Estrogen Receptor Content Research Articles

Immunohistochemical study of hormone receptors in endometrium with abnormal uterine bleeding

In the case of AUB, estrogen and progesterone exert their effects by acting on specific nuclei. Receptor proteins such as estrogen receptor (ER) and progesterone receptor (PR). And periodic changes in its expression have been shown in human uterine tissue. Immunohistochemistry (IHC). Immunohistochemistry using specific monoclonal antibodies estimate the content of the receptor at the cellular level and interpret their exact location and distribution.A two-year descriptive study was conducted. There were a total of 50 cases clinically diagnosed with AUB who underwent hysterectomy and endometrial curettage, and 20 non-AUB controls were included in this study. IHC was performed for ER and PR receptors in all cases and controls.The age range was 29 to 65 years and the average age of the patients was 43.21 years. Out of 50 patients, 26 were pre-menopausal and 24 were post-menopausal, and the mean hemoglobin, fertility and bleeding duration were 10.34 g/dL, 2.8 and 8.4 days per cycle. The average thickness of the endometrium was 10.5 mm in the cases and 6.25 mm in the control group. Increasing endometrial thickness was statistically significantly related to AUB and increased ER and PR HSCORE in stroma and glands (p<0.001). In terms of histomorphology, the highest number of cases was endometrial hyperplasia without atypia (27 cases), followed by early proliferative phase (7 cases). ER expression was more abundant in glands and PR expression was more in stroma. In endometrial hyperplasia with atypia, the expression of ER and PR was higher than in other endometrial stages.In addition to pelvic ultrasound and histology, immunohistochemistry. ER and PR have the advantage of allowing these hormone receptors to localize to the tissue.Endometrial aspiration samples and anxiliary tests in patient treatment allong with AUB. Endometrial hyperplasia is more common in patients with AUB. The concentration of estrogen and progesterone receptors is significantly higher. Therefore, the location and concentration of ER and PR receptors increases and this makes it possible for targeted therapies and avoid invasive surgical procedures using selective progesterone receptor modulators and progesterone antagonists in the treatment of patients with AUB and endometrial hyperplasia.

Reasons and Mechanisms of Development of Mastopathy (Review of Literature)

This review article examines the relationship between various etiological factors in the development of mastopathy. Multiple exogenous and endogenous risk factors disrupt the normal ratio of sex hormones, causing hyperestrogenism and hyperproliferation of the epithelium in breast tissue, which leads to the development of mastopathy. In the presence of polymorphism of coding genes (ESR1 and PRG), mastopathy also increases the risk of developing breast cancer. Disturbances in the ESR1 receptor gene (Estrogen Receptor 1) lead to changes in the sensitivity of receptors to hormones and even to its loss. According to the literature, polymorphism in the intronic region of the ESR1 gene is associated with the risk of fibrocystic mastopathy. The presence of polymorphism of the PRG (Progesterone Receptor Genes) gene may cause the occurrence of defective forms of PR (Progesterone Receptor) with disruption of the ligand- and hormone-binding zones, which is confirmed by the effects of progesterone in tissues. Thus, progesterone deficiency makes possible excessive proliferation of not only epithelial cells, but also the connective tissue component of the mammary gland. In this case, the expression of the progesterone receptor gene is regulated by the level of estrogen. The association of various combinations of predisposition genes indicates the presence of a specific combination of genotypes for each clinical and pathogenetic variant. In proliferative forms of mastopathy, foci with an excess content of estrogen receptors in the epithelium and high activity of cell proliferation and apoptosis are formed in the breast tissue; there is an increase in the content of alpha-estrogen receptors, a change in the relative content of alpha- and beta-estrogen receptors, A- and B-progesterone receptors; the sensitivity of the epithelium to estradiol increases; a complex of cyclin-dependent kinases, alpha-transforming and epidermal growth factors is activated; the proliferative activity of the epithelium increases; apoptosis processes are disrupted; vascularization of the parenchyma increases. All these pathological mechanisms that increase the risk of breast cancer have not yet been sufficiently studied both at the experimental and clinical level. In the treatment of mastopathy, diet therapy, psychological correction, hormonal drugs, non-hormonal drugs, such as vitamins, minerals, hepatoprotectors, diuretics, enzyme drugs, immunocorrectors, adaptogens, non-steroidal anti-inflammatory drugs, herbal remedies are used. The general disadvantages of most of the remedies used today are attempts to eliminate the causes of mastopathy rather than conducting pathogenetic treatment.

Toxicity and Estrogenicity of Bisphenol TMC in Oryzias melastigma via In Vivo and In Silico Studies.

Bisphenol 4-[1-(4-hydroxyphenyl)-3,3,5-trimethylcyclohexyl] phenol (BPTMC), as a substitute for bisphenol A, has been detected in environments. However, the ecotoxicological data of BPTMC are extremely scarce. Here, the lethality, developmental toxicity, locomotor behavior, and estrogenic activity of BPTMC at different concentrations (0.25-2000 μg/L) in marine medaka (Oryzias melastigma) embryos were examined. In addition, the in silico binding potentials of O. melastigma estrogen receptors (omEsrs) with BPTMC were assessed by docking study. Low-concentration BPTMC exposure (including an environmentally relevant concentration, 0.25 μg/L) resulted in stimulating effects, including hatching rate, heart rate, malformation rate, and swimming velocity. However, elevated concentrations of BPTMC led to an inflammatory response, changed heart rate and swimming velocity in the embryos and larvae. In the meantime, BPTMC (including 0.25 μg/L) altered the concentrations of estrogen receptor, vitellogenin, and endogenous 17 β-estradiol as well as the transcriptional levels of estrogen-responsive genes in the embryos or/and larvae. Furthermore, elaborate tertiary structures of omEsrs were built by ab initio modeling, and BPTMC exerted potent binding potential with three omEsrs with -47.23, -49.23, and -50.30 kJ/mol for Esr1, Esr2a, and Esr2b, respectively. This work suggests that BPTMC has potent toxicity and estrogenic effects in O. melastigma.

Evaluation of Serum Hormones and Biochemical Tumor Markers among Breast Cancer Patients in the South-South Region, Nigeria

Breast cancer remains one of the most frequent malignancies in women and the primary cause of cancer-related death in women all over the world. Hormones and tumor biomarkers have been implicated as possible causes and prognostic biomarkers of breast cancer. The present study investigated the use of serum hormonal levels and tumor biomarkers; Carcinoembrvonic antigen (CEA), and cancer antigen (CA 15-3) for the diagnosis of Breast Cancer, and generation of different subtypes using Estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). This study is a cross-sectional study comprising 120 subjects; sixty breast cancer patients, and sixty apparently healthy women who served as control. One hundred and twenty blood samples were collected and analyzed for estrogen, progesterone, Carcinoembrvonic antigen, and cancer antigen. The results showed that the peak age of incidence of breast cancer was 40-49 years. Majority (56.7%) of the cases were menopausal women, while 43.3% were in their premenopausal period. Serum estrogen, cancer antigen, and Carcinoembrvonic antigen concentrations were elevated significantly (p<0.05) in the breast cancer patients than the control group. Premenopausal patients have higher progesterone, estradiol, Carcinoembrvonic antigen, and cancer antigen levels than those during the postmenopausal period. HER2 negative subtype (50.0%) is the most prevalent subtype of breast cancer. 8.4% were triple negative (ER–/PR–/HER2–) cases, while 1.7% were triple positive (ER+/PR+/HER2+) cases. Breast cancer is strongly associated with hormonal changes especially estrogen and progesterone during the premenopausal and postmenopausal periods in women. Thus, the determination of the concentration of estrogen, progesterone, Carcinoembrvonic antigen, cancer antigen, and human epidermal growth factor receptor 2 statuses could help in the prognosis of breast cancer.

Investigation of the infertility structure and outcomes of ART programs in patients of late reproductive age

Purpose: comparative study of the structure of infertility and outcomes of ART programs among patients of different age groups. Materials and methods: the case histories of 180 patients with infertility were studied: Group I ≥ 35 years old; Group II <35 years old. Laboratory and immunohistochemical studies were carried out, and the outcomes of ART were assessed. Results: in group I, a shortened menstrual cycle was determined in comparison with group II (27,15 ± 3,39 days vs 29,57 ± 2,38 days, p = 0,001). Also, in group I, the following were more often found: inflammatory diseases (p = 0,05); history of unilateral tubectomy (p = 0,019); increased FSH levels (9,73 ± 2,43 vs 8,74 ± 2,50); decrease in the concentration of LH, progesterone, AMH (p <0,05). In the cells of the aspirate of the uterine cavity in patients of group I, there was an increased content of progesterone receptors and a decreased concentration of estrogen receptors (p <0,05). Patients of group I received a smaller number of oocytes (8,34 ± 3,51 vs 10,78 ± 4,37) and quality embryos by the 5th day of cultivation (82,7% vs 87,97%; p <0,05). The number of pregnancies in groups I and II was 22,22% and 36,67%, respectively, and live births – 14,44% and 27,78% (p <0,05). Patients who gave birth had increased progesterone levels, greater endometrial thickness, more oocytes with transvaginal puncture, and high-quality embryos. Conclusion: the factors that reduce fertility were: genital pathology, inhibition of ovarian function, depletion of the follicular reserve. Fewer live births are associated with defects in embryonic and implantation factors.

Short-Term Supplementation With Fermented Red Clover Extract Reduces Vascular Inflammation in Early Post-menopausal Women.

The decline in estrogen at menopause poses a critical challenge to cardiovascular and metabolic health. Recently, a growing interest in the role of phytoestrogens, with a particular focus on isoflavones, has emerged as they can bind to estrogen receptors and may mimic the roles of endogenous estrogen. Fermented red clover extract (RC) contains isoflavones with superior bioavailability compared to non-fermented isoflavones, however little is known regarding the impact of isoflavones on cardiovascular and metabolic health. We assessed markers of vascular health in plasma and skeletal muscle samples obtained from healthy but sedentary early post-menopausal women (n = 10; 54 ± 4 years) following 2 weeks of twice daily treatment with placebo (PLA) or RC (60 mg isoflavones per day). The two interventions were administered using a randomized, double-blind, crossover design with a two-week washout period. Plasma samples were utilized for assessment of markers of vascular inflammation. There was a statistically significant reduction (~5.4%) in vascular cell adhesion molecule 1 (VCAM-1) following 2 weeks of RC supplementation compared to PLA (p = 0.03). In contrast, there was no effect of RC supplementation compared to PLA on skeletal muscle estrogen receptor content and enzymes related to vascular function, and angiogenesis. Supplementation with RC reduces vascular inflammation in early post-menopausal women and future studies should address the long-term impact of daily supplementation with RC after menopause.

Androgen and oestrogen receptors in the growing antlers velvet of adult and yearling pampas deer (Ozotoceros bezoarticus) males.

To determine the presence, quantity and distribution of androgen (AR) and oestrogen receptors (ER) in the antler velvet (AV), samples were collected from adult and yearling pampas deer males, as well as skin from the ventrolateral abdominal area (ASK). Samples were analysed with ligand-binding assays for AR and ER activity and processed for AR and ERβ immunohistochemistry. There was more content of AR in adults than in yearling males (p=0.02), as well as a category and type of tissue interaction (p=0.03). There was more ER content in adults than in yearling males (p=0.005) and in the AV than in the skin (p=0.0001). The AR-positive score (PS) was greater in AV than in ASK in the surface stroma (p=0.0007). In the intermediate epidermis, the AR PS was greater in adults than in yearling males (p=0.04) and in the ASK than in the AV (p<0.0001). There was a male category and type of tissue interaction for AR PS in the sebaceous glands (p=0.014). The ERβ PS in the surface stroma was greater in ASK than in AV (p=0.004) and tended to be greater in yearling than in adult males (p=0.093). The ERβ PS in the intermediate epithelium and the sebaceous glands was greater in adults than in yearlings (p=0.004 and p=0.007, respectively). In conclusion, we reported for the first time the presence of AR and ER in the velvet skin of growing antlers in pampas deer males. Therefore, the velvet skin is sensitive to both androgens and oestrogens. Furthermore, the greater content of oestrogen receptors in the velvet of adult males suggests that adults are more sensitive to this hormone than yearlings, and thus, oestrogens have greater importance in velvet activity regulation in adult males.

Does and duration of exposure time dependent regulation of proteins in the urinary bladder of hamsters exposed to sodium arsenite

Does and duration of exposure time dependent regulation of proteins in the urinary bladder of hamsters exposed to sodium arsenite

Regulation of transgelin and GST-pi proteins in the tissues of hamsters exposed to sodium arsenite

Regulation of transgelin and GST-pi proteins in the tissues of hamsters exposed to sodium arsenite

Resting skeletal muscle PNPLA2 (ATGL) and CPT1B are associated with peak fat oxidation rates in men and women but do not explain observed sex differences

What is the central question of this study? What is the relationship between proteins in skeletal muscle and adipose tissue determined at rest and at peak rates of fat oxidation in men and women? What is the main finding and its importance? The resting contents of proteins in skeletal muscle involved in triglyceride hydrolysis and mitochondrial lipid transport were more strongly associated with peak fat oxidation rates than proteins related to lipid transport or hydrolysis in adipose tissue. Although females displayed higher relative rates of fat oxidation than males, this was not explained by the proteins measured in this study, suggesting that other factors determine sex differences in fat metabolism. We explored key proteins involved in fat metabolism that might be associated with peak fat oxidation (PFO) and account for sexual dimorphism in fuel metabolism during exercise. Thirty-six healthy adults [15 women; 40±11years of age; peak oxygen consumption 42.5±9.5ml(kg body mass)-1 min-1 ; mean±SD] completed two exercise tests to determine PFO via indirect calorimetry. Resting adipose tissue and/or skeletal muscle biopsies were obtained to determine the adipose tissue protein content of PLIN1, ABHD5 (CGI-58), LIPE (HSL), PNPLA2 (ATGL), ACSL1, CPT1B and oestrogen receptorα (ERα) and the skeletal muscle protein content of FABP 3 (FABPpm), PNPLA2 (ATGL), ACSL1, CTP1B and ESR1 (ERα). Moderate strength correlations were found between PFO [in milligrams per kilogram of fat-free mass (FFM) per minute] and the protein content of PNPLA2 (ATGL) [rs =0.41 (0.03-0.68), P<0.05] and CPT1B [rs =0.45 (0.09-0.71), P<0.05] in skeletal muscle. No other statistically significant bivariate correlations were found consistently. Females had a greater relative PFO than males [7.1±1.9 vs. 4.5±1.3 and 7.3±1.7 vs. 4.8±1.2mg(kg FFM)-1 min-1 in the adipose tissue (n=14) and skeletal muscle (n=12) subgroups, respectively (P<0.05)]. No statistically significant sex differences were found in the content of these proteins. The regulation of PFO might involve processes relating to intramyocellular triglyceride hydrolysis and mitochondrial fatty acid transport, and adipose tissue is likely to play a more minor role than muscle. Sex differences in fat metabolism are likely to be attributable to factors other than the resting content of proteins in skeletal muscle and adipose tissue relating to triglyceride hydrolysis and fatty acid transport.

Sex Difference in the Tolerance of Hepatic Ischemia-reperfusion Injury and Hepatic Estrogen Receptor Expression According to Age and Macrosteatosis in Healthy Living Liver Donors.

Hepatic estrogen signaling, which is important in liver injury/recovery, is determined by the level of systemic estrogen and hepatic estrogen receptor. We aimed to evaluate whether females' advantage in the tolerance of hepatic ischemia-reperfusion injury decreases according to the age of 40 y (systemic estrogen decrease) and macrosteatosis (hepatic estrogen receptor decrease). We included 358 living liver donors (128 female and 230 male individuals). The tolerance of hepatic ischemia-reperfusion injury was determined by the slope of the linear regression line modeling the relationship between the duration of intraoperative hepatic ischemia and the peak postoperative transaminase level. Estrogen receptor content was measured in the biopsied liver samples using immunohistochemistry. In the whole cohort, the regression slope for aspartate transaminase was comparable between female and male individuals (P = 0.940). Within the subgroup of donors aged ≤40 y, the regression slope was significantly smaller in female individuals (P = 0.031), whereas it was comparable within donors aged >40 y (P = 0.867). Within the subgroup of nonmacrosteatotic donors aged ≤40 y, the regression slope was significantly smaller in female individuals in univariable (P = 0.002) and multivariable analysis (P = 0.006), whereas the sex difference was not found within macrosteatotic donors aged ≤40 y (P = 0.685). Estrogen receptor content was significantly greater in female individuals within nonmacrosteatotic donors aged ≤40 y (P = 0.021), whereas it was not different in others of age >40 y or with macrosteatosis (P = 0.450). The tolerance of hepatic ischemia-reperfusion injury was greater in female individuals than in male individuals only when they were <40 y and without macrosteatosis. The results were in agreement with the hepatic estrogen receptor immunohistochemistry study.

Abstract PS5-46: Introduction and clinical validation of metrology standards for immunohistochemistry (IHC); New tool for standardization of estrogen receptor (ER) IHC assay in breast cancer

Abstract Traceability of measurement to a higher order reference standard is a foundation of laboratory testing. There is as yet no method for creating reference standards for cellular proteins in situ in an analogous fashion as for soluble analytes. At present, IHC laboratories produce results for breast cancer hormone receptors without connection to a reference standard. Not surprisingly, high rates of testing variation as well as discrepancies among IHC laboratories have been reported. To address this need, we developed a system of measurement traceability using a linked fluorescein tag for creating reference standards for any cellular analyte and, as a first test, validate it for estrogen receptor (ER) testing. In this study, the newly developed ER standard defines and compares the thresholds separating “high positive”, “low positive”, and “negative” tests according to updated ASCO/CAP guidelines as detected by clinical IHC laboratories in a national external quality assessment survey. This reference standard utilizes NIST Standard Reference Material (SRM) 1934 as a universal IHC standard. We calculated ER concentration based on a linked fluorescence measurement traceable to NIST SRM 1934 as each ER is linked to a single fluorescein, and fluorescein concentration equals ER concentration. Each laboratory’s lowest detected ER concentration (i.e. “limit of detection”, LOD) was compared to their results with 80 tumor samples enriched for triple negative breast cases. For the Canadian Immunohistochemistry Quality Control (CIQC) ER proficiency testing run, calibrator sets with peptides for the SP1, EP1, and 6F11 epitopes were created. The various concentrations were pipetted onto histology slides used by CIQC to place its 80-case breast cancer tissue microarray. These slides were stained by participating laboratories using their routine ER IHC protocols and returned to the CIQC. For the purpose of this study, Histology Score and ASCO/CAP categorical scoring recommendation was used for the readout. Results with SP1 clone are reported here because it was employed by overwhelming majority of laboratories. A total of 3,038 readouts were included in the analysis. Most IHC laboratories had a LOD between 10,000 – 25,000 molecules ER per microbead. Highly sensitive ER assays (low LODs) detected more positive cases while those with poorly sensitive assay detected fewer. The LOD correlated with the percent positive cases (R2= -.767, p &amp;lt; 0.0001, Spearman Correlation) as well as with cumulative individual laboratory Histology Score (R2= -.612, p &amp;lt; 0.0001, Spearman Correlation). The tumor samples accounting for this difference were principally “low positive” (ASCO/CAP classification) while ER-high positive tumors were less affected. Although the concept of a non-quantitative IHC LOD has been introduced with critical assay performance controls (iCAPCs, e.g. germinal center cells in the tonsil for ER), there were no tools until now to actually measure LOD for ER IHC testing. With the calibrators introduced here, it is now possible to measure the LOD for ER IHC assays. Furthermore, our data show that ER low positive and some cases of ER high positive breast cancers are highly affected by the variability of analytic sensitivity between clinical IHC laboratories. These data argue for the urgent need to standardize testing to a defined analytic sensitivity range, which is now possible for the first time. Beyond inter-laboratory standardization, known measured LOD also enables: i) methodology transfer from clinical trials to clinical laboratories; ii) determination of optimal analytical sensitivity to produce assays with highest clinical and analytical validity, and iii) daily monitoring of analytical sensitivity for each tested IHC slide. Citation Format: Emina Emilia Torlakovic, Seshi R. Sompuram, Kodela Vani, Lili Wang, Anika K Schaedle, Paul C. DeRose, Steven A. Bogen. Introduction and clinical validation of metrology standards for immunohistochemistry (IHC); New tool for standardization of estrogen receptor (ER) IHC assay in breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS5-46.

38P Frequent changes in the environment can lead to heterogeneity of tumour cells

The mouse mammary tumors developed on the background of changes in the hormonal environment during several cycles of estrus differ from tumors of mice developed during a stable high or stable low hormonal environment. The difference is that the changing hormonal background leads to heterogeneity in the level of estrogen and progesterone receptors in tumors. Female C3H/Sn mice were used in this study. Transplantable mammary tumors were received from spontaneous mammary tumors C3H/Sn mice. ER-positive (ER +) adenocarcinomas in 90% of cases were obtained from mice at the age of 12 months with intensive reproduction (giving birth at least 4-5 times). The level of ER was determined by means of the dextran-coated charcoal technique with the use of 3H-estradiol. The level of receptors protein in mice tumors developed with stable high level of hormones reaches up the range higher than 20,0 fmol/mg of protein up to 110 fmol/mg. The transplanted mice mammary tumors which developed against the environment of high level of estrogens (frequent parturition without mammary feeding of off-springs) contains high level of estrogen receptors. The hormonal environment of the organism determines the level of receptors in the tumors at the end of their development independent of the level of receptors in tumor before transplantation. The wavy development of tumor depending from oestrus phase of cycles during 3-5 cycles is observed in transplanted mice tumors with normal oestrus cycle (cycle changes of hormonal environment). The change of hormones level which is observed during 4-daily of mice oestrus cycle is capable to cyclic influence on change of volume under progressive growth of receptor-positive tumors. The growth loses the wavy feature after 5-th cycle. At the end of tumor development approximately half of cells lose the receptors. Thus, the population become heterogeneous in the content of estrogen receptors. Heterogenization of tumors indicates on the one hand the harmful properties of tumors to adapt to new conditions. On the other hand, the high sensitivity of tumors to changes in the environment gives hope for multi-stage therapy.

The content of gonadotropin-releasing hormone (GnRH), kisspeptin, and estrogen receptors (ERα/ERβ) in the anteromedial hypothalamus displays daily variations throughout the rat estrous cycle.

The content of gonadotropin-releasing hormone (GnRH), its mRNA, and estrogen receptor alpha (ERα) and beta (ERβ) in the hypothalamus varies throughout the estrous cycle. Furthermore, the abundance of these molecules displays asymmetry between the right and left side. In the present study, we investigated the changes in the content of ERα, ERβ, kisspeptin, and GnRH by western blot in the left and right anteromedial hypothalamus, at four different times during each stage of the rat estrous cycle. The serum levels of the follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were also measured. ERα and ERβ levels changed depending on the stage of the estrous cycle, meanwhile that of kisspeptin was modified according to both the hour of the day and the stage of the cycle. Except in estrus day, ERβ was higher in the right hypothalamus, while ERα was similar in both sides. During both proestrus and estrus, the content of kisspeptin and GnRH was higher in the right hypothalamus. The highest levels of FSH and LH occurred at 17:00h of proestrus. But at estrus, the highest FSH levels were observed at 08:00h and the lowest at 17:00h. Thus, the current results show that the content of ERα, ERβ, kisspeptin, and GnRH in the anteromedial hypothalamus are regulated as a function of the stage of the estrous cycle and the hour of the day. Furthermore, the content of these proteins is regularly higher in the right anteromedial hypothalamus, regardless of the stage of the cycle or time of the day.

Molecular Mechanisms Underpinning The Regulation Of Peak Fat Oxidation Rates During Exercise

PURPOSE: The molecular regulation of peak fat oxidation (PFO) during exercise remains poorly characterized. The aim of this study was to examine the relationship between the content of key proteins involved in adipose tissue and skeletal muscle fat metabolism with PFO. METHODS: Thirty-six healthy men and women adults [15 females; mean (SD) age 40 (11) years; V̇O2peak 42.5 (9.5) mL·kg BM-1·min-1; body fat %: 21.8 (8.2) %] completed two incremental exercise tests (separated by 7-28 days) to determine PFO via indirect calorimetry. A DEXA scan and adipose tissue and/or skeletal muscle biopsies were obtained 2-7 days after the second exercise test to determine the protein content of PLIN1, CGI-58, ATGL, HSL, ACSL1, and oestrogen receptor α (ERα) in adipose tissue, and FABPpm, ATGL, ACSL1, CTP1b and ERα in skeletal muscle. Sex comparisons were performed on sub-groups of males and females matched for aerobic capacity relative to fat free mass and classifications of the physical activity level index and fat mass index (n = 14 and 12 for adipose tissue and skeletal muscle comparison sub-groups, respectively). RESULTS: Moderate strength correlations were found between PFO (mg·kg FFM-1·min-1) and the protein content of ATGL [r = 0.41 (0.05 - 0.68), p < 0.05] and CPT1b [r = 0.41 (0.05 - 0.68), p < 0.05] in skeletal muscle. No other statistically significant bivariate correlations were found between PFO and the content of proteins in adipose tissue or skeletal muscle. Females had a greater PFO compared to males when expressed relative to fat-free mass [mean (SD): 7.1 (1.9) and 7.3 (1.7) vs 4.5 (1.3) and 4.8 (1.2) mg·kg FFM-1·min-1 in the adipose tissue and skeletal muscle-sub-groups, respectively, p < 0.05]. No statistically significant sex differences were found in the content of any of the measured proteins involved in lipid metabolism in adipose tissue or skeletal muscle. CONCLUSIONS: The molecular regulation of PFO may primarily lie within skeletal muscle rather than adipose tissue, involving processes relating to intramyocellular triglyceride hydrolysis (ATGL) and mitochondrial fatty acid transport (CPT1b). Future studies should explore alternative molecular mechanisms that may account for sexual dimorphism in exercise fuel metabolism.

The relationship between estrogen-induced phenotypic transformation and proliferation of vascular smooth muscle and hypertensive intracerebral hemorrhage.

BackgroundTo explore the effect of estrogen on human cerebral vascular smooth muscle cells (VSMCs) and to clarify the molecular mechanism of estrogen inhibition of VSMC proliferation, which could provide an important reference basis for the clinical treatment of hypertensive intracerebral hemorrhage.MethodFirstly, the effects of different concentrations of estradiol and estrogen receptor (ESR) blocker (tamoxifen) on the proliferation of human VSMCs and the expression of estrogen-related receptor gene (ESR: ESR1, ESR2, GPER), myocardin (MYOCD), serum reaction factor (SRF), and apoptosis gene caspase-3 were measured to discover the effect and mechanism of tamoxifen on the proliferation and apoptosis of VSMCs. Secondly, the effects of estradiol on human VSMCs treated with angiotensin II (Ang II) were observed by measuring the expression of vascular smooth muscle markers, α-smooth muscle actin (α-SMA), SM22α, FLN, MCP-1, and TLR4.ResultsEstradiol inhibited the proliferation of VSMCs by upregulating the expression of ESR1, ESR2, and GPER and downregulating the expression of caspase-3, MYOCD, and SRF, thereby inhibiting the apoptosis of vascular smooth muscle. At the same time, tamoxifen had opposite effects. Angiotensin II decreased the expression of α-SMA and SM22α and promoted the expression of FLN, MCP-1, and TLR4 protein, while estrogen had the opposite effects.ConclusionsEstrogen suppresses apoptosis by inhibiting the proliferation of human VSMCs and preventing it from changing from contractile to synthetic. Estrogen can further prevents vascular damage and regulate peripheral inflammatory reaction, thereby producing a protective effect on cardiovascular and cerebrovascular.

Development of bivalent triarylalkene- and cyclofenil-derived dual estrogen receptor antagonists and downregulators

Up to 80% of mammary carcinoma initially exhibit estrogen-dependent growth, which can be treated by aromatase inhibitors or SERMs/SERDs. To increase the options after failure of the hormonal therapy with these drugs, the search for alternatives with a different mode of action to prevent estrogen action is of high relevance. Therefore, this study focused on the inhibition of coactivator recruitment at the estrogen receptor (ER) by targeted attachment of bivalent compounds at the coactivator binding site besides the primary binding at the ligand binding domain. Eight homodimeric 4-[1-(4-hydroxyphenyl)-2-phenyl-1-butenyl]cinnamic acid (GW7604)- or cyclofenilacrylic acid-based ER ligands with diaminoalkane linkers (C2–C5) were synthesized and their effects on the ER subtypes were assessed in vitro. All compounds possessed full antagonistic potency at ERα/β as determined in a transactivation assay. Furthermore, they exerted medium downregulatory effects dependent on the spacer length and did not stimulate the ER expression as observed for 4-hydroxytamoxifen. The cyclofenil-derived dimer with C4 spacer (15b) showed the highest binding affinity to ERα (RBA = 79.2%) and downregulated the ER content in MCF-7 cells with an efficiency of 38% at 1 μM.

Comparison of Clinico-Pathological Presentations of Triple-Negative versus Triple-Positive and HER2 Iraqi Breast Cancer Patients.

BACKGROUND:Breast cancer remains the most common malignancy among the Iraqi population. Affected patients exhibit different clinical behaviours according to the molecular subtypes of the tumour.AIM:To identify the clinical and pathological presentations of the Iraqi breast cancer subtypes identified by Estrogen receptors (ER), Progesterone receptors (PR) and HER2 expressions.PATIENTS AND METHODS:The present study comprised 486 Iraqi female patients diagnosed with breast cancer. ER, PR and HER2 contents of the primary tumours were assessed through immunohistochemical staining; classifying the patients into five different groups: Triple Negative (ER/PR negative/HER2 negative), Triple Positive (ER/PR positive/HER2 positive), Luminal A (ER/PR positive/HER2 negative), HER2 enriched ((ER/PR negative/HER2 positive) and all other subtypes.RESULTS:The major registered subtype was the Luminal A which was encountered in 230 patients (47.3%), followed by the Triple Negative (14.6%), Triple Positive (13.6%) and HER2 Enriched (11.5%). Patients exhibiting the Triple Negative subtype were significantly younger than the rest of the groups and presented with larger size tumours. A significant difference in the distribution of the breast cancer stages was displayed (p < 0.05); the most advanced were noted among those with HER2 enriched tumours who exhibited the highest frequency of poorly differentiated carcinomas and lymph node involvement.CONCLUSION:The most significant variations in the clinicopathological presentations were observed in the age and clinical stage of the patients at diagnosis. Adoption of breast cancer molecular subtype classification in countries with limited resources could serve as a valuable prognostic marker in the management of aggressive forms of the disease.

Kisspeptin/Neurokinin B/Dynorphin (KNDy) cells as integrators of diverse internal and external cues: evidence from viral-based monosynaptic tract-tracing in mice

Neurons in the hypothalamic arcuate nucleus (ARC) that co-express kisspeptin, neurokinin B and dynorphin (KNDy cells) are essential for mammalian reproduction as key regulators of gonadotropin-releasing hormone (GnRH) secretion. Although multiple endogenous and exogenous signals act indirectly via KNDy neurons to regulate GnRH, the identity of upstream neurons that provide synaptic input to this subpopulation is unclear. We used rabies-mediated tract-tracing in transgenic Kiss1-Cre mice combined with whole-brain optical clearing and multiple-label immunofluorescence to create a comprehensive and quantitative brain-wide map of neurons providing monosynaptic input to KNDy cells, as well as identify the estrogen receptor content and peptidergic phenotype of afferents. Over 90% of monosynaptic input to KNDy neurons originated from hypothalamic nuclei in both male and female mice. The greatest input arose from non-KNDy ARC neurons, including proopiomelanocortin-expressing cells. Significant female-dominant sex differences in afferent input were detected from estrogen-sensitive hypothalamic nuclei critical for reproductive endocrine function and sexual behavior in mice, indicating KNDy cells may provide a unique site for the coordination of sex-specific behavior and gonadotropin release. These data provide key insight into the structural framework underlying the ability of KNDy neurons to integrate endogenous and environmental signals important for the regulation of reproductive function.

Androgenic regulation of sexually dimorphic expression of RNA binding motif protein 48 in the developing mouse cortex and hippocampus

To further reveal the molecular mechanism underlying sexual differentiation of the mouse cerebral cortex and hippocampus, we reanalyzed our previous microarray study with Gene Ontology (GO) term enrichment and found that the GO term “RNA binding” was over-represented among the 89 sexually dimorphic candidate genes. Thus, we selected 16 autosomal genes annotated to the term RNA binding and profiled their mRNA expression in the developing male and female mouse cortex/hippocampus. During the first three weeks after birth, sex differences in mRNA levels of Khdrbs2, Nanos2, Rbm48, and Tdrd3 were observed in the mouse cortex/hippocampus. Of these genes, only the female-biased expression of Rbm48 in neonates was abolished by prenatal exposure to testosterone propionate (TP), while postnatal treatment of TP three weeks after birth increased Rbm48 and Tdrd3 mRNA levels in both sexes. Regardless of sex, the postnatal cortex/hippocampus also showed a marked increase in the content of androgen receptor (Ar) and estrogen receptor β (Esr2), but a decrease in estrogen receptor α (Esr1) and aromatase (Cyp19a1), which might confer the different responses of Rbm48 to prenatal and postnatal TP. Our results suggest that androgen-regulated, sexually dimorphic Rbm48 expression might present a novel molecular mechanism by which perinatal androgens control development of sexual dimorphism in cortical and hippocampal structure and function.