important components of a successful embryo transfer program in cattle are the superovulatory response and embryo production. Anti-Müllerian hormone (AMH) and antral follicle count (AFC) have been proposed as biomarkers of the reproductive response in superovulated cattle; however, their predictive ability appears to be influenced by climate. Objective herein was to evaluate AMH and AFC as predictors of the number of ovarian follicles, corpora lutea, collected embryos and transferable embryos in cows managed in northwest Mexico under different climates. Multiparous Simmental beef donor cows were assigned in two experimental climatic groups, cool (January 15– May 14; n=66) and warm (May 15– September 14; n=62). All cows were given a superovulatory treatment, and then subjected to artificial insemination and non-surgical embryo collection. Transrectal ultrasonography was performed to determine AFC. Blood samples were collected to measure blood AMH concentrations. A mixed effects model was used to analyze superovulatory and embryo production responses in cattle including AMH, AFC and climate as fixed effects, age as covariate and herd as random effect. All fixed effects resulted as significant (P<0.05) sources of variation for the evaluated traits, and interactions between AMH/AFC with climate were observed but only for transferable embryos. Pearson correlations were calculated between AMH/AFC with superovulatory and embryo production variables for cool and warm experimental groups, and a regression analyses was performed to quantify these relationships. Cows with either AMH>400 pg/mL or intermediate to higher AFC increased (P<0.05) mean values for superovulatory and embryo production variables, suggesting both AMH and AFC as predictors. Climate also influenced cow responses as number of follicles, corpora lutea, as well as collected and transferable embryos increased 33, 38, 40 and 35%, respectively, during cool- (8-23 °C) compared to warm-climate (24-34 °C; P<0.05). Both AMH and AFC were moderately correlated with number of follicles, corpora lutea and collected embryos (P<0.05) and highly correlated with transferable embryos (P<0.01) during cool climate; conversely, these correlations were low (P>0.05) for donor cows managed in the warm months. Regression analyses suggested that total number of transferable embryos increased 0.005 per 1 pg/mL of AMH and improved 0.484 per 1 unit of AFC. In conclusion, superovulatory and embryo production responses in Simmental beef cows appear to be closely associated with AMH and AFC, which are proposed as predictors for transferable embryos; however, such relationships were only detectable in absence of heat stress.

MO162IMPACT OF THE IMMUNOSUPPRESSIVE TREATMENT, X-RAY EXPOSITION AND RENAL PATHOLOGY ON OVARIAN RESERVE IN YOUNG WOMEN

Abstract Background and Aims Cyclophosphamide treatment and X-ray exposition in the area of the ovaries are proven to be the gonadotoxic factors in childbearing age women. Anti-Müllerian hormone (AMH) is regarded as a biomarker for ovarian reserve. Method The study included 167 consecutive premenopausal women attending Nephrology Clinic who gave informed consent and met exclusion criteria (past ovarian surgical procedure, PCOS, eGFR &amp;lt;30ml/min, irregular menstrual cycles). Clinical (renal pathology, smoking, X-rays exposition, eGFR standardized MDRD4, contraceptives) and demographic characteristics, as well as previous and current immunosuppressive therapies were recorded. Serum AMH levels were measured by Beckman Coulter’s Gen II enzyme linked immunosorbent assay (ELISA) kit; AMH levels were classified as low or normal/above age-adjusted reference levels (published norms). Statistical analysis included both regressive and non-regressive relationships between the studied clinical features. Due to the presumption of the remaining non-regression relationships between clinical factors, an original taxonomic method by Marczewski &amp; Steinhaus was used instead of general linear modeling. Based on patient age, eGFR, renal pathology an ‘optimal’ segregation of patients was performed following the created classification tree (dendrogram). Results Median age of the patients was 33 (range 18-44). Median AMH concentration was 2,66 ng/ml; AMH levels were classified in 64 females as low (median 1,015) and in 103 as normal/above age-adjusted reference (median 4,04). Sixty one patients were treated with cyclophosphamide in the past and 45 underwent kidney transplantation. 92 women suffered from glomerulonephritis, 44 from lupus nephritis, 12 from interstitial kidney disease, 5 from ADPKD and 14 from other kidney diseases. Age, eGFR, pregnancies in the past as well as being the kidney transplant recipient were the coefficients strongly correlated with AMH level. Presence of lupus nephritis was correlated with lower levels of AMH in comparison to other renal diseases and in contrast to cyclophosphamide. X-ray exposition measured in cumulative lifetime dose was not correlated with AMH levels. Conclusion Although previous researchers suggested cyclophosphamide to be the strong gonadotoxic factor, our statistical analysis approach shows that lupus as the disease often treated with cyclophosphamide may be the lowering ovarian reserve factor itself. Further studies on this subject are still necessary.

Vitamin D and anti-Müllerian hormone concentration in human follicular fluid individually aspirated from all patient follicles

Objective The main aim of this prospective study was to investigate the relationship between intrafollicular vitamin D and anti-Müllerian hormone (AMH) concentration and its impact on oocyte quality and developmental competence. Methods The analysis was performed on 208 follicular fluid (FF) samples obtained from 33 patients undergoing ovarian stimulation as part of in vitro fertilization (IVF) treatment that included intracytoplasmic sperm injection. Results Our study shows that vitamin D concentration in FF varies according to the developmental stage of the oocyte and corelates with embryo development status on day 3, while AMH concentration in FF is not correlated with the developmental potential of an oocyte. We demonstrated that the levels of vitamin D and AMH were higher in FF than in serum. Moreover we showed that AMH and vitamin D levels were positively correlated in FF but not in serum. Conclusion FF-AMH levels do not appear to be a suitable as noninvasive test of the developmental potential of an oocyte, while FF-vitamin D level can be used to evaluate whether embryos obtained from particular oocytes have potential of reaching the third day of culture. However, our results encourage further research to be carried out on a larger number of patients and testing additional components found in FF such as androgens.

The association between the levels of anti-Müllerian hormone (AMH) and dietary intake in Iranian women.

This paper aimed to assess the association between AMH with dietary intake of adult women referred to the infertility clinic of Al-Zahra Hospital in Rasht, Iran. This cross-sectional study was performed on 234 adult women referred to the infertility clinic of Al-Zahra Hospital, Rasht, Iran. The participants were categorized into two groups based on their AMH levels. Participants' data on lifestyle and anthropometry as well as blood samples were collected. Dietary intakes were assessed by a validated food frequency questionnaire (FFQ). Serum AMH concentration was negatively associated with the intake of fast foods (P = 0.002) and saturated fats (P = 0.040). These associations remained significant after adjustments for age, rural or urban location, and education. Additional adjustments for body mass index (BMI) and physical activity did not change the results. The results remained significant after further adjustments for menstrual age, menstrual pattern, and oral contraceptive pills. Fast foods and saturated fats were significantly associated with lower AMH concentrations and modifying the amount of these dietary components may be an important strategy for increasing the reservation of ovaries in women. Future longitudinal studies are warranted to confirm these findings and to identify the underlying mechanisms.

Early-life Farm Exposure and Ovarian Reserve in a US Cohort of Women.

In a previous exploratory study, we reported lower concentrations of the ovarian reserve biomarker anti-Müllerian hormone (AMH) in adulthood with prenatal farm exposure. We now examine this association as well as childhood farm exposure using enrollment data from the Sister Study, a large US cohort of women. We collected prenatal and childhood farm exposure data by questionnaire and telephone interview. However, serum AMH data were available only for a nested subset: premenopausal women ages 35-54 subsequently diagnosed with breast cancer (n = 418 cases) and their matched controls (n = 866). To avoid potential bias from restricting analyses to only premenopausal controls, we leveraged the available cohort data. We used data from both premenopausal cases and controls as well as postmenopausal women ages 35-54 (n = 3,526) (all presumed to have undetectable AMH concentrations) and applied weights to produce a sample representative of the cohort ages 35-54 (n = 17,799). The high proportion of undetectable AMH concentrations (41%) was addressed using reverse-scale Cox regression. An adjusted hazard ratio (HR) <1.0 indicates that exposed individuals had lower AMH concentrations than unexposed individuals. Prenatal exposure to maternal residence or work on a farm was associated with lower AMH concentrations (HR 0.66; 95% confidence intervals [CI] = 0.48 to 0.90). Associations between childhood farm residence exposures and AMH were null or weak, except childhood contact with pesticide-treated livestock or buildings (HR 0.69; 95% CI = 0.40 to 1.2). Replication of the prenatal farm exposure and lower adult AMH association raises concern that aspects of prenatal farm exposure may result in reduced adult ovarian reserve.

ENDOMETRIOSIS-ASSOCIATED INFERTILITY: THE ROLE OF HORMONES AND ITS CORRECTION

Background. Endometriosis-associated infertility (EAI) has a number of specific features, which are crucial in the choice of medical treatment.&#x0D; Objective. The aim of the study is to analyze endocrine profile in women with EAI before and after sclerotherapy and pregravid preparation (PP), which includes a vitamin complex FT 500 plus with inositol and vitamin D3. &#x0D; Methods. The study involved 70 women aged 21-40 years with endometriosis-associated infertility. The comparison group included 30 women with tuboperitoneal infertility. ELISA was used to determine concentrations of Anti-Mullerian hormone (AMH), follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone and estradiol in blood serum using a standard kit by Diagnostic Systems Laboratories, Inc (USA). During two menstrual cycles the FT 500 plus was prescribed once a day from the 2nd/3rd day of the cycle, vitamin D3 was prescribed at the dose of 2,000 IU for women without its deficiency and in therapeutic doses in case of hypovitaminosis. The sclerotherapy with 95% ethanol solution was performed on the 6th-8th day of menstrual cycle.&#x0D; Results. It was established that in women with EAI undergoing PP and sclerotherapy the level of AMH was lower (by 12.90%) as well as progesterone (by 9.84%), while FSH (by 14.47%), LH (by 21.14%) and estradiol (by 35.55%) was higher compare to the comparison group. At the same time, FSH (by 21.98%), LH (by 32.89%) and estradiol (by 32.23%) concentrations were significantly lower compare to their primary indices before sclerotherapy.&#x0D; Conclusions. Sclerotherapy and PP with a vitamin complex, inositol and vitamin D3 has a positive effect on endocrine profile in women with endometriosis-associated infertility

Anti-Müllerian hormone concentration regulates activin receptor-like kinase-2/3 expression levels with opposing effects on ovarian cancer cell survival.

Anti-Müllerian hormone (AMH) type II receptor (AMHRII) and the AMH/AMHRII signaling pathway are potential therapeutic targets in ovarian carcinoma. Conversely, the role of the three AMH type I receptors (AMHRIs), namely activin receptor-like kinase (ALK)2, ALK3 and ALK6, in ovarian cancer remains to be clarified. To determine the respective roles of these three AMHRIs, the present study used four ovarian cancer cell lines (COV434-AMHRII, SKOV3-AMHRII, OVCAR8, KGN) and primary cells isolated from tumor ascites from patients with ovarian cancer. The results demonstrated that ALK2 and ALK3 may be the two main AMHRIs involved in AMH signaling at physiological endogenous and supraphysiological exogenous AMH concentrations, respectively. Supraphysiological AMH concentrations (25 nM recombinant AMH) were associated with apoptosis in all four cell lines and decreased clonogenic survival in COV434-AMHRII and SKOV3-AMHRII cells. These biological effects were induced via ALK3 recruitment by AMHRII, as ALK3-AMHRII dimerization was favored at increasing AMH concentrations. By contrast, ALK2 was associated with AMHRII at physiological endogenous concentrations of AMH (10 pM). Based on these results, tetravalent IgG1-like bispecific antibodies (BsAbs) against AMHRII and ALK2, and against AMHRII and ALK3 were designed and evaluated. In vivo, COV434-AMHRII tumor cell xenograft growth was significantly reduced in all BsAb-treated groups compared with that in the vehicle group (P=0.018 for BsAb 12G4-3D7; P=0.001 for all other BsAbs). However, the growth of COV434-AMHRII tumor cell xenografts was slower in mice treated with the anti-AMRII-ALK2 BsAb 12G4-2F9 compared with that in animals that received a control BsAb that targeted AMHRII and CD5 (P=0.048). These results provide new insights into type I receptor specificity in AMH signaling pathways and may lead to an innovative therapeutic approach to modulate AMH signaling using anti-AMHRII/anti-AMHRI BsAbs.

Ovarian reserve and recurrence 1 year post-operatively after using haemostatic sealant and bipolar diathermy for haemostasis during laparoscopic ovarian cystectomy

Research questionIs there a difference in the ovarian reserve 1 year post-operatively in those who used a haemostatic sealant or bipolar diathermy for haemostasis during laparoscopic ovarian cystectomy for ovarian endometriomas? DesignThis was an extended follow-up observational study of a previous randomized controlled trial where women aged 18 to 40 years with 3–8 cm unilateral or bilateral endometriomas were randomized to receive haemostasis by a haemostatic sealant or bipolar diathermy following ovarian cystectomy. The primary outcome was the ovarian reserve as assessed by antral follicle count (AFC) 1 year post-operatively. Secondary outcomes included the recurrence rate of ovarian endometrioma, the change in anti-Müllerian hormone (AMH) and FSH concentrations, and reproductive outcomes. ResultsThe significant increase in AFC at 3 months after initial surgery (P = 0.025) in the haemostatic sealant group compared with the diathermy group was sustained at 1 year (P = 0.024) but there was no difference in AMH or FSH concentrations between the groups throughout the follow-up period. The recurrence rate in the FloSeal group was 7.7% (n = 3/39) compared with 22.2% (n = 8/36) in the diathermy group (P = 0.060). The recurrence rate in women who had bilateral lesions was significantly higher than those with unilateral lesions (risk ratio 5.33, interquartile range 1.55–18.38). No difference in reproductive outcomes was found between the two groups. ConclusionsApplying haemostatic sealant after laparoscopic cystectomy of ovarian endometriomas produces a significantly greater improvement in AFC, which was apparent at 3-month follow-up, and was sustained at 1-year follow-up without compromising the recurrence rate.

PCOCheck AMH ELISA: A Clinical Case Report to Resolve Miss-Matched Antral Follicle Counts (AFC) to Serum AMH

Introduction: At times clinicians wonder why a subject’s Anti-Mullerian Hormone (AMH) does not correlate to the antral follicle count (AFC). AMH is secreted as a full-length protein and undergoes proteolytic cleavage at amino acid 451 to become biologically active. Additional proteolytic processing takes place at aa229. This processing, which may differ between individuals with different clinical conditions, exposes new antigenic sites which affect AMH measurements. Moreover, AMH epitopes might be masked by protein interaction in the circulation. Clinical Case: A 24-year-old woman gravida 0 presented with metrorrhagia for several months. The bleeding pattern was unresponsive to a trial of oral contraceptive pills, and they were discontinued. The patient’s history is remarkable for a microprolactinoma, managed with dopamine agonist. She has facial acne, but no hirsutism. Her BMI is 24 kg/m2. Laboratory studies revealed normal testosterone (27 ng/mL) and DHEAS (64 mcg/dL). Her prolactin was 1.1 ng/mL Thyroid function tests were in the normal range. hCG was negative. A random progesterone was 12 ng/mL, consistent with an ovulatory cycle. Her ovarian assessment showed FSH of 6.6 IU/L, LH of 6.7 IU/L, estradiol of 51 pg/mL, Inhibin B of 139 pg/mL, all in the normal range. Transvaginal pelvic ultrasound determined the ovarian volumes to be 8 and 6 mL, respectively, with a basal antral follicle count of 15 per side. This would have predicted a normal AMH level in this patient. However, AMH levels were repeatedly found to be low (0.05 to 0.1 ng/mL) using the pro-Mature AMH assay. As this patient was interested in future fertility, additional studies to further investigate her AMH production were carried out. AMH Measurement: Novel AMH ELISAs with antibody epitopes specific to Pro-Mature, Mature-Mature and Pro-Pro were used to analyze the sample. The observed AMH concentration by Pro-AMH ELISA, Mature-Mature ELISA and PCOCheck ELISAs were 0.1 ng/mL, 0.14 ng/mL, and 4.99 ng/mL, respectively. AMH Pro-Pro assay (PCOCheck) uses a linear epitope two-sided antibody that is not impacted by post-translational modifications, known AMH mutations or conformational changes due to thermal instability. Conclusions: The AMH measurements indicate that the patient’s serum may not contain pro-mature associated form of AMH or the C-fragmented AMH in circulation. This may be due to mutated AMH in the C-terminus and can result in C-truncation. This subject’s AMH is mostly fragmented and contains enriched pro-region AMH, which was measured by PCOCheck ELISA. The Teaching: Tests such as PCOCheck ELISA, targeted to specific regions of AMH will provide clinicians better tools for patient management. Such tests will also help resolve the observed AMH discrepancies with AFC using legacy AMH tests. PCOCheck ELISA can be a valuable tool for assessment of ovarian reserve or polycystic ovary syndrome (PCOS).

Factors affecting the accuracy and reliability of the measurement of anti-Müllerian hormone concentration in the clinic.

ObjectiveWe aimed to identify the factors that influence serum anti-Müllerian hormone (AMH) concentration measurements.MethodsWe collected serum samples between May and September 2018 and compared the effect on AMH concentration measured by ELISA of conditions including venepuncture, storage time, storage temperature, locations of the reaction microplate, and the use of the oral contraceptive pill and gonadotrophin-releasing hormone (GnRH).ResultsAMH concentration was not affected by food intake but was affected by haemolysis. It was also much higher in samples on the edge of the ELISA microtitre plate. AMH concentration increased after incubation at room temperature for 1 day, 4°C for 3 days, −20°C for 1 month and −40°C for 4 months, but no change occurred during storage at −80°C for 9 months. AMH concentration was high in patients following GnRH agonist treatment but was not affected by oral contraceptives.ConclusionsNo fasting is required prior to AMH measurement. Placement of serum samples on the edge of microtitre plates affects the results of the AMH ELISA. If serum samples cannot be assayed immediately, it is best to store them at −80°C. Basal AMH concentration cannot be used as a measure of ovarian reserve after GnRH agonist treatment.