High E2 Research Articles

Correlation between serum and saliva sex hormones in young female athletes.

Saliva sampling provides a practical noninvasive way for biological monitoring of steroid hormones but few studies have focused on saliva sex hormones in female athletes, and it is still unknown whether salivary concentrations are an accurate reflection of blood concentrations in this population. The aim of this study was therefore to evaluate the correlation between serum and saliva estradiol (E2) and progesterone (PG) in 10 young female athletes with normal menstrual cycle. Thirty blood and saliva samples were taken at rest in a fasting state in order to analyze E2 and PG concentrations, during 3 different hormonal periods: Status 1: low E2 and low PG (end luteal/early follicular); Status 2: high E2 and low PG (end follicular/peri-ovulatory); Status 3: high PG (mid-luteal). A strong correlation was found between serum and saliva PG (r=0.914, P<0.01), whereas a weaker but still significant relationship was observed between serum and saliva E2 (r=0.755, P<0.01). The most significant correlations obtained for E2 were during Status 2 (r=0.941, P<0.01) and during Status 3 for PG (r=0.817, P<0.01). Although it remains challenging to produce precise identification with only saliva samples for each menstrual cycle phase, results seem to show that saliva can provide a convenient substitute to serum for sex hormones in female athletes during longitudinal monitoring. This is of particular interest in this population, with the number of female athletes suffering from an altered menstrual cycle, sometimes even during regular menstruation.

The High Estradiol Environment after IVF Causes the Increased Risk of Glucose Metabolic Dysfunction in Offspring.

Assisted reproductive technology (ART) is associated with increased metabolic risks in offspring. The effect of high maternal estradiol (E2) levels during early pregnancy on the glucose metabolism of offspring remains unclear. To evaluate glucose metabolism in in vitro fertilization (IVF)-conceived children and assess whether high E2 exposure during early pregnancy is associated with metabolic alterations. This retrospective analysis included 500 singletons aged 3-10 years born after fresh embryo transfer (ET) (n=200), frozen ET (n=100), and natural conception (NC) (n=200) from a university hospital. Children underwent anthropometric measurements and examinations for fasting glucose, insulin, and lipid levels. A mouse model of high E2 exposure during early pregnancy was established to study glucose and insulin tolerance, and insulin secretion. Compared with NC, children born after fresh ET showed higher fasting glucose/insulin levels, increased insulin resistance, and higher incidence of impaired fasting glucose, which might be associated with a higher maternal E2 levels. Frozen ET showed intermediate results. In mice, offspring exposed to high E2 levels during gestation exhibited impaired glucose/insulin tolerance and defects in insulin secretion. High maternal E2 levels in early pregnancy are associated with altered glucose metabolism and increased metabolic risks in IVF-conceived children. Further studies are needed to elucidate the underlying mechanisms.

Can ultrasound elastography be useful in the diagnosis of poor ovarian response?

This study aims to compare the elastographic properties of the ovaries of patients with poor ovarian response (POR) with healthy controls by using real-time ultrasound tissue elastography and to investigate the predictive value of ovarian strain ratio (OSR) in the diagnosis of POR. A cross-sectional study was conducted among infertility patients. OSR measurements of 46 women diagnosed with POR and 41 healthy controls with male infertility were performed by real-time ultrasound tissue elastography. Ovarian volume, OSR, antral follicle count (AFC), age, body mass index (BMI), anti-mullerian hormone (AMH), follicle stimulating hormone (FSH), and estradiol (E2) values, were compared between the groups. A receiver operating characteristic curve was used to compare the sensitivity and specificity of OSR, AFC, AMH, FSH, and E2 for POR diagnosis. Patients with POR were significantly older, had a lower duration of infertility, lower AMH, higher FSH, higher E2, lower AFC and ovarian volume (p < 0.01, p = 0.02, p < 0.01, p < 0.01, p < 0.01, p < 0.01, p < 0.01, respectively). The mean OSR of both ovaries was similar between the groups. For the diagnosis of POR, the sensitivity and specificity for AMH were 97.5% and 100%, for AFC were 86.7% and 97.6%; for FSH were 66.7% and 80.5%, for E2 were 53.3% and 90.2%, respectively. The significant independent predictors of the POR were AMH, AFC, and FSH, with no additional significant contribution from OSR. However, considering the relationship between POR and ovarian fibrosis, it is recommended to investigate this issue in more comprehensive studies.

Rapid nongenomic estrogen signaling controls alcohol drinking behavior.

Ovarian-derived estrogen is a key modulator of numerous physiological processes via genomic and nongenomic mechanisms, including signaling non-canonically at membrane-associated estrogen receptors in the brain to rapidly regulate neuronal function. However, the mechanisms mediating estrogen regulation of behaviors such as alcohol consumption remain unclear. Early alcohol drinking confers greater risk for alcohol use disorder in women than men, and binge alcohol drinking is correlated with high circulating estrogen levels, but a causal role for estrogen signaling in driving alcohol drinking in gonadally-intact animals has not been established. We found that female mice displayed greater binge alcohol drinking and reduced avoidance behavior when circulating estrogen was high during the proestrus phase of the estrous cycle than when it was low, contributing to sex differences in these behaviors. The pro-drinking, but not anxiolytic, effect of high endogenous estrogen state occurred via rapid estrogen signaling at membrane-associated estrogen receptor alpha in the bed nucleus of the stria terminalis, which promoted synaptic excitation of corticotropin-releasing factor neurons and facilitated their activity during alcohol drinking behavior. This study is the first to demonstrate a rapid, nongenomic signaling mechanism for ovarian-derived estrogen signaling in the brain controlling behavior in gonadally intact females, and it establishes a causal role for estrogen in an intact hormonal context for driving alcohol consumption that contributes to known sex differences in this behavior.

Abstract Mo021: Role of Estrogen and Testosterone in promoting Sex Dimorphism in Doxorubicin Cardiotoxicity.

Background: Doxorubicin (DOX), a cancer chemotherapeutic agent, causes sex-specific cardiotoxicity. Lower cardiotoxicity in females compared to males is typically attributed to the cardioprotective effects of estrogen (E2). However, the role of the sex hormones, E2 and testosterone (T), in modulating DOX induced cardiac dysfunction and its underlying mechanisms are not known. Methods: C57BL/6J mice were divided into four groups, 1) Control, 2) DOX (5 mg/kg/week for 6 weeks), 3) Reverse hormone profile (RevH, castrated males+E2 and ovariectomized females+T), and 4) RevH + DOX. Mouse morbidity was tracked daily. Total body weight and cardiac function was assessed biweekly for 6 weeks by echocardiography, ECG and optical mapping. Dependence on fatty acid versus glucose for metabolism was measured by seahorse analysis. Results: DOX treatment reduced total body weight (11%) in males, and increased morbidity in males and females. This was associated with reduced mechanical function (15% decrease in stroke volume) in males and electrical remodeling (29% action potential duration prolongation) in females. In RevH mice, no changes in morbidity or cardiac function were observed compared to Control. In RevH+DOX males, the onset of morbidity was delayed by two weeks compared to DOX males. This cardioprotective effect was associated with preserved mechanical and electrical function. On the other hand, in RevH+DOX females, morbidity was increased, and this correlated with mechanical (20% reduction in cardiac output) and electrical remodeling (22% increase in action potential duration). Comparing RevH+DOX males to females, males had greater fatty acid dependency while females had greater glucose dependency for metabolism. Conclusions: These results suggest that sex hormones play a significant role in modulating specific aspects of DOX cardiotoxicity. While E2 has a critical role in preserving cardiac mechanics during DOX therapy, high T levels were associated with mechanical dysfunction in both males and RevH females. Switching of metabolic substrates with greater preference for fatty acid metabolism in mice with higher E2 could underlie cardiac mechanics preservation. Finally, DOX induced electrical remodeling was not altered by sex hormones.

Associations of endogenous estrogens, plasma Alzheimer's disease biomarkers, and APOE4 carrier status on regional brain volumes in postmenopausal women.

Women carrying the APOE4 allele are at greater risk of developing Alzheimer's disease (AD) from ages 65-75 years compared to men. To better understand the elevated risk conferred by APOE4 carrier status among midlife women, we investigated the separate and interactive associations of endogenous estrogens, plasma AD biomarkers, and APOE4 carrier status on regional brain volumes in a sample of late midlife postmenopausal women. Participants were enrolled in MsBrain, a cohort study of postmenopausal women (n = 171, mean age = 59.4 years, mean MoCA score = 26.9; race = 83.2% white, APOE4 carriers = 40). Serum estrone (E1) and estradiol (E2) levels were assessed using liquid chromatography-tandem mass spectrometry. APOE genotype was determined using TaqMan SNP genotyping assays. Plasma AD biomarkers were measured using single molecule array technology. Cortical volume was measured and segmented by FreeSurfer software using individual T1w MPRAGE images. Multiple linear regression models were conducted to determine whether separate and interactive associations between endogenous estrogen levels, plasma AD biomarkers (Aβ42/Aβ40, Aβ42/p-tau181), and APOE4 carrier status predict regional brain volume (21 regions per hemisphere, selected a priori); and, whether significant interactive associations between estrogens and AD biomarkers on brain volume differed by APOE4 carrier status. There was no main effect of APOE4 carrier status on regional brain volumes, endogenous estrogen levels, or plasma AD biomarkers. Estrogens did not associate with regional brain volumes, except for positive associations with left caudal middle frontal gyrus and fusiform volumes. The interactive association of estrogens and APOE4 carrier status on brain volume was not significant for any region. The interactive association of estrogens and plasma AD biomarkers predicted brain volume of several regions. Higher E1 and E2 were more strongly associated with greater regional brain volumes among women with a poorer AD biomarker profile (lower Aβ42/40, lower Aβ42/p-tau181 ratios). In APOE4-stratified analyses, these interactions were driven by non-APOE4 carriers. We demonstrate that the brain volumes of postmenopausal women with poorer AD biomarker profiles benefit most from higher endogenous estrogen levels. These findings are driven by non-APOE4 carriers, suggesting that APOE4 carriers may be insensitive to the favorable effects of estrogens on brain volume in the postmenopause.

P-445 High serum estradiol on the day of frozen blastocyst transfer is associated with higher early miscarriage rates in hormonal replacement therapy cycles using transdermal estrogens

Abstract Study question Do serum estradiol(E2) levels on the day of frozen blastocyst transfer(FBT) affect the live birth rate (LBR) in hormonal replacement therapy(HRT) cycles using transdermal estrogens? Summary answer E2 levels ≥ 313 pg/mL on the day of FBT are associated with increased early miscarriage rates, but no significant impact on LBR. What is known already E2 plays a crucial role in endometrial receptivity during the periconceptional period and a massive role in placentation. However, the impact of serum E2 levels measured around the time of a FBT in HRT cycles outcomes remains controversial, with some studies showing a detrimental impact of high levels, and others underlining no difference. To this date, there is no study focusing on HRT cycles with transdermal estrogens only, though it is the safest way of estrogen administration regarding thrombo-embolic complications. Study design, size, duration Observational cohort study with 2364 patients undergoing HRT-FBT cycles at a university hospital, between January 2019 and December 2022. Women were only included once during the study period. Participants/materials, setting, methods Patients undergoing single autologous FBT under HRT using transdermal estrogens and vaginal micronized progesterone. The serum E2 level was measured in the morning of the FBT, in a single laboratory. The primary outcome was the LBR, secondary outcomes included clinical pregnancy rates, early miscarriage rates(EMR), and obstetrical outcomes. Patients were divided into three groups based on the 25th and the 75th percentiles of E2 levels and compared using univariate and multivariate logistic regression models. Main results and the role of chance A total of 2364 patients were divided into 3 groups: group A &amp;lt; 122 pg/mL (n = 590), group B 122-312 pg/mL (n = 1184), and group C ≥ 313 pg/mL (n = 590). Mean serum E2 level on the day of FBT was 247.8 pg/mL ± 209.01 pg/ml. The LBR was 32.0% (756/2364), and the early miscarriage rate was 26.3% (280/1075) in the overall population. Multivariate logistic regression showed a negative impact of high E2 levels (≥ 313 pg/mL) on EMR: an increase in the EMR was noted between group C and referent group B (30.3% versus 24.6%, OR 1.5 95%CI (1.06-2.17)). However, no significant impact of E2 levels was noted regarding the LBR (group C vs group B, adjusted OR 0.9 95%CI (0.69-1.10)), the clinical pregnancy rates, or the obstetrical outcomes (birthweight and term of delivery Limitations, reasons for caution The main limitation of our study is linked to its observational design. Extrapolation of our results to other laboratories, or other routes and/or doses of administering estrogens also needs to be validated. Wider implications of the findings Limiting high serum E2 levels on the day of FBT in HRT cycles may improve ART outcomes. Further studies are needed to evaluate if modifications of estrogen doses may improve pregnancy chances, in an approach to individualize the management of ART patients. Trial registration number not applicable

Reproductive hormones in relation to white matter hyperintensity volumes among midlife women.

Although reproductive hormones are implicated in cerebral small vessel disease in women, few studies consider measured hormones in relation to white matter hyperintensity volume (WMHV), a key indicator of cerebral small vessel disease. Even fewer studies consider estrone (E1), the primary postmenopausal estrogen, or follicle-stimulating hormone (FSH), an indicator of ovarian age. We tested associations of estradiol (E2), E1, and FSH to WMHV among women. Two hundred twenty-two women (mean age=59) underwent hormone assays (E1, E2, FSH) and 3T brain magnetic resonance imaging. Associations of hormones to WMHV were tested with linear regression. Higher E2 (B[standard error (SE)]=-0.17[0.06], P=0.008) and E1 (B[SE]=-0.26[0.10], P=0.007) were associated with lower whole-brain WMHV, and higher FSH (B[SE]=0.26[0.07], P=0.0005) with greater WMHV (covariates age, race, education). When additionally controlling for cardiovascular disease risk factors, associations of E1 and FSH to WMHV remained. Reproductive hormones, particularly E1 and FSH, are important to women's cerebrovascular health. Despite widespread belief that sex hormones are important to women's brain health, little work has considered how these hormones in women relate to white matter hyperintensities (WMH), a major indicator of cerebral small vessel disease. We considered relations of estradiol (E2), estrone (E1), and follicle-stimulating hormone (FSH) to WMH in midlife women. Higher E2 and E1 were associated with lower whole-brain WMH volume (WMHV), and higher FSH with higher whole-brain WMHV. Associations of E1 and FSH, but not E2, to WMHV persisted with adjustment for cardiovascular disease risk factors. Findings underscore the importance of E2 and FSH to women's cerebrovascular health.

Abstract PO1-14-09: Age-related remodeling of the systemic and breast microenvironment promotes a tumor-permissive locale for ER+ breast cancer in older women

Abstract Introduction: The peak incidence of ER+ breast cancer occurs in women around the age of 70. Compared to younger cohorts, we and others have shown that older patients with ER+ breast cancer have an enrichment of luminal disease and experience fewer recurrences, suggesting overtreatment. Differences in the biological phenotypes of tumors in older women remain poorly understood and treatment remains challenging as older patients are underrepresented in clinical trials. In this study, we hypothesized that aging alters both local and systemic hormones and inflammatory cytokines, creating a permissive environment for tumor formation and growth. To address this hypothesis, we employed a systems biology approach across multiple model systems and tissue samples. Methods: Our study included specimens from two cohorts of patients: (1) n = 90 women without breast cancer obtained from the Komen Tissue Bank, from whom we obtained matched plasma and non-cancerous breast tissue (n = 30 across age groups comprising of young donors (35-45yo), middle-aged donors (55-69yo), and older donors (≥ 70yo); and (2) n = 115 women with ER+ breast cancer (n = 25 young patients, n = 57 middle-aged patients, and n = 33 older patients) obtained from our institutional biobank, including matched plasma, tumor tissue, and tumor-adjacent tissue. From each of these sets of specimens, we measured estrogen disposition using mass spectrometry, a 22-plex panel of inflammatory markers, and transcriptomic changes using RNA-seq. Using scRNA-seq and multiplexed IHC, we further characterized the immune changes that occur with age. Lastly, we used an aged, carcinogen-induced ER+ tumor model in F344 rats (obtained from the NIA) to model tumor development and growth. Results: In the plasma, estradiol (E2) drastically decreased in post-menopausal women, leaving estrone (E1) as the predominant circulating estrogen in older women. However, the breast tumor microenvironment (TME) showed comparable levels of E1 and E2 across age groups. Multi-class concordance analysis of breast cancer tissue from RNA revealed significant increases in gene expression of the HSD17B7 enzyme and decreases in HSD17B2 enzyme across age, likely elevating local E1-to-E2 conversion and yielding high local E2 levels despite low circulating levels specifically in older women. In older patients with ER+ breast cancer, there was a significant increase in a chemokine network characterized by CCL2 and CXCL9 within the TME. scRNA-seq revealed enrichment of inflammatory M2-like macrophages in the aged TME. Immune dysfunction and decreased immunosurveillance in older patients manifested with widespread decreases in the presence of cytotoxic lymphocytes and decreased pathway enrichment of key immune pathways, such as JAK/STAT and IFNy signaling. mIHC analysis revealed decreases in CD4 and CD8 T cells and increases in M2-like macrophages in older patients in both tumor and stromal regions. Lastly, in the carcinogen-induced F344 rat model, aged rats (~ 24mo, human equivalent 60-70yo) had a shorter tumor-free interval than the younger rats (~ 4-6mo, human equivalent 20-30yo), likely due to increases in tumor-promoting inflammation and decreased immune surveillance. Ongoing mechanistic work is focused on macrophage-patient derived organoid co-cultures evaluating the functional relevance of E2, CCL2, and CXCL9 in shaping the aged TME. Conclusions: Our study of systemic and local breast hormones and cytokines in both healthy individuals and breast cancer patients showed that older women with ER+ breast cancer harbor different systemic and local environments compared to younger women. The aged TME is characterized by high E2, cytotoxic lymphocyte depletion, and immune dysfunction, creating a permissive environment for tumor formation. Future translational work should be aimed at chemo-preventative strategies to reduce the age-related chronic inflammation and immune dysfunction. Citation Format: Neil Carleton, Jian Zou, Sanghoon Lee, Dixcy Jaba Sheeba John Mary, Ruxuan Li, Jennifer Atkinson, Ziyu Huang, Hatice Osmanbeyoglu, Peter Lucas, Emilia Diego, Michael Lotze, George Tseng, Jagmohan Hooda, Ioannis Zervantonakis, Priscilla McAuliffe, Steffi Oesterreich, ADRIAN LEE. Age-related remodeling of the systemic and breast microenvironment promotes a tumor-permissive locale for ER+ breast cancer in older women [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO1-14-09.

Sex-Specific Effects of Neonatal Stress on Brainstem Expression of Orexin1-Receptors

Introduction and hypothesis: Located within the hypothalamus, orexin-producing neurons (ORX) regulate arousal, feeding, and cardiorespiratory function. Panic disorders (PD) affects ~5% of the North American population. Stress-related “hyperactivation” of the ORX system is a leading hypothesis to explain the pathophysiology of PD. One important sub-group of PD patients show significant respiratory symptoms, including an excessive ventilatory response to CO2 inhalation. Manifestations of PD are sex-specific: the prevalence of PD is ~2 times higher in women compared to men and the intensity of the ventilatory response to CO2 fluctuates with the hormonal cycle. Early life stress is a risk factor for PD and we recently demonstrated that in rats, neonatal maternal separation (NMS) augments the ventilatory response to CO2 (5%; 10 min) of adult females (but not males); this effect peaks with the rise of 17-b estradiol (E2) during proestrus. Because NMS-related enhancement of the hyperventilatory response to CO2 can be prevented by a selective ORX1-antagonist, we tested the hypothesis that sex, hormonal status and NMS interact to influence the expression of ORX-1 receptors in brainstem regions that regulate breathing. Methods: Following birth, pups were either exposed to NMS (3h/day from postnatal days 3 to 12) or raised under standard conditions. Rats were then raised until adulthood (8 weeks); experiments were performed on males, females in proestrus (high E2), and ovariectomised (low E2) females (2 weeks post-surgery). We used in situ hybridization histochemistry to quantify the expression of mRNA for ORX-1 receptors. The ORX-1 receptors signal was revealed by autoradiography emulsion and slides were examined under darkfield microscopy. Expression of ORX-1 receptors was quantified by measuring the pixel density of the hybridization signal in selected brainstem regions that regulate CO2 response. Results: The locus coeruleus (LC) contained the highest expression of ORX-1 receptors amongst the structures studied. In males, NMS was associated with significant decreases in ORX-1 expression in the LC and nucleus of the solitary tract (NTS). By comparison with OVX females, proestrus augmented ORX-1 expression in the LC of controls, but not NMS. By comparison with OVX, proestrus augmented ORX-1 expression in the raphe obscurus, especially in NMS females. Expression of ORX-1 was unchanged in the NTS of females. Conclusion and perspectives: We conclude that sex and the female’s hormonal status are important determinants of ORX-1 expression and that NMS can alter these relationships; however, those effects are structure-specific. In light of the evidence indicating that 5-HT dysfunction contributes to PD, the results observed in the raphe obscurus point to a plausible mechanism. This study was supported by ‘the Québec Heart and Lung Institute Research Foundation’. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Cross Sex Steroid Therapy and Baseline Cardiac Function in a Rodent Model of Gender Affrming Hormone Therapy in the Male Rat

Transgender females (TGF) are individuals who are born biologically male, but identify as female. Gender-Affrming Hormone Therapy (GAHT), consisting of 17β-Estradiol (E2) supplementation with androgen suppression [i.e. castration (CTX)], is used in TGFs to match their physical characteristics with their perceived gender. Previously, we reported that E2+CTX in the normotensive male Sprague-Dawley (SD) rat is associated with a significant increase in E2 and decrease in testosterone, body weight, and lean mass. Whether GAHT enhances cardiac morbidity and mortality in the transgender population on GAHT relative to cisgender individuals is still unclear. Moreover, the sustained effect of high E2 coupled to androgen suppression in male rats on their cardiac morphology and function is unknown. Thus, this study tested the hypothesis that GAHT in the adult male SD rat is associated with impaired cardiac function under baseline conditions. Methods: Adult male SDs (13 weeks of age) were randomly assigned to either: Control or E2+CTX therapy (E2 at 5 mg/day, pellet sc, replaced every 3 weeks, with CTX performed at 15 weeks of age) (n=8/grp). Baseline cardiac function was measured via echocardiogram (VEVO 3100), body composition via EchoMRI. Results: at 28 weeks of age, total lean mass and body weight were significantly decreased in the E2+CTX group, as previously observed; total fat mass did not differ between groups. Heart rate was significantly decreased; yet, other indices of cardiac function were not altered at baseline. Although no major differences in baseline cardiac function were observed, the long-term impact of GAHT on CV health and the ability of the heart to respond to secondary challenges are unknown. This study is ongoing and further studies will determine the ability of the heart to recover from secondary insults in order to determine if GAHT in males is associated with increased CV risk. [Formula: see text] Funding sources: HL143459, P20-GM-104357, P20-GM121334, T32-HL105324. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

New liquid chromatography tandem mass spectrometry reference data for estradiol show mini-puberty in both sexes and typical pre-pubertal and pubertal patterns.

Reliable estradiol (E2) reference intervals (RIs) are crucial in pediatric endocrinology. This study aims to develop a sensitive ultra-performance liquid chromatographic tandem mass spectrometry (UPLC-MS/MS) method for E2 in serum, to establish graphically represented RI percentiles and annual RIs for both sexes, and to perform a systematic literature comparison. First, a UPLC-MS/MS method for E2 was developed. Second, graphically represented RI percentiles and annual RIs covering 0-18 years were computed (cohort of healthy children [1181 girls and 543 boys]). Subsequently, RIs were compared with published data by systematic searches. Lower limit of quantification was 11 pmol/L, indicating high sensitivity. Estradiol first peaked during mini-puberty in both sexes (girls up to 192 pmol/L; boys up to 225 pmol/L). As could be expected, girls showed higher pubertal E2 (up to 638 pmol/L). However, boys' RIs (up to 259 pmol/L) overlapped considerably. We found 4 studies in the literature that also used LC-MS/MS to determine E2 and published RIs for the complete pediatric age range. Reference intervals varied considerably. Pre-pubertal and pubertal phases were present in all studies. Higher E2 during the time of mini-puberty in both sexes was documented in 3 studies including ours. Variability of RIs for E2 between studies illustrates the importance of laboratory-specific RIs despite using a LC-MS/MS reference method. In boys, the striking E2 peak during mini-puberty as well as high pubertal E2 without phenotypic estrogenization in regular male puberty indicates that the role of E2 in children and, especially in boys, requires better functional understanding.

Endogenous Hormones and Cognitive Decline in Women: Unveiling the Complex Interplay

This narrative review delves into the area of endogenous hormones and their impact on cognitive function, with a focus on women transitioning through perimenopause. While artificial intelligence technologies have revolutionized cognitive research, the inclusion of hormonal biomarkers remains sparse. The review synthesizes findings from diverse studies exploring the relationships between estrogen, progesterone, testosterone, other sex hormones, and cognitive performance. The research question explores the potential for monitoring endogenous hormonal levels during perimenopause to predict cognitive decline and inform preventive strategies. An analysis of relevant studies reveals a complex relationship, with varying impacts on cognitive domains. Thus, high E2 levels correlate positively with verbal memory and retrieval efficiency, contrasting with lower levels associated with enhanced visual memory, and testosterone shows positive links to verbal fluency. The limitations of existing research, including heterogeneous methodologies and a dearth of premenopausal representation, emphasize the necessity for future studies. To achieve this objective, it is important to leverage data from studies implementing standardized methodologies for tracking endogenous hormonal levels while accounting for cycle phases and menopausal transition stages. Additionally, employing standardized assessments for cognitive decline and analyzing extensive datasets derived from real-world sources, such as hospital or outpatient clinic chains, and digital apps, is crucial.

Hashimoto’s Thyroiditis Negatively Influences Intracytoplasmic Sperm Injection Outcome in Euthyroid Women on T4 Substitution Therapy: A Retrospective Study

Objective: The objective of this study was to analyze the impact of thyroid autoimmunity (TAI) on reproductive outcome parameters of intracytoplasmic sperm injection (ICSI) cycles as compared to TAI-negative ICSI cycles. Design: In this single in vitro fertilization (IVF) center retrospective study, 86 infertile women with elevated thyroid peroxidase or TGAb levels, but euthyroid after thyroxine replacement (study group), were compared to 69 female patients with no thyroid abnormalities (controls). Following ICSI treatment fertilization rate (FR), clinical pregnancy rate (CPR), miscarriage rate (MR), and live birth rate (LBR) were analyzed. Materials, Setting, Methods: All subjects with various infertility factors were treated with ICSI in university-based IVF center. Patients in the study group received thyroxine replacement and were euthyroid at IVF treatment. Before the IVF cycles, endocrinological parameters were uniformly assessed: thyroid function and antibodies, reproductive hormones (anti-Müllerian hormone [AMH], follicular stimulating hormone [FSH], luteinizing hormone, E2, PRL, testosterone, DHEAS, 17-OHP, AD) and OGTT (0–60–120 min glucose and insulin). Following descriptive comparison of laboratory parameters, age-adjusted analyses of FR, CPR, MR, and LBR were performed. Results: TAI-positive women were older (mean age 35.31 ± 4.95 vs. 32.15 ± 4.87 years; p = 0.002), had higher FSH (8.4 ± 3.4 vs. 7.4 ± 2.32 U/L; p = 0.024), higher E2 (53.94 ± 47.61 vs. 42.93 ± 18.92 pg/mL; p = 0.025) levels, while AMH (2.88 ± 2.62 vs. 3.61 ± 1.69 ng/mL; p = 0.0002) was lower. There were no differences in TSH levels (1.64 ± 0.96 vs. 1.66 ± 0.65 µIU/mL; p = 0.652) between the two groups. FT3 (2.63 ± 0.58 vs. 2.98 ± 0.55 pg/mL; p = 0.002) was lower and FT4 (1.3 ± 0.29 vs. 1.13 ± 0.21 ng/dL; p = 0.0002) was higher in the TAI-positive group, reflecting clinically irrelevant differences. Egg cell counts (6 ± 3.8 vs. 7.5 ± 3.95; p = 0.015) were lower in TAI and remained so following age adjustment. Although the overall ICSI FR did not differ (62.9% vs. 69.1%, p = 0.12), it was lower for patients under 35 with TAI showing decreasing differences in line with age. The CPR (36.04% vs. 69.56%; p < 0.001) and LBR (23.25% vs. 60.86%; p < 0.001) were lower, the MR (35.48% vs. 12.5%; p = 0.024) was higher in the TAI group, and these differences remained after age adjustment. Limitations: Since the higher age of the study group may interfere with the effect of TAI, age adjustment calculations were necessary to perform to eliminate this confounding factor. Conclusion: Despite optimal thyroid supplementation in clinical or subclinical hypothyroidism, the presence of TAI negatively influences CPR and is connected to a higher MR, thus resulting in a lower LBR after ICSI. Decreased FR with ICSI in TAI patients may also contribute to poorer outcomes, especially in younger women.

Abstract A043: Building a mammary-specific microenvironment for investigating sex hormones in microvessels and tumors

Abstract The mammary gland is highly vascularized and is exposed to alterations in sex hormones, including estrogen (E2) and progesterone (P4), which also play a role in breast cancer development and progression. Yet, our understanding of hormonal changes on the microvasculature in mammary tissue is limited to studies in rodents and simple 2D cultures. Advances in organ-on-chip technology now allow for models of 3D human microvessels to be generated. However, most microvascular models lack tissue-specific cell types, limiting their physiologic relevance. Thus, we first aimed to generate a tissue-specific breast microvasculature, and next used this model to study the impact of sex hormones. Our final goal is to generate a combined vascularized model of luminal A tumor on-chip. In our recently archived work (doi: 10.1101/2023.04.21.537664), we achieved the first in vitro perfusable breast-specific microvessels. Optimal conditions were achieved to co-culture human mammary vascular endothelial cells HMVEC and mammary fibroblasts on-chip. Through image-based quantification, microvessel morphology, not barrier function, was shown to be strongly influenced by the abundance of fibroblasts, which were critical to achieve lumenized (patent) vessels. Attaining perfusable microvessels allowed the study of discreet phases of the menstrual cycle (period, follicular, ovular, and luteal). The most striking results were observed with high concentrations of E2 (mimic of ovulation phase), which presented with increased vascular growth, and in contrast with high concentrations of P4 (mimic of luteal phase) that resulted in vascular rarefaction (pruning). Effects from combined E2 and P4 were dose-dependent, as demonstrated by significant branching and remodeling in the follicular phase. Hormonal effects were also tissue-specific, as shown by HUVEC-derived microvessels with significantly different morphologic phenotypes in comparison to HMVEC. Interestingly, fibroblasts expressed significantly high levels of receptors for both hormones and likely are major contributors to the hormone-responsiveness of microvessels. With mammary vessels now established, model tumors, spheroids, were generated from luminal-like breast cancer cells. Namely, MCF-7 and T47D, were treated by the same E2 and P4 conditions as in the microvessels. For both tumor spheroids, phase-dependent responses were observed, with high E2 resulting in the most significant growth. Ongoing work has shown the capacity to incorporate these tumor spheroids into our microvascular model, which results in varied local changes in vascular density. Future work involves investigating the role of hormones in the combined tumor-microvascular model. This complex humanized model will be useful in highlighting potential cancer treatment timelines (relevant to female menstrual cycle) and may be used to uncover novel therapeutic targets for luminal A breast cancers. Citation Format: Carmen Moccia, Marta Cherubini, Guido Frigieri, Kristina Haase. Building a mammary-specific microenvironment for investigating sex hormones in microvessels and tumors [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr A043.

Influence of repeated estrus synchronization treatment on hormone secretion, growth, and development of dairy goats.

In large-scale intensive farms, dairy goats often undergo frequent estrus synchronization (ES) treatment, which may result in a decline in reproductive performance; however, the underlying mechanism remains unclear. The present study aimed to investigate the effect of pregnant mare serum gonadotropin (PMSG) and progesterone (P4)-mediated ES treatment on fertility in dairy goats, while also identifying key metabolic and endocrine mechanisms that influence reproductive performance in does subjected to repeated ES treatment. Forty-eight Saanen does were randomly assigned to two groups (24 goats each) that received ES treatments either thrice fortnightly (3-PMSG) or once (1-PMSG) simultaneously with the third ES treatment of the 3-PMSG group during the breeding season. ES treatment was performed via the intravaginal insertion of a controlled internal drug release (CIDR) device impregnated with 300 mg P4, followed by 300 IU PMSG injections 48 h before CIDR withdrawal. Blood was collected to detect the level of hormones and blood biochemical indices. Additionally, estrus rate, fecundity rate, body weight, size, and lactation performance were measured. The results showed that repeated ES treatment markedly decreased the estrus rate and fecundity rate of goats. Among the does in all groups, there was no substantial difference in follicle stimulating hormone, luteinising hormone, gonadotropin-releasing hormone, melatonin, growth hormone, PMSG, total cholesterol, total protein, and glucose levels, as well as the body weight, body size, and lactation performance. Repeated ES treatment elevated estrogen (E2) levels 36, 48, and 72 h post-CIDR removal; increased P4 upon CIDR insertion; and raised PMSG antibody levels 24, 48, and 72 h post-CIDR removal. The results suggest that elevated anti-PMSG levels are the primary reason for the decline in ES efficiency, and that high E2 and P4 levels at some time points also impair reproductive performance. These findings provide novel insights into the metabolic effects of repeated PMSG stimulation in goats, guiding future reproductive hormone use in breeding practices.

AVPV Kiss1 neuron-specific knockdown of purinergic P2X2 receptor suppresses LH surge and ovulation in Kiss1-Cre rats.

Ovulation disorders are a major cause of low pregnancy rates and infertility in humans and livestock. Kisspeptin neurons located in the anteroventral periventricular nucleus (AVPV) are responsible for the generation of the gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) surge and the consequent ovulation in female rodents. The present study aimed to examine whether purinergic neurons are direct upstream stimulators of AVPV kisspeptin neurons that trigger the GnRH/LH surge and consequent ovulation in Kiss1-Cre rats. We specifically knocked down the mRNA expression of the P2rx2 purinergic receptor in AVPV kisspeptin neurons by administering an adeno-associated virus (AAV) vector containing Cre-dependent P2rx2 short hairpin RNA (shRNA) into the AVPV region of ovariectomized (OVX) Kiss1-Cre rats treated with a proestrus level of estradiol-17β (OVX + high E2) or ovary-intact Kiss1-Cre rats. The E2-induced afternoon LH surge was significantly suppressed by AVPV kisspeptin neuron-specific knockdown of P2rx2 in OVX + high E2 Kiss1-Cre rats compared with scrambled shRNA-treated control OVX + high E2 Kiss1-Cre rats. Furthermore, the specific knockdown of P2rx2 in AVPV kisspeptin neurons largely disrupted the estrous cycle, spontaneous LH surge, and ovulation in ovary-intact Kiss1-Cre rats. These findings suggest that purinergic neurons directly stimulate AVPV kisspeptin neurons via P2X2 receptors (P2RX2) to induce the GnRH/LH surge and consequent ovulation in female rats.

The effect of serum estrogen level on male infertility &amp; semen parameters on Sudanese males attending Khartoum Dermatology Teaching Hospital

Background Infertility is defined as the inability to achieve pregnancy after one year of unprotected intercourse. As estimated 10–15% of couples were considered infertile by W.H.O. One of the major causes of non-obstructive infertility is hormonal disturbance. Testosterone and Estrogen, traditionally, have been regarded as male and female sex hormones respectively. However, Estradiol, the predominant form of Estrogen, also plays a pivotal role in male sexual function. Estradiol in men is quintessential in many sexual physiological processes. Objectives To evaluate the effect of Estrogen levels on infertile men and their semen parameters Method In this prospective case-control hospital based study, a total number of 50 infertile men with primary and secondary infertility (case group) and 50 fertile men (control group) were included. The medical histories of all the participants were taken and physical examinations were conducted thoroughly. Serum Estradiol (E2) levels were measured for both infertile and fertile group. Seminal analysis was done for the infertile group. Serum E2 was measured by Radio-Immunosorbent Assay (RIA) using Estradiol kits for measurement. Standard semen analysis was done by computer-assisted seminal analysis (CASA) using the WHO manual 2010. The collected data were analyzed using Statistical Package for the Social Sciences (SPSS) version 25. The results were obtained and presented in tables and diagrams. The significance of testing was assessed by Chi-Square T test with P value ≤0.05 was considered statistically significant. Results A significant correlation was observed between the fertility status and serum E2 levels. 18% (n=9) of the subfertile patients were found to have serum E2 levels more than 55 pg/ml. on the other hand, only 6% (n=3) of the fertile group (control group) were above 55 pg/ml (P value less than 0.05). Azoospermia was found to be the most semen parameter among patients with elevated serum E2 level (P value less than 0.05). It is also observed that 46% of the infertile patients were found to be overweight (BMI=25–29.9) &amp; they have significantly high serum E2 levels (P value less than 0.05). Conclusion High serum Estradiol levels were found significantly correlated with male infertility; Azoospermia was the most commonly observed abnormal semen parameter. Overweight patients have significantly elevated Serum E2 levels.

Lipopolysaccharide-binding protein in follicular fluid is associated with the follicular inflammatory status and granulosa cell steroidogenesis in dairy cows.

Metabolic stress and subsequent hepatic dysfunction in high-producing dairy cows are associated with inflammatory diseases and declining fertility. Lipopolysaccharide (LPS)-binding protein (LBP) is produced by hepatocytes and controls the immune response, suggesting that it is involved in the pathophysiology of inflammation-related attenuation of reproductive functions during metabolic stress. This study investigated the effect of LBP on the inflammatory status, oocyte quality, and steroidogenesis in the follicular microenvironment of dairy cows. Using bovine ovaries obtained from a slaughterhouse, follicular fluid and granulosa cells were collected from large follicles to evaluate the follicular status of metabolism, inflammation, and steroidogenesis. Cumulus-oocyte complexes were aspirated from small follicles and subjected to in vitro embryo production. The results showed that follicular fluid LBP concentrations were significantly higher in cows with fatty livers and hepatitis than in those with healthy livers. Follicular fluid LBP and LPS concentrations were negatively correlated, whereas LPS concentration showed a positive correlation with the concentrations of non-esterified fatty acids (NEFA) and β-hydroxybutyric acid in follicular fluid. The blastulation rate of oocytes after in vitro fertilization was impaired in cows in which coexisting large follicles had high NEFA levels. Follicular fluid NEFA concentration was negatively correlated with granulosa cell expression of the estradiol (E2) synthesis-related gene (CYP19A1). Follicular fluid LBP concentration was positively correlated with follicular fluid E2 concentration and granulosa cell CYP19A1 expression. In conclusion, follicular fluid LBP may be associated with favorable conditions in the follicular microenvironment, including low LPS levels and high E2 production by granulosa cells.

Mammary Microvessels are Sensitive to Menstrual Cycle Sex Hormones.

The mammary gland is a highly vascularized organ influenced by sex hormones including estrogen (E2) and progesterone (P4). Beyond whole-organism studies in rodents or cell monocultures, hormonal effects on the breast microvasculature remain largely understudied. Recent methods to generate 3D microvessels on-chip have enabled direct observation of complex vascular processes; however, these models often use non-tissue-specific cell types, such as human umbilical vein endothelial cells (HUVECs) and fibroblasts from various sources. Here, novel mammary-specific microvessels are generated by coculturing primary breast endothelial cells and fibroblasts under optimized culture conditions. These microvessels are mechanosensitive (to interstitial flow) and require endothelial-stromal interactions to develop fully perfusable vessels. These mammary-specific microvessels are also responsive to exogenous stimulation by sex hormones. When treated with combined E2 and P4, corresponding to the four phases of the menstrual cycle (period, follicular, ovular, and luteal), vascular remodeling and barrier function are altered in a phase-dependent manner. The presence of high E2 (ovulation) promotes vascular growth and remodeling, corresponding to high depletion of proangiogenic factors, whereas high P4 concentrations (luteal) promote vascular regression. The effects of combined E2 and P4 hormones are not only dose-dependent but also tissue-specific, as are shown by similarly treating non-tissue-specific HUVEC microvessels.