anti-Müllerian Hormone Concentrations Research Articles

Correlation between melatonin and vitamin D3 deficiency in Iraqi female patients with polycystic ovarian syndrome

Polycystic ovary syndrome (PCOS) is the most common hyperandrogenic disorder in women and one of the leading causes of anovulatory infertility. Depletion of vital hormones in the organism, such as in the case of PCOS, has been identified as one factor contributing to poor pregnancy outcomes and associated issues in women. Vitamin D level in PCOS patients regulates fertility through membrane-bound receptors in oocytes, regulating luteinizing hormone (LH), and progesterone surges.The objective. To examine the correlation between melatonin and vitamin D3 deficiency in Iraqi pregnant women with PCOS under the age of 40.Material and methods. Two hundred women were recruited for the study and divided into two groups: patients with PCOS (n=100) and healthy females without PCOS (n=100). All participants were administered medical questionnaires, and their body mass index (BMI) was determined. Blood samples from participants were obtained for biochemical analysis to assess follicle-stimulating hormone (FSH), luteinizing hormone (LH), anti-Müllerian hormone (AMH), melatonin, and vitamin D levels.Results. The results revealed that 65% of PCOS patients were between 20 and 30 years old and healthy control persons did not show any variations based on age. Patients with PCOS had significantly (p<0.001) higher levels of LH and FSH (1.98±0.07 nmol/mL and 11.15±0.25 mIU/L, respectively) compared with healthy controls (1.06±0.02 nmol/mL and 6.67±0.25 mIU/L, respectively). The PCOS group had significantly (p≤0.01) reduced mean vitamin D3 level related to the control group. There was no significant correlation between vitamin D3 and melatonin levels in the PCOS group. The study showed that BMI was significantly higher in women with PCOS compared to healthy women. The study also found a positive relationship between BMI and AMH concentration, as well as between FSH and vitamin D3 levels in women with PCOS. However, there was no association between vitamin D3 and melatonin or BMI. Conclusion: The study found that D3 deficiency is a significant component in female PCOS patients that might be a new biomarker for PCOS in Iraqi women.

Evaluation of Anogenital Distance and Anti-Müllerian Hormone Plasmatic Concentration as Potential Phenotypes to Predict Reproductive Performance in Holstein Heifers.

Anogenital distance (AGD) is a marker of the degree of prenatal exposure to androgens in multiple species, and it has been suggested that there is an inverse association between AGD and fertility. Therefore, the aim of this study was to determine the usefulness of AGD and anti-Müllerian hormone (AMH) concentrations, an indirect marker of the follicular population, as predictors of future reproductive potential in Holstein cattle. The AGD was measured in 566 females from 9 dairy farms in Galicia (Spain). A group of 172 females underwent a second measurement 9 months after. Additionally, data on the age at first insemination (1stAI age), number of AI (AI-PREG), age at first pregnancy (1stPREG age), age at first calving (1stCAL age), and calving-pregnancy (CAL-PREG) and calving-calving (CAL-CAL) intervals were collected. Blood samples were collected from 80 heifers to determine AMH concentrations. Our results showed that AGD varied minimally with age, and that cows with short AGD had earlier 1stAI age, 1stPREG age, and 1stCAL age (p < 0.05) than cows with long AGD. No significant differences were observed for the CAL-PREG and CAL-CAL intervals. Additionally, no significant association was found between AMH concentration and AGD or reproductive parameters. Consequently, the results suggest the possibility of using AGD as a marker of future reproductive performance in Holstein heifers. However, there was insufficient evidence to associate AMH concentrations and reproductive performance, underlining the need for further studies with larger sample sizes.

12559 Objective Versus Subjective Measures Of Reproductive Development And Psychological Well-Being In Peri-Menarcheal Adolescents: A Pilot Study

Abstract Disclosure: K.C. McCormick: None. M.E. Lujan: None. J. Chang: None. L. Ipp: None. A. Alladeen: None. H. Lamar: None. J. Mendle: None. H. Vanden Brink: None. Introduction: Risk of psychopathology emerges during the pubertal and peri-menarcheal years; however objective physiological measures of maturation often differ from adolescents’ own reports of their development. Understanding how psychological symptoms emerge in relation to both objective and subjective evaluations of reproductive development can provide critical insights into mental health risk and variability in psychological symptom severity during the pre-menarcheal and early post-menarcheal years. Methods: 52 females 9-15 years old (n=16 pre-menarcheal, n=36 &amp;lt;2 years post-menarche) underwent a non-fasting blood draw for reproductive hormones (estradiol, anti-mullerian hormone (AMH), luteinizing hormone (LH), follicle stimulating hormone (FSH)), anthropometry, detailed reproductive and menstrual history, and a series of surveys to evaluate depression (CES-DC), anxiety (MASC), perceptions of pubertal-related change (POPS), and transdiagnostic psychological processes (rumination, emotional clarity) which can contribute to, and are risk factors for, psychopathology. Whether psychological symptoms are associated with objective measures (menarche, gynecological age, reproductive hormone concentrations) versus subjective measures (self-reported tanner stage, severity of dysmenorrhea) were contrasted using t-tests and Pearson Correlation Coefficient analyses (JMP Pro (v17)). Results: The majority of participants (26/36 post-menarcheal and 11/16 pre-menarcheal) exceeded the standard cut-off of 15 for depressive symptoms, consistent with research on mental health symptomatology in this cohort of youth. POPS scores were higher (39±10.9) in post-menarcheal versus pre-menarcheal (31±12.6) adolescents indicating post-menarcheal adolescents reported more puberty-linked change in their lives (Pttest=0.04). Lower emotional clarity was associated with higher AMH concentrations (r= -0.27, p=0.0944). Depressive symptoms tended to be greater (27+14.8) in those with moderate dysmenorrhea versus mild/no dysmenorrhea (19±11.1, Pttest=0.08). Depression (r=0.30, p=0.032) and anxiety symptoms (r=0.32, p=0.027), and rumination (r=0.30, p=0.033) were associated with higher self-reported Tanner scores; POPS was trending (r=0.25, p=0.0776). Neither estradiol nor FSH were associated with any measures of psychological well-being. Conclusion: Both objective measures of reproductive development and adolescent self-reports may help providers assess individual vulnerability to internalizing psychopathology, such as depression and anxiety. Additionally, given population level increases in depressive symptoms in adolescents, researchers should avoid utilizing simple cutoffs for depression and anxiety, which lack specificity and overlook critical variability which can further elucidate mental and physical health dynamics. Presentation: 6/2/2024

B-093 Analytical performances of a novel fluorescent immunoassay of anti-Müllerian hormone and establishment of the reference intervals in Chinese children

Abstract Background Anti-Müllerian Hormone (AMH) plays a important role in children growth and the concentrations change with age as well as gender. This study aimed to evaluate the analytical performances of a novel AMH assay based on the cycle enhanced fluorescence immunoassay (CEFIA) and establish the pediatric reference intervals of AMH Methods The experiments on imprecision, sensitivity, linearity, linear range and comparison were carried out to evaluate the analytical performances. Furthermore, the concentrations of serum AMH in 238 females and 346 males aged 0-18 years were measured by the Pylon AMH assays and the reference ranges for ages group were calculated using robust methods. Results The repeatability and the within-laboratory imprecision CVs of the assay were 3.7% and 6.4% at at 2.25 ng/mL, and 4.6% and 6.4% at 15.49 ng/mL, respectively. The sensitivity of the assay (LoB =0.05 ng/mL, LoD= 0.1 ng/mL and LoQ= 0.3 ng/mL) was verified. The AMH Pylon assays showed good linearity within the detection range (between 0.1 and 24 ng/ml) and can report up to 480 ng/mL with 20x pre-dilution of samples. The AMH concentrations of serum samples measured by the Pylon assays correlated to those measured by the Elecsys assays. In the males under 18 years old, the AMH levels were high at birth (0-1 month: median 95.10 ng/mL) and continued to increase to a peak between 7 months to one year old (median 158.80 ng/mL) before they started to decrease with age to the lowest level between 15 and 18 years old (median 6.31 ng/mL). On the other side, the serum AMH concentrations in females were low at birth (0-1 month: median 0.20 ng/mL) and increased with age (15-18 years old: median 3.03 ng/mL). The reference intervals were provided for each gender and age groups,respectively. Conclusions The Pylon AMH assays showed good analytical performances, and the AMH reference intervals in chinese children determined may provide a basis for the use in clinical diagnosis and treatment of related diseases.

Macro-FSH is a rare cause of inappropriately high FSH concentrations.

Assessment of hormone concentrations can be subjected to laboratory pitfalls. Macro-hormones are hormone-autoantibody complexes which are cleared slowly from circulation and cause a false elevation in hormones' concentrations. Macro-prolactin and macro-thyroid-stimulating hormone (TSH) are most frequently encountered while macro-follicle-stimulating hormone (FSH) has been rarely reported. We describe the case of a 30-year-old woman who had a gynaecological consultation due to failure in achieving pregnancy after 8 months of unprotected intercourse. She had regular menses, did not complain of climacteric symptoms and her medical history was unremarkable. Antral follicle count and anti-mullerian hormone concentrations were normal, and regular ovulation was documented. Unexpectedly, high early follicular phase FSH concentrations were confirmed on two occasions (57 and 51 IU/L), raising the suspicion of primary ovarian insufficiency. After excluding Turner's syndrome and autoimmune oophoritis, a laboratory artifact was hypothesized. Following polyethylene glycol precipitation, FSH levels dropped from 41.1 IU/L to 6.54 IU/L (recovery 16%) and the presence of macro-FSH was concluded. Laboratory interference can lead to misdiagnosis and unnecessary treatments. A laboratory artifact should be suspected when inconsistency exists between clinical presentation and laboratory results. Only five other cases of macro-FSH have been reported to date. Although macro-hormones generally have low biological activity and do not require treatment, the role of anti-FSH antibodies has been hypothesized in primary ovarian insufficiency and in vitro fertilization failure. Hormone quantification is a cornerstone in the diagnostic workup of endocrine disorders, but it can be subjected to laboratory interferences which can lead to unnecessary investigations and inappropriate treatments. A laboratory artifact should be suspected when a discrepancy is observed between clinical presentation and laboratory results, when extremely unusual analyte concentrations are observed and when inconsistent results are obtained by different analytical methods. Macro-hormones are hormone-autoantibody complexes which are cleared slowly from circulation and cause a false elevation in hormone concentrations. Macro-prolactin and macro-TSH are most frequently encountered, while macro-FSH has been rarely reported. Macro-hormones can be detected by polyethylene glycol precipitation, gel filtration chromatography, or by using protein G or protein A columns. Although macro-hormones generally have low biological activity and do not require treatment, the role of anti-FSH antibodies has been hypothesized in primary ovarian insufficiency and in vitro fertilization failure.

Relationship Between Anti-Mullerian Hormone (AMH) and Other Influencing Factors with in Vitro Fertilization (IVF) Outcome of Iraqi Infertile Women

The clinical application of Anti-Mullerian Hormone (AMH) measurement has been proposed in the prediction of qualitative and quantitative aspects of assisted reproductive technologies (ART). The aim of this study is to investigate the role of AMH level and other influencing factors in the outcomes of in vitro fertilization (IVF) treatment. The study focused on AMH and its association with infertile women undergoing IVF. The population included in this study was 131 infertile women who sought therapy for infertility at Kamal Al-Samarra'ay Specialized Hospital. They were divided into three groups according to the level of AMH hormone; group A (0.5-1.5 ng/mL, n=52), group B (1.6-4 ng/mL, n=70), and group C (&gt; 4 ng/mL, n=9). Data collection was performed by means of a specially designed questionnaire, independent of demographic data. It included questions related to infertility. Statistical analysis was performed with SPSS 26. Clinical parameters of infertile women, infertility hormones and their relationship with AMH, as well as FMS-like Tyrosine kinase 1 levels, were all investigated. The study findings revealed a significant difference in the concentration of AMH between infertile women (p&lt;0.01). Our analysis further demonstrated that the level of AMH reduces with advancing age in infertile women. Additionally, a positive correlation was observed between the number of oocytes obtained and AMH levels in infertile women. Furthermore, the study found that AMH levels had an effect on clinical pregnancy outcomes. Specifically, the highest success rates for IVF were observed in women with normal AMH levels falling within the ranges of group B (6-4 ng/mL(. Additionally, the study demonstrated significant differences in AMH and TSH levels among infertile groups (p&lt;0.05). Our study concluded that there is a significant correlation between clinical parameters and IVF outcomes, which can provide information for future clinical interventions aimed at improving success rates. Our analysis highlighted the role of FMS-like Tyrosine kinase 1 in infertility, where higher FMS-like Tyrosine kinase 1 levels were associated with higher success rates in IVF. The results indicate that the FMS-like Tyrosine kinase 1 level is a valuable marker for confirming the success or failure of IVF and can help determine the appropriate treatment plan.

Anti-Müllerian hormone: biology and role in endocrinology and cancers.

Anti-Müllerian hormone (AMH) is a peptide belonging to the transforming growth factor beta superfamily and acts exclusively through its receptor type 2 (AMHR2). From the 8th week of pregnancy, AMH is produced by Sertoli cells, and from the 23rd week of gestation, it is produced by granulosa cells of the ovary. AMH plays a critical role in regulating gonadotropin secretion, ovarian tissue responsiveness to pituitary hormones, and the pathogenesis of polycystic ovarian syndrome. It inhibits the transition from primordial to primary follicles and is considered the best marker of ovarian reserve. Therefore, measuring AMH concentration of the hormone is valuable in managing assisted reproductive technologies. AMH was initially discovered through its role in the degeneration of Müllerian ducts in male fetuses. However, due to its ability to inhibit the cell cycle and induce apoptosis, it has also garnered interest in oncology. For example, antibodies targeting AMHR2 are being investigated for their potential in diagnosing and treating various cancers. Additionally, AMH is present in motor neurons and functions as a protective and growth factor. Consequently, it is involved in learning and memory processes and may support the treatment of Alzheimer's disease. This review aims to provide a comprehensive overview of the biology of AMH and its role in both endocrinology and oncology.

PSX-1 Genomic insights into reproduction traits of Nellore cattle: A meta-analysis of genomic regions

Abstract Enhancing reproductive efficiency could lead to economic benefits for Nellore producers. However, several factors limit our understanding of the main genetic factors controlling reproduction traits in Nellore cattle, including genetic differences between populations and statistical limitations. A meta-analysis can assist in pinpointing the most likely functional candidate genes, quantitative trait loci, and biological processes simultaneously associated with several traits in a group. Hence, the goal of this study was to perform a meta-analysis of genomic regions previously associated with reproduction traits in Nellore cattle, to improve our comprehension of the main genetic mechanisms regulating these traits within the breed. This study incorporated a total of 549 genomic regions sourced from 21 scientific publications, published between 2014 to 2024. The reproduction traits included in this study are scrotal circumference at various ages (365, 450, and 550 d), early pregnancy (&amp;lt; 31 mo of age), early puberty (defined as pregnancy at 18 mo), age at first calving, antral follicle count, calving interval, calving ease, testicular hypoplasia, stayability, heifer rebreeding, gestation length, anti-mullerian hormone concentrations, preweaning calf mortality, and post-natal mortality. All genomic regions identified in the literature were updated to the ARS-UCD1.2 genome assembly, and the Gallo R package was used to identify positional candidate genes associated with each genomic region. A total of 8,441 positional candidate genes were annotated and used in an overrepresentation analysis performed in meshr. The significant genes (adjusted P-values &amp;lt; 0.05) identified in the overrepresentation analysis underwent prioritization analyses using the GUILDify and ToppGene software. Subsequently, enrichment analysis of the prioritized overrepresented candidate genes was performed using the ClusterProfiler R package. The 8,441 annotated genes generated 1,757 overrepresented genes, of which 17 candidate genes were prioritized (FDR P -value ≤ 10-3). The overrepresented candidate genes that were prioritized are: SERTAD2, RFWD3, C1QTNF9B, SLFN14, CATSPERE, GFRA4, ZNF408, NOP58, ADGRG3, KIZ, REM2, FSCN3, DCAF11, ACSM4, ANKRD34B, CMKLR2-AS, and TRV-AAC1-1. The functional enrichment analysis highlighted the following genes: FSCN3 (exhibits highly specific expression in testis), CATSPERE (has a vital role in cellular processes related to reproduction and is necessary for normal fertility), ANKRD34B (associated with cytoplasm found in oocytes or early embryos), and RFWD3 (regulates the transition between phases of the mitotic cell cycle). All other prioritized genes also have roles in regulating metabolic pathways or biological processes related to reproduction, such as sperm capacitation, sperm motility, cilium or flagellum-dependent cell motility, mitotic cell cycle phase transition, and RNA endonuclease activity. The enrichment of the functional candidate genes identified highlights that these genes have an important role in the expression of reproduction traits in Nellore cattle. Candidate genes identified in this study can be used as target genes in gene expression studies, which are necessary to validate our findings.

Population-based anti-Müllerian hormone reference intervals help define gonadal status in the bitch.

To establish statistically valid, population-based reference intervals (RIs) for canine anti-Müllerian hormone (AMH) and define changes in AMH and inhibin-B in bitches during breeding cycles. A homologous canine ELISA was used to measure AMH in serum samples (collected between May 2019 and July 2024) from 102 intact and 78 reportedly ovariohysterectomized (OVH) bitches and 8 bitches before and after ovariohysterectomy, and in longitudinal samples from 24 bitches undergoing breeding management. Established 95% RIs were used in a retrospective assessment of 3,193 clinical submissions. Cyclic variation of AMH and inhibin-B (heterologous ELISA) were regressed with time and normalized to the rise in progesterone in samples from breeding bitches. Intact and OVH RIs for AMH were calculated with and without inclusion of 7 samples from reportedly OVH bitches that had AMH concentrations in the intact RI. Anti-Müllerian hormone and inhibin-B were positively correlated, and AMH was 3 times higher in proestrus than in estrus. Retrospectively, of 3,193 samples submitted for clinical AMH testing, 41% to 56% were in or above the intact AMH interval, 37% to 44% were within the OVH interval, and < 10% were inconclusive, depending on how RIs were defined. Statistically valid, population-based RIs establish a sound basis for interpreting results of clinical submissions requesting AMH to assess gonadal status in the bitch. Confirmation of cyclic variation in AMH (and, for the first time, inhibin-B) reaffirms proestrus as the optimum time to draw samples, and ≤ 10% of samples submitted for determination of gonadal status are expected to fall in an inconclusive AMH RI.

Physiological Anti-Müllerian Hormone Concentrations in Male and Female Dogs and Cats before and around Puberty.

In recent years several studies established the diagnostic value of anti-Müllerian hormone (AMH) in companion animals. However, less is known about physiological AMH concentrations in young individuals highlighting the necessity to apply the diagnostic findings to this group. The aim of this study was to determine the AMH values of healthy male and female dogs between the age of 8 and 48 weeks, tomcats under 8 weeks and up to 48 weeks of age and queens between 2 to 12 weeks of age. In total, 96 blood samples were collected. Anti-Müllerian hormone was measured in all samples and testosterone was measured in the oldest age group of the males in both species. The hormones were analyzed using a human based chemiluminescence immune assay. Overall, AMH concentrations were higher in males than in females (p < 0.001). According to the AMH concentration there was no difference in males, but queens had significant higher AMH concentrations than bitches (p < 0.001). AMH remained high in males up to week 24 and decreased significantly thereafter (tomcats: p = 0.015; male dogs: p = 0.013), which correlated with an increase in testosterone levels for male dogs only. In bitches, AMH remained below the detection limit until the week 16 and slightly increased subsequently. In queens, AMH was detectable from the beginning with a significant increase in the older age group (p = 0.003). Half of the cats in the older age group even approached the chemiluminescence immune assay's upper limit. The results show that female cats secrete AMH much earlier than female dogs in which AMH secretion begins just shortly before the start of the puberty. In the male animals, the decrease in AMH concentration around puberty was similar in dogs and cats, but a correlation with the increase of testosterone was only observed in dogs. Further research is required to determine the origin of the high AMH concentrations in female kittens and the lack of correlation between testosterone and AMH concentrations in male kittens.

Effect of AMH on primordial follicle populations in mouse ovaries and human pre-pubertal ovarian xenografts during doxorubicin treatment.

Survival rates of the childhood cancer patients are improving, however cancer treatments such as chemotherapy may lead to infertility due to loss of the primordial follicle (PMF) reserve. Doxorubicin (DXR) is a gonadotoxic chemotherapy agent commonly used in childhood cancers. Anti-Müllerian Hormone (AMH) has been reported to have a protective effect on the mouse ovarian reserve against DXR in vivo. However, whether AMH can prevent PMF loss in conjunction with DXR in human ovarian tissue in vivo has not been determined. In order to investigate this, we first established an optimum dose of DXR that induced PMF loss in cultured mouse ovaries and investigated the efficacy of AMH on reducing DXR-induced PMF loss in mice in vitro. Second, we investigated the effects of DXR on pre-pubertal human ovarian tissue and the ability of AMH to prevent DXR-induced damage comparing using a mouse xenograft model with different transplantation sites. Mouse ovaries treated with DXR in vitro and in vivo had reduced PMF populations and damaged follicle health. We did not observe effect of DXR-induced PMF loss or damage to follicle/stromal health in human ovarian cortex, this might have been due to an insufficient dose or duration of DXR. Although AMH does not prevent DXR-induced PMF loss in pre-pubertal and adult mouse ovaries, in mouse ovaries treated with higher concentration of AMH in vitro, DXR did not cause a significant loss in PMFs. This is the first study to illustrate an effect of AMH on DXR-induced PMF loss on pre-pubertal mouse ovaries. However, more experiments with higher doses of AMH and larger sample size are needed to confirm this finding. We did not observe that AMH could prevent DXR-induced PMF loss in mouse ovaries in vivo. Further studies are warranted to investigate whether AMH has a protective effect against DXR in xenotransplanted human ovarian tissue. Thus, to obtain robust evidence about the potential of AMH in fertility preservation during chemotherapy treatment, alternative AMH administration strategies need to be explored alongside DXR administration to fully interrogate the effect of DXR and AMH on human xenografted tissues.

Conditional loss of Brca1 in oocytes causes reduced litter size, ovarian reserve depletion and impaired oocyte in vitro maturation with advanced reproductive age in mice

An estimated 1 in 350 women carry germline BRCA1/2 mutations, which confer an increased risk of developing breast and ovarian cancer, and may also contribute to subfertility. All mature, sex steroid-producing ovarian follicles are drawn from the pool of non-renewable primordial follicles, termed the 'ovarian reserve'. The clinical implications of early ovarian reserve exhaustion extend beyond infertility, to include the long-term adverse health consequences of loss of endocrine function and premature menopause. We aimed to determine whether conditional loss of Brca1 in oocytes impacts ovarian follicle numbers, oocyte quality and fertility in mice with advancing maternal age. We also aimed to determine the utility of AMH as a marker of ovarian function, by assessing circulating AMH levels in mice and women with BRCA1/2 mutations, and correlating this with ovarian follicle counts. In this study, we addressed a longstanding question in the field regarding the functional consequences of BRCA1 inactivation in oocytes. To recapitulate loss of BRCA1 protein function in oocytes, we generated mice with conditional gene deletion of Brca1 in oocytes using Gdf9-Cre recombinase (WT: Brca1fl/flGdf9+/+; cKO: Brca1fl/flGdf9cre/+). While the length of the fertile lifespan was not altered between groups after a comprehensive breeding trial, conditional loss of Brca1 in oocytes led to reduced litter size in female mice. Brca1 cKO animals had a reduced ovarian reserve and oocyte maturation was impaired with advanced maternal age at postnatal day (PN)300, compared to WT animals. Serum anti-Müllerian hormone (AMH) concentrations (the gold-standard indirect marker of the ovarian reserve used in clinical practice) were not predictive of reduced primordial follicle number in Brca1 cKO mice versus WT. Furthermore, we found no correlation between follicle number or density and serum AMH concentrations in matched samples from a small cohort of premenopausal women with BRCA1/2 mutations. Together, our data demonstrate that BRCA1 is a key regulator of oocyte number and quality in females and suggest that caution should be used in relying on AMH as a reliable marker of the ovarian reserve in this context. This work was made possible through Victorian State Government Operational Infrastructure Support and Australian Government NHMRC IRIISS. This work was supported by funding from the Australian Research Council (ALW - DE21010037 and KJH - FT190100265), as well as the National Breast Cancer Foundation (IIRS-22-092) awarded to ALW and KJH. LRA, YML, LT, EOKS and MG were supported by Australian Government Research Training Program Scholarships. LRA, YML and LT were also supported by a Monash Graduate Excellence Scholarship. YC, SG and XC were supported by Monash Biomedicine Discovery Institute PhD Scholarships. LRA was also supported by a Monash University ECPF24-6809920940 Fellowship. JMS was supported by NHMRC funding (2011299). MH was supported by an NHMRC Investigator Grant (1193838).

P-652 Chronic kidney disease and female reproductive hormonal profile: A multi-centre cohort study

Abstract Study question Does Chronic Kidney Disease (CKD) impact female reproductive hormone profiles? Summary answer Women with CKD had significantly lower AMH and testosterone, and significantly higher LH levels. There were no differences in FSH, oestradiol, prolactin and progesterone. What is known already Chronic diseases, such as CKD, is rising in women of reproductive age. It is expected that the prevalence of CKD in adults will increase to 16.7% in 2030.Coupled with improvements in obstetric care and advancement in assisted reproductive technology, more women with CKD are seeking to conceive with ART. It is known that CKD reduces fertility, but underlying mechanisms are not fully understood. It is proposed that CKD leads to dysregulation of the hypothalamus-pituitary-ovarian (HPO) axis, but studies are small, use outdated assays, are done at different times in the menstrual cycle and do not compare with healthy controls. Study design, size, duration Observational cohort study involving women with CKD and healthy women of reproductive age without underlying medical conditions was conducted between August 2021-September 2022.Women with CKD were identified from four units to attend a study visit at a tertiary fertility unit.Healthy controls were recruited via word of mouth and from couples within the unit undergoing treatment for male factor infertility.For the study to be powered 100 women with CKD and 50 healthy controls were recruited. Participants/materials, setting, methods Ethical approval was provided by the Health Research Authority (IRAS 285546) and written informed consent was obtained from participants.Serum Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), prolactin, testosterone, oestradiol, progesterone, Anti Mullerian Hormone (AMH) and b-HCG were taken between the first and fifth day of menstrual cycle. If there was amennorhoea, bloods were taken on a day suiting them. Comparisons between controls and CKD Stage utilised t-test and Mann Whitney U tests according to data-distribution. Main results and the role of chance 101 women with CKD and 63 healthy women were recruited. The mean age of women in the CKD group was 35.1± 6.7 years and of controls was 33.44 ±4.8 years, median BMI for CKD women was 25.45 kg/m2 (IQR 22.7,29.4) and controls was 24.5 kg/m2 (IQR 22,26.7).The renal cohort had similar numbers in each of the four CKD stages. 20.8% women had renal transplants.The two cohorts were matched for age and BMI. Median AMH (13.6(IQR 3,4,28.7) vs 18.4 (IQR 11.2,33.6) pmol/L;p=0.03) was significantly lower in the renal group than controls. Median testosterone concentrations (0.6 (IQR 0.5,0.9) vs 0.5 (IQR 0.4,0.7)nmol/L; p = 0.01) were significantly lower in women with CKD even from stage 1 CKD. LH (4.4 (IQR 3.2,5.5) vs 5.9 (IQR 3.8,9.4) IU/L;p=0.001) concentrations were significantly higher in women with CKD compared to controls but there were no differences in FSH, oestradiol, prolactin and progesterone. LH increased and testosterone and AMH concentrations fell with CKD severity. Interestingly, b-HCG was &amp;gt;6 IU/L in 3.0% (3/101) patients with CKD. When stratified by CKD stage; CKD stage 2 and onwards had significantly higher LH concentrations (p = 0.03).CKD stage 3 and above participants had significant lower AMH concentrations (p = 0.002).This is at a later stage of CKD than previously published. Limitations, reasons for caution Although this is the largest study of its kind known to explore female hormonal profiles in women with CKD, the numbers are still limited. In addition, our population had a mixed ethnic background but due to the small numbers we were unable to analyse the results per ethnic category. Wider implications of the findings This is the largest prospective study comparing fertility hormonal profiles in women with CKD and healthy controls. Unfortunately, there remains a paucity of evidence on the effects of CKD on female fertility. Future work includes exploring longitudinal changes which will better facilitate patients to make informed and timely fertility-related decisions. Trial registration number 285546

P-620 Oocyte competence in Vitro Fertilization: relation between serum anti-Müllerian hormone (AMH) concentrations and complement C4 protein expression in follicular fluid

Abstract Study question To investigate if serum AMH concentration variations are correlated with protein expression in follicular microenvironment affecting IVF outcomes. Summary answer Serum AMH concentrations were associated with altered levels of intrafollicular protein involvement of the complement pathway in immunity response in follicular fluid (FF). What is known already Anti-Müllerian hormone (AMH) plays a crucial role in regulating growth and development of ovarian follicles. The follicular fluid is produced during folliculogenesis and contains a variety of proteins that play important roles in follicle development and oocyte maturation. AMH concentration in follicular fluid can be a marker of ovarian reserve status and oocyte competence. AMH elevated levels may indicate a higher ovarian reserve and are sometimes associated with a greater ovarian response to ovarian stimulation used in IVF. However, the exact relationship between AMH levels in follicular fluid and IVF outcomes may vary among women and depend on various factors. Study design, size, duration The study included FF collected from 75 patients underwent IVF from December 2018 to June 2019.All patients enrolled underwent GnRH antagonist stimulation protocol with r-FSH according to clinical/ovarian features.Patients were sorted into three groups according to serum AMH concentrations: AMH≤1.0 ng/ml, n = 25 (A group); AMH=1.1-2.0 ng/ml, n = 29 (B Group); AMH&amp;gt;2.0 ng/ml, n = 21 (C group).Cutoffs for defining low(A), average(B), and high(C) AMH serum concentrations corresponded to round values of 30th and 70th centiles of each measurement. Participants/materials, setting, methods Exclusion criteria: endometriosis, metabolic/endocrinology diseases and oligoasthenoteratozoospermia. FF aspirates from individual follicles (diameter ≥18 mm) were collected on the day of oocyte retrieval. The FF was analyzed by two-dimensional gel electrophoresis (2-DE). Oocyte number and quality, fertilization rate are IVF outcome measures. Data were analyzed using the unpaired Student’s t-test by IBM SPSS-STATISTICS versions 20.0 and considered significant if p-value ≤ 0.05. Main results and the role of chance 180 protein spots were highlighted in three groups: 80% is represented in all the samples analyzed, whereas 19 protein spots out of 180 are group-specific. Five unique upregulated proteins were identified in the A group and five in C group respect to control group (B). These were identified IG, CO4, ALBU, TRFE, IGLC in A group, while A1AT, ANT3 A2HS, HPT and CO4 in C group. A and C group showed worse oocyte quality associated with a lower fertilization rate than B group. On the basis of the proteome analysis, the component of the complement cascade C4 was found more plentiful in patient with low and high AMH serum concentration compared to average AMH serum concentration ones. The presence of high levels of the unprocessed precursor C4 correlated to serum AMH fluctuations is an indication of non-activation of the complement pathways fundamental for follicle maturation and ovulation compromising IVF outcome. Limitations, reasons for caution Limited number of patients. Age and lifestyle factors (i.e. smoking, diet) are not considered. Wider implications of the findings Results showed an increase of CO4 in FF of patients with low and high AMH serum concentrations. CO4 FF high levels could be indicative of defective complement activation during ovulation with consequent impaired processes and lower standard oocyte competence. Thus, CO4 could be identify as biomarker to predict IVF success. Trial registration number Not Applicable

P-697 The effect of smoking and vaping on hormonal ovarian reserve markers in women of reproductive age in the United Kingdom

Abstract Study question What impact do smoking and vaping have on ovarian reserve markers Anti-Mullerian Hormone (AMH) and Follicle-Stimulating Hormone (FSH)? Summary answer Smoking and vaping negatively affect AMH and FSH levels in women, with significant age-related differences in ovarian reserve markers observed. What is known already Previous studies have established a link between smoking and accelerated ovarian follicular depletion, evidenced by lower serum AMH levels and reduced antral follicle count (AFC). Smoking is known to increase FSH levels indirectly by affecting pituitary function. However, the impact of vaping, a growing trend especially among younger individuals, on ovarian reserve markers like AMH and FSH remains under-researched and requires comprehensive investigation to understand its full implications on reproductive health. Study design, size, duration This cross-sectional study analysed medical history and serum AMH and FSH concentrations from capillary blood samples of 8,340 women aged 21-45. Participants were stratified by 5-year age intervals and excluded if they had reproductive health conditions or used hormonal contraception. Samples were analysed from individuals using an at-home hormone testing service, with statistical analysis including unpaired t-tests and Cohen’s d for effect size, focusing on smokers/vapers versus non-smokers/non-vapers. Participants/materials, setting, methods The study included UK women aged 21-45 providing capillary blood samples for serum AMH and FSH measurement using an at-home testing service. Exclusions applied to those with reproductive conditions or on hormonal contraception. Smoking and vaping status were self-reported through an online health assessment. AMH and FSH concentration differences were analysed via unpaired t-test; a p-value of &amp;lt; 0.05 was considered statistically significant and mean values±SD were reported. Main results and the role of chance Among 8,340 participants, 312 smoked and 653 vaped. Lower serum AMH values were observed in smokers and vapers in all age groups, with significant differences at ages 31-35 for smokers (20.73±16.23 vs 25.37±19.80 pmol/L in smokers and non-smokers respectively; p = 0.003) and 36-40 for vapers (13.65±13.34 vs 16.88±14.32 pmol/L in vapers and non-vapers respectively; p = 0.01). No significant differences in serum FSH concentration were found among vapers, however, FSH levels were higher in smokers across all ages. This was significant in individuals aged 21-25 (8.01±2.52 vs 6.87±3.33 IU/L in smokers and non-smokers respectively; p = 0.02) and 31-35 (8.33±3.31 vs 7.70±3.51 IU/L smokers and in non-smokers respectively; p = 0.04). These results suggest a detrimental effect of smoking and vaping on ovarian reserve markers, with statistical significance indicating the role of chance is minimal in the observed differences. Limitations, reasons for caution Self-reported smoking/vaping status may introduce bias. The study does not report the duration or frequency of participants’ smoking and vaping habits. The study’s cross-sectional design limits causality inference, however it provides a valuable insight for future longitudinal research to explore these relationships over time and explore underlying mechanisms. Wider implications of the findings These findings highlight the potential risks of smoking and vaping on women’s reproductive health, emphasising the need for awareness and preventive measures, especially among younger populations and the increased use of vaping. Trial registration number not applicable

Associations between a normal-range free thyroxine concentration and ovarian reserve in infertile women undergoing treatment via assisted reproductive technology

BackgroundSome recent studies have shown that female subclinical hypothyroidism (SCH) is associated with diminished ovarian reserve (DOR). In this study, we aimed to investigate whether serum-free thyroxine (fT4) concentrations within the reference range are associated with ovarian reserve in women.MethodsThis cross-sectional study included 4933 infertile women with normal-range fT4 concentrations who received assisted reproductive technology treatment in our clinic. The data of women in different fT4 concentration tertiles (namely 12–15.33, 15.34–18.67, and 18.68–22 pmol/L) were compared with ovarian reserve markers, namely the anti-Müllerian hormone (AMH) concentration, the antral follicle count (AFC), and the number of aspirated oocytes. The primary outcomes were the AMH concentration and the risk of DOR, diagnosed as an AMH concentration < 1.1 ng/mL.ResultsThe average ages of women in the low-normal, middle-normal, and high-normal fT4 tertiles were 33.20 (standard deviation [SD]: 5.11), 32.33 (SD: 5.13), and 31.61 (SD: 5.10) years, respectively (p < 0.0001). AMH concentrations (adjusted mean: 3.32 [95% confidence interval {CI}: 3.16 to 3.50] vs. 3.51 [3.40 to 3.62] vs. 3.64 [3.50 to 3.80] ng/mL, p = 0.022) were significantly different between the fT4 concentration tertiles. The risk of DOR was significantly increased in the low-normal (adjusted odds ratio: 1.61 [95% CI: 1.01 to 2.58]) and middle-normal (1.47 [95% CI: 1.00 to 2.16]) tertiles compared with the high-normal tertile. Subgroup analysis showed that AMH concentrations were significantly different among the fT4 concentration tertiles in women aged < 35 years (adjusted mean: 3.94 [95% CI: 3.70 to 4.20] vs. 4.25 [4.11 to 4.39] vs. 4.38 [4.18 to 4.58], p = 0.028), whereas this difference was not significant in women aged ≥ 35 years (p = 0.534). The general additive models using fT4 as a continuous variable indicated that a lower fT4 concentration within the normal range was significantly associated with a lower AMH concentration (p = 0.027), a lower AFC (p = 0.018), a lower number of aspirated oocytes (p = 0.001), and a higher risk of DOR (p = 0.007).ConclusionLow-normal fT4 concentrations are associated with lower ovarian reserve in infertile women.

Effect of serum anti-Müllerian hormone levels on the superovulation response in Holstein heifers.

Anti-Müllerian hormone (AMH) holds potential as a biomarker for assessing the superovulation (SO) response in cattle. Nonetheless, there exists scant information regarding this aspect in the literature concerning dairy heifers. Given this gap, our objective is to explore the viability of AMH as an indicator for gauging the SO response specifically in Holstein heifers. Furthermore, our aim encompasses examining the variations in AMH levels within the same individuals before and after undergoing SO. The study included 41 Holstein heifers. All heifers were superovulated and blood samples were taken both before and after the SO protocol. The findings revealed that the mean values of serum AMH concentrations before and after SO were 0.122ng/mL (0.093-0.248ng/mL) and 0.119ng/mL (0.084-0.170ng/mL), respectively. AMH concentrations in heifers were stratified into low (<0.106ng/mL), medium (0.107-0.126ng/mL) and high (>0.127ng/mL) categories both before and after SO. There was no significant correlation between AMH levels in the heifers both before and after SO treatment with the number of follicles, corpora lutea, total embryos collected or embryos transferred (p>0.05). Furthermore, this study showed that serum AMH concentrations in Holstein heifers did not change after SO treatment. In this study, as AMH levels in Holstein heifers were in a narrow range, a relationship between AMH and SO response could not be determined. In future studies, we believe that it would be more useful to plan more studies in Holstein donor heifers, taking into account the number of animals and AMH levels.

Serum anti-Müllerian hormone is an indirect predictor of ovarian reserve in domestic cats

Anti-Müllerian hormone (AMH) serves as an indirect marker for predicting primordial follicles that are representative of ovarian reserve. In this study the possibility of using AMH and age to predict the ovarian reserve in domestic cats. Ovaries and blood were collected from 30 cats undergoing routine ovariohysterectomy. The animals were divided into three age groups: prepubertal (<4 mo, n = 10), adult (1–5 y, n = 10), and senior (>5 y, n = 10). Blood was collected at surgery for serum AMH measurements using the AMH Gen II ELISA kit. The intra-assay coefficient of variation (CV) and inter-assay CV were 3.56 % and 7.68 %, respectively. One side of the ovary was processed to determine AMH localization using immunohistochemistry and for a histological count of follicles, which is the gold standard. The expression of AMH protein was quantified from the contralateral ovary by Western blot analysis. Primordial follicles exhibited the most pronounced inverse relationship with age (rho = −0.779, P < 0.05), followed by a positive association with serum AMH concentration (rho = 0.490, P < 0.05), indicating that both age and AMH are potential markers indicative of primordial follicles. Furthermore, secondary (rho = 0.651, P < 0.05) and small antral follicles (rho = 0.648, P < 0.05) were identified as the major sources of circulating AMH, as indicated by the stronger correlation with serum AMH concentrations compared with primary follicles. However, there was no significant correlation between the expression of AMH protein and other factors, including age, primordial follicles, primary follicles, secondary follicles, small antral follicles, and serum AMH concentration. A model for predicting primordial follicle number using serum AMH concentration (AIC = 672.66, P < 0.05) and age (AIC = 668.93, P < 0.05) was established. In conclusion, both serum AMH concentration and age may serve as comparable markers of ovarian reserve in domestic cats. Moreover, AMH is particularly useful in situations where age information is not available.

The effect of dienogest treatment on anti-Mullerian hormone in patients with endometrioma: a 12-month follow-up study.

To assess the effect of dienogest treatment on endometrioma (OMA) size, serum anti-Mullerian hormone (AMH) levels and associated pain over a 12-month follow-up period. A longitudinal cohort study of 104 patients with OMA who were treated with dienogest, between January 2017 and January 2020. Of the included patients, each had a 12-month follow-up period with transvaginal or pelvic ultrasound and measurement of serum AMH concentration at the sixth and twelfth months of follow-up. The alteration in OMA size in the sixth and twelfth months of treatment was the primary outcome measure and the alteration in AMH concentration over the same period was the secondary outcome measure. The only exclusion criterion was having surgical intervention for OMA during the follow-up period (n=44). In patients with bilateral OMA (n=21), the change in size of the largest OMA was considered in the analysis. A total of 60 patients with a mean ± standard deviation (SD) age of 31.5±8.0 years were included. The mean ± SD OMA size on the day the dienogest was started was 46.3±17.4 mm and the mean AMH level was 3.6±2.4 ng/mL. After six months, the mean OMA size had decreased to 38.6±14.0 mm, with a median difference of 7.8 mm [95% confidence interval (CI): 3.0 to 12.6; p=0.003]. The mean AMH level was 3.3±2.7 ng/mL at 6 months follow-up (95% CI: -0.2 to 0.8; p=0.23) and the average difference was 0.3 ng/mL. At the 12th-month visit, when compared with the beginning of the treatment, OMA size had again significantly decreased by a median of -8.9 mm (95% CI: -2.9 to -14.9; p=0.005), and the decline in median AMH was also significant (-0.9 ng/mL, 95% CI: -0.1 to -1.7; p=0.045). The initial mean ± SD visual analog scale pain score at the commencement of dienogest treatment was 6.3±3.4. The mean values at the sixth and twelfth months of dienogest therapy were 1.08±1.8 and 0.75±1.5, respectively (both p<0.001 compared to baseline). At the sixth and twelfth months of dienogest treatment a significant decrease in OMA size and reported pain scores were observed, whereas the AMH concentrations did not change significantly.

Limited association between stallion-like behavior and hormonal indicators of testicular remnants in geldings

Persistent stallion-like behavior is a common sign of cryptorchidism in supposed geldings. The presence of testicular tissue can be evaluated by analyzing hormones such as testosterone and anti-Müllerian hormone (AMH). Here, we used hormonal analysis to investigate relationships between the likely presence of testicular tissue and stallion-like behavior in samples submitted from presumptive geldings (n = 1,202), retrospectively. Most geldings with stallion-like behaviors had serum concentrations of testosterone (851/1,056; 80.6 %) and AMH (682/877; 77.8 %) below the laboratory reference range for cryptorchids (< 60 pg/mL and ≤ 0.15 ng/mL for testosterone and AMH, respectively). A total of 13 samples (13/716; 1.8 %) showed AMH concentrations typical for geldings but testosterone above the cryptorchid range. Conversely, 31 samples (31/716; 4.3 %) had high AMH, suggesting cryptorchidism, but testosterone concentrations implied no testicular tissue. Among the cryptorchid stallions, the AMH and testosterone concentrations did not vary based on the season. However, age categories affected the concentration of both hormones among the presumptive true cryptorchid stallions. The results of this study demonstrate that undesirable behavior in geldings is rarely associated with the presence of testicular tissue, as assessed by these two hormonal biomarkers. This information highlights the complexity of behavior and demonstrates that persistent stallion-like behavior in geldings could be related to factors other than the presence of testicular tissue.