Higher E2 Levels Research Articles

Modulation of spatial and response strategies by phase of the menstrual cycle in women tested in a virtual navigation task

Different memory systems are employed to navigate an environment. It has been consistently shown in rodents that estrogen impacts multiple memory system bias such that low estradiol (E2) is associated with increased use of a striatal-mediated response strategy whereas high E2 increases use of a hippocampal-dependent spatial memory. Low E2 also enhances performance on a response-based task whereas high E2 levels improve learning on a spatial task. The purpose of the present cross-sectional study was to investigate navigational strategies in young, healthy, naturally cycling women. Participants were split into either an early follicular (i.e., when E2 levels are low), ovulatory (i.e., when E2 levels are high) or mid/late luteal (i.e., end of the cycle, when E2 levels decrease and progesterone levels rise) phase group, using self-reported date of the menstrual cycle. Serum hormone level measurements (E2, progesterone, testosterone) were used to confirm cycle phase assignment. Participants were administered a verbal memory task as well as a virtual navigation task that can be solved by using either a response or spatial strategy. Women tested in the ovulatory phase, under high E2 conditions, performed better on a verbal memory task than women tested during the other phases of the cycle. Interestingly, women tested in the mid/late luteal phase, when progesterone is high, predominantly used a spatial strategy, whereas the opposite pattern was observed in the early follicular and ovulatory groups. Our data suggest that the specific memory system engaged differs depending on the phase of the menstrual cycle and may be mediated by both E2 and progesterone, rather than E2 alone.

The effects of Estrogen on the IFN signature in Lupus

Abstract Dendritic cells (DCs) are pivotal immune regulators and are important in the pathogenesis of Systemic Lupus Erythematosus (SLE). We have previously found that DCs from female lupus-prone mice constitutively over-express Type I Interferon (IFN) responsive genes resembling the IFN Signature found in SLE patients. Since this Signature is present in pre-diseased mice, it may contribute to disease pathogenesis. Nucleic acids, major auto-antigens in lupus, are potent stimulators of innate immunity and may contribute to the IFN signature by stimulating DCs through TLR7 and 9. As SLE is ~9 times more likely to affect women than men and the onset of SLE is most often in the reproductive years of women, estrogens have been suggested to affect the IFN response. To investigate the effects of estrogen on the IFN signature, we studied conventional DCs (cDCs), from male and female Sle1,2,3 lupus prone mice or from wild type C57BL/6 mice, generated in media void of sterols with supplemental titrations of 17-beta-estradiol (E2). We have found that DCs from male pre-diseased lupus-prone mice express the IFN Signature found previously in female DCs. Furthermore, E2 stimulation, regardless of the sex of the DCs, modulates the activation of cDCs in response to TLR stimulation. Interestingly, while high levels of E2 in hormone replete conditions were capable of exacerbating the IFN signature, the difference seen between wild-type and Sle DCs in standard conditions indicates both an E2 and E2-independent exacerbation in the activation of Sle DCs.

A Quantification of the Injury-Induced Changes in Central Aromatase, Oestrogenic Milieu and Steroid Receptor Expression in the Zebra Finch.

In songbirds and mammals, brain injury results in the up-regulation of aromatase (oestrogen synthase) expression in astroglia. The resulting presumed synthesis of neural oestradiol (E2 ) has neuroprotective effects including a decrease in neurodegeneration, neuroinflammation and apoptosis. The development of therapeutic tools that exploit oestrogenic neuroprotection in the treatment of neurotrauma requires a precise quantification of the endogenous changes in neural aromatase and E2 following brain injury. Surprisingly, the expected increase in neural oestrogens following brain injury has not been demonstrated. Furthermore, we are just beginning to unravel the mechanisms behind the protective effects of centrally synthesised E2 . In the present study, levels of aromatase immunoprotein, neural E2 and steroid receptor mRNA were quantified in adult male and female zebra finches 48h following a unilateral penetrating brain injury. Both aromatase and E2 were up-regulated in the injured hemisphere of the brain compared to the uninjured hemisphere, demonstrating for the first time a robust increase in neural E2 levels following injury. We did not detect an effect of injury on mRNA expression of the oestrogen receptors (ER)-α, ER-β or GPER-1, but observed a significant decrease in androgen receptor transcription in the injured lobe relative to the contralateral uninjured hemisphere. We conclude that mechanical damage causes a dramatic increase in local aromatisation, and the resultant high levels of central E2 are available to modulate steroid sensitive targets. Studies using alternate methods of receptor detection and/or time points may be necessary to understand the complete suite of mechanisms underlying the neuroprotective effects of induced oestrogen synthesis in this animal model.

Correlation of serum estradiol level on the day of ovulation trigger with the reproductive outcome of intracytoplasmic sperm injection.

BACKGROUND:Serum estradiol (E2) levels are measured in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI), to assess the ovarian response and to predict ovarian hyperstimulation syndrome. The impact of peak E2 levels on IVF-ICSI outcome was found to be inconsistent in the previous studies.AIM:To evaluate the impact of the serum E2 levels on the day of ovulation trigger with the reproductive outcome of ICSI.SETTINGS AND DESIGN:Retrospective observational study. ART Center, at a Tertiary Care University Teaching Hospital.SUBJECTS AND METHODS:Eighty-nine infertile women, who underwent ICSI with fresh embryo transfer over a period of 3 years, were included in the study. The study subjects were grouped based on the serum E2 level on the day of ovulation trigger:- Group I - <1000 pg/ml, Group II - 1000–2000 pg/ml, Group III – 2000.1-3000 pg/ml, Group IV – 3000.1–4000 pg/ml, and Group V >4000 pg/ml. The baseline characteristics and controlled ovarian hyperstimulation (COH) outcome were compared among the study groups.STATISTICAL ANALYSIS USED:Chi-square test, Student's t-test, ANOVA, and logistic regression analysis.RESULTS:The study groups were comparable with regard to age, body mass index, ovarian reserve. Group V had significantly higher number of oocytes retrieved than I and II (18.90 vs. 11.36 and 11.33; P = 0.009). Group IV showed significantly higher fertilization rate than I, III, and V; (92.23 vs. 77.43, 75.52, 75.73; P = 0.028). There were no significant differences in the implantation rates (P = 0.368) and pregnancy rates (P = 0.368).CONCLUSION:Higher E2 levels on the day of ovulation trigger would predict increased oocyte yield after COH. E2 levels in the range of 3000–4000 pg/ml would probably predict increased fertilization and pregnancies in ICSI cycles.

Localization of P450 Aromatase in The Brain of Adult Catfish Heteropneustes fossilis and Regional Distribution of Estradiol-17β and Testosterone: Gender and Seasonal Differences

P450 aromatase is the key enzyme that converts aromatizable androgens to estrogens. It is coded by the gene that exists in two forms: ovarian type and brain type. In the present study, we localized aromatase protein and gene (brain type) by immunocytochemical and in situ hybridization techniques, respectively, and the gene expression by RT-PCR. Aromatase immunoreactivity and transcript signals were observed in basophils of the proximal pars distalis of the pituitary, purported to be the gonadotrophs. The transcript signals were detected in the periventricular grey zone of optic tectum (PGZ), ventral telencephalic region, and radial glial cells lining the telencephalon, hypothalamus and olfactory bulb. Transcript levels of cyp19a1b showed sex, seasonal and regional differences. We measured testosterone (T) and estradiol-17β (E2) levels by specific ELISA kits. In the brain, T level is low in comparison to E2 level. Testosterone level is higher in postspawning phase and lower in spawning phase. The dimorphic expression varied with seasons, being higher in males in the preparatory and prespawning phases. E2 level is higher in female brain than in male brain especially in the preparatory and prespawning phases. E2 level is the highest in prespawning phase and lowest in postspawning phase. The steroids showed brain regional distribution with significant seasonal and dimorphic variations. Olfactory bulb has the highest T: E2 ratio in the resting, preparatory and prespawning phases in comparisons to other brain regions. All brain regions elicited a high level of E2, which showed dimorphic and seasonal variations, indicating high rate of aromatization. Differential levels of aromatase transcript, E2 and T in brain regions suggest a high level of neuroestrogen formation and E2 involvement in various neuroendocrine functions such as regulating spawning behaviour, plasticity, neurogenesis, etc.

Why more is less and less is more when it comes to ovarian stimulation

The purpose of the present study is to describe the possible mechanisms which may explain the apparent paradox of "less is more." Mild ovarian stimulation for in vitro fertilization (IVF) minimizes ovarian hyperstimulation syndrome (OHSS) and multiple gestations without compromising the pregnancy rate (PR). The pertinent English literature (PubMed) addressing mild stimulation for IVF/assisted reproductive technology (ART) and publications addressing "mild" or "soft" controlled ovarian stimulation (COS) vs conventional COS for IVF, OHSS, natural cycle IVF, and IVF outcome in association with COS was searched. Four possible mechanisms can be put forward to explain the apparent paradox of "less is more." (1) In the natural or mild stimulation cycles, the healthiest follicles are selected by the principle of "quality for quantity"; (2) high estradiol (E2) in the late follicular phase significantly correlated with higher rates of small for gestational age (SGA) and low-birth-weight (LBW) neonates; (3) anti-Mullerian hormone (AMH), LH, testosterone, and E(2) are significantly higher in natural cycle (NC)-IVF than in stimulated IVF follicles, suggesting an alteration of the follicular metabolism in stimulated cycles; and (4) supraphysiological E(2) may increase the growth hormone-binding protein (GH-BP) bio-neutralizing GH and diminishing the resultant insulin-like growth factor (IGF) levels, necessary for optimal synergism with follicle-stimulating hormone (FSH). It is suggested to aim at the retrieval of around eight to ten eggs. Mild stimulation should be the common practice for IVF. In cases where more than ten ova are retrieved or high E(2) levels are reached, either intentionally or unintentionally, "freeze-all policy" should be considered and embryo transfer (ET) done in a subsequent natural cycle.

Estrogen as an antioxdant in the lens

SummaryFemale gender is a known risk factor for age‐related cataract, although the explanation for this is still largely unknown. We have demonstrated that physiologic concentrations of 17b‐estradiol (E2) protect cultured human lens epithelial cells (HLECs) from oxidative stress induced by H2O2, as evident by decreased production of reactive oxygen species (ROS) and a stabilization of the mitochondrial membrane potential. Total superoxide dismutase (SOD) activity was increased by these concentrations of E2 but no change in gene or protein expression of SOD was seen. In cataract patients and controls, a significant correlation between higher age and decreasing serum levels of E2 was found but no correlation between serum levels of E2 and SOD was demonstrated. Men exhibited higher E2 levels compared to postmenopausal women. To conclude, estrogen seems to exert antioxidative effects on human lens cells, possibly by affecting SOD activity, which may explain the difference in risk of cataract between genders. The finding that men have higher levels of E2 than postmenopausal women further supports the hypothesis that it is the decline in estrogen at menopause that causes the higher risk of cataract in women.

Emergence of a metalloproteinase/phospholipase A2 axis of systemic inflammation

We review select aspects of the biology of matrix metalloproteinases (MMPs) with a focus on the modulation of inflammatory responses by MMP-2. MMP-2 is a zinc- and calcium-dependent endoprotease with substrates including extracellular matrix proteins, vasoactive peptides and chemokines. Humans and mice with MMP-2 deficiency exhibit a predominantly inflammatory phenotype. Recent research shows that MMP-2 deficient mice display elevated activity of a secreted phospholipase A2 in the heart. Additionally, MMP-2 deficient mice exhibit abnormally high prostaglandin E2 levels in various organs (i.e., the heart, brain and liver), signs of inflammation and exacerbated lipopolysaccharide-induced fever. We briefly review the biology of sPLA2 enzymes to propose the existence of a heart-centric MMP-2/sPLA2 axis of systemic inflammation. Moreover, we postulate that PLA2 activation is induced by chemokines, whose ability to signal inflammation is regulated in a tissue-specific fashion by MMPs. Thus, genetic and pharmacologically induced MMP-deficiencies can be expected to perturb PLA2-mediated inflammatory mechanisms.

High Maternal Serum Estradiol Levels Induce Dyslipidemia in Human Newborns via a Hepatic HMGCR Estrogen Response Element.

While the intrauterine environment is essential for the health of offspring, the impact of high maternal serum estradiol (E2) on lipid metabolism in offspring and the mechanisms are unknown. We found that ovarian stimulation (OS) could result in high E2 levels in women throughout pregnancy. Strikingly, their newborns showed elevated total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) levels that were positively related with E2 in newborns. In vitro, E2 dose-dependently stimulated TC and LDL-C secretion, and increased expression of the cholesterol synthesis rate-limiting enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) in HepG2 cells and mouse fetal hepatocytes. In vivo, high maternal E2 was detected and fetal livers also showed significantly higher HMGCR expression in an OS mouse model. Notably, an estrogen response element (ERE) was identified in the HMGCR promoter, indicating that high maternal serum E2 could up-regulate HMGCR expression in fetal hepatocytes via an ERE that in turn induces elevated levels of TC and LDL-C in offspring. Conclusion: OS can induce a high maternal E2 environment, which up-regulates HMGCR expression in fetal hepatocytes via an ERE in the promoter, and induces elevated levels of TC and LDL-C in newborns that may be related to increased risk of metabolic disease in adulthood.

Modulation of immune-response in female aP2-Cre ERalpha-knockout mice due to high-fat diet feeding and high estradiol levels

Previously reported abnormalities in the reproductive-tract of female adipose-tissue-specific (ap2-Cre) ERα-knockout (ERaKO)-mice were linked with a 40% decrease of ERα-gene-expression in the hypothalamus of these animals, when compared to their control-mice (ERα-floxed;WT). Subsequently, ERaKO-mice showed a pronounced upregulation of estradiol (E2)-plasma levels (KO: 45.9 vs. WT: 19.8pg/ml;p < 0.01) resulting from diminished ERα-mediated feedback in the hypothalamus. When fed with high-fat-diet (HFD), KO-mice developed a pronounced inflammation of uteri. To investigate the cause underlying this inflammatory process we performed H&E-, cd3-, Mac387- and Ly6G-stainings, a semiquantitative scoring of the inflammatory grade and microbiological analysis of the uteri. As HFD-feeding seems to accelerate the inflammatory response in those mice, we also analysed the plasma fatty-acid-profile of the ERaKO-mice fed with a normal-chow-diet (NCD) and HFD. We identified a massive accumulation of neutrophils in the uterus wall, while macrophages and lymphocytes were rarely present. The grade of inflammation (scored from 0 = no inflammation to 3 = most severe form) in HFD-fed KO-mice was significantly higher than in NCD-fed KO-mice (HFD: 2.44 vs. NCD: 1.76 arbitrary units). In accordance, we found that HFD-feeding resulted in increased stearic (c18:0; 983 vs. 567 µg/ml;p < 0.001) and arachidonic (c20:4 n-6; 976 vs. 594 µg/ml;p < 0,001) acid-levels in plasma. Moreover, PCR-analysis of bacterial DNA identified E. coli and Enterococcus sp. as main pathogenic bacteria present in tissue and uterine-fluid of ERaKO-mice. Taken together, our data suggest that high E2-levels in combination with a dysregulated fatty-acid-plasma profile modulate the immune-response of neutrophils and/or macrophages to bacterial infections, resulting in the inability of terminating the inflammatory process.

Endocrine determinants of incident sarcopenia in middle-aged and elderly European men.

BackgroundIn men, the long-term consequences of low serum levels of sex steroids, vitamin D metabolites, and insulin-like growth factor 1 (IGF-1) on the evolution of muscle mass, muscle strength, or physical performance are unclear. Moreover, there are no data about the relationship between these hormones and incident sarcopenia defined as low muscle mass and function. The aim of this study was to determine whether the baseline levels of sex hormones, vitamin D metabolites, and IGF-1 predict changes in muscle mass, muscle strength, physical performance, and incident sarcopenia.MethodsIn 518 men aged 40–79 years, recruited for participation in the European Male Ageing Study, total, free, and bioavailable testosterone (T), oestradiol (E), sex hormone-binding globulin, IGF-1, 25-hydroxyvitamin D (25OHD), 1,25-dihydroxyvitamin D (1,25(OH)2D), and parathyroid hormone were assessed at baseline. Appendicular lean mass (aLM), gait speed, and grip strength were measured at baseline and after a mean follow-up of 4.3 years. Sarcopenia was defined by the definition of Baumgartner (relative aLM ≤7.26 kg/m2), the International Working Group on Sarcopenia (IWGS), and the European Working Group on Sarcopenia in Older People (EWGSOP).ResultsaLM significantly decreased from age 50 years, while gait speed and grip strength significantly decreased from age 70 years. The incidence of sarcopenia by the definitions of Baumgartner, IWGS, and EWGSOP was 8.1%, 3.0%, and 1.6%, respectively. After adjustment for age, centre, body mass index, smoking, and number of comorbidities at baseline, baseline levels of T and vitamin D metabolites were not associated with change in aLM, gait speed, and/or grip strength, while a high baseline level of total E2 was associated with a greater decrease in aLM. In men aged ≥70 years, low IGF-1 was associated with a greater decrease in gait speed. Baseline endocrine variables were not independently associated with an increased risk of incident sarcopenia by any definition.ConclusionsLow levels of T and 25OHD do not predict loss of muscle mass, gait speed, or grip strength in middle-aged and elderly community-dwelling European men. Low IGF-1 predicts change in gait speed in men aged ≥70 years.

Lipoxins exert antiangiogenic and anti-inflammatory effects on Kaposi's sarcoma cells

Lipoxin A4 (LXA4) is an endogenously produced host molecule with anti-inflammatory resolution effects. Previous studies demonstrated it to be involved in anti-vascular endothelial growth factor (VEGF)-mediated angiogenesis and in a possible anticancer role via interaction with its receptor, lipoxin A 4 receptor (ALXR). Here, we examined the effects of LXA4 and its epimer 15-epi-LXA4 in inhibiting proinflammatory and angiogenic functions in a human Kaposi's sarcoma tumor-derived cell line (KS-IMM). KS-IMM cells expressed increased levels of inflammatory cyclooxygenase 2 (COX-2) and 5-lipoxygenase (5-LO) pathway enzymes when compared with human microvascular dermal endothelial cells (HMVEC-d). KS-IMM cells secreted high levels of prostaglandin E2 (PGE2) and chemotactic leukotriene B4 (LTB4). Treatment with LXA4 or 15-epi-LXA4 effectively reduced the levels of COX-2, 5-LO proteins, and secretion of PGE2 and LTB4 in KS-IMM cells. LXA4 or 15-epi-LXA4 treatment also decreased secretion of proinflammatory interleukin 6 (IL-6) and IL-8 cytokines but induced the secretion of anti-inflammatory IL-10. LXA4 treatment reduced the phosphorylation of VEGF receptor (VEGFR) and ephrin family receptor tyrosine kinases. LXA4 treatment effectively induced dephosphorylation of multiple cellular kinases such as Focal Adhesion Kinase, Protein kinase B, nuclear factor kappa-light-chain-enhancer of activated B cells, and Extracellular signal-regulated kinases (ERK)1/2, and reduced angiogenic factor VEGF-C secretion in KS cells. LX treatment drastically induced the Src-homology 2 domain-containing phosphatase tyrosine (Y542) phosphatase and reduced VEGFR-2 phosphorylation at sites Y1059, Y1175, and Y1212. Treatment of KS-IMM cells with LXA4 resulted in selective localization of VEGFR-2 in nonlipid raft (non-LR) and ALXR to LR fractions. These results demonstrated that LXA4 or 15-epi-LXA4 induce anti-inflammatory and antiangiogenic effects in KS cells and suggest that treatment with LXs is an attractive novel strategy against KS.

Effects of a luteinizing hormone-releasing hormone agonist on cognitive, sexual, and hormonal functions in patients with prostate cancer: relationship with testicular and adrenal androgen levels.

PurposeTo assess the cognitive and sexual/hormonal functioning of prostate cancer patients treated with a luteinizing hormone-releasing hormone (LH-RH) agonist, and the relationships thereof with adrenal and residual testicular androgen levels.Materials and methodsPreviously, we reported the effect of a luteinizing hormone-releasing hormone (LH-RH) agonist on testicular and adrenal androgen production in patients with prostate cancer. A 6-month treatment with an LH-RH agonist significantly reduced testicular androgens by 90–95% and adrenal androgens by 26–40%. This study evaluated the changes in cognitive and sexual/hormonal functions in the same cohort using the Mini-Mental State Evaluation (MMSE) and Expanded Prostate Cancer Index Composite (EPIC) questionnaire, respectively. In addition, the associations of each function with the serum testosterone (T), dihydrotestosterone (DHT), estradiol (E2), dehydroepiandrosterone-sulfate (DHEA-S), dehydroepiandrosterone (DHEA), androstenedione (A-dione), and cortisol levels were studied.ResultsCognitive functions did not change significantly during the treatment. Sexual functions were relatively low before treatment and worsened significantly after 6 and 12 months of treatment. Interestingly, sexual bothers were improved with the treatment. The treatment significantly worsened hormonal functions and bothers. Regarding specific items in the hormonal domains, hot flashes and body weight changes were the main effects of worsened hormonal function. Low levels of T and E2 and high levels of A-dione were associated with low MMSE scores at 6 months. Regarding sexual and hormonal functions, A-dione, E2, T, cortisol, and DHEA-S were associated with poorer functioning and bother. Especially, low T levels and high E2 levels were the most significant factors associated with worse sexual and hormonal bothers.ConclusionThe LH-RH agonist monotherapy worsened sexual and hormonal functions and hormonal bothers, but not sexual bothers or cognitive functions. The changes in these functions were related to the testicular and adrenal androgens levels.

The Effects of Vitamin A Administration to Heifer and Pregnant Cow on the Changes of Hormonal and Body Weight

The objective of this study was to investigate the interrelationship of weight and hormonal changes through the administration of vitamin A to first time pregnant heifers. A total of 28 Korean heifers was used for this study. The heifers were divided into two groups - with vitamin A (n=14) and without vitamin A (control) (n=14) in the feed. Body weight increased in vitamin A treated heifers starting 9 months until 15 months. After pregnancy, vitamin A treated heifers were found to maintain higher body weights than the control group. Pre-pregnancy and post pregnancy progesterone levels were not different between the two groups. Serum estradiol levels of heifers at different growth stages showed relatively higher E2 levels than the control. Also, the control during pregnancy may show higher serum E2 levels than the vitamin A treated heifers. The growth phase serum estradiol levels in heifers may be relatively higher than the control. During pregnancy it showed a similar trend. Serum levels of vitamin A treated heifers did not differ from pregnant heifers at 5 months of age. However, after 5 months from conception until 8 months of treatment it showed a high level. Serum cholesterol in pregnant cows was higher in the control group than the treatment from beginning until the end of pregnancy. This is considered to be related to fetus development during pregnancy, as well as the mother's health.

Estradiol replacement enhances fear memory formation, impairs extinction and reduces COMT expression levels in the hippocampus of ovariectomized female mice

Females experience depression, posttraumatic stress disorder (PTSD), and anxiety disorders at approximately twice the rate of males, but the mechanisms underlying this difference remain undefined. The effect of sex hormones on neural substrates presents a possible mechanism. We investigated the effect of ovariectomy at two ages, before puberty and in adulthood, and 17β-estradiol (E2) replacement administered chronically in drinking water on anxiety level, fear memory formation, and extinction. Based on previous studies, we hypothesized that estradiol replacement would impair fear memory formation and enhance extinction rate. Females, age 4weeks and 10weeks, were divided randomly into 4 groups; sham surgery, OVX, OVX+low E2 (200nM), and OVX+high E2 (1000nM). Chronic treatment with high levels of E2 significantly increased anxiety levels measured in the elevated plus maze. In both age groups, high levels of E2 significantly increased contextual fear memory but had no effect on cued fear memory. In addition, high E2 decreased the rate of extinction in both ages. Finally, catechol-O-methyltransferase (COMT) is important for regulation of catecholamine levels, which play a role in fear memory formation and extinction. COMT expression in the hippocampus was significantly reduced by high E2 replacement, implying increased catecholamine levels in the hippocampus of high E2 mice. These results suggest that estradiol enhanced fear memory formation, and inhibited fear memory extinction, possibly stabilizing the fear memory in female mice. This study has implications for a neurobiological mechanism for PTSD and anxiety disorders.

Adipose tissue, estradiol levels, and bone health in obese men with metabolic syndrome.

Visceral adipose tissue (VAT) is associated with an increased risk of metabolic syndrome (MetS). Recent studies have suggested that VAT negatively affects bone. However, MetS has also been associated with higher estradiol (E2) levels, which is bone protective. We therefore investigated the impact of VAT and E2 levels on bone density, structural parameters, and strength estimates. A cross-sectional study was conducted in 72 obese men with MetS to investigate the impact of VAT and E2 levels on bone. Bone parameters were assessed by dual-energy X-ray absorptiometry (DXA), quantitative computed tomography (QCT), and high-resolution peripheral QCT (HRpQCT) at lumbar spine, proximal femur, radius, and tibia. VAT volume was measured by magnetic resonance imaging (MRI) and sexual hormones were measured in blood samples. Men with high VAT had a lower bone density at the hip (P<0.05), lower cortical thickness, and higher buckling ratio at femoral neck (FN) (P=0.008 and P=0.02), compared with men with low VAT, despite a similar body weight and BMI. Generally, E2 levels were low (median 43 pmol/l), and men with E2 levels below median had reduced bone density at lumbar spine (P=0.04), and impaired structural parameters at radius and tibia, compared with men with E2 levels above median. At the hip, VAT volume and E2 levels affected bone density independently and additively, and 50% of men with high VAT and low E2 levels had osteopenia with significantly lower T-score at FN (P=0.004). High VAT and low E2 negatively affect bone in obese men with MetS. VAT and E2 affect bone density at the hip independently and additively, revealing an unexpected high prevalence of osteopenia in middle-aged men with MetS.

Prenatal alcohol exposure alters response of kisspeptin-ir neurons to estradiol and progesterone in adult female rats.

Prenatal alcohol exposure (PAE) has adverse effects on reproductive function and hypothalamic-pituitary-gonadal (HPG) activity. Kisspeptin neurons play a role in mediating feedback effects of estradiol (E2 ) and progesterone (P4 ) on the HPG axis. We hypothesized that PAE will have long-term effects on the response of kisspeptin neurons to E2 and P4 . Adult female rats (53 to 58days) from prenatal ad libitum-fed control (C), pair-fed (PF), and alcohol-exposed (PAE) groups were subjected to Sham ovariectomy (OVX) or OVX without or with replacement with low or high physiological levels of E2 and P4 , and terminated under basal conditions. E2 and P4 levels, and the response of kisspeptin-ir neurons in the arcuate (ARC) and anteroventral periventricular (AVPV) nuclei to these hormones, were measured. As the E2 signal is conveyed to kisspeptin neurons via estrogen receptor-α (ER-α), we investigated PAE effects on the number of kisspeptin-ir/ER-α-ir neurons. To determine whether PAE alters interactions between kisspeptin and gonadotropin-releasing hormone (GnRH) neurons, close contacts between kisspeptin-ir fibers and GnRH-ir cell bodies were examined. Our data present the novel finding that kisspeptin-ir neurons in the ARC of PAE females show differential responses to E2 and to the combined treatment with E2 and P4 compared with controls: (i) OVX increased the number of kisspeptin-ir neurons in C and PF, but not PAE females compared with their Sham counterparts; (ii) E2 replacement restored kisspeptin-ir cell numbers to Sham levels in C and PF females but caused a robust down-regulation of kisspeptin-ir neurons below Sham levels in PAE females; (iii) OVX and replacement with high physiological concentrations of E2 resulted in fewer kisspeptin-ir cells in PAE than C females; (iv) OVX and replacement with high levels of both E2 and P4 markedly decreased the number of kisspeptin-ir neurons, below levels observed following E2 alone, in PF and C females, but had no significant effect in PAE females. These data suggest that a possible mechanism underlying adverse effects of PAE on HPG function involves actions of alcohol on the kisspeptin system.

Postmenopausal patients with endometrial cancer of type 1 have elevated serum estradiol levels in the ovarian vein.

Type 1 endometrial cancer (EC) is typically sex hormone sensitive; however, most women diagnosed with EC have already gone through menopause. Several studies have reported that the postmenopausal ovary is hormonally active, and estradiol (E2) production from the ovaries persists for as much as 10 years beyond menopause. The aim of this study was to evaluate whether sex steroid production from the ovaries contributes to the pathogenesis of type 1 EC. This was a prospective study of 53 women treated for EC (28 cases of type 1 disease and 25 cases of type 2 disease). Serum specimens were collected from the peripheral and ovarian veins of participants undergoing bilateral oophorectomy. The sex steroid hormone levels and hormonal milieu on cervical cytology were evaluated as maturation value (MV). In addition, the degree of stromal hyperplasia of the ovary was evaluated histologically. Although the E2 levels of the peripheral veins did not show any significant differences [8.2 (5.1-12.4) vs 7.4 (5.1-11.7) pg/mL, respectively; P < 0.05], the patients with type 1 EC had a higher E2 level in the ovarian vein than those with type 2 EC [25 (13.8-42.5) vs 15 (10.0-23.0) pg/mL, respectively; P < 0.05]. There were also no significant differences in the rate of moderate to marked hyperplasia of the ovarian stroma between the groups; however, the thickness of the ovarian cortex demonstrated a correlation with the ovarian E2 level. In addition, the MV displayed a strong correlation with the ovarian E2 level, but not the peripheral E2 level. The postmenopausal ovary is hormonally active, especially in patients with type 1 EC. The degree of ovarian stromal hyperplasia may (at least in part) contribute to the progression of type 1 EC, and MV may predict the level of E2 production from the ovaries in postmenopausal women.

269P - The Relationship Between Serum E2 Level and Recurrence During Endocrine Therapy for Er Positive Pre-Menopausal Early Breast Cancer Patients

ABSTRACT Aim: 10-year tamoxifen as adjuvant endocrine therapy become common for pre-menopausal ER-positive breast cancer patients according to the report from ATLAS trial (SABCS 2012) or aTTom study (ASCO 2013). On the other hand, it is generally believed that exposure to estradiol is strongly associated in breast cancer, however there are different opinions among experts voting in the duration of using LHRH agonist. The aim of this study is to evaluate the recurrence factor for endocrine therapy in ER positive pre-menopausal breast cancer patients. Methods: We chose 193 cases of premenopausal ER-positive early-stage breast cancer patients who received LHRH agonist and tamoxifen with 45 year-old or more younger onset. The median age was 40 years old (24-45) when the treatment started. They included 98 cases who were received pre-operative or post-operative chemotherapy as adjuvant systemic therapy. We studied about the relationship of local recurrence or distant metastasis and the duration of administration of the drug. We monitored of E2, FSH concentrations in blood before, during and after treatment. The median follow up period was 68 months (9-324 months). Results: We obtained the data of E2 dynamics during endocrine therapy from patients' sample. During LHRH agonist and tamoxifen therapy, serous E2 level decreased to the postmenopausal level, less than 31.0 pg/ml. However E2 concentration with tamoxifen and without LHRH agonist elevated to the average 58.1 pg/ml (10.0-5248.0 pg/ml). The duration of LHRH agonist was not the most important factor of breast cancer recurrence. There was a relation between continuous high serum level of E2 after LHRH agonist and metastasis regardless of the menstruation state. This phenomenon could be explained by the similar mechanism of low dose estrogen therapy after AI therapy for metastatic breast cancer. Conclusions: There might be association with breast cancer recurrence and E2 concentration in premenopausal ER-positive early-stage breast cancer. Disclosure: All authors have declared no conflicts of interest.

Systemic and localized seminal plasma hypersensitivity patients exhibit divergent immunologic characteristics

Systemic and localized seminal plasma hypersensitivity patients exhibit divergent immunologic characteristics