Ansh Labs Research Articles

7701 Comparative Analysis of Dried Blood Spot and Serum ELISAs for AMH, FSH, and LH in Patients Seeking IVF Treatment

Abstract Disclosure: T. Kumar: None. R. Chauhan: None. B. Kalra: None. A.S. Patel: None. S. Chauhan: None. A. Kumar: None. Relevance: Dried Blood Spot (DBS) technology has not been widely implemented in the diagnostic industry due to its limitations in obtaining reliable, quantitative data from small blood samples. However, recent advancements in methods, i.e., sensitive antibodies, homogeneous and reproducible paper quality, and painless needles have renewed the interest in the DBS collection technique. Objective: The aim of the study is to validate the hormone measurements using DBS technology and compare the correlation matrix between AMH, LH, FSH, inhibin B, Antral Follicle Counts (AFC), age, and BMI in patients seeking infertility workup. Materials and Methods: Matched serum and DBS samples from thirty-nine patients between the ages of 18 and 45 years were collected on day 3 of the cycle. The subjects were visiting the clinic for initial fertility workup and subjects on medications for ovarian suppression were excluded. The samples were tested on Ansh Labs DBS AMH (AL-129), FSH (AL-187), and LH (AL-190), PCOCheck AMH (N-N terminus, AL-196), picoAMH (C-N terminus, AL-124-r), US Inhibin B (AL-195) ELISAs. Baseline AFC obtained via transvaginal ultrasound, BMI, and age were collected. Spearman’s correlations, and regression analysis using Passing and Bablok were calculated to examine the relationship between AFC, AMH, inhibin B, FSH LH, age, and BMI. Results: DBS samples were highly correlated to their matched serum specimens. The slope and Spearman’s rank correlation (rs) between matched serum and DBS specimens for AMH, FSH, and LH were 1.17x, 1.07x, 1.07x, and 0.98, 0.93, and 0.97, respectively. DBS AMH strongly correlated with AFC. The Spearman’s rank correlation (rs) between AFC and DBS AMH, DBS LH, and DBS FSH were 0.80, -0.11, and -0.31, respectively. A strong correlation was observed between age and AMH, LH and FSH. A weak correlation was observed between BMI and AMH, LH and FSH. The Spearman’s rank correlation (rs) between age and DBS AMH, LH, and FSH were -0.34, 0.28, and 0.25, respectively. DBS AMH, LH, and FSH correlation with BMI were -0.06, 0.05, and 0.09, respectively. Inhibin B had a strong correlation to age (rs -0.33), BMI (rs -0.67), AFC (rs 0.44), DBS AMH (rs 0.40), DBS FSH (rs -0.19), DBS LH (rs-0.11), respectively. AMH method comparison between N-N and N-C terminal ELISA using 39 serum samples yielded slopes of 0.85x and Spearman’s rank correlation of 0.97. Conclusions: DBS and serum measurements for AMH, LH, and FSH are very accurate and precise. They can be used interchangeably. The strong relationship between BMI and Inhibin B will require further investigation and may open new avenues in obesity and reproductive endocrinology. Presentation: 6/1/2024

Associations of bisphenol and phthalate exposure and anti-Müllerian hormone levels in women of reproductive age

In women, exposure to endocrine disrupting chemicals might accelerate the depletion of the ovarian reserve and might be associated with accelerative reproductive aging and fertility. We examined the longitudinal associations of exposure to bisphenols and phthalates with anti-Müllerian hormone concentrations. Pregnant women of 18 years or older that resided in Rotterdam between 2002 and 2006 were eligible for participation in this longitudinal prospective cohort study. We measured urinary bisphenol and phthalate concentration at three time-points in pregnancy among 1405 women, of whom 1322 women had serum Anti-Müllerian Hormone (AMH) measurements 6 and/or 9 years postpartum. We performed linear regression models to assess the association of urinary bisphenol and phthalate metabolites with AMH after 6 and 9 years, and linear mixed-effect model to assess the association with AMH over time. Models were adjusted for sociodemographic and lifestyle factors. In our multivariable linear regression models we observed associations of higher urinary pregnancy-averaged mono-isobutyl phthalate (mIBP), mono-(2-ethyl-5-oxohexyl) phthalate (mEOHP), and monobenzyl phthalate (mBzBP) with lower serum AMH after both 6 and 9 years. However, these associations did not remain after adjustment for multiple testing. No significant associations of bisphenol A with AMH were present in our study sample. In our linear mixed-effects models, higher mIBP, mono-(2-ethyl-5-hydroxyhexyl) phthalate (mEHHP), mEOHP, and mBzBP were associated with lower overall AMH levels (differences-0.07 (95% CI -0.13,-0.02),-0.09 (-0.15,-0.02),-0.08 (95% CI -0.14,-0.02), and-0.08 (-0.13,-0.03) μg/L per doubling in mIBP, mEHHP, mEOHP, and mBzBP respectively) (all False Discovery Rate adjusted p-values < 0.05). We identify decreases in indices of ovarian reserve in relationship to prenatal phthalate exposures. Studies are needed replicating our results among large multi-ethnic non-pregnant populations and assessing transgenerational effects of exposure on ovarian reserve. This study was supported by the Erasmus Medical Center and Erasmus University Rotterdam, the Netherlands Organisation for Health Research and Development, the European Research Council, the Dutch Heart Foundation, the Dutch Diabetes Foundation, the European Union's Horizon 2020 Research and Innovation Program, the National Institutes of Health, Ansh Labs Webster, and the Royal Netherlands Academy of Arts and Sciences.

P-646 Prognostic statistical model of age-specific anti-Müllerian hormone (AMH) decrease based on hormone values of 23,528 women

Abstract Study question The analysis of anti-Müllerian (AMH) levels, can a useful statistical model be constructed to predict the rate of decrease of AMH levels at different ages? Summary answer A determination of AMH levels can be an objective marker of a ovarian age and a prognostic marker of the rate of ovarian function decline. What is known already The anti-Müllerian hormone is a dimeric glycoprotein produced by granulosa cells of the antral follicles in the ovaries, and AMH participates in the formation of the dominant follicle during folliculogenesis. Serum AMH levels are relatively constant and show little variability between and within the menstrual cycle. The level of serum AMH is proportional to the number of antral follicles and therefore most suitable for determining ovarian reserve. For this reason, AMH is a convenient marker for the biological age of the ovary and for the response to controlled ovarian stimulation (COS). Study design, size, duration A nationwide cross-sectional retrospective study was conducted between 2017 and 2023 and included 23,528 women the ages of 20 - 50, in which AMH levels were measured once. We performed an analysis of the different AMH levels at different ages and constructed a predictive statistical model for the rate of AMH decline. Results were analyzed in subgroups with AMH values &amp;lt; 1 ng/mL, between 1 and 3,5 ng/mL, and &amp;gt; 3,5 ng/mL, at one-year intervals. Participants/materials, setting, methods All women who visited the laboratory for AMH testing were included in the analysis. The samples were collected in 38 points throughout the country and the analysis was carried out in one centralized laboratory using the Ansh Labs (Webster, TX, USA) manual assays, including the ultra-sensitive AMH enzyme-linked immunosorbent assay (ELISA), with a management range of 0,08 - 14,2 ng/ml. The intra-assay variations were 1,97% and 4,00%. The inter-assay variations were 4,63% and 1,98%, respectively. Main results and the role of chance When analyzing all samples in each consecutive year, we found that the rate of AMH decline was approximately linear and inversely proportional to the woman’s age. However, when analyzing the results for the rate of AMH reduction in the different subgroups, we found out that: In women with AMH &amp;lt; 1 ng/ml, the rate of decline with advancing age changes as follows: until age 34, the decline in AMH is very slow, between ages 35 and 45 there is a rapid acceleration of the process.. In women with AMH between 1 and 3,5 ng/ml, the rate of decline with age was approximately constant. In women with AMH &amp;gt; 3,5 ng/ml, the rate of hormone decline is approximately constant but the year-on-year decline rate is decreasing. Based on our analysis and clinical experience, it is our opinion that the rate of AMH decline for the low AMH group has a non-linear characteristic, while ingeneral different groups exhibit different dynamics for the rate of decline. On this basis, we created a statistical model to predict the decrease in AMH levels at different ages. Limitations, reasons for caution Women of only one ethnicity were included in the study. The studied group is heterogeneous, which does not exclude the inclusion of women with a certain pathology and infertility. In some cases, a single determination of AMH could not predict individual differences in rates of hormone reduction. Wider implications of the findings Determination of reference values for AMH at different ages, based on the large volume of examined samples may facilitate the application of AMH in clinical practice. Based on our statistical analysis, creating a prognostic model for decreasing ovarian function and time to menopause appears feasible. Trial registration number NA

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, as included in the recently updated international guideline. Although different evaluations of age- and assay-specific reference ranges have been published in the past few 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 3 different assays. Retrospective cross-sectional study. A total of 2725 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 percentile 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 Lambda-Mu-Sigma method for all the assays. AMH levels were significantly different between PCOS phenotypes. AMH levels were positively correlated to LH, LH/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 women with PCOS. These findings may help to interpret AMH levels in patients with PCOS and facilitate the use of AMH as a diagnostic tool across age ranges.

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.

FRI423 Obesity And Adiposity Are Associated With Decreased Anti-Mullerian Hormone (AMH) Levels In A Reproductive Aged Latina/LatinX Population

Abstract Disclosure: S. Schon: None. C. Jiang: None. E.L. Ferrell: None. F. Valbuena: None. L. Neff: Other; Self; Eli Lilly &amp; Company, Dr. Neff is currently an employee and shareholder of Eli Lilly and Company, which has no affiliation with this work. At the onset of the study, she was employed by Northwestern University Feinberg Sch. M. Carnethon: None. E.E. Marsh: Consulting Fee; Self; Pfizer, Inc. Objective: Research suggests that obesity has an adverse effect on ovarian reserve, as assessed by AMH. Prior studies have explored this association predominantly in non-Hispanic Caucasian and African American cohorts. In addition, few studies have utilized metrics beyond BMI to characterize adiposity. The objective of this study was to examine the association of obesity/adiposity with AMH in a Latina/LatinX population using multiple measures of obesity/adiposity. Materials &amp; Methods: This cross-sectional study utilized data from the Environment, Leiomyomas, Latinas, and Adiposity Study (ELLAS). ELLAS is a prospective longitudinal cohort study following Latina/LatinX females for 5 years. ELLAS used community based participatory research principles for engagement. Participants were between the ages of 21-50 at time of enrollment. Data from the first study visit were utilized for analysis. Assessment included anthropometrics, measurement of body composition using bioelectrical impedance analysis (BIA) (Tanita MC-280U) and serum AMH (pico-AMH assay, Ansh Labs). The association between BMI on AMH was assessed as a continuous and categorical outcome. Adiposity based on body fat %, and visceral adiposity index (VAI) was studied in relation to AMH using linear regression models. Statistical associations were determined using Chi-square, Wilcoxon rank-sum and linear or logistic regression as appropriate. Results: 621 women completed the first study visit and had BMI and AMH data available. BIA data was available on 591 participants. The mean age of participants was 37.5 ± 7.0 years. The mean BMI was 30.1 ± 6.8 kg/m2, with 261 (42%) of participants classified as having obesity by BMI (≥30 kg/m2). 214 women (34.5%) reported irregular menses and 89 reported currently taking hormonal contraception (HC) (14.3%). BMI was negatively associated with AMH (β=-0.055 p=0.014), however, this association was no longer significant after adjusting for age. Similarly, body fat % and VAI were also negatively associated with AMH (β=-0.058 p= 0.013, and β=-0.256 p&amp;lt;0.001), however this association was attenuated after adjusting for age. When the analysis was restricted to women not taking HC and with regular menses (n=290), higher BMI remained negatively associated with AMH after adjusting for age (β=-0.04, p=0.003). Similarly, higher body fat % and VAI were also negatively associated with AMH after adjusting for age (β=-0.04, p=0.005, and β=-0.090 p=0.009). Highest lifetime BMI was also associated with AMH after adjusting for age (β=-0.03, p=0.03). Conclusions: Among a cohort of Latina/LatinX females with normal menses, obesity and adiposity as assessed by BMI, body fat % and visceral adiposity index were negatively associated with AMH. This suggests that excess adiposity may compromise ovarian reserve and demonstrates the importance of assessing baseline gynecologic characteristics. Presentation: Friday, June 16, 2023

FRI420 Prevalence Of Ovarian Hormone Suppression Is High Among Regularly Cycling Reproductive-aged Women With Overweight Or Obesity

Abstract Disclosure: A. Caldwell: None. A.J. Polotsky: None. A.P. Bradford: None. H. Smyth: None. K. Kuhn: None. J. Siegart: None. N.F. Santoro: Advisory Board Member; Self; Astellas Pharma, Menogenix Inc., Que Oncology, Project Ember/Amazon. Grant Recipient; Self; Menogenix Inc. Other; Self; Ansh Labs Scientific Consultant. V.A. Catenacci: None. Endo Abstract Background: The prevalence of ovarian hormone suppression in regularly cycling, pre-menopausal women with overweight or obesity (termed relative hypogonadotropic hypogonadism of obesity) is not fully appreciated, as there are no overt symptoms aside from subfertility in women trying to conceive. In addition to impacting fertility, ovarian hormones play an important role in energy homeostasis and fat distribution. Other states characterized by ovarian hormone suppression (polycystic ovarian syndrome, PCOS and menopause) are associated with increased cardiometabolic disease risk. It is therefore critical to have a better understanding of the prevalence of ovarian hormone suppression in women with overweight or obesity. Methods: We compared cycle length and ovarian hormones in pre-menopausal women (age 18-40) who had regular menstrual cycles, no history of PCOS or diabetes, and were not using hormonal birth control (currently or in the past 3 months) and were either with overweight or obesity (OW/O, BMI 27-40 kg/m2, n=23) or healthy weight (HW, BMI 18-25kg/m2, n=15). Both groups provided daily, first-morning voided urine samples over a complete menstrual cycle. Progesterone and estrogen metabolites were assayed (pregnanediol-3-glucuronide [Pdg], estrone-1 conjugates [E1c]; Centaur CP; Siemens) and adjusted for creatinine. Hormones were assessed using whole cycle area under the curve (AUC) measurement. Ovarian hormone suppression was defined a priori as Pdg AUC &amp;lt; 25th percentile of the HW control group. T-tests and Spearman correlations were used to compare groups and examine associations between hormones and BMI. Mean±SD are presented. Results: Cycle length was not different between groups, but BMI (OW/O: 34.3±5.3, HW: 21.6±1.3 kg/m2) and age (OW/O: 34.6±5.3, HW: 29.2±4.2 years) were (p &amp;lt; 0.01), thus all analyses controlled for age. Pdg was significantly lower (p&amp;lt;0.01) in OW/O (AUC = 21.29±13.98 ug/mg Cr) relative to HW (AUC = 39.13±22.20 ug/mg Cr) and the majority (82.6%) of the OW/O women exhibited ovarian hormone suppression. E1c was also significantly lower (p&amp;lt;.0001) in the OW/O (AUC = 384.8±211.4 ng/mg Cr) group relative to HW (AUC = 1740.00±627.20 ng/mg Cr). Across the full sample, there were negative correlations between BMI and Pdg AUC (rpartial = -0.34, p = .03) and BMI and E1c AUC (rpartial = -0.61, p &amp;lt; .0001). Conclusion: Ovarian hormone suppression was observed in a majority of our sample of pre-menopausal women with overweight or obesity. Despite the difficulty of identifying this phenotype clinically, it may have important implications for weight management and cardiometabolic disease risk, in addition to fertility, and warrants further investigation to improve women’s health during the reproductive lifespan and beyond. Presentation: Friday, June 16, 2023

THU171 Reference Intervals For Bioavailable IGF-I Values In Dutch Healthy Children, Adolescents And Young Adults

Abstract Disclosure: K. Pellikaan: None. M. Elizabeth: None. A.C. Hokken-Koelega: None. S.A. van den Berg: None. L. de Graaff-Herder: None. Background: Immunoreactive ‘total’ IGF-I is the main biochemical marker for titration of growth hormone (GH) treatment. However, in several endocrine disorders like Prader-Willi syndrome (PWS) and Silver-Russell syndrome, total IGF-I levels are often higher than expected based on GH dose and clinical effects. When patients reach total IGF-I concentrations above +2SDS, fear of negative long-term effects makes clinicians reduce GH dose. This dose reduction often leads to a deterioration in body composition, growth (in children) and quality of life, which suggests that bioavailability of IGF-I is reduced in patients with these disorders. Measurement of bioavailable IGF-I could improve GH dose titration. However, use of bioavailable IGF-I assays has been hindered by the lack of proper reference intervals. We report reference intervals of bioavailable IGF-I in a large group of healthy children, adolescents and adults. Methods: Serum was collected of 260 Dutch healthy controls aged 1-21 years. Total IGF-I and unbound, bioavailable IGF-I were measured manually using Ansh Labs total and free IGF-I assays. We calculated male and female age-related reference intervals for total and bioavailable IGF-I. Results: We present male and female age-related reference intervals. Both total and bioavailable IGF-I increased with age and were significantly higher in females than in males (p &amp;lt; 0.001). Conclusions: the reported reference intervals for bioavailable IGF-I concentrations will facilitate the use of bioavailable IGF-I levels as an alternative marker for GH dose titration in patients with disorders in which total IGF-I levels are unreliable. Presentation: Thursday, June 15, 2023

Sexual dysfunction in women with PCOS: a case control study.

What is the relationship of sex steroid levels with sexual function in women with and without polycystic ovary syndrome (PCOS)? Women with PCOS reported more sexual dysfunction and more sexual distress compared to those without PCOS, but only few and weak associations between androgen levels and sexual function were observed. The literature shows that women with PCOS report lower levels of sexual function and sexual satisfactionand more sexual distress. Contributing factors seem to be obesity, alopecia, hirsutism, acne, infertility, anxiety, depression, and low self-esteem. In women with PCOS clinical and/or biochemical hyperandrogenism is common; its relationship with sexualfunction is, however, inconclusive. This observational prospective case control study with 135 women (68 PCOS, 67 control) was conductedfrom March 2017 until March 2020. Heterosexual women with and without PCOS, aged 18-40 years, in a steady relationshipand without any comorbidities, underwent an extensive medical and endocrine screening using liquid chromatography-tandem mass spectrometry and validated sexual function questionnaires. Women with PCOS reported significantly lower sexual function (Female Sexual Function Index (FSFI) P < 0.001, partial η2 = 0.104), higher levels of sexual distress (Female Sexual Distress Scale-Revised P < 0.001, partial η2 = 0.090), and they more often complied with the definition of sexual dysfunction (41.2% vs 11.9%, P < 0.001, Phi V = 0.331) and clinical sexual distress (51.5% vs 19.4%, P < 0.001, Phi V = 0.335). Regression analysis adjusted for confounders showed only few and weak associations between androgen levels and sexual function, with each model explaining a maximum of 15% sexual function. Following significant Group × Hormone interactions, analyses for both groups separately showed no significant associations in the PCOS group. The control group showed only weak negative associations between testosterone and FSFI pain (β = -6.022, P = 0.044, Adj R2 = 0.050), between FAI and FSFI orgasm (β = -3.360, P = 0.023, Adj R2 = 0.049) and between androstenedione and clinical sexual distress (β = -7.293, P = 0.036, exp(β) = 0.001). The focus of the study on sexual functioning potentially creates selection bias. Possibly women with more severe sexual disturbances did or did not choose to participate. Differences between women with PCOS and controls in relationship duration and hormonal contraceptive use might have skewed the sexual function outcomes. Sexual function is impaired in women with PCOS. However, endocrine perturbations seem to have minimal direct impact on sexual function. Addressing sexuality and offering psychosexual counseling is important in the clinical care for women with PCOS. This study was funded by the departments of the participating centers: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Erasmus University Medical Center, Rotterdam, the Netherlands; Department of Psychosomatic OBGYN and Sexology, Leiden University Medical Center, Leiden, the Netherlands; and Department of Sexology and Psychosomatic OBGYN, Amsterdam University Medical Center, University of Amsterdam, Amsterdam, the Netherlands. J.S.E.L. received unrestricted research grants from the following companies (in alphabetical order): Ansh Labs, Ferring, Merck Serono and Roche Diagnostics. He also received consultancy fees from Ansh Labs, Ferring, Titus Healthcare and Roche Diagnostics. The other authors have no conflicts of interest. CCMO register, registration number: NL55484.078.16, 10 March 2016. https://www.toetsingonline.nl/to/ccmo_search.nsf/Searchform?OpenForm.

Recommendations From the 2023 International Evidence-based Guideline for the Assessment and Management of Polycystic Ovary Syndrome.

Recommendations From the 2023 International Evidence-based Guideline for the Assessment and Management of Polycystic Ovary Syndrome.

Recommendations from the 2023 International Evidence-based Guideline for the Assessment and Management of Polycystic Ovary Syndrome†.

Recommendations from the 2023 International Evidence-based Guideline for the Assessment and Management of Polycystic Ovary Syndrome†.

Associations of reproductive and breastfeeding history with anti-Müllerian hormone concentration among African-American women of reproductive age

Are gravidity, parity and breastfeeding history associated with anti-Müllerian hormone concentration among African-American women of reproductive age? This study included baseline data from the Study of the Environment, Lifestyle and Fibroids, a 5-year longitudinal study of African-American women. Within this community cohort, data from 1392 women aged 25-35 years were analysed. The primary outcome was serum anti-Müllerian hormone concentration measured using the Ansh Labs picoAMH assay, an enzyme-linked immunosorbent assay. Multivariable linear regression models were used to estimate mean differences in anti-Müllerian hormone concentration (β) and 95% CI by self-reported gravidity, parity and breastfeeding history, with adjustment for potential confounders. Of the 1392 participants, 1063 had a history of gravidity (76.4%). Of these, 891 (83.8%) were parous and 564 had breastfed. Multivariable-adjusted regression analyses found no appreciable difference in anti-Müllerian hormone concentration between nulligravid participants and those with a history of gravidity (β = -0.025, 95% CI -0.145 to 0.094). Among participants with a history of gravidity, there was little difference in anti-Müllerian hormone concentration between parous and nulliparous participants (β = 0.085, 95% CI -0.062 to 0.232). There was also little association between anti-Müllerian hormone concentration and breastfeeding history (ever versus never: β = 0.009, 95% CI -0.093 to 0.111) or duration of breastfeeding (per 1-month increase: β = -0.002, 95% CI -0.010 to 0.006). Gravidity, parity and breastfeeding history were not meaningfully associated with anti-Müllerian hormone concentration in this large sample of the Study of the Environment, Lifestyle and Fibroids cohort.

PAPP-A as a screening tool for assessment of gestational age before medication abortion in an intended-use population.

Introduction: Prior studies indicate that PAPP-A could serve as a marker of gestational age (GA) with the potential to determine eligibility for medication abortion. The authors validated the relationship between PAPP-A and GA in an actual-use population. Materials & methods: The authors collected blood samples, medical histories and ultrasound-determined GA from patients presenting for abortion services. They measured PAPP-A using two immunoassays and assessed diagnostic accuracy for predicting GA ≥71 days. Results: The Ansh Labs and R&D Systems immunoassays produced an area under the ROC curve of 0.982 (95% CI: 0.958-0.994) and 0.986 (95% CI: 0.963-0.996), respectively, for predicting GA ≥71 days. Conclusion: This validation study in an intended-use population confirmed that PAPP-A has a strong ability to distinguish pregnancies above and below 71 days' gestation. Clinical trial registration: NCT04232189 (ClinicalTrials.gov).

Association between an AMH promoter polymorphism and serum AMH levels in PCOS patients.

STUDY QUESTIONDo polymorphisms in the anti-Müllerian hormone (AMH) promoter have an effect on AMH levels in patients with polycystic ovary syndrome (PCOS)?SUMMARY ANSWERWe have identified a novel AMH promoter polymorphism rs10406324 that is associated with lower serum AMH levels and is suggested to play a role in the mechanism of regulation of AMH gene expression in women.WHAT IS KNOWN ALREADYFollicle number is positively correlated with serum AMH levels, reflected by elevated AMH levels in women with PCOS. In addition, it is suggested that AMH production per follicle is higher in women with PCOS than in normo-ovulatory women, implying an altered regulation of AMH in PCOS.STUDY DESIGN, SIZE, DURATIONA discovery cohort of 655 PCOS women of Northern European ancestry and both an internal and external validation PCOS cohort (n = 458 and n = 321, respectively) were included in this study. Summary-level data of an AMH genome-wide association study meta-analysis including 7049 normo-ovulatory women was included as a control cohort. A genetic approach was taken through association analysis and in silico analysis of the associated variants in the AMH promoter. In vitro analysis was performed to investigate the functional mechanisms.PARTICIPANTS/MATERIALS, SETTING, METHODSAll common two-allelic single-nucleotide polymorphisms (SNPs) in the region Chr19:2 245 353–2 250 827 bp (Build 37) were selected for the analysis. Linear regression analyses were performed to determine the association between SNPs in the AMH promoter region and serum AMH levels. For the in silico analysis, the webtools ‘HaploReg’ v4.1 for ENCODE prediction weight matrices and ‘atSNP’ were used. In vitro analysis was performed using KK1 cells, a mouse granulosa cell line and COV434 cells, a human granulosa tumor cell line. Cells were transfected with the reference or the variant human AMH promoter reporter construct together with several transcription factors (TFs). Dual-Glo® Luciferase Assay was performed to measure the luciferase activity.MAIN RESULTS AND THE ROLE OF CHANCEPolymorphism rs10406324 was significantly associated with serum AMH levels in all three PCOS cohorts. Carriers of the minor allele G had significantly lower log-transformed serum AMH levels compared to non-carriers (P = 8.58 × 10−8, P = 1.35 × 10−3 and P = 1.24 × 10−3, respectively). This result was validated in a subsequent meta-analysis (P = 3.24 × 10−12). Interestingly, rs10406324 was not associated with follicle count, nor with other clinical traits. Also, in normo-ovulatory women, the minor allele of this variant was associated with lower serum AMH levels (P = 1.04 × 10−5). These findings suggest that polymorphism rs10406324 plays a role in the regulation of AMH expression, irrespective of clinical background. In silico analysis suggested a decreased binding affinity of the TFs steroidogenenic factor 1, estrogen-related receptor alpha and glucocorticoid receptor to the minor allele G variant, however in vitro analysis did not show a difference in promoter activity between the A and G allele.LIMITATIONS, REASONS FOR CAUTIONFunctional analyses were performed in a mouse and a human granulosa cell line using an AMH promoter reporter construct. This may have limited assessment of the impact of the polymorphism on higher order chromatin structures. Human granulosa cells generated from induced pluripotent stem cells, combined with gene editing, may provide a method to elucidate the exact mechanism behind the decrease in serum AMH levels in carriers of the −210 G allele. We acknowledge that the lack of follicle number in the external validation and the control cohort is a limitation of the paper. Although we observed that the association between rs10406324 and AMH levels was independent of follicle number in our discovery and internal validation PCOS cohorts, we cannot fully rule out that the observed effects on serum AMH levels are, in part, caused by differences in follicle number.WIDER IMPLICATIONS OF THE FINDINGSThese results suggest that variations in serum AMH levels are not only caused by differences in follicle number but also by genetic factors. Therefore, the genetic context should be taken into consideration when assessing serum AMH levels in women. This may have clinical consequences when serum AMH levels are used as a marker for the polycystic ovarian morphology phenotype.STUDY FUNDING/COMPETING INTEREST(S)No external funding was used. J.S.E.L. has received consultancy fees from the following companies: Ferring, Roche Diagnostics and Ansh Labs and has received travel reimbursement from Ferring. J.A.V. has received royalties from AMH assays, paid to the institute/lab with no personal financial gain. The other authors declare no competing interests.TRIAL REGISTRATION NUMBERN/A.

Genome-wide association study meta-analysis identifies three novel loci for circulating anti-Müllerian hormone levels in women.

Genome-wide association study meta-analysis identifies three novel loci for circulating anti-Müllerian hormone levels in women.

LONGITUDINAL EXAMINATION OF THE ASSOCIATION BETWEEN HORMONAL CONTRACEPTIVE USE AND ANTI-MÜLLERIAN HORMONE (AMH) LEVELS IN REPRODUCTIVE-AGED AFRICAN-AMERICAN WOMEN

To determine the impact of hormonal contraceptive (HC) use on trends in AMH levels in a longitudinal cohort of reproductive-aged African American women (AAW). This study used an established cohort of AAW from Detroit, MI aged 25 to 35 years, who were recruited as a part of the Study of the Environment, Lifestyle and Fibroids. Anthropometric measurements, self-reported health history, and serum AMH levels (picoAMH assay, Ansh Labs, Webster, TX) were collected from participants at 4 visits over 5 years. All types of HC were included. Percent changes in AMH levels were calculated from baseline to final visit and compared across groups with different patterns of HC use using linear regression models, adjusting for age, BMI, history of abnormal menstrual bleeding, thyroid disorder, and a history of infertility. The effect of starting, continuing, or stopping HC on rate of change in AMH for all follow up visits was also assessed using linear mixed models adjusted for age. A total of 1531 women were included with a mean age of 29.2 ± 3.4 years and a median AMH of 4.07 (IQR 2.29 - 6.70) ng/mL at baseline. At the final follow-up visit, mean age was 34.1 ± 3.4 years and median AMH was 2.97 (IQR 1.52 - 5.15) ng/mL, with a 19.8% average decline in AMH. Women who started and continued HC during the study period had significantly greater % decrease in AMH levels (-23.1%, 95% CI -40.3%, -5.9%) compared with women who never used HC. When compared with women who never used HC, the % change in AMH levels among women who started and stopped HC during the study was -14.4% (95% CI -29.3%, 0.5%), among women who only used HC prior to the study was -13.5%, (95% CI -26.9%, -0.1%) , and among long-term users of HC (on HC before and throughout study period) was -8.7% (95% CI -24.6%, 7.2%). We next sought to determine whether the rate of decline in AMH was impacted by the recency of HC start or discontinuation. Women who started HC between study visits had a faster rate of decline in AMH levels compared with women not on HC at their previous or current visit, (-11.9%; 95% CI -15.5%, -8.0%), while women who used HC at a previous visit but discontinued prior to the most recent visit had a slower rate of decline in AMH (10.7% 95% CI 6.7%, 15.0%). There was no difference in rate of decline among women who continued HC between visits (2.3%, 95% CI -0.6%, 5.7%) compared to those who were not on HC between visits. In this study, the % decline in AMH levels with age was greatest among those who started and continued HC during the study period. This finding, along with the non-significant difference in % decrease with age in AMH levels among long-term HC users, suggests that the impact of HC use on AMH may be most significant after initiation with recovery of AMH following stoppage of HC suggested by the smaller decline with age in this subset of women. Thus, the impact of HC on AMH levels appears reversible. In addition, being on HC neither increases nor protects against the age related rate of decline.

ASSOCIATION BETWEEN ANTI-MULLERIAN HORMONE AND MOTHER'S AGE AT MENOPAUSE

To assess the association between anti-müllerian hormone (AMH) and mother’s age at natural menopause. We hypothesized that AMH levels in the daughter would be directly associated with the mother’s age at menopause. In this secondary analysis of data from the Time to Conceive (TTC) study, which was a prospective time-to-pregnancy study of women aged 30 to 44, women provided a serum sample on cycle day 2, 3, 4, or 5 that was analyzed for AMH (Ultrasensitive AMH ELISA, Ansh Labs). Time to mother’s age at menopause was the primary outcome. Censoring occurred at time of hysterectomy, endometrial ablation or uterine embolization, chemotherapy, or at age of interview if premenopausal. AMH was utilized as both a continuous variable and categorical: low (<10th percentile, 0.62 ng/ml), normal, or high (≥90th percentile, 8.86 ng/ml). A Cox proportional hazards model was used to assess the independent association between AMH and mother’s age at menopause while controlling for age, race, body mass index, education, birth control use, coffee and alcohol intake, and marijuana use. Adjusted hazard ratios (aHR) and 95% confidence intervals (CI) were presented. A HR <1 suggested a likelihood of later age at menopause while a HR >1 indicated an earlier age at menopause. Of the 689 women with a blood sample who were asked about mother’s age at menopause, 352 (51%) were not aware of their mother’s age at last menstrual period, leaving 337 women in the analysis. There were no significant differences in characteristics between those women included in the analysis and those that were not included. Median maternal age at menopause was 52 years (95% CI 52-53). There was no significant association between AMH (continuous) and mother’s age at menopause (aHR 0.97; 95% CI 0.93-1.02, p=0.21). Compared to normal AMH, a low AMH was associated with an earlier age at menopause in the mother (aHR 1.78; 95% CI 0.99-3.19, p=0.05). There was not a statistically significant association between high AMH and mother’s age at menopause (aHR 0.76; 95% CI 0.46-1.27, p=0.30). Among women in their thirties and early forties, ovarian reserve, as measured by continuous AMH, is not associated with mother’s age at menopause. However, low ovarian reserve (AMH <.62) is associated with an earlier age of menopause in the mother.

GROUP BASED TRAJECTORY MODELING (GBTM) OF ANTI-MULLERIAN HORMONE (AMH) IN A COHORT OF REPRODUCTIVE AGE AFRICAN AMERICAN WOMEN (AAW)

AMH has been used as a marker of ovarian reserve. Limited data are available to observe longitudinal changes in AMH over time. Our objective was to evaluate group based AMH trajectory models in a group of reproductive age AAW over the course of 5 years. This longitudinal study used an established cohort of AAW from Detroit, MI aged 23-35 years, who were recruited as a part of the Study of the Environment, Lifestyle and Fibroids. Anthropometric measurements, self reported medical history, and serum AMH levels (picoAMH assay, Ansh Labs, Webster, TX) were collected from participants at 4 visits over 5 years. GBTM was used to establish clusters of women with similar AMH trajectories using age as the time variable. The number of groups and form of temporal trend (linear, quadratic, etc.) for each group trajectory was determined using Bayes Information Criteria. Time varying BMI and hormonal contraception (HC) use were tested to determine their effect on group trajectories. AMH is not normally distributed so analyses were carried out on the logarithmic scale and back transformed to report predicted AMH. 1551 women with AMH for at least 2 visits were included with mean baseline age of 29.2 ± 3.4 years and median AMH of 4.08ng/mL (IQR 2.28 - 6.70). At visit 4, mean age was 34.2 ± 3.4 years and median AMH was 3.00ng/mL (1.52 - 5.15). GBTM model results indicate 5 different groups of AMH trajectories in this population. Group 1 (0.9% of participants) has a linear decline in log AMH with predicted AMH of 0.03ng/mL at age 23 and 0.001ng/mL at age 35 for a normal weight woman not using HC. Group 2 (6.7% of participants) has a cubic trajectory with an AMH value of 1.80ng/mL at age 23 and 0.31ng/mL at age 35. Groups 3, 4, and 5 (comprising 27.8%, 45.4%, and 19.3% of participants respectively) all had a quadratic trajectory for log AMH, although with different slopes. In group 3, at age 23 the predicted AMH is 2.87ng/mL while at age 35 is 1.37ng/mL; for group 4 the predicted AMH at age 23 is 6.13ng/mL while at age 35 is 3.61ng/mL, and for group 5 the predicted AMH at age 23 is 13.8ng/mL and at age 35 is 8.7ng/mL. Time varying HC use and overweight or obese status is associated with AMH trajectories in some groups. In groups 3, 4, and 5, HC use is associated with 15.4% (p<0.001), 15.5% (p<0.001), and 23.9% (p<0.001) lower AMH compared to not using HC, respectively. Being obese is associated with 10.2% (p=0.016) and 12.1% (p=0.029) lower AMH compared to normal weight in groups 4 and 5, respectively. Group 1, with very low AMH at all 4 visits, is associated with 846.6% (p<0.001) higher AMH in HC users, 258.8% (p<0.001) higher AMH in overweight women, and 611.4% (p<0.001) higher AMH in obese women. We identified five groups of AMH trajectories over time. HC and overweight or obese status were found to influence AMH trajectories in some, but not all, groups. Further study is needed to understand other factors that may impact AMH trajectories or risk factors for group membership.

Obese Women Exhibit Reduced Inhibin B and Estradiol SecretionFollowing Pulsatile Intravenous FSH Administration

Introduction: Maternal obesity is an independent risk factor for reduced reproductive fitness. Decreased secretion of FSH in women with obesity is well documented but poorly understood. Furthermore, obese women secrete less protein and steroid hormones from their ovaries. In mice, prior studies have demonstrated that pulsatile release of FSH enhances ovarian function and fertility. Hypothesis: We hypothesize that insufficient FSH pulsatility, as seen in women with obesity, results in inadequate folliculogenesis and reduced ovarian steroid production. We attempt to correct pulsatile FSH secretion in obese women by administering exogenous FSH to compensate for the suppressed circulating ovarian hormones. Our primary outcome is the change in peak inhibin B between pre- and post-treatment. We present results from our interim analysis. Methods: Reproductive aged, regularly menstruating, normal weight (NW) (BMI 18.5-24.9) and obese (OB) (BMI >30) women were recruited for a 26hr study during the early follicular phase. Frequent blood sampling (q10min) for 10h was performed to obtain baseline hormone levels. At 10h, 3 mg of cetrorelix, a gonadotropin hormone antagonist, was given followed by a secondary dose (0.25mg) 6h later. At this time, hourly IV recombinant (r)FSH (30IU) was initiated and frequent blood sampling continued for 10h. LH, FSH, estradiol (E2) were measured by immunoassay (Advia Centaur XP, Siemens). Inhibin B was measured using an ELISA kit (Ansh labs). Differences between groups were modeled by linear regression, adjusted for age and cycle day (continuous). The relationship between change in peak inhibin B and change in peak E2 was estimated in a linear regression. Results: A total of 36 participants (19 NW and 17 OB) were included in our interim analysis. There were no differences in age, cycle day of study, race, and waist/hip ratio. Inhibin B and E2 rises following the intervention were statistically significant within each group. Peak Inhibin B and E2 levels following intervention were lower in obese women compared to normal weight (133.4 vs 202.5 pg/mL and 85.8 vs 126.4 pg/mL, respectively). The difference in pre and post peak inhibin B levels trended lower in the obese group (-40.1 (95%CI: -86.2, 6.1, p=0.087). No difference was seen in maximal E2 response. There was no relationship between inhibin B and E2 response [0.08 (95%CI -0.26, 0.42), p=0.634]. Conclusions: These early results suggest obese women may have a lower response to pulsatile rFSH as compared to normal weight counterparts even with intravenous administration. We speculate this may be due to decreased uptake of rFSH in obese patients or a sign of ovarian dysfunction in obese women. Additional subjects are recruited to detect these differences.

Myelin basic protein and its diagnostic value in HIV-infected individuals with 4th clinical stage and neuroinfections

It was shown that in HIV-infected patients, pathomorphological changes in the white matter in the form of demyelinization are already observed in the early stages of the disease. The most studied marker of this process is myelin basic protein that can be detected in cerebrospinal fluid or serum immediately after acute myelin breakdown.&#x0D; The aim. To assess the diagnostic value of myelin basic protein content in serum and cerebrospinal fluid of HIV-infected individuals with 4th clinical stage and central nervous system opportunistic infections.&#x0D; Materials and methods. Using ELISA with diagnostic kit “MBP ELISA” (Ansh Labs, USA), we studied the myelin basic protein content in serum and cerebrospinal fluid of 53 HIV-infected patients with 4th clinical stage and central nervous system opportunistic infections depending on its etiology, the outcome of the diseases and according to Glasgow coma scale score. As well correlation analysis with some laboratory and clinical indicators was performed.&#x0D; Results. We found significantly increased myelin basic protein content in both cerebrospinal fluid and serum of HIV-infected patients 4th clinical stage with central nervous system opportunistic infections compared to control (p˂0.01), which indicate the presence of active demyelinization in central nervous system. The highest cerebrospinal fluid myelin basic protein was registered in patients with an unfavourable outcome of the disease, as death or residual neurologic deficit, and patients with cerebral toxoplasmosis. The cerebrospinal fluid myelin basic protein had an association with the size of white matter lesions on magnetic resonance imaging and serum myelin basic protein content.&#x0D; Conclusions. Myelin basic protein detection in cerebrospinal fluid as well as in serum can serve as an additional quantitative marker of myelin disruption, which can be used along with magnetic resonance imaging for the diagnosis improvement and prognosis of central nervous system opportunistic infections in HIV-infected individuals with 4th clinical stage