Stimulating Estradiol Research Articles

Linear growth and endocrine function in children with ataxia telangiectasia.

Introduction:Ataxia telangiectasia (AT) is a rare, genetic, primary immune deficiency disease characterized by immunodeficiency and neurological manifestations, with an increased tendency to infection, malignancy, and autoimmune diseases. Both growth delay and endocrine abnormalities are occasionally reported in these patients.Patients and Methods:We studied growth parameters height (Ht), weight, body mass index (BMI) and calculated the Ht standard deviation scores (HtSDS) of 13 patients (age 7.7 ± 3.5 years-age range: 3–14.5 years) with AT in relation to their mid-parental Ht SDS (MPHtSDS). We measured their serum calcium (Ca), phosphorus (PO4), alkaline phosphatase, alanine transferase (ALT), serum ferritin, creatinine and albumin concentrations. Endocrine investigations included the assessment of serum free thyroxine (FT4), thyrotropin (TSH), insulin-like growth factor-I (IGF-I) and morning cortisol. Complete blood count and serum immunoglobulins (IgG, IgM and IgA antibodies) were also measured. Growth data were correlated to hormonal and immune data.Results:About 31% of patients with AT had short stature (HtSDS <−2). However, their MPHtSDS denoted that their short stature was familial because four out of 13 had MPHtSDS <−2. They had low BMI, and two of them had low serum albumin and IGF-I, denoting malnutrition or disturbed growth hormone secretion. Elevated serum ALT and ferritin in some patients suggest immune-related inflammation in the liver. 30% of patients had high TSH, two of them had low FT4 diagnosing overt (15%) and sub-clinical (15%) hypothyroidism. Anti-thyroid peroxidase antibodies were high in two out of 13 patients denoting immune-related thyroid aggression. Eight out of 13 patients had Vitamin D deficiency (<20 ng/ml) however, their serum Ca and PO4 levels were in the normal range. One adolescent girl (14.5 years) had hyper-gonadotropic hypogonadism (low estradiol and high follicle stimulating hormone). All patients had normal 8 AM cortisol and renal function. None of the growth parameters were correlated with the IgG, IgM or IgA levels.In summary:Patients with AT had a high prevalence of growth retardation and endocrine dysfunction in the form of low IGF-I, overt and subclinical hypothyroidism and hypogonadism. Physicians should be aware of these possible endocrinopathies for an early diagnosis and proper treatment.

Role of plasma membrane estrogen receptors in mediating the estrogen induction of progesterone receptors in hypothalamic ventromedial neurons

Progesterone is well known for its role in the modulation of sexual behavior. In the ventromedial nucleus (VMN), a part of the mediobasal hypothalamus that regulates sexual behavior in female rodents, estrogens induce the expression of progesterone receptors (PRs). This effect is known to be dependent on the activation of nuclear estrogen receptors (ERs). However, recent studies have documented estrogen activation of genomic transcription triggered by protein-protein phosphorylation cascades initiated at membrane receptors. The aim of this study was to examine if membrane-initiated estradiol (E2 ) stimulation is able to induce PR expression in the VMN or, at least, to modulate nuclear ER action. To achieve this goal, 2-month-old ovariectomized Wistar rats were injected bilaterally, in the vicinity of VMN, with free E2 and with E2 conjugated with bovine serum albumin (E2 BSA), alone or in sequence, by using a two-pulse injection paradigm. Stereological methods and western blot analysis were used to estimate the total number of PR-immunoreactive neurons in the VMN and the PR protein content of the VMN, respectively. The results showed that the administration of E2 BSA alone increases the number of PR-immunoreactive neurons and the expression level of PR protein to values similar to those resulting from E2 administration. They also showed that the sequential administration of E2 and E2 BSA potentiates the effects resulting from the injection of E2 or E2 BSA alone. These data provide the first evidence that membrane-initiated E2 stimulation is able to induce and to potentiate the genomic activation of PR expression in the VMN.

Oral Bisphenol A (BPA) given to rats at moderate doses is associated with erectile dysfunction, cavernosal lipofibrosis and alterations of global gene transcription.

IntroductionBisphenol A (BPA), a suspected reproductive biohazard and endocrine disruptor released from plastics is associated with erectile dysfunction (ED) in occupationally exposed workers. However, in rats, despite the induction of hypogonadism, apoptosis of the penile corporal smooth muscle, fat infiltration into the cavernosal tissue, and changes in global gene expression with the intraperitoneal administration of high dose BPA, ED was not observed.AimsWe investigated whether BPA administered orally rather than intraperitoneally to rats for longer periods and lower doses will lead to ED.Main Outcomes MeasuresED, histological, and biochemical markers in rat penile tissues.Methods2.5-month old rats were given drinking water daily without and with BPA at 1 and 0.1 mg/kg/day. Two months later, erectile function was determined by cavernosometry (DIC) and electrical field stimulation (EFS) and serum levels of testosterone (T), estradiol (E2), and BPA were measured. Penile tissue sections were assayed by Masson (smooth muscle (SM)/collagen), Oil Red O (fat), TUNEL (apoptosis), immunohistochemistry for Oct 4 (stem cells), and α-SM actin/ calponin (SM and myofibroblasts), applying quantitative image analysis. Other markers were assayed by western blots. DNA microarrays/microRNA assays defined transcription profiles.ResultsOrally administered BPA did not affect body weight, but: 1) decreased serum T and E2; 2) reduced the EFS response and increased the DIC drop rate; 3) increased within the corporal tissue the presence of fat, myofibroblasts and apoptosis; 4) lowered the contents of SM and stem cells, but not nerve terminals; and 5) caused alterations of the transcriptional profiles for both mRNA and microRNAs within the penile shaft.ConclusionsLong-term exposure of rats to oral BPA,caused a moderate corporal veno-occlusive dysfunction (CVOD), possibly due to alterations within the corporal tissue that pose gene transcriptional changes related to inflammation, fibrosis and epithelial/ mesenchymal transition (EMT).

Study On The Fertility Status Of Females With Primary Mediastinal B-Cell Lymphoma Following Treatment With Dose-Adjusted EPOCH-Rituximab

BackgroundIn young females with lymphoma, the impact of immunochemotherapy on fertility status has been poorly studied. Primary mediastinal B-cell lymphoma (PMBL) predominantly affects young females and is highly curable with the dose-adjusted EPOCH-rituximab (DA-EPOCH-R) regimen (Dunleavy et al., NEJM 2013:368;1408). We therefore identified this group of PMBL patients as an excellent population to study fertility status and evaluated the impact of DA-EPOCH-R on gonadal function of females aged 50 years or less. MethodsA questionnaire was first mailed to 27 patients, and this was followed by a semi-structured phone interview to obtain information on patients' medical and gynecologic/obstetric histories before PMBL diagnosis, during treatment and after its completion. Serum estradiol (E2), luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels were analyzed before commencing chemotherapy, following 5-6 cycles of DA-EPOCH-R, and 10-18 months after treatment completion. ResultsAll patients included in the study completed 6-8 cycles of DA-EPOCH-R and were in complete remission from PMBL at a median follow-up of 63 months. Median age at diagnosis was 30.5 years (r 21-50). Twenty-three patients participated in the questionnaire. Prior to chemotherapy, twenty of twenty-three were menstruating, three patients were not. Of the twenty menstruating patients treated with DA-EPOCH-R three were unsure of their menstrual histories during chemotherapy, fourteen (70%) developed amenorrhea and three (15%), all younger than 25 years, continued to menstruate. After completion of chemotherapy, fifteen of the twenty patients (75%) had menstrual periods. All were 35 years or younger. Five patients (25%) remained amenorrheic following completion of chemotherapy, however one had a hysterectomy shortly after treatment completion. To date six of twenty (30%) patients have conceived naturally and delivered healthy children with nine not yet desiring motherhood.On laboratory evaluations performed before treatment, after 5-6 cycles of DA-EPOCH-R and 10-18 months after completion of chemotherapy, median serum LH levels were 2.5, 33.4 and 4.6 U/L respectively, median FSH values at the same time points were 3.6, 69.5 and 8 U/L, while median E2 levels were 24.4, <5 and 16.7 pg/mL, time courses consistent with transient ovarian damage from chemotherapy, but with recovery at a median follow-up of 12 months. Eight patients, who were 40 years or older were treated with DA-EPOCH-R. The median endocrine laboratory parameters of six of these patients, who had clinically normal ovarian function prior to treatment were examined at the indicated time points and were as follows: LH: 2.6, 36.4, 26.7 U/L; FSH: 6.4, 102.8, 83.9 U/L; E2: 68.8, <5, <5 pg/mL, a pattern consistent with premature menopause. ConclusionFertility appears to be preserved in female PMBL patients receiving DA-EPOCH-R, particularly in those less than 40 years old. The risk of premature menopause seems to be confined to patients over 40 years of age. Further analysis of ovarian reserve in patient younger than 40 years, measuring Anti-Muellerian hormone and inhibin-B pre- and post-treatment, is ongoing. Disclosures:No relevant conflicts of interest to declare.

Characterization of follicle and CL development in beef heifers using high resolution three-dimensional ultrasonography

The aim was to characterize dominant follicle (DF) and CL development through the estrous cycle of cattle using three-dimensional (3D) ultrasonography while making a comparison with conventional two-dimensional (2D) B-mode ultrasound (US) and to relate the measures taken to systemic concentrations of steroid hormones and gonadotropins. After synchronization of estrus, the ovaries of crossbred beef heifers (N = 5) were assessed using daily US with a GE Voluson i US scanner until the end of the first follicle wave, then every other day until emergence of the final (ovulatory) wave, when daily US resumed until ovulation. Follicle and CL growth were recorded and mapped. Measures of diameter (2D) and volume (3D) of the DF from the first and ovulatory waves of the cycles; and CL development were captured and stored for further analysis. Blood flow to the DF and CL were assessed using 3D power Doppler US measuring vascularization index (VI; %), vascularization flow index (0/100) and flow index (0/100). Jugular blood samples were collected every 24 hours for progesterone from the first estrus until the second ovulation. Concentrations of estradiol (E2) and follicle stimulating hormone (FSH) were measured every 8 hours from estrus to second follicle wave emergence; then, E2 only was measured from final follicle wave emergence until ovulation. Data were analyzed using PROC MIXED and PROC REG in SAS. Dominant follicle blood flow tended to decrease during follicle wave emergence and DF VI increased (P < 0.05) 24 hours before ovulation after peak E2. Measures of the DF and CL volume (3D) were highly predictive of 2D diameter measures throughout the cycle (P < 0.0001). Predictive values (r2) for day of wave emergence and day from ovulation were similar for 2D and 3D measures; however, 2D measures had higher repeatability when compared with 3D measures. There was no relationship between CL VI and progesterone early in the cycle (r2 = 0.12; P = 0.1); however, there was a strong positive relationship approaching ovulation (r2 = 0.77; P < 0.0001). In conclusion, 3D power Doppler measures of blood flow appears to be representative of vascular changes in the DF and CL throughout the estrous cycle. However, the extra time required to acquire and analyze a 3D image and the relatively little additional information obtained over that achievable with 2D imaging in terms of follicle and CL development might preclude its widespread use other than for detailed research purposes.

English

Medicinal plants contain physiologically active principles that over the years had been exploited in traditional medicine for the treatment of various ailments. The present study was undertaken to investigate the effects of ethanolic extract of&nbsp;Hibiscus sabdariffa&nbsp;calyx on rat reproductive hormones. The effects on the basal levels of estradiol, testosterone, prolactin and follicle stimulating hormone were conducted in experimental animals.&nbsp;H. sabdariffa&nbsp;calyx extract at a dose of 250 mg/kg produced minor effects on rat reproductive hormones, namely testosterone and estradiol while no change was observed on both prolactin and follicle stimulating hormone levels. Moreover, no histological changes were detected on both testes and ovaries of the experimental animals after 28 days of administration. It can be concluded that&nbsp;H. sabdariffa&nbsp;calyx extract at a dose of 250 mg/kg caused mild effects on rat reproductive hormones. &nbsp; Key words: Estrogenic effect, reproductive hormones,&nbsp;Hibiscus sabdariffa&nbsp;extract.

Cycle cancellation and pregnancy after luteal estradiol priming in women defined as poor responders: a systematic review and meta-analysis

Does a luteal estradiol (LE) stimulation protocol improve outcomes in poor responders to IVF? LE priming is associated with decreased cycle cancellation and increased chance of clinical pregnancy in poor responders Poor responders to IVF are one of the most challenging patient populations to treat. Many standard protocols currently exist for stimulating these patients but all have failed to improve outcomes. Systematic review and meta-analysis including eight published studies comparing assisted reproduction technology (ART) outcomes in poor responders exposed to controlled ovarian hyperstimulation with and without LE priming. A search of the databases MEDLINE, EMBASE and PUBMED was carried out for studies in the English language published up to January 2012. Studies evaluating women defined as poor responders to ART were evaluated. These studies were identified following a systematic review of the literature and data were analyzed using the DerSimonian-Laird random effects model. The main outcomes of interest were cycle cancellation rate and clinical pregnancy. Although the definition of clinical pregnancy varied between studies, the principal definition included fetal cardiac activity as assessed by transvaginal ultrasonography after 5 weeks of gestation. A total of 2249 publications were identified from the initial search, and the bibliographies, abstracts and other sources yielded 11 more. After excluding duplications, 1227 studies remained and 8 ultimately met the inclusion criteria. Compared with women undergoing non-LE primed protocols (n = 621), women exposed to LE priming (n = 468) had a lower risk of cycle cancellation [relative risk (RR): 0.60, 95% confidence interval (CI): 0.45-0.78] and an improved chance of clinical pregnancy (RR: 1.33, 95% CI: 1.02-1.72). There was no significant improvement in the number of mature oocytes obtained or number of zygotes obtained per cycle. These findings are limited by the body of literature currently available. As the poor responder lacks a concrete definition, there is some heterogeneity to these results, which merits caution when applying our findings to individual patients. Furthermore, the increased clinical pregnancy rate demonstrated when using the LE protocol may be principally related to the decreased cycle cancellation rate. The LE protocol may be of some utility in the poor responder to IVF and may increase clinical pregnancy rates in this population by improving stimulation and thereby decreasing cycle cancellation. NIH K12 HD063086 (ESJ, MGT), NIH T32 HD0040135-11 (KAR), F32 HD040135-10 NIH (KRO), 5K12HD000849-25 (PTJ). No competing interests.

Sex differences in noninvasive vascular function in the community

The relation of noninvasive vascular function to sex, sex hormones, and reproductive history in the general population is little understood. We simultaneously assessed flow-mediated dilation (FMD) and peripheral arterial tonometry in 454 women (mean age 40.4±16.1 years, age range 19-78 years) and 100 men (mean age 44.7±15.3 years) in a community-based cohort. Plasma estradiol, progesterone, luteinizing hormone, and follicle stimulating hormones were measured, and menstrual cycle and reproductive history were recorded. Vascular function was blunted in men as compared to women irrespective of menopausal status and adjustment for classical cardiovascular risk factors and hormones. Vascular reactivity changed during the menstrual cycle and correlated with estradiol concentrations for FMD, r=0.13 and inversely with progesterone for pulse amplitude, r=-0.14, and brachial artery diameter, r=-0.10. Multivariable-adjusted regressions showed a relation of estradiol with FMD, β 0.658, 95% confidence interval (CI) 0.084/1.232, P=0.025 in women. Age at menarche (β 0.070, 95% CI 0.039/0.101, P<0.0001) and breastfeeding duration (β -0.006, 95% CI -0.011/-0.001, P=0.036) were related to brachial artery diameter, age at menarche also to FMD (β -0.455, 95% CI -0.886/-0.023, P=0.039). Sex differences in noninvasive conduit and peripheral arterial function with better vascular reactivity in women were not fully explained by female sex hormones and menopausal status. Age at menarche and duration of breastfeeding were also related to vascular function and need further investigation.

A randomized trial of chinese medicine lirukang granule (利乳康颗粒) combined with psychological intervention for cyclomastopathy and menoxenia

To observe the influence of therapy with Chinese medicine Lirukang Granule (, LRKG) combined with psychological intervention on anxiety states and sex hormones in patients with cyclomastopathy and menoxenia. A total of 470 subjects were randomly assigned to three groups by the net-central randomization system, the treatment group (161 patients, treated with LRKG and psychological intervention), the Chinese medicine group (157 patients, treated with LRKG), and the psychological intervention group (152 patients, treated with psychological intervention). The dose of LRKG was 12 g three times per day; psychological intervention included establishing relations, cognitive intervention and psychological persuasion, 30-40 min per session, once a week. The therapy duration for all groups was three months. The efficacy was compared and anxiety state/State-Trait Anxiety Invertory (STAI) scoring was measured before and after treatment. The serum estradiol (E2), progesterone (P), prolactin (PRL) and follicle stimulating hormone (FSH) levels of 60 patients selected randomly from each group during the luteal phase were measured before and after treatment, and a group of 20 healthy women were evaluated for comparison. A follow-up was arranged for one year after treatment. Thirty subjects were lost to follow-up. (1) Comparison of efficacy: the markedly effective rate and the total effective rate of the treatment group were 86.67% (131/150) and 98.00% (147/150), respectively; of the Chinese medicine group, 64.58% (93/144) and 90.27% (130/144), respectively; and of the psychological intervention group, 0% (0/146) and 3.42% (5/146), respectively. The markedly effective rate and the total effective rate in the treatment group were significantly higher than those in the Chinese medicine and psychological intervention groups (P < 0.05). (2) Comparison of STAI scoring: STAI scoring was decreased dramatically in the treatment group after treatment compared with that of the Chinese medicine group (P < 0.01), but there was no significant difference compared with the psychological intervention group. (3) Comparison of levels of sex hormones: E2, P, PRL and FSH of the three patient groups were disordered before treatment, and significantly different from healthy women (P < 0.01). After treatment, the levels of P and FSH of the treatment group were significantly increased (P < 0.01), E2 and PRL were significantly reduced, which were also significantly decreased compared with the psychological intervention groups (P < 0.01). (4) FOLLOW-UP: the markedly effective rate and the total effective rate of the treatment group remained higher than those of the other two groups after one year of treatment (P < 0.05). (5) Adverse reactions: no obvious adverse reactions were found among the three groups. Therapy with Chinese medicine combined with psychological intervention was effective for short-term and long-term treatment of cyclomastopathy and menoxenia. The mechanism might be related to the regulation of sex hormones.

Abstract P6-05-09: Unravelling the global effect of estrogen on breast cancer cell proteome using quantitative proteomics.

Abstract Estrogens exert their function through genomic and non-genomic pathways mediated mainly by estrogen receptors. Estrogen signalling is highly complex and once activated initiates cancer cell proliferation and survival, playing a pivotal role in breast cancer development and progression. The identification of estrogen regulated genes is the first step towards elucidating the mechanisms of estrogen function. Although numerous studies have investigated the effect of estradiol at the mRNA level, assessment of global changes at the protein level has been limited mainly due to technological limitations. Here, we present the most extensive proteomic study in the quest of proteins whose expression is regulated or associated with estradiol action. To facilitate accurate quantification of proteins, we first developed and optimized a proteomic protocol for cell lysis and sample preparation, based on a mass spectrometry-compatible detergent. Following our protocol coupled to stable isotope labelling with amino acids in cell culture (SILAC) of MCF-7 breast cancer cells, we quantified approximately 4,000 proteins and identified 153 proteins differentially expressed upon estradiol stimulation. Notably, known estrogen regulated proteins such as trefoil 1(TFF1) and progesterone receptor (PGR) showed a four- and three-fold increase respectively, at 48 hours after estradiol stimulation. Forty eight of 153 differentially expressed proteins have been found to contain estrogen receptor elements (EREs) indicating direct regulation by estradiol. Protein-protein network analysis (STRING) revealed an extensive protein network related to cell proliferation. Differential expression of proteins was verified in three estrogen receptor positive breast cancer cell lines (MCF-7, HCC-1428, BT-483) using a targeted mass spectrometry-based quantitative approach; selected reaction monitoring (SRM). A multiplex SRM assay was developed for the simultaneous quantification of 56 proteins. The same assay was utilized to study the kinetics of these proteins in time and in presence of inhibitors of estrogen receptor facilitating the distinction between direct and indirect protein targets. To our knowledge, such an extensive network of estrogen-regulated genes has never been previously studied at the protein level. Interestingly, numerous differentially expressed proteins identified in the present study have not been connected to estrogen signalling or breast cancer before. The role of these proteins in breast carcinogenesis and their potential as therapeutic targets warrants further investigation. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-05-09.

Serum levels of estradiol and follicle stimulating hormone,and vasomotor symptoms in Thai women receivingGnRH analogue before laparoscopic enucleationof endometriotic cysts

Serum levels of estradiol and follicle stimulating hormone,and vasomotor symptoms in Thai women receivingGnRH analogue before laparoscopic enucleationof endometriotic cysts

Quantitative Analysis of Energy Metabolic Pathways in MCF-7 Breast Cancer Cells by Selected Reaction Monitoring Assay

To investigate the quantitative response of energy metabolic pathways in human MCF-7 breast cancer cells to hypoxia, glucose deprivation, and estradiol stimulation, we developed a targeted proteomics assay for accurate quantification of protein expression in glycolysis/gluconeogenesis, TCA cycle, and pentose phosphate pathways. Cell growth conditions were selected to roughly mimic the exposure of cells in the cancer tissue to the intermittent hypoxia, glucose deprivation, and hormonal stimulation. Targeted proteomics assay allowed for reproducible quantification of 76 proteins in four different growth conditions after 24 and 48 h of perturbation. Differential expression of a number of control and metabolic pathway proteins in response to the change of growth conditions was found. Elevated expression of the majority of glycolytic enzymes was observed in hypoxia. Cancer cells, as opposed to near-normal MCF-10A cells, exhibited significantly increased expression of key energy metabolic pathway enzymes (FBP1, IDH2, and G6PD) that are known to redirect cellular metabolism and increase carbon flux through the pentose phosphate pathway. Our quantitative proteomic protocol is based on a mass spectrometry-compatible acid-labile detergent and is described in detail. Optimized parameters of a multiplex selected reaction monitoring (SRM) assay for 76 proteins, 134 proteotypic peptides, and 401 transitions are included and can be downloaded and used with any SRM-compatible mass spectrometer. The presented workflow is an integrated tool for hypothesis-driven studies of mammalian cells as well as functional studies of proteins, and can greatly complement experimental methods in systems biology, metabolic engineering, and metabolic transformation of cancer cells.

Is there any correlation between amh and obesity in premenopausal women?

To evaluate the correlation between the levels of anti-mullerian hormone and body mass index between obese and non-obese premenopausal women. Serum anti-mullerian hormone levels of women younger than 45years admitted to our reproductive endocrinology clinic for investigation of infertility were examined in this cross-sectional study. Body mass indices were lower than 30kg/m(2) in 222 patients and equal to or higher than 30kg/m(2) in 37 patients. Levels of antimullerian hormone were analyzed in each group. Blood samples obtained from study subjects were assayed for levels of anti-mullerian hormone, follicle-stimulating hormone, luteinizing hormone, estradiol, prolactin and thyroid stimulating hormone. There was no significant difference in terms of mean age between the two groups. There was no statistically significant difference between these two groups in terms of FSH, LH, estradiol and prolactin levels. Anti-mullerian hormone levels were 3.46±2.79ng/ml and 3.79±2.93ng/ml in non-obese and obese participants, respectively. No statistically significant correlation was found between Anti Müllerian Hormone (AMH) levels and BMI levels in either group (P>0.05). Body mass index does not have an effect on serum AMH levels in women of reproductive age. Obesity has no association with levels of serum follicle stimulating hormone, luteinizing hormone, estradiol, prolactin and thyroid stimulating hormone. Obesity is unlikely to affect ovarian reserve in the premenopausal age group.

Serum Anti-müllerian hormone, follicle stimulating hormone and antral follicle count measurement cannot predict pregnancy rates in IVF/ICSI cycles

To investigate whether serum anti-müllerian hormone (AMH), follicle stimulating hormone (FSH), or antral follicle count (AFC) are predictive for clinical pregnancy in in vitro fertilization (IVF) patients. Serum AMH, inhibin B, FSH, luteinizing hormone (LH), estradiol (E2), prolactin, and thyroid stimulating hormone (TSH) levels and AFC of 189 women under 40 years of age were investigated. Pregnant and non-pregnant women were compared. Forty-seven (24.8 %) clinical pregnancies were observed in 189 women. There was no significant difference in terms of mean age, duration of infertility, body mass index, AMH, LH, FSH, E2, TSH, Inhibin B, AFC and total oocyte number between women who did and who did not become pregnant. Additionally, there was no significant difference in clinical pregnancy rates between the quartiles of AMH, FSH and AFC. (P values were 0.668, 0.071, and 0.252, respectively.) Serum AMH and FSH, and AFC cannot predict clinical pregnancy in IVF patients under 40; the pregnancy rate tends to increase as AMH increases, although this remains non-significant.

Role of LH in luteolysis and growth of the ovulatory follicle and estradiol regulation of LH secretion in heifers

The role of LH in luteolysis and development of the ovulatory follicle and the involvement of GnRH receptors in estradiol (E2) stimulation of LH secretion were studied in heifers. A pulse of PGF2α, as indicated by a metabolite, was induced by E2 treatment on Day 15 (Day 0 = ovulation) and LH concentration was reduced with a GnRH-receptor antagonist (acyline) on Days 15, 16, and 17. Blood samples were collected every 6 h on Days 14–17 and hourly for 10 h beginning at the Day-15 treatments. Four groups were used (n = 6): control, acyline, E2, and E2/acyline. The number of LH pulses/heifer during the 10 h posttreatment was greater (P < 0.0002) in the E2 group (2.3 ± 0.4, mean ± SEM) than in the acyline group (0.2 ± 0.2) and was intermediate in the E2/acyline group (1.4 ± 0.2). Concentrations of progesterone in samples collected every 6 h on Day 15 showed a group-by-hour interaction (P < 0.02); concentrations decreased in the acyline group but not in the control group. The 12 heifers in the combined acyline and E2/acyline groups had three follicular waves compared to two waves in 10 of 12 heifers in the combined control and E2 groups. Results (1) supported the hypothesis that LH delays the progesterone decrease associated with luteolysis, (2) supported the hypothesis that LH has a positive effect on the continued development and growth of the selected ovulatory follicle, and (3) indicated that E2 stimulates LH production through an intracellular pathway that involves GnRH receptors on the gonadotropes and a pathway that does not involve the receptors.

Pro-survival Effects of 17β-Estradiol on Osteocytes Are Mediated by Nitric Oxide/cGMP via Differential Actions of cGMP-dependent Protein Kinases I and II

Estrogens promote bone health in part by increasing osteocyte survival, an effect that requires activation of the protein kinases Akt and ERK1/2, but the molecular mechanisms involved are only partly understood. Because estrogens increase nitric oxide (NO) synthesis and NO can have anti-apoptotic effects, we examined the role of NO/cGMP signaling in estrogen regulation of osteocyte survival. Etoposide-induced death of MLO-Y4 osteocyte-like cells, assessed by trypan blue staining, caspase-3 cleavage, and TUNEL assays, was completely prevented when cells were pre-treated with 17β-estradiol. This protective effect was mimicked when cells were pre-treated with a membrane-permeable cGMP analog and blocked by pharmacological inhibitors of NO synthase, soluble guanylate cyclase, or cGMP-dependent protein kinases (PKGs), supporting a requirement for NO/cGMP/PKG signaling downstream of 17β-estradiol. siRNA-mediated knockdown and viral reconstitution of individual PKG isoforms demonstrated that the anti-apoptotic effects of estradiol and cGMP were mediated by PKG Iα and PKG II. Akt and ERK1/2 activation by 17β-estradiol required PKG II, and cGMP mimicked the effects of estradiol on Akt and ERK, including induction of ERK nuclear translocation. cGMP induced BAD phosphorylation on several sites, and experiments with phosphorylation-deficient BAD mutants demonstrated that the anti-apoptotic effects of cGMP and 17β-estradiol required BAD phosphorylation on Ser(136) and Ser(155); these sites were targeted by Akt and PKG I, respectively, and regulate BAD interaction with Bcl-2. In conclusion, 17β-estradiol protects osteocytes against apoptosis by activating the NO/cGMP/PKG cascade; PKG II is required for estradiol-induced activation of ERK and Akt, and PKG Iα contributes to pro-survival signaling by directly phosphorylating BAD.

P3-04-03: Identification of Hormone-Responsive Genes as Biomarkers for Menstrual Cycle Phases and Menopausal Status.

Abstract Background The definition and validation of biomarkers in archived breast tissue samples for prognostic research is limited by the fact that the exact menstrual cycle phases and menopausal status at the time of tissue sampling is often unknown or not accurate by patient survey. Biomarkers that vary with menstrual cycle phases in premenopausal women would be difficult to standardize. There are also significant differences in gene expression in pre- and post-menopausal women. Therefore, menstrual cycle fluctuation and menopausal status need to be considered for all candidate biomarkers as part of the validation process. The aim of this study is to identify genes responsive to different hormones to accurately define menstrual cycle phases and menopausal status. Methods: We studied gene expression profiles of 18 random fine-needle aspirate (rFNA) samples from unaffected contralateral breast (8 pre-menopausal, mean age 44.5; 10 post-menopausal, mean age 58.8) and investigated the correlation between gene expression and serum hormone levels of estradiol (E2), progesterone (P4) and follicle stimulating hormone (FSH). Genes that were highly correlated with the serum levels of each hormone (Pearson correlation coefficient r &amp;gt; 0.60) were considered as specific hormone-responsive genes (P &amp;lt; 0.0085). The combined gene profiles of hormone-responsive genes were used to dissect samples in different menstrual cycle phases and menopausal status. Selected genes related to mammary gland development and hormone regulation based on gene function and gene network analysis were validated using qRT-PCR in 18 original rFNA samples and in 28 independent samples. Results: From 35,964 genes and 12,838 undesignated transcripts, we identified genes/transcripts highly correlated with E2 (1091 genes), P4 (127 genes) or FSH (58 genes). The most significantly correlated genes in each group were selected to define four panels of genes: Panel A-21genes stimulated by E2 (r &amp;gt; 0.78); Panel B-22 genes stimulated by P4 (r &amp;gt; 0.75); Panel C-7 genes stimulated by FSH (r &amp;gt; 0.65); and Panel D-10 genes suppressed by FSH (r &amp;lt; −0.65). Hierarchical clustering analysis using the combination of gene panels dissected the samples into four clusters based on three phases of menstrual cycles and post-menopausal status. Specifically, high panel C and low panel D expression segregated post- from pre-menopausal samples. Low expression of panel A and B genes dissected early follicular phase from late follicular and luteal phases, while higher expression of panel B genes discriminated luteal phase from late follicular samples. Conclusion: Our results indicate that the menstrual cycle phases and menopausal status determined by age, patient survey and serum hormone concentrations are reflected in the expression of specific gene sets in the normal breast. The combination of hormone-responsive gene panels would allow the classification of breast samples regarding to the menstrual phases and menopausal status at the time of sampling. It would also facilitate the selection and validation of breast cancer biomarkers that are independent of menstrual cycle fluctuation and menopausal variation for clinical use. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-04-03.

VIP expression in vessels of the vaginal wall: relationship with estrogen status

ABSTRACTObjective Vasoactive intestinal peptide (VIP) is a neuropeptide with elevated expression in regions that control urogenital functions. Estrogen appears to modulate VIP expression in various organs, but this effect has not been demonstrated in the vaginal wall. The aim of this study was to evaluate the influence of estrogen status on VIP expression in vessels of the vaginal wall.Methods Surgical specimens were removed from the vaginal walls of 18 premenopausal women and 12 postmenopausal women who were given surgery for genital prolapse grade I or II. Vaginal specimens were stained with estrogen receptor-alpha (ER-α) and VIP antibodies. Levels of follicle stimulating hormone (FSH), estradiol, prolactin, fasting glucose and serum thyroxine stimulating hormone were also measured. Estrogen status was assessed on the basis of FSH and ER-α scores.Results The vaginal walls of premenopausal women had significantly higher ER-α scores than those of menopausal women (premenopausal group, 3.6 ± 2.2; menopausal group, 1.4 ± 1.8; p = 0.01). Premenopausal women also had significantly higher levels of VIP in the vaginal wall than menopausal women (p = 0.02). Increasing age was associated with lower level of VIP staining (odds ratio 0.88; 95% confidence interval 0.78–0.99).Conclusion Levels of ER-α and VIP expression in the posterior vaginal wall were higher in premenopausal than in menopausal women, but VIP expression was not associated with estrogen status. Age was an independent predictor of VIP staining in vaginal wall biopsies.

Hypothyroidism caused by phenobarbital affects patterns of estrous cyclicity in rats

We found that repeated treatment with phenobarbital (PB), a thyroid modulator, resulted in a persistent estrous stage in the present study. Although the effects of PB in blocking the surge release of luteinizing hormone (LH), inducing anovulation and prolonging the diestrous period has been well established, there is still no research describing the appearance of persistent estrous states in normal cycling rats dosed with PB. To further study this phenomenon, female rats exhibiting regular estrous cycle were administered an oral dose of PB for 14 consecutive days. Consecutively, vaginal smears were observed and rats from all the groups were sacrificed and serum hormone levels for prolactin, progesterone, estradiol, triiodothyronin (T3), thyroxine (T4) and thyroid stimulating hormone (TSH) were measured. Pituitary, thyroid, liver, uteri and ovaries were excised, weighed and further subjected to histological observations. We found that PB induced irregular estrous cycles, especially persistent estrus in rats. Histopathologically, the persistent estrous stages are characterized by persistent vaginal cornification in the vagina, cystic follicles and anovulation in the ovaries. Endocrinologically, serum T3 and T4 levels were significantly lower, and TSH was higher in treated-female rats compared to control females. The serum estradiol level and the estradiol/progesterone ratio tend to increase in treated-females. Furthermore, PB-treated animals with irregular estrous cycle were reduced by T4 replacement. Our data indicate that treatment with PB resulted in hypothyroidism and irregular estrous cycle, particularly a persistent estrous stage in normal cycling female rats.

Estradiol stimulation of prolactin secretion in horses: Interaction with type of secretagogue

s / Journal of Equine Veterinary Science 31 (2011) 230-356 308 [7] Bilby TR, Block J, do Amaral BC, Sa Filho O, Silvestre FT, Hansen PJ, Staples CR, Thatcher WW. Effects of dietary unsaturated fatty acids on oocyte quality and follicular development in lactating dairy cows in summer. J Dairy Sci 2006;89:3891-903. Estradiol stimulation of prolactin secretion in horses: Interaction with type of secretagogue D.L. Thompson, Jr., S.E. Clavier, P.B. Mitcham, and L.R. Earl LSU Agricultural Center, Louisiana Agricultural Experiment Station, Baton Rouge, LA, U.S.A Introduction: Combining estradiol pretreatment with administration of a dopamine antagonist, such as sulpiride or domperidone, greatly enhances prolactin secretion in horses relative to treatment with either estradiol or dopamine antagonist alone [1-3]. Kelley et al. [2] first used this combination to stimulate ovarian activity and ovulation in seasonally anovulatory mares. Given that both day-to-day secretion (blood levels) of prolactin are stimulated in the long-term (weeks), as well as the immediate release of hormone in response to a testing dose of sulpiride, it is concluded that production rates of prolactin in the pituitary gland must be greatly enhanced by the combined treatment. The current experiments were designed to determine whether secretagogues other than dopamine antagonists would interact with estradiol pretreatment in the same manner. Materials and Methods: Two experiments were conducted. The first (July, 2009) tested the effects of estradiol pretreatment in geldings on the prolactin responses to exercise, epinephrine injection, prostaglandin-F2 a (Lutalyse; animalhealth.pfizer. com) injection, and sulpiride injection. Six geldings were administered estradiol cypionate (ECP; betlabs.com) and six geldings were administered vegetable oil on day 0. All geldings were exercised (trotting for 5 min at hourly intervals for a total of 4 bouts) on day 5, were administered epinephrine at 5 mg/kg BW i.v. on day 7, were administered 10 mg Lutalyse i.v. on day 10, and were administered sulpiride (0.1 mg/kg BW i.v.) on day 12. Prolactin was measured in blood samples collected immediately before and at frequent intervals after each treatment. The second experiment (February, 2010) tested the effects of daily administration of thyrotropin releasing hormone (TRH) and(or) ECP on daily prolactin concentrations in a 2 x 2 factorial design with 4 geldings per subgroup. Half the geldings received 50 mg ECP i.m. on day 0 and the rest received vegetable oil. Starting on day 10, half of each of those groups received either daily TRH injections (1.5 mg i.m) or saline injections of similar volumes. The TRH-saline injections were repeated through day 30. Prolactin was measured in all samples with previously validated RIA [4]. Data in each experiment were analyzed via ANOVA in SAS in repeatedmeasures designs. Differences between groups within time periods were assessed by the LSD test. Results and Discussion: In the first experiment, ECP treatment increased (P 1⁄4 .07) plasma prolactin concentrations before the onset of exercise, and exercise stimulated (P < .001) plasma prolactin concentrations after each bout. There was no interaction with estradiol pretreatment. Epinephrine did not alter prolactin concentrations. Lutalyse administration stimulated (P < .001) prolactin concentrations, but there was no interaction with ECP pretreatment. Sulpiride administration stimulated (P < .001) prolactin concentrations, and there was a greater (P 1⁄4 .038) response in ECP-treated geldings relative to controls. In the second experiment, ECP administration did not affect prolactin concentrations. Injection of TRH stimulated prolactin concentrations on the first day of injection, but had no effect on prolactin concentrations otherwise. There was no interaction of ECP and TRH administration either on the prolactin response on the first day of TRH injection, or across the 20 days of treatment. Conclusion: Unlike the synergistic interaction of estradiol pretreatment and dopamine antagonist administration on prolactin secretion, there was no interaction of ECP and other types of prolactin secretagogues in the first experiment. Even though exercise and Lutalyse stimulated prolactin secretion, it was equally stimulated in ECP-treated and control geldings; subsequent administration of sulpiride revealed that the estradiol-dopamine antagonist interaction existed in the same geldings 2 days later. In some way, the mechanism of release of prolactin from estradiol-stimulated lactotropes differs for dopamine antagonists and other prolactin-releasing stimuli. This would indicate that the stimulus for prolactin secretion after exercise or Lutalyse is not a reduction in dopamine input to the pituitary gland. Unlike sulpiride or domperidone, daily TRH injection after ECP pretreatment did not stimulate long-term prolactin secretion. Thus, even though TRH is a secretagogue for prolactin, it is unable to enhance prolactin production in combination with estradiol stimulation. The practical impact of these data is that stimulation of prolactin concentrations after ECP treatment in winter, in an effort to stimulate ovarian activity in seasonally anovulatory mares, appears to be limited to dopamine antagonists.