anti-Müllerian Hormone Measurement Research Articles

Abstract PO2-11-06: Anti-Müllerian hormone in Young women with breast CAncer to predict permanent loss of ovarian function after chemotherapy and anti-HER2 therapy (AMYCA): a biomarker analysis of the BETH and KAITLIN trials

Abstract Background: The ability to predict and identify ovarian function loss accurately after anticancer treatment is important for appropriate oncofertility counseling and to aid in therapy decision making for young women with early breast cancer (eBC). Anti-Müllerian hormone (AMH) has been proposed as both a pre- and post-treatment predictor of subsequent permanent loss of ovarian function. The present biomarker analysis conducted within two randomized controlled trials (RCTs) in a large and well-characterized patient population with HER2+ eBC receiving homogeneous treatment aimed to assess AMH use, alone and in combination with other hormonal biomarkers, for predicting loss of ovarian function at 36 months from randomization, i.e. approximately 2 years after end of therapy. Methods: BETH (NCT00625898) and KAITLIN (NCT01966471) were RCTs investigating adjuvant chemotherapy (CT) plus anti-HER2 therapy in high-risk HER2+ eBC patients. For the purpose of the present analysis, only samples of patients receiving currently adopted treatments were used, i.e. cohort 1A (6 cycles of docetaxel/carboplatin+trastuzumab (T)) and cohort 2A (3 cycles of docetaxel plus T followed by 3 cycles of anthracycline- and cyclophosphamide-based CT) of BETH trial, and arm 1 (3-4 cycles of anthracycline- and cyclophosphamide-based CT followed by 3-4 cycles of taxane plus T plus pertuzumab (P)) of KAITLIN trial. Women selected for this analysis were ≤45 years with known premenopausal status, available serum samples at baseline (pre-treatment) and/or end of therapy and 36 months (± 6 months) from randomization (i.e. min of 2 samples in all cases). Selected samples were centrally tested measuring AMH, follicle stimulating hormone (FSH) and estradiol (E2) using Roche Elecsys assays. Permanent ovarian loss was defined as FSH >25 IU/L and E2 < 110 pmol/L at 36 months. Results: Of 194 included patients, 62 were from BETH and 132 from KAITLIN. 139 (71.6%) received both taxane and anthracycline- and cyclophosphamide-based CT and 55 (28.4%) taxane only. Pertuzumab was used in addition to trastuzumab in 132 (68.0%) patients. AMH values declined from baseline median 8.44 pmol/L to undetectable levels in many women at end of therapy (median 0.08 pmol/L), with partial recovery at 36 months (median 0.14 pmol/L). Ovarian loss at 3 years was observed in 56 women with AMH available at baseline (29.5%) and in 52 women with AMH available at end of therapy (29.7%). For prediction of ovarian loss, in ROC analysis, AMH measured at baseline showed AUC 0.784 (internally validated using bootstrap: 0.783). While age influenced risk of ovarian loss, addition to AMH only slightly improved AUC to 0.800 (internal validation: 0.795) while addition of baseline FSH and E2 did not improve prediction. AMH measured at the end of therapy showed AUC 0.741 (internal validation: 0.742), which increased slightly to 0.785(internal validation: 0.781) with addition of age but without improved prediction with the addition of FSH and E2. The combination of AMH at baseline and end of therapy increased prediction slightly to 0.808 (internal validation 0.806) and with addition of age added to 0.820 (internal validation: 0.806). Conclusions: These results support the use of pretreatment measurement of AMH in predicting loss of ovarian function in women withHER2+ eBC aged ≤45 receiving CT and anti-HER2 therapy. Measurement of AMH at end of treatment was comparably valuable to pretreatment and added slightly to the value of pretreatment sampling. Inclusion of ovarian function assessment by measuring AMH before commencing treatment can inform subsequent risk of ovarian function loss with value for fertility preservation counselling and treatment decisions. Further validation is required in larger cohorts. Citation Format: Matteo Lambertini, Deirdre Allegranza, Ruediger Laubender, Nadia Harbeck, Sandra Swain, Charles Geyer, Dennis Slamon, Gabriella Bobba, Chiara Lambertini, Sanne Lysbet De Haas, Eleonora Restuccia, Ines Vaz Luis, David Cameron, Ian Krop, Eric Winer, Richard Anderson. Anti-Müllerian hormone in Young women with breast CAncer to predict permanent loss of ovarian function after chemotherapy and anti-HER2 therapy (AMYCA): a biomarker analysis of the BETH and KAITLIN trials [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-11-06.

Do the blood anti-Müllerian hormone concentrations in young females correlate with the ovarian follicular population later in life in water buffalo?

In this longitudinal study, the anti-Müllerian hormone (AMH) levels in blood were determined in 32 Murrah buffalo females at 8, 10, 12, 16 and 19 months of age when females were synchronized and the antral follicular population (AFP) was estimated. Correlations of AFP to the AMH level at 19 months of age and retrospectively to younger ages were investigated. Then females were split into high and low AFP, and their AMH levels were compared for all ages and tested as predictors of AFP categories. The highest AMH level (p < .05) was detected at 8 months, reducing but not differing (p > .05) at 10, 12 and 16 months then reducing again (p < .05) at 19 months of age. The mean AFP was 17.6 ± 6.3 follicles, and it was positively correlated with AMH in all ages tested. High AFP females had approximately two times more antral follicles than low AFP (p < .05) and their AMH levels were higher (p < .01) than in low AFP ones in all ages. Only at 8 months, AMH levels can be used to precociously detect high AFP heifers (a cut-off point of 464.7 pg/mL; p < .05), while low AFP heifers could be detected by AMH measurements at 8, 10, 12 and 16 months of age (p < .05). We conclude that AMH of buffalo calves correlates with AFP of heifers later in life and depending on the age, its levels could be used to identify future females with low or high AFP.

Age-related curves of AMH using the Gen II, the picoAMH and the Elecsys assays in women with polycystic ovary syndrome.

Several challenges still exist to adopt the anti-Müllerian hormone (AMH) as a marker of polycystic ovary morphology (PCOM), as included in the recently updated international guideline. Although different evaluations of age- and assay-specific reference ranges have been published in the last years, these studies have mainly been conducted in normo-ovulatory or infertile women. To develop an age-specific percentile distribution of AMH in patients with polycystic ovary syndrome (PCOS) measured by three different assays. Retrospective cross-sectional study. 2,725 women aged 20 to 40 years with PCOS diagnosis were included. Serum AMH measurement by the Gen II (Beckman Coulter), the picoAMH (Ansh Labs), and the Elecsys (Roche) assays. Age-specific centile curves for all the assays and correlations between AMH, clinical, hormonal, and ultrasound characteristics. Age-related nomograms for the 5th, 10th, 25th, 50th, 75th, 90th, and 95th percentiles of AMH were calculated using the LMS method for all the assays. AMH levels were significantly different between PCOS phenotypes. AMH levels were positive correlated to luteinizing hormone (LH), LH/follicular stimulating hormone (FSH) ratio, testosterone, androstenedione, free androgen index, mean follicular number, and mean ovarian volume. To our knowledge this is the first study reporting age specific percentile nomograms of serum AMH levels measured by the Gen II, the picoAMH and the Elecsys assays in a large population of PCOS women. These findings may help to interpret AMH levels in PCOS patients and facilitate the use of AMH as a diagnostic tool across age ranges.

Fertility and sexual activity in patients with Triple A syndrome.

Triple A syndrome, caused by autosomal recessively inherited mutations in the AAAS gene is characterized by alacrima, achalasia, adrenal insufficiency, and neurological impairment. To the best of our knowledge, no patients of both sexes have been reported to have offspring. Our aim was to assess the causes of infertility in male patients with this multisystemic syndrome, and to present a female patient that spontaneously conceived a child. Cross-sectional study. Six males aged 19-48 years were included. Gonadotropins, testosterone, DHEAS, androstenedione, inhibin B, anti-Mullerian hormone measurements and testicular ultrasound were performed. All six male patients had impaired general health and neurological symptoms including erectile and ejaculatory dysfunction. None of them had an offspring. The only demonstrated cause of infertility in our male patients was erectile and ejaculatory dysfunction which precludes sexual intercourse. Our patients had normal libido but were sexually abstinent. Except for low adrenal androgen levels, the concentrations of all measured hormones as well as testicular ultrasound were normal which may indicate the possibility of spermatogenesis in male patients with triple A syndrome. Little is known about fertility in female patients, but based on our observations spontaneous pregnancies seem to be possible. Our results contribute to still scarce knowledge on fertility in patients with Triple A syndrome and as well represents a foundation for further research on causes of infertility and possible treatment options.

Minimally Invasive Method of Measuring Anti-Müllerian Hormone Level in Dried Blood Spots of Indonesian Infertile Population

This study aims to evaluate the certainty of anti-Müllerian hormone (AMH) levels in dried blood spots (DBS) compared to serum AMH. A total of 109 infertile women who planned to undergo their controlled ovarian stimulation cycle were recruited. Serum and DBS samples were collected from each patient for the quantification of AMH levels. Serum AMH was measured with an automated clinical chemistry assay (Roche) while DBS AMH was measured using the Ansh Labs DBS AMH ELISA reagent kit. Mean AMH level in DBS was [Formula: see text][Formula: see text]ng/mL, which ranged from 0.01 to 17.3[Formula: see text]ng/mL, while the mean AMH in serum samples was [Formula: see text][Formula: see text]ng/mL (0.01–10.8[Formula: see text]ng/mL). Comparable accuracy for low AMH quantification was observed between the two methods over basal antral follicle counts (AFC) as a clinical comparator (AUC 86%, cutoff [Formula: see text][Formula: see text]ng/mL, [Formula: see text], [Formula: see text] vs. 85%, cutoff [Formula: see text][Formula: see text]ng/mL, [Formula: see text], [Formula: see text], respectively for DBS and serum sample). Likewise, DBS AMH achieved high concordance for low AMH discrimination in comparison to serum AMH method (AUC 99% vs. 93%, respectively). A strong linear Spearman regression ([Formula: see text], [Formula: see text]) and sufficient agreement (94.5%, mean difference of −1.6, 2.7 to −5.9) was demonstrated among studied groups with mathematical Passing and Bablok regression equation of [Formula: see text]. The confidence interval of intercept (−0.12, 95% CI [−0.29 to −0.01]) and slope (1.80, 95% CI [1.66–1.94]) indicated a constant and proportional difference between the two methods but it was not significant ([Formula: see text]). AMH measurement in DBS could be useful as an alternative approach to screening ovarian reserve through a more convenient remote sample collection.

A systematic review of the association between modifiable lifestyle factors and circulating anti-Müllerian hormone.

Levels of anti-Müllerian hormone (AMH) are known to be associated with lifestyle determinants such as smoking and oral contraception (OC) use. When measuring AMH in clinical practice, it is essential to know which factors may influence circulating levels or ovarian reserve in general. To date, there is no systematic review or summarizing consensus of the nature and magnitude of the relation between AMH and modifiable lifestyle factors. The purpose of this review was to systematically assess the evidence on association of lifestyle behaviors with circulating AMH levels. We performed a pre-registered systematic review of publications in Embase and PubMed on the lifestyle factors BMI, smoking, OC use, alcohol consumption, caffeine consumption, physical activity, and waist-hip ratio (WHR) in relation to circulating AMH levels up to 1 November 2023. The search strategy included terms such as 'Anti-Mullerian hormone', 'lifestyle', and 'women'. Studies were considered eligible if the association between at least one of the lifestyle factors of interest and AMH was assessed in adult women. The quality of included studies was assessed using the Study Quality Assessment Tools of the National Heart, Lung, and Blood Institute. The results were presented as ranges of the most frequently used association measure for studies that found a significant association in the same direction. A total of 15 072 records were identified, of which 65 studies were eligible for inclusion, and 66.2% of the studies used a cross-sectional design. The majority of studies investigating BMI, smoking, OC use, and physical activity reported significant inverse associations with AMH levels. For WHR, alcohol, and caffeine use, the majority of studies did not find an association with AMH. For all determinants, the effect measures of the reported associations were heterogeneous. The mean difference in AMH levels per unit increase in BMI ranged from -0.015 to -0.2 ng/ml in studies that found a significant inverse association. The mean difference in AMH levels for current smokers versus non-smokers ranged from -0.4 to -1.1 ng/ml, and -4% to -44%, respectively. For current OC use, results included a range in relative mean differences in AMH levels of -17% to -31.1%, in addition to a decrease of 11 age-standardized percentiles, and an average decrease of 1.97 ng/ml after 9 weeks of OC use. Exercise interventions led to a decrease in AMH levels of 2.8 pmol/l to 13.2 pmol/l after 12 weeks in women with polycystic ovary syndrome or a sedentary lifestyle. Lifestyle factors are associated with differences in AMH levels and thus should be taken into account when interpreting individual AMH measurements. Furthermore, AMH levels can be influenced by the alteration of lifestyle behaviors. While this can be a helpful tool for clinical and lifestyle counseling, the nature of the relation between the observed differences in AMH and the true ovarian reserve remains to be assessed. PROSPERO registration ID: CRD42022322575.

Anti-Müllerian hormone: A novel biomarker for detecting bovine freemartinism.

The anti-Müllerian hormone (AMH) indicates ovarian reserve in cattle, maintaining a consistent trajectory post-puberty. In heterosexual pregnancies, the development of the Müllerian duct in female foetuses is inhibited, resulting in an anticipated minimal or absent ovarian reserve capacity. This investigation aimed to compare AMH levels in healthy Holstein heifers that had reached puberty with those of freemartin animals of the same breed and age. The study incorporated Holstein heifers reaching puberty between 11 and 15 months of age in Group 1 (G1, n = 20) and freemartin animals in Group 2 (G2, n = 19, 16). AMH measurements (AMH-1/AMH-2) were recorded at 12-day intervals for the study participants. Notably, AMH levels in three freemartin animals could not be detected, prompting statistical analysis based on measurements from the remaining 16 freemartin animals in G2. A statistically significant correlation was observed between two separate measurements in G1 and G2 (p < .001). Furthermore, AMH-1 and AMH-2 levels were statistically higher in G1 than in G2 (p < .001). In G1, AMH-1 levels ranged from 227 to 677 pg/mL, with an average of 367.3 ± 25.5 pg/mL, and AMH-2 levels ranged from 234 to 645 pg/mL, with an average of 380.8 ± 24.4 pg/mL. Conversely, in G2, AMH-1 levels ranged from 10 to 72 pg/mL, with an average of 26.8 ± 4.44 pg/mL, and AMH-2 levels ranged from 12 to 68 pg/mL, with an average of 28.75 ± 4.18 pg/mL. The mean AMH levels in G1 were approximately 14 times higher than in G2 (p < .001). Consequently, ROC analysis utilizing AMH-1 and AMH-2 data established cut-off values of ≤72 and ≤ 68 pg/mL respectively for distinguishing freemartin animals. In conclusion, AMH could be used as a reliable biomarker for identifying Holstein freemartin animals.

Correlation of serum anti-Mullerian hormone and Inhibin-B levels with osteoporosis of menopausal woman in Chinese Daur ethnic group.

To investigate the diagnostic value of anti-Mullerian hormone (AMH) and Inhibin B (InhB) in menopausal women with osteoporosis from the Chinese Daur ethnic group. A total of 175 menopausal women were selected and divided into the osteoporosis group (N = 90) and the control group (N = 85). BMD was measured by dual-energy X-ray absorptiometry, and laboratory indicators of osteoporosis, for example, serum osteocalcin (OC), β-collagen special sequence (β-CTX), and procollagen type I amino-terminal propeptide (PINP), bone alkaline phosphatase (BALP), AMH, and InhB were measured by commercial kits. The relationship between osteoporosis and AMH or InhB was analyzed. The predictive values of AMH and InhB were reflected by the ROC curve and logistic regression. The level of BMD was decreased and the levels of OC, β-CTX, PINP, and BALP of the menopausal osteoporosis group were increased. The concentration of AMH and InhB in the menopausal osteoporosis group was decreased and they had connections with each other. AMH and InhB could be used as independent indicators for the occurrence of osteoporosis in menopausal women and their combination had a higher diagnostic value. AMH and InhB measurements in menopausal women had a certain clinical significance in the detection of osteoporosis. The occurrence of osteoporosis was related to BMD, OC, β-CTX, BALP, AMH, and InhB.

Serum anti-Müllerian hormone is lower in patients with multiple radioiodine dose for treatment of pediatric thyroid cancer.

Treatment of patients with pediatric differentiated thyroid cancer (DTC) often involves radioiodine (RAI), which is associated with increased risks of short- and long-term adverse outcomes. The impact of RAI treatment on the female reproductive system remains uncertain. Anti-Müllerian hormone (AMH) is a marker of ovarian reserve and is related to fertility. The aim was to analyze the association between RAI and serum AMH level in women treated with RAI. We evaluated women with pediatric DTC treated with RAI at the age of ≤19 years. Serum AMH was measured. The study included 47 patients with a mean age of 25.1 years (12.4-50.8) at AMH measurement and follow-up of 11.8 ± 8.4 years. The mean RAI administered was 235 mCi (30-1150). Sixteen (34%) received multiple RAI doses (471 ± 215 mCi). Mean AMH level was 2.49 ng/mL (0.01-7.81); the level was 1.57 ng/mL (0.01-7.81) after multiple RAI doses and 2.99 ng/mL (0.01-6.63) after a single RAI dose (P = 0.01). Patients who received a cumulative RAI lower than 200 mCi had higher AMH levels (2.23 ng/mL, 0.39-7.81) than those who received more (1.0 ng/mL, 0.01-6.63; P = 0.02). In patients with similar cumulative RAI activities, administration of multiple RAI doses was significantly and independently associated with AMH level lower than the reference range for age (HR: 5.9, 1.55-52.2, P = 0.014) after age adjustments. Levels of AMH were lower after multiple RAI doses, especially after a cumulative RAI dose above 200 mCi. More studies are needed to clarify the impact of RAI on fertility considering its cumulative activity and treatment strategy.

Heavy Metals and Trajectories of Anti-Müllerian Hormone During the Menopausal Transition.

Experimental and epidemiological studies have linked metals with women's reproductive aging, but the mechanisms are not well understood. Disrupted ovarian folliculogenesis and diminished ovarian reserve could be a pathway through which metals impact reproductive hormones and outcomes. The study aimed to evaluate the associations of heavy metals with anti-Müllerian hormone (AMH), a marker of ovarian reserve. The study included 549 women from the Study of Women's Health Across the Nation with 2252 repeated AMH measurements from 10 to 0 years before the final menstrual period (FMP). Serum AMH concentrations were measured using picoAMH ELISA. Urinary concentrations of arsenic, cadmium, mercury, and lead were measured using high-resolution inductively coupled plasma mass spectrometry. Multivariable linear mixed regressions modeled AMH as a function of time before the FMP interaction terms between metals and time to the FMP were also included. Adjusting for confounders, compared with those in the lowest tertile, women in the highest tertile of urinary arsenic or mercury concentrations had lower AMH concentrations at the FMP (percent change: -32.1%; 95% CI, -52.9 to -2.2, P-trend = .03 for arsenic; percent change: -40.7%; 95% CI, -58.9 to -14.5, P-trend = .005 for mercury). Higher cadmium and mercury were also associated with accelerated rates of decline in AMH over time (percent change per year: -9.0%; 95% CI, -15.5 to -1.9, P-trend = .01 for cadmium; -7.3%; 95% CI, -14.0 to -0.1, P-trend = .04 for mercury). Heavy metals including arsenic, cadmium, and mercury may act as ovarian toxicants by diminishing ovarian reserve in women approaching the FMP.

Concordance of hemoglobin A1c and reproductive hormone levels in menstrual and venous blood

ObjectiveTo explore whether menstrual blood collected via a modified menstrual pad is a surrogate for venous blood draw in analyzing HbA1c and fertility-associated hormones DesignCross-sectional study SubjectsStudy included 152 female participants who have regular menses, aged 19-50 years old. InterventionsParticipants collected menstrual effluent using a menstrual pad, modified with a removable dried blood spot (DBS) strip. Peripheral blood samples were collected by venipuncture within 60 hours of menstrual pad use. Main Outcome MeasuresMenstrual pad and venous blood draw samples were analyzed for levels of HbA1c, thyroid stimulating hormone (TSH), Follicle Stimulating Hormone (FSH), anti-Mullerian hormone (AMH), and Luteinizing hormone (LH). Correlation between menstrual pad and venipuncture samples was performed by Deming linear regression, and r coefficients were measured using Pearson correlation. ResultsInter-assay variability of menstrual pad DBS sample measurements was <6%. Menstrual HbA1c values were stabilized in the DBS strips through 53 days, and menstrual hormone levels remained stable through 15 days. Menstrual HbA1c levels were highly correlated with venipuncture samples (r=0.96). TSH (r=0.94), AMH (r=0.94), FSH (r=0.91), and LH (r=0.91) also showed high correlation between menstrual strip and venipuncture samples. ConclusionsHbA1c, TSH, AMH, FSH, and LH measurements in menstrual effluent showed high correlation to venous blood samples, supporting use of menstrual effluent as a surrogate sample for hormone testing.

Association of exposure to ozone and fine particulate matter with ovarian reserve among women with infertility

Evidence linking diminished ovarian reserve, a significant cause of female infertility, and exposure to particulate matter with aerodynamic diameters ≤2.5 μm (PM2.5) or O3 exposure remains a critical knowledge gap in female fertility. This study investigated the association between ambient PM2.5, O3 pollution, and anti-Müllerian hormone (AMH), a sensitive marker of ovarian reserve, in reproductive-aged Chinese women. We enrolled 2212 women with spontaneous menstrual cycles who underwent AMH measurements at a reproductive medicine center between 2018 and 2021. The daily mean concentrations of outdoor PM2.5 and O3 were estimated using a validated spatiotemporal model, followed by matching the participants' residential addresses. Three exposure periods were designed according to AMH expression patterns during follicle development. A generalized linear model was used to investigate changes in AMH associated with air pollution. The results showed a mean AMH level of 3.47 ± 2.61 ng/mL. During the six months from primary to early antral follicle stage (Period 1), each 10 μg/m3 increase in PM2.5 and O3 exposure was associated with AMH changes of −0.21 (95% confidence interval [CI]: −0.48, 0.06) ng/mL and −0.31 (95% CI: −0.50, −0.12) ng/mL, respectively. Further analyses indicated that the reduced ovarian reserve measured by AMH level was only significantly associated with PM2.5 exposure during follicle development from the primary to preantral follicle stage (Period 2) but was significantly associated with O3 exposure during Periods 1, 2, and 3. These observations were robust in the dual-pollutant model considering co-exposure to PM2.5 and O3. The results indicated an inverse association between ovarian reserve and ambient O3 exposure and suggested distinct susceptibility windows for O3 and PM2.5 for reduced ovarian reserve. These findings highlight the need to control ambient air pollution to reduce invisible risks to women's fertility, especially at high O3 concentrations.

Evaluation of serum anti-Müllerian hormone concentrations following treatment with vitamin D in Holstein Friesian heifers.

Anti-Müllerian hormone (AMH) levels in the blood of dairy cattle are associated with the count of antral follicles (AFC), ovarian function. Numerous studies, particularly in human medicine, have shown that vitamin D3 (Vit D3) has a positive effect on AMH levels. To our knowledge, the effects of Vit D3 on serum AMH levels in heifers have not been investigated. The goal of the current study was to evaluate the concentrations of serum AMH in dairy heifers following Vit D3 treatment. The study included 20 healthy non-pregnant Holstein Friesian heifers. These animals came to estrus at least once. All heifers received a single intramuscular dosage of 5 million IU Vit D3 (Cholecalciferol). Blood samples were collected from the coccygeal veins of all animals before and after Vit D3 injection (days 7, 14, and 28) for the measurement of AMH and 25(OH)D concentrations. In our analysis, we observed that after exogenous Vit D3 administration, circular 25(OH)D increased constantly (up to day 28 after Vit D3 injection) in all animals. On day 28, AMH concentrations saw a 10% increase in comparison with those measured the day before the Vit D3 injection. However, a comparison of AMH concentrations measured across days did not reveal statistically significant differences between Day 0, 7, 14, and 28 levels (p = .10). Furthermore, no statistical correlation was observed between the pairs (age-AMH) and (weight-AMH). Considering all times, no correlation was found between 25(OH)D and blood AMH levels. These findings demonstrated that exogenous Vit D3 did not affect serum AMH in Holstein Friesian heifers.

Anti-Müllerian hormone in female dromedary camel and its association with super-ovulatory response in embryo donors

Anti-Müllerian hormone (AMH) has a conserved role in regulating the reproductive cycle in several species. Its circulating concentration reflects the size of the growing primordial follicle reserve and is a reliable predictor of superovulation response in embryo/oocyte donors. This study investigated the possible application of AMH measurement in dromedary camels (Camelus dromedarius) multiple ovulation embryo transfer programs. In experiment 1, the follicular cycle of synchronized and naturally cycling camels (n = 12) was monitored. Blood was collected at 6 timepoints in 2 consecutive cycles corresponding to emergence, mid-cycle, and dominance in both group and hormonal fluctuations were evaluated for repeatability of measurements within and between cycles. In experiment 2, the correlation between circulating AMH concentrations prior to initiation of superovulation and the outcome of superovulation was evaluated. The results were compared between donors with higher (n = 7) and lower than median (n = 8) AMH values. Mean AMH concentrations in synchronized and non-synchronized camels were 1.46 ± 0.15 and 0.95 ± 0.09, respectively. Intercycle and intracycle values of AMH showed high repeatability in camels of both groups (>96.4% and >92.74%, respectively) with significant correlations between values at different stages of the ovarian cycle (Emergence and mid-cycle: R2 = 0.82; emergence and. dominance: R2 = 0.86; Mid-cycle and dominance: R2 = 0.93, P < 0.05). Total follicles, CLs, and recovered embryos were highly correlated with AMH values prior to superovulation (R2 = 0.64, R2 = 0.77, and R2 = 0.64, respectively, P < 0.05). A greater number of developed follicles prior to mating (17.00 ± 2.09 vs. 7.62 ± 1.06), CLs (12.58 ± 1.36 vs. 5.12 ± 0.93), transferable (10.85 ± 1,31 vs. 3.37 ± 0.82), and spherical embryos (8.14 ± 1.07 vs. 2.62 ± 0.7) were observed in camels with higher than median concentrations of AMH (P < 0.05). Fluctuations in estradiol and progesterone did not affect variations in mean AMH values (r2 < 0.19 and r2 < 0.24, respectively, P > 0.05). In conclusion, highly consistent AMH values in dromedary camels are a reliable predictor of superovulation response and outcome in dromedary camels.

Abstract 019: Perimenopause Exacerbates Hypertension In Doca-salt Rats

Blood pressure (BP) is lower in premenopausal women than in age-matched men, but after menopause BP rates and the incidence of hypertension (HTN) show opposite tendencies. The mechanisms by which fluctuating ovarian hormone environment during that period impacts the onset and development of HTN are poorly understood, and strategies to reduce HTN risk in peri- and postmenopausal females are lacking. An ovary-intact rat model of perimenopause using the chemical, 4-vinylcyclohexene diepoxide (VCD), has been developed to accelerate the natural process of slow follicular loss. Daily dosing with VCD for 15 days selectively destroys primordial and primary follicles in ovaries of mice and rats by accelerating atresia processes, thus inducing precocious perimenopause/menopause. Hypothesis: The induction of perimenopause by VCD will increase susceptibility to the development of HTN in DOCA-salt rats. Female Sprague-Dawley rats at post-natal day 28 received subcutaneous injection of VCD (160mg/kg) or corn oil (control) for 15 days. 57 days after the first injections, nephrectomy and telemetric pressure transducer implantation were performed. After 2 weeks of recovery, a 100 mg/kg DOCA silicone or placebo pellet was implanted subcutaneously. The rats that received DOCA were provided 0.9% saline during the 21 days of DOCA protocol and mean arterial pressure (MAP) was continuously evaluated. Rats were housed weekly in metabolic cages for 24 hours to measure saline intake and urine volume. After protocol completion the animals were decapitated, and blood was collected for the measurement of anti-Mullerian hormone (AMH) and estradiol (E2) by ELISA. The AMH plasma levels were reduced in the periestropause rats (VCD + DOCA, n=9) compared to oil (Oil + DOCA, n=6) confirming the VCD-induced follicular depletion. The steady state increase in MAP (day 21 of DOCA-salt) was higher in periestropause (±45.7mmHg) compared to oil (±28.6 mmHg). Urine volume was higher in periestropause rats compared to oil. No differences were observed in the saline intake between the groups. Our data demonstrate that periestropause rats showed greater susceptibility to the development of HTN in a DOCA-salt rat model.

Using serum anti-Müllerian hormone levels to predict the chance of live birth after spontaneous or assisted conception: a systematic review and meta-analysis.

Is serum anti-Müllerian hormone (AMH) level predictive of cumulative live birth (CLB) rate after ART or in women trying to conceive naturally? Serum AMH level is linked to CLB after IVF/ICSI but data are lacking after IUI or in women trying to conceive without ART. Serum AMH level is a marker of ovarian reserve and a good predictor of ovarian response after controlled ovarian stimulation. It is unclear whether AMH measurement can predict CLB in spontaneous or assisted conception. A systematic review and meta-analysis was undertaken to assess whether serum AMH level may predict chances of CLB in infertile women undergoing IVF/ICSI or IUI and/or chances of live birth in women having conceived naturally. A systematic review and meta-analysis was performed using the following keywords: 'AMH', 'anti-mullerian hormone', 'live-birth', 'cumulative live birth'. Searches were conducted from January 2004 to April 2021 on PubMed and Embase. Two independent reviewers carried out study selection, quality, and risk of bias assessment as well as data extraction. Odds ratios were estimated using a random-effect model. Pre-specified sensitivity analyses and subgroup analyses were performed. The primary outcome was CLB. A total of 32 studies were included in the meta-analysis. Overall, 27 articles were included in the meta-analysis of the relation between AMH and CLB or AMH and LB after IVF/ICSI. A non-linear positive relation was found in both cases. A polynomial fraction was the best model to describe it but no discriminant AMH threshold was shown, especially no serum AMH level threshold below which live birth could not be achieved after IVF/ICSI. After IVF-ICSI, only four studies reported CLB rate according to AMH level. No statistically significant differences in mean serum AMH levels were shown between patients with and without CLB, but with a high heterogeneity. After exclusion of two studies with high risks of bias, there was no more heterogeneity [I2 = 0%] and the mean AMH level was statistically significantly higher in women with CLB. There were not enough articles/data to assess the ability of AMH to predict CLB rate or find an AMH threshold after IUI or in women without history of infertility trying to conceive without ART. The systematic review and meta-analysis had some limitations owing to the limits and bias of the studies included. In the present meta-analysis, heterogeneity may have been caused by different baseline characteristics in study participants, different stimulating protocols for ART, different serum AMH level thresholds used and the use of various assays for serum AMH. This could explain, in part, the absence of a discriminating AMH threshold found in this analysis. Serum AMH level is linked to CLB rate after IVF/ICSI but no discriminating threshold can be established, therefore low serum AMH level should not be used as the sole criterion for rejecting IVF treatment, especially in young patients. Data are lacking concerning its predictive value after IUI or in women trying to conceive without ART. Our findings may be helpful to counsel candidate couples to IVF-ICSI. No external funding was obtained for this study. There are no conflicts of interest. PROSPERO CRD42021269332.

P-622 Age-specific reference intervals for anti-müllerian hormone in Mexico: A population-based study. implications for Latin America

Abstract Study question To determine the age-specific normal range of antimullerian hormone (AMH) levels in a female Latin American population. Summary answer Age-related nomograms for general population for the 3rd-97th percentiles were produced. These nomograms could provide a reference guide for clinicians to consult women with infertility. What is known already The variation of AMH levels among different races and ethnicities has been well documented (Seifer et al., 2009). Despite the wide use of AMH measurements in clinical assessment of ovarian reserve, a population-based estimate for its reference range in Latin America is not available. Study design, size, duration A longitudinal, observational, retrospective, real-world study was carried out, including all AMH measurements performed from January 2017 to December 2018 at a large clinical laboratory network in the Mexico City area, encompassing 35 locations. A total of 6444 AMH measurements were included. Participants/materials, setting, methods AMH was assayed using either the cobas or ELISA methodologies. All AMH measurements from women 25-45 years were included. The Kolmogorov-Smirnov test was used to confirm that AMH levels between patients assayed with cobas were comparable to those assayed with ELISA at every age. Main results and the role of chance In total, 6444 measurements were identified and available for analysis. Age-related normograms for the 3rd, 10th, 25th, 50th, 75th, 90th, and 97th percentiles of AMH were produced. The overall rate of AMH decline accelerated after 40 years of age. In women 25-30 years, 11.4% (63/554) had an AMH &amp;lt;1.0ng/mL. In women 30-35 years, 18.6% (276/1484) had an AMH &amp;lt;1.0ng/mL. In women 35-40 yeras, 35.8% (830/2319) had an AMH &amp;lt;1.0ng/mL. In women 40-45 years, 58.0% (1120/1932) had an AMH &amp;lt;1.0ng/mL. Limitations, reasons for caution The lack of medical history for patients precluded identification and exclusión of patients with iatrogenically abnormal AMH levels (e.g. due to oophorectomy). Patients undergoing measurement of AMH were primarily referred through fertility clinics, limiting the potential for extrapolation to the general population. Wider implications of the findings The study reports age-specific AMH levels drawn from a large sample of women around Mexico City. The study includes the largest dataset to date. Latin American patients considering fertility preservation should be aware of the high prevalence of diminished ovarian reserve in this large cohort, including among women 35-39. Trial registration number Not Applicable

Zonulin as a potential biomarker for diminished ovarian reserve: A prospective study.

The purpose of this research is to investigate the relationship between zonulin levels and diminished ovarian reserve (DOR), and to evaluate the potential role of autoimmunity in the development of DOR. The study contributes to the understanding of the pathogenesis of DOR, which can be an unexpected diagnosis often associated with infertility and unpleasant physical symptoms in women. This cross-sectional study was conducted by scanning 224 patients. The demographic characteristics of the patients were recorded. Antral follicle counts of the patients were determined by ultrasound, and Anti-Mullerian hormone (AMH) levels were examined. Follicle-stimulating hormone (FSH), luteinizing hormone, estradiol, AMH measurement, and antral follicle counts were made on the 2nd or 3rd day of menstrual bleeding. The zonulin levels of the participants were measured by the ELISA method. The patients were divided into two groups according to the presence of DOR. The patients' demographic characteristics and hormone levels were compared between these two groups, serum zonulin levels were examined, and the relationship between other hormone parameters and zonulin was investigated. When the median ages of the patients in both groups were compared, the median age of patients with DOR was 38 years, significantly higher (p<0.001) than the median age of those without DOR, which was 27 years. The median zonulin levels of both groups were compare; it was observed that it was 19.71 ng/mL in the group with DOR and 11.03 ng/mL without DOR, and a statistically significant difference was found between the zonulin levels of the patients in both groups (p<0.001). A moderate inverse correlation (p<0.001) between patients' zonulin and AMH levels, and a moderate correlation between FSH levels (p<0.001). In conclusion, zonulin levels of patients with DOR were higher than women without DOR. Evaluation of zonulin levels may also be considered during the diagnosis of DOR.

Measurement of serum anti-Müllerian hormone by revised Gen II or automated assay: Reproducibility under various blood/serum storage conditions.

We investigated the agreement between anti-Müllerian hormone (AMH) levels measured with revised Gen II (rev-Gen II) and automated AMH (Access) assays and evaluated the reproducibility of each method under various blood/serum storage conditions. AMH levels in blood samples from 74 volunteers were measured by rev-Gen II and Access assays under various conditions: immediate serum separation and AMH measurement (fresh control); serum stored at -20 °C and AMH measured after 48 hours, 1 week, and 2 years; serum stored at 0 to 4 °C and AMH measured after 48 hours and 1 week; and blood kept at room temperature and delayed serum separation after 48 hours and 1 week, with immediate AMH measurement. In fresh controls, all rev-Gen II-AMH values were higher than comparable Access-AMH values (difference, 8.3% to 19.7%). AMH levels measured with the two methods were strongly correlated for all sample conditions (r=0.977 to 0.995, all p<0.001). For sera stored at -20 °C or 0 to 4 °C for 48 hours, Access-AMH values were comparable to control measurements, but rev-Gen II-AMH values were significantly lower. AMH levels in sera stored at -20 °C or 0 to 4 °C for 1 week were significantly lower than in fresh controls, irrespective of method. Across methods, long-term storage at -20 °C for 2 years yielded AMH measurements significantly higher than control values. When serum separation was delayed, rev-Gen II-AMH values were significantly lower than control measurements, but Access-AMH values varied. The rev-Gen II and Access-AMH assays showed varying reproducibility across blood/serum storage conditions, but automated Access yielded superior stability to rev-Gen II.

Low ovarian reserve and risk of miscarriage in pregnancies derived from assisted reproductive technology.

Do low levels of anti-Müllerian hormone (AMH) or antral follicle count (AFC) properly predict miscarriage in young women conceiving with ART? Low ovarian reserve, as indicated by AMH or AFC, is not associated with miscarriage in young women conceiving with ART. Presently, the impact of low ovarian reserve on the risk of miscarriage remains controversial. Some studies have reported an association between serum AMH levels and AFC and miscarriage, but others have failed to confirm these findings. The main limitation that undermines the reliability and consistency of the results is the confounding effect of female age. Indeed, after 35 years of age, on the one hand, the risk of miscarriage starts increasing because of impaired oocyte quality while, on the other, the physiological decline in AMH and AFC levels continues, thus hampering the possibility to properly explore the real effects of reduced ovarian reserve. Indeed, the two processes, i.e. the gradual loss of resting primordial follicles and the loss of oocyte quality, progress in parallel. In other words, the older the woman becomes, the higher is the risk of miscarriage, but one cannot distinguish between the effects of biological aging on oocyte quality and those mediated by a lower ovarian reserve. The present retrospective monocentric cohort study was carried out at Fondazione IRCSS Ca Granda Ospedale Maggiore Policlinico, Milan. All women referred to the ART Unit between 2014 and 2021 and who underwent either conventional IVF (c-IVF), ICSI, or IUI were reviewed. Only women younger than 35 were eligible because, up to this age, the risk of miscarriage is steady and not strictly related to age. Women younger than 35 who achieved a singleton clinical pregnancy with c-IVF, ICSI, or IUI were selected. Women with patent causes of recurrent miscarriage were excluded, as well as those undergoing pregnancy termination for fetal or medical causes. Women who did and did not have a pregnancy loss before 20 weeks' gestation were compared. Detailed information was obtained from charts of the consulting patients. ART procedures were performed according to the standardized policy of our Unit. All women underwent serum AMH measurement and a transvaginal assessment of AFC prior to initiation of treatment. AMH levels were measured by a commercially available ELISA assay. To assess AFC, all identifiable antral follicles 2-10 mm in diameter at ultrasound were recorded. The primary outcome was the risk of miscarriage for women with serum AMH levels below 5 pmol/l. There were 538 women were included, of whom 92 (17%) had a miscarriage. The areas under the ROC curves for prediction of miscarriage based on AMH levels and AFC were 0.51 (95% CI: 0.45-0.58) and 0.52 (95% CI: 0.45-0.59), respectively. The odds ratio (OR) of miscarriage for women with serum AMH levels below 5.0 pmol/l was 1.10 (95% CI: 0.51-2.36); the adjusted OR was 1.12 (95% CI: 0.51-2.45). Analyses were repeated considering other thresholds for AMH (2.9, 3.6 and 7.9 pmol/l) and for AFC (thresholds of 7 and 10). No associations emerged. The retrospective design of the study hampered the collection of more precise but potentially relevant clinical information of the couples. We did not exclude women suffering from PCOS, a condition possibly associated with miscarriage. Moreover, the baseline characteristics of women who did and did not have a miscarriage differed in some characteristics. Thus, we adjusted the OR using a multivariate analysis, but we cannot fully exclude residual confounding effects. Finally, our results cannot be inferred to women older than 35. The mechanisms causing premature exhaustion of ovarian reserve may be different in younger and older women and this may lead to a different impact on the risk of miscarriage. Women embarking on ART with low ovarian reserve should be informed of their likely poor response to ovarian stimulation but can be reassured that, if conception occurs, their risk of miscarriage is not increased. This study was partially funded by Italian Ministry of Health-Current research IRCCS. E.S. reports grants from Ferring and honoraria for lectures from Merck-Serono and Gedeon-Richter. All the other authors do not have any competing interest to declare. N/A.