Number Of Progesterone Receptors Research Articles

Association between pathological characteristics and recurrence score by OncotypeDX in resected T1-3 and N0-1 breast cancer: a real-life experience of a North Hungarian regional center.

Introduction: The 21-gene analysis (OncotypeDX) is validated test for pT1-3, pN0-1 with hormone receptor (HR) positive and normal expression of human epidermal growth factor receptor-2 (HER2) breast cancer (BC) to determine the aggressiveness of the disease based on the calculation of Recurrence Score (RS). Methods: In this retrospective study the authors correlated pathological characteristics and Recurrence Score (RS) by traditional statistical methods and Observed Oriented Modeling (OOM) in a realistic cohort of BC patients. Results: OncotypeDX tests were performed in 94 tumour specimens of 90 BC patients. >83% of node-negative (pN0) and >72% of node-positive (pN1) cases could avoid chemotherapy. For pN0 cases, non-parametric correlation and tests demonstrated significant association in eight types of characteristics [progesterone receptor (PR) expression, Ki-67 value, Ki-67 group, PR group, grade, estrogen receptor (ER) expression, Nottingham Prognostic Index (NPI) and Clinical Risk]. For pN1 cases, parametric correlation and tests showed significant association in six characteristic types (number of positive nodes, ER and PR expression, PR group, Ki-67 group and NPI). Based on OOM for pN0 cases, significant associations were established in three characteristics (Ki-67 group, grade and NPI group). For pN1 cases OOM found significant associations in seven characteristics (PR group, PNI, LVI, Ki-67 group, grade, NPI group and number of positive nodes). Conclusion: First in oncology, OOM was applied, which found some other significant characteristics associated with RS than traditional statistical methods. There were few patients, where no clinical associations were found between characteristics and RS contrary to statistically significant differences. Therefore, the results of these statistical analyses can be neither applied for individual cases nor able to provide the bases for screening patients, i.e., whether they need for OncotypeDX testing or not. OncotypeDX still provides a personalised approach in BC.

Locoregional recurrences of breast cancer after reconstructive plastic and organ-preserving surgery

Objective. To analyze locoregional relapses (LRR) after reconstructive plastic and organ-preserving operations in breast cancer (BC)Materials and methods. A retrospective analysis of the results of the treatment of 303 patients with BC who underwent reconstructive plastic and organ-preserving operations in the period from 2014 to 2019 in the Oncological Department of General Oncology and Rehabilitation of Gomel Regional Clinical Oncological Dispensary was carried out. The median age was 44 years (25-70 years). Statistical data processing was carried out using the application software package “Statistica”, 10.0. To characterize the surveyed groups, a standard methodological approach based on the calculation of descriptive statistics data was used. In order to statistically assess the relationship between the frequency of relapses depending on the stage and the molecular biological type of neoplasm, the Spearman correlation coeffi cient was used. The accepted level of statistical signifi cance (p) was 0.05.Results. Locoregional relapses were recorded in 11 (3.6%) patients, of which local — in 8 (2.6%), regional — in 3 (1.0%). The minimum period of development of LRR was 15 months, the maximum was 74 months; the median period of occurrence of LRR was 47 months. Depending on the stage of breast cancer, relapses developed: at stage I — in 4 (3.7%) cases, II — 5 (3.5%), III — 2 (4.2%). Depending on the molecular biological subtype of the neoplasm, LRR were distributed as follows: with luminal A — 1 (1.3%) case, luminal In HER2-negative — 1 (0.9%), luminal In HER2-positive — 6 (9.7%), non-luminal HER2-positive 1 (6.3%), three times negative — 2 (5.6%). Discordance of the receptor status was detected in 8 (72.7%) patients with recurrent tumor, most often due to the loss or decrease in the number of progesterone receptors (PR). The level of tumor-infi ltrating lymphocytes (TILs) in primary neoplasm ranged from 4 to 12%, in recurrent tumors it remained low: 5-10%.Conclusion. Given the heterogeneity of BC, the risk of developing LRR depends on many factors. LRR developed in luminal HER2-positive cancer in 9.7% of cases (p 0.05), stage III breast cancer — 4.2% (p 0.05). One of the important predictive factors is the evaluation of TILs. We noted a low level of TILs in both primary and recurrent tumors. There is a high discordance in the receptor status — 72.7%, which is important to take into account when prescribing systemic therapy.

Abstract 3162: Macrophage-derived C/EBPb regulates adult mammary gland homeostasis

Abstract Throughout mammary gland development, there are distinct stages that are “hot spots” of breast cancer risk. During these critical windows, tissue resident macrophages function to maintain the homeostatic balance within the stromal microenvironment and the ductal epithelium, and alterations in these interactions disrupt tissue homeostasis and impact breast cancer risk. However, the signaling and mechanisms by which macrophages regulate mammary gland homeostasis are poorly understood. Our recent studies (and others) have identified transcriptional profiles of distinct tissue resident macrophage subpopulations in the mammary gland. We found that Cebpb, a transcription factor important for myeloid cell differentiation and mammary gland development, is highly expressed in Cx3cr1+ ductal macrophages and localizes to macrophages in close contact with the epithelial ducts. Genetic ablation of C/EBPβ in CSF1R-expressing macrophages results in deficient alveolar budding during diestrus, while branching morphogenesis was normal. Interestingly, this phenotype is also seen in macrophage null models and suggests that C/EBPβ in macrophages plays a crucial role in signaling to the mammary epithelium. Analysis of serum hormone levels show that circulating estrogen and progesterone levels are not changed in CebpbΔMφ mice, however, the number of progesterone receptor (PR)-expressing cells are significantly increased in the ductal epithelium, suggesting altered local progesterone signaling. Further examination of the mammary epithelium shows alterations in a number of Wnt family members, including Wnt1, Wnt2b, Wnt3a and Axin2, indicative of decreased canonical Wnt signaling. Importantly, canonical Wnt signaling has been shown to be critical for regulating mammary stem cell expansion during diestrous budding. Analysis of the C/EBPβ-null macrophages show decreased expression of TGFβ, which is known to signal to basal epithelial stem cells to induce Wnt expression. Finally, other signals, such as Notch 2, which have important roles in the mammary gland development are altered. Ongoing studies are aimed at delineating how macrophage-derived C/EBPβ facilitates mammary stem cell function during the estrous cycle in a progesterone-independent manner. The results of these studies will have critical implications for understanding how the signaling between macrophages and the ductal epithelium regulate tissue homeostasis and the inherent dysregulations that lead to tumorigenesis. Citation Format: Michelle D. Rojo, Caitlin M. Burke, Alexa M. Dow, Kathryn L. Schwertfeger, Heather L. Machado. Macrophage-derived C/EBPb regulates adult mammary gland homeostasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3162.

RETROSPECTIVE STUDY CORRELATION BETWEEN GRADE AND AMOUNT OF MITOSIS WITH ER, PR, AND HER2 EXPRESSION IN INVASIVE BREAST CARCINOMA OF NO SPECIAL TYPE

Background: In 2018 there were about 24.2% new cases of breast carcinoma worldwide. Molecular breast carcinoma, like expression estrogen receptor (ER), progesterone receptor (PR), and Human Epidermal Growth Factor Receptor2 (HER2) have a higher sensitivity than morphological feature, like tumour grade and mitotic count as prognostic factors. The grade of the tumor reflects how similar these cells are to normal breast epithelial cells. The dominant growth pathway in normal breasts predominantly involves ER and PR, so tumours with ER and PR positivity show lower grade and proliferation activity. In contrast, tumours with ER and PR negativity often have more severe mutation, like overexpression of HER2. Objective: To analyze the correlation between grade and mitotic count with ER, PR, and HER2 expression. Methods: Analytical observational study with a retrospective design. This study used medical record data from ER, PR, and HER2 immunohistochemical examinations as well as histopathological grade and mitotic data for invasive breast carcinoma of no special type in the Anatomical Pathology Laboratory of Dr. Soetomo Surabaya for the period January 1, 2017 - December 31, 2019. Furthermore, the analysis of the relationship between grade and mitotic count was carried out with the expression of ER, PR, and HER2. Results: There was a significant negative relationship between grade and ER with rs = -0.170 (p = 0.000), the number of mitosis and ER with rs = -0.010 (p = 0.001), grade and PR with rs = -0.168 (p = 0.000), and the number of mitosis and PR with rs = -0.110 (p = 0.000). There was no significant relationship between grade and HER2 (p = 0.470) and the number of mitosis and HER2 (p = 0.279). Conclusion: There is a negative significant relationship between grade and amount of mitosis with ER and PR expression. There is no significant relationship between grade and amount of mitosis with HER2 expression.

The Influence of Pregnancy on Female Prostate Morphophysiology in Gerbils (Meriones unguiculatus)

Morphophysiological changes of the female prostate during pregnancy are still little known. Considering that this gland is highly influenced by steroid hormones, the aim of this study was to evaluate the impact of the pregnancy on female prostate morphophysiology in gerbils. Pregnant females were timed, and the prostates were analyzed at pregnancy days 6 (P6), 12 (P12), 18 (P18), and 24 (P24). Virgin females were used as the control group (C). We observed a profound change in the hormonal profile during gestation, which was marked by a high oscillation of the progesterone (P4) hormone. P4 serum levels increased, peaking at the middle of gestation, and decreased to the end of the pregnancy. The morphology of the gland in pregnant females also changed, being marked by an increase of acini lumen, and a decrease in stroma. Indeed, the acinar changes during pregnancy were followed by a significant reduction of the epithelial height, besides a change of the smooth muscle cells' morphology that became more relaxed. The number of progesterone receptor (PR) and androgen receptor (AR)-positives cells decreased with the increase of progesterone serum levels, showing an inverse relationship. Finally, we observed a reduction of epithelial proliferation and asignificant increase of gland PAS-positive secretion at the end of pregnancy. Altogether, these results showed, for the first time, that the female prostate morphophysioloy is profoundly influenced by the gestational period, suggesting that the fluctuation of the P4 serum levels is the main factor influencing the gland during this period.

Результати лікування хворих з поліпами ендометрія при застосуванні диференційованого підходу

The problem of treatment of endometrial polyps remains relevant, because it has high risks of malignancy and a steady tendency to increase the frequency of relapses and is inherent from 26 to 78% in women of different age categories. The objective: is to reduce the frequency of relapses of endometrial polyps by introducing a differentiated treatment strategy for patients with endometrial polyps based on the study of new pathogenesis links. Materials and methods. Clinical and laboratory examinations and treatment of 66 women diagnosed with endometrial polyp at the age of 24–43 years were carried out, which were further divided into two statistically equivalent groups: A (n=34) and B (n=32). All women received treatment according to the 4-stage algorithm of the current order of the Ministry of health of Ukraine No. 676 dated 31.12.2004. The difference in the management of group A patients was that their treatment was supplemented by immunomodulate therapy. At the first stage of the study, additional hysteroresectoscopy was performed, endometrial samples were obtained to determine its type of pathology, develop an immunohistochemical profile of the endometrium with the establishment of a receptor phenotype, identify the inflammatory process, and determine the state of the apoptosis system and the APUD system of the endometrium. Results. Active histological screening of the endometrial condition showed that signs of chronic endometritis (positive reaction of CD-138 and CD-68 markers) in women with endometrial polyp with physiological background endometrium were observed in 26.5% of group A patients after 3 months of treatment, which required a repeat course of treatment based on the results of viral and bacteriological examination. In group B, this rate was 46.9%. All the examined women, against the background of the use of gestagens, showed changes in the endometrial receptor phenotype, which were manifested by a decrease in the number of progesterone receptors in the glandular epithelium by an average of 1.4 times. However, the expression levels of Bcl-2 protein and EC cells had no statistical differences, since there were no signs of proliferation in the background endometrium, including local ones. Viral-bacterial screening of the study showed a sharp decrease in the number of pathogens in the endometrium, but in studies of patients of both groups, viral-bacterial associations were determined. Bacterial screening revealed the presence of anaerobic microflora. The study of the level of tumor necrosis factor in flushes from the uterine cavity indicated a unidirectional trend with the dynamics of the CD-138 index. The results of treatment of patients with endometrial polyp with physiological endometrium showed that the full effect of treatment in group A (antibacterial therapy with gestagens + immunomodulatory therapy) was determined after a year in 85.2% of patients, 14.8% of women had a relapse of endometrial polyp, in the group where patients received only antibacterial therapy with gestagens – a relapse was diagnosed in 37.5% of patients. Conclusion. Determining the tactics of differentiated treatment of endometrial polyps, it is necessary to take into account the results of immunohistochemical research and viral-bacterial control of the endometrial condition, which allows improving the results of treatment of endometrial polyps from 62.5% to 85.2 %, and is a prevention of relapse of endometrial polyp. Key words: endometrial polyps, hysteroscopy, endometrial immunohistochemical examination, apoptosis system, endometrial APUD system, viral-bacterial screening.

Abstract P3-08-40: Prognostic factors associated with clinical outcomes in HR+, HER2- advanced breast cancer: Systematic literature review

Abstract Background Advanced breast cancer (ABC) is a heterogeneous disease with several well-defined subtypes, among which, HR+, HER2- is the most prevalent. While clinical factors and genomic signatures have clear prognostic significance in the early breast cancer setting, this is less clear in the advanced disease setting. The aim of this systematic literature review was to identify the strength and consistency of evidence for prognostic factors in HR+, HER2-, ABC patients. Methods A comprehensive search was conducted of the major electronic databases (MEDLINE, EMBASE and Cochrane Controlled Register of Trials) in November 2018 for primary research clinical studies published since 2010 in HR+, HER2- ABC patients. Endpoints of interest were tumor response, progression-free survival (PFS), overall survival (OS), and breast-cancer specific survival (BCSS). Studies were screened by two independent reviewers for eligibility. Results Seventy-nine studies (72 full-text publications and 7 conference abstracts) were included wherein all patients were diagnosed with ABC and ≥50% of the population were HR+, HER2-. The majority were observational studies (n=71). Among the four endpoints, OS was the most commonly (n=67) assessed. Negative progesterone receptor (PR) status, higher tumor grade, higher CTC count, higher Ki67 level, number of metastatic sites (multiple vs. single) and sites of metastases (e.g., presence of liver metastases vs. absence), patients with relapsed BC compared with de novo metastatic BC, shorter time to recurrence or progression to ABC, poor performance status, prior therapy attributes in the early or metastatic setting (type of therapy, treatment line, response of prior therapy), and race (black vs. white) were consistently (>50% of studies found significant association) associated with worse OS; the relationship was inconsistent for tumor size, histological type (lobular vs. ductal), lymph node involvement and age. Directionality of relationship was consistent for all factors except lymph node involvement. Strength of association with OS was moderate [hazard ratio (HR) between 1.5-2.9] for PR status, tumor grade, CTC count, and Ki67 level, number and site (e.g., bone, liver, lung) of metastases, time to recurrence or progression to ABC, performance status, prior therapy attributes, and weak (HR<1.5) for de novo metastatic BC and race. Heterogeneity was observed across the studies in the composition of the patient population, definition and categorization for a few prognostic factors (e.g., number and sites of metastases, prior therapy, age). Similar results from fewer number of studies were observed for tumor response, PFS, BCSS. Conclusions Based on consistency and strength of the data PR status, tumor grade, CTC count, number and sites metastases, time to recurrence or progression to ABC, performance status, and prior therapy attributes were identified as the prognostic factors that had the strongest evidence. The application of these factors may be able to inform future research and clinical decision-making to improve outcomes in patients with HR+/HER2- ABC. Association between prognostic factors and overall survivalPrognostic factor Total no. of studies that assessed associationTotal no. of studies reporting significant association No. studies with significant multivariate analysis PR status1085Tumor Grade211411CTC count1097Ki67542No. of metastatic sites262313Site of metastasis332117De novo metastatic BC543Tumor size1255Lymph node1141Histological type 511Time to disease recurrence or progression18138Performance status14118Prior therapy352720Age371713Race1376 Citation Format: Gebra C Carter, Keri Stenger, Maitreyee Mohanty, Amy L Chong, Pradeep Basa, Shivaprasad Singuru, Sheena Singh, Vanita Tongbram, Sherko Kümmel, Valentina Guarneri, Sara M Tolaney. Prognostic factors associated with clinical outcomes in HR+, HER2- advanced breast cancer: Systematic literature review [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-08-40.

Progesterone in vitro increases growth, motility and progesterone receptor expression in third stage larvae of Toxocara canis

The in vitro effect of progesterone in T. canis larvae on their enlargement and motility were evaluated, together to the possible presence of progesterone receptors (PRs). T. canis larvae were cultured in RPMI-1640 with different concentrations of progesterone (0, 20, 40, 80, 400 and 800 ng/mL). Enlargement and increases in motility were dependent on the concentration only from 0 to 80 ng/mL (p < 0.05). The mean percentage of PR + cells in newly obtained larvae as measured by flow cytometry was 8.16 ± 0.4. The number of PR + cells increased depending on concentration from 0 to 80 ng/mL (p < 0.001). Cells obtained from larvae stimulated at any of the studied hormone concentrations showed greater mean fluorescence intensity when compared to non-stimulated cells. Additionally, the expression and location of PR + cells were determined in the larvae. The sequence of an amplicon (420-bp) obtained by PCR from T. canis larvae showed 100% homology with a gene fragment that codes for the PR of the dog. PR + cells were immunolocated using confocal microscopy in the intestinal region of the larvae that had been recently obtained. The results of this study show that T. canis larvae can recognize and respond to the presence of progesterone through a molecule possibly able to bind it. Since we previously observed a similar response to prolactin, we suggest that both hormones could participate sequentially in the reactivation of T. canis larvae in pregnant bitches.

Correlation between the Expression of Topo IIα and Ki67 in breast cancer and its clinical Pathological characteristics.

Objective:To investigate the expression of Topo IIα and Ki67 and its clinical significance.Methods:The clinical pathological data of one hundred and sixteen invasive breast cancer patients who were admitted into our hospital from July 2013 to December 2015 and underwent radical mastectomy were retrospectively analyzed. The expression of topoisomerase (Topo) IIα and Ki67 was detected using immunohistochemical method, and the correlation between the two kinds of proteins and the general clinical pathological characteristics of the patients was analyzed.Results:The positive expression rates of Topo IIα and Ki67 in breast cancer were 58.6% and 75% respectively. The expression of Topo IIα was in no apparent correlation with the age, tumor size, estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER-2) (P>0.05), but in a correlation with the number of metastatic lymph glands (P<0.05). The expression of Ki67 was in no apparent correlation with the age, tumor size, EP and HER-2, but in a correlation with the number of metastatic lymph glands and PR (P<0.05). The multi-factor logistic regression analysis results suggested that the number of metastatic lymph glands was the independent predictive factor of Topo IIα positive expression and the number of metastatic lymph glands and PR protein expression state are the independent predictive factors of Ki67 positive expression.Conclusion:Topo IIα and Ki67 can be regarded as the indicators for reflecting the proliferation activity of tumor cells, and the detection of Topo IIα and Ki67 expression is of great significance to the prognosis evaluation of breast cancer patients and clinical treatment.

Insulin Induced Down Regulation of the Progesterone Receptor Number in Neutrophils in the Synthesis of Maspin in Breast Cancer

Purpose: The binding of either progesterone or insulin to their specific receptors on neutrophils has been reported to stimulate nitric oxide (NO) induced maspin synthesis in these cells. Experiments were carried out to determine the role of progesterone receptor interaction in the nitric oxide induced maspin synthesis in neutrophils that was preincubated with insulin. Methods: progesterone receptor positive (PR+), progesterone receptor negative (PR–) neutrophils were isolated from the blood cancer subjects. Maspin was determined by enzyme linked immunosorbent assay after in vitro translation of maspin mRNA. NO was determined by methemoglobin method. Results: Immunohistochemical studies of progesterone receptor (PR) demonstrated the presence of progesterone receptor in the normal peripheral neutrophils and less in number in PR+ breast cancer neutrophils. In contrast, PR– breast cancer neutrophils lacked the progesterone receptor, suggesting pathophysiological defects in the synthesis of PR protein in peripheral PR– neutrophils. It was also found that as a result of incubation of neutrophils with insulin the binding affinity for progesterone to its receptor in normal neutrophils remained essentially unchanged which demonstrated Kd = 47.619 nM compared to Kd of the binding of progesterone is 46.08 nM in the normal neutrophils that were not pretreated with insulin. The progesterone receptors which were 11.5×1010/cell in the untreated cells was found to be decreased to 8.2×1010/cell (p<0.005, n=6) after the same cell were treated with 200μU of insulin. The reduction of PR number on normal neutrophils due to the pretreatment with insulin resulted in the decreased NO induced maspin synthesis from 2.329 ± 0.012 nM to 1.410 ± 0.002 nM. Decreased PR number in PR+ breast cancer neutrophils due to disease condition and pretreatment with insulin reduced the maspin synthesis from 1.138 ± 0.024 nM to 0.555 ± 0.003 nM compared to normal control. Conclusion: These results suggested that insulin down regulated maspin synthesis in normal and in breast cancer neutrophils by decreasing the progesterone receptor number in both cases.

Estrogen receptors α and β have different roles in the induction and trafficking of progesterone receptors in hypothalamic ventromedial neurons.

Progesterone receptor (PR) activation in the ventrolateral division of the hypothalamic ventromedial nucleus (VMNvl) is essential for promoting female sexual behavior. Estrogen receptor (ER) α, in contrast to ERβ, has been implicated in the induction of PRs. The simultaneous activation of ERα and ERβ, although not increasing the number of PR-immunoreactive neurons in the VMNvl, facilitates lordosis, which suggests that ERβ and/or the ERα-ERβ interaction might play a role in PR dynamics and/or PR expression by individual neurons. To address this question, we used western blot and immunohistochemical studies to determine the amounts and subcellular distributions of both PR isoforms in VMNvl neurons of ovariectomized rats injected with estradiol benzoate or with specific agonists of ERα and ERβ, alone or in association. The present data show that ERα activation does not change PR expression in individual neurons, but increases the number of PRs in the VMNvl, because it increases the number of neurons expressing PRs. Conversely, ERβ activation does not change the total number of PRs in the VMNvl, but increases the labeling intensity of the perikaryal cytoplasm, which suggests that it promotes the transport of PRs from neurites into cell bodies. In addition, the simultaneous activation of ERα and ERβ increases the expression of PRs by individual neurons and, consequently, increases the total number of PRs in the VMNvl. Our findings reveal that individual and simultaneous activation of ERα and ERβ have different effects on the levels and subcellular location of PRs in VMNvl neurons.

Estradiol Upregulates Progesterone Receptor and Orphanin FQ Colocalization in Arcuate Nucleus Neurons and Opioid Receptor-Like Receptor-1 Expression in Proopiomelanocortin Neurons That Project to the Medial Preoptic Nucleus in the Female Rat

Background: Ovarian steroids regulate sexual receptivity in the female rat by acting on neurons that converge on proopiomelanocortin (POMC) neurons in the arcuate nucleus of the hypothalamus (ARH) that project to the medial preoptic nucleus (MPN). Estradiol rapidly activates these neurons to release β-endorphin that activates MPN μ-opioid receptors (MOP) to inhibit lordosis. Lordosis is facilitated by the subsequent action of progesterone that deactivates the estradiol-induced MPN MOP activation. Orphanin FQ (OFQ/N; also known as nociceptin) infusions into the ARH, like progesterone, deactivate MPN MOP and facilitate lordosis in estradiol-primed rats. OFQ/N reduces the activity of ARH β-endorphin neurons through post- and presynaptic mechanisms via its cognate receptor, ORL-1. Methods: We tested the hypotheses that progesterone receptors (PR) are expressed in ARH OFQ/N neurons by immunohistochemistry and ORL-1 is expressed in POMC neurons that project to the MPN by combining Fluoro-Gold injection into the MPN and double-label fluorescent in situ hybridization (FISH). We also hypothesized that estradiol increases coexpression of PR-OFQ/N and ORL-1-POMC in ARH neurons of ovariectomized rats. Results: The number of PR- and OFQ/N-immunopositive ARH neurons was increased as was their colocalization by estradiol treatment. FISH for ORL-1 and POMC mRNA revealed a subpopulation of ARH neurons that was triple labeled, indicating these neurons project to the MPN and coexpress ORL-1 and POMC mRNA. Estradiol was shown to upregulate ORL-1 and POMC expression in MPN-projecting ARH neurons. Conclusion: Estradiol upregulates the ARH OFQ/N-ORL-1 system projecting to the MPN that regulates lordosis.

Effects of progesterone and RU486 on the development and expression of adult male sexual behaviour and gene expression in the amygdala and preoptic area of the hypothalamus

The number of progesterone receptors is greater in the male than female neonatal rat hypothalamus. The aims of the present study were to determine developmental effects of progesterone on the expression of adult male sexual behaviour and whether changes in behaviour were reflected by altered gene expression within the hypothalamic preoptic area (POA) or medial amygdala. Male rats were treated with progesterone (40 µg kg(-1), i.p.), the progesterone receptor antagonist RU486 (40 µg kg(-1), i.p.) or an equal volume of vehicle (10% ethanol, 90% corn oil) on postnatal Days 1-5. Treatment with either progesterone or RU486 inhibited (P ≤ 0.07) the initial expression of consummatory sexual behaviour at 10.5 weeks of age without influencing growth or serum concentrations of testosterone. Sexual interest, as measured by latency to exhibiting mounting behaviour or the number of mounts achieved, was not influenced by treatment with either progesterone or RU486. The effects of treatment with progesterone or RU486 on sexual behaviour were diminished by experience. Microarray analysis of the POA indicated 61 genes that were upregulated and 49 that were downregulated (P ≤ 0.01) following RU486 treatment of male rats. However, the altered expression of selected genes was not confirmed by real-time reverse transcription-polymerase chain reaction. The expression of targeted genes within the amygdala was not influenced by treatment with either progesterone or RU486. Neonatal treatment with RU486, but not progesterone, decreased testes weight (P=0.02) without affecting testes morphology. The results indicate that altering the progesterone environment during a critical developmental period affects the expression of behaviour, but that changes in behaviour are not mirrored by the altered expression of selected genes.

Possible role of the nuclear factor kappa B (NF‐kappa B) (NF) in the development of retained placenta (RP)

The low NF level seen in the serum of buffalos (B) (P = 0.01) may initiate RP through: a. Myometrial quiescence (MQ) which could be due the inhibited expression of target genes causing increased myometrial contractility e.g. COX‐2 catalyzing prostanoids synthesis. This may account for the abrogated increase in PGE2 level in BFR. The low NF level may cause an inhibition of progesterone withdrawal; decrease in number of progesterone receptors(PR); that accompany parturition. This increases the sensitivity of myometrium to progesterone. A mutual antagonism exists between PR and NF transcriptional activities. A decrease in oxytocin (OX) receptors numbers(OTR) may arise due to the low NF level since NF is involved in the up regulation of OTR. This may cause a low sensitivity of myometrium to OX thus mediating a negative feed back inhibition of the expected decrease in serum OX level accounting for the unchanged OX level in BRP. The low TNF‐alpha (P = 0.02) and NO (P = 0.001) levels in BRP may contribute to MQ since TNF‐alpha activates NF, also the expression of NO‐2 involves the activation of NF. b. Immunosuppression (IM) in which immune response that affects placental rejection at parturition is inhibited in BRP. This hypothesis is based on reduced NF and TNF‐alpha levels in BRP. NF activation induces cell adhesion molecules, cytokines and iNOS expression. And, NF is highly inducible by cytokines including TNF‐alpha.

The number of ERα and PR in the mammary glands of bitches with and without tumor mass using immunohistochemical assay

The objectives of this study were to evaluate the expressions of estrogen receptors alpha (ERα) and progesterone receptors (PR) in histologically normal and tumoral canine mammary tissues within the same bitch and to correlate between the immunohistochemical staining scores of both receptors and duration of tumor growth. Twelve tumoral and contralateral normal mammary tissues, both determined by histology were surgically obtained from 12 dogs. The expressions of ERα and PR were evaluated by avidin–biotin–peroxidase complex (ABC) method and ERα and PR scores were calculated. The expressions of the ERα and the PR between normal and tumoral mammary tissues were not significantly different. The PR expression and duration of tumor growth showed a significantly inverse correlation. A positive correlation was found between the number of the ERα and the PR in both normal and tumoral mammary tissues which may indicate that either ERα or PR could be used as a prediction of hormonal therapy parameter in dogs. However, the information of ERα and PR status as well as hormonal therapy is not routinely available in veterinary medicine and it appears that half of the mammary tumors from the present study are hormone-independent tumors (ERα and PR negative). Therefore, the treatment of canine mammary tumor by hormonal therapy should be carefully considered.

Expression of Ovarian Steroid Hormone Receptors in Tuberoinfundibular Dopaminergic Neurones During Pregnancy and Lactation

During late-pregnancy, tuberoinfundibular dopaminergic (TIDA) neurones, a critical component of the negative-feedback loop regulating prolactin secretion, become unresponsive to the stimulatory effects of prolactin. The change in TIDA responsiveness to prolactin at this time results in a decrease in dopamine secretion and a prolactin surge. As the onset of parturition and the antepartum prolactin surge depend on the withdrawal of progesterone in the presence of oestrogen, it is likely that ovarian steroid hormones mediate this change in TIDA responsiveness. To determine whether ovarian steroids can directly modulate TIDA activity, and whether changes of receptor numbers might contribute to overall steroid-regulation of these neurones, we investigated the level of oestrogen receptor alpha (ERalpha) and progesterone receptor (PR) expression within TIDA neurones during pregnancy and lactation. Animals were sacrificed on dioestrous, days 12, 19 and 21 of pregnancy and day 5 of lactation, and the proportion of TIDA neurones expressing ERalpha or PR, as well as the total number of PR expressing cells within the arcuate nucleus, was determined. Approximately 75% and 55% of tyrosine hydroxylase neurones expressed ERalpha and PR, respectively. Levels of steroid receptor expression within TIDA neurones remained fairly constant, except for an increase in ERalpha on days 12 and 19 of pregnancy compared to dioestrous and lactation day 5. The presence of steroid receptors on TIDA neurones during pregnancy and lactation supports the concept of a direct effect of steroid hormones on these neurones at this time. Thus, steroid hormones may directly act on TIDA neurones to regulate maternal prolactin secretion. The relatively stable level of expression during late pregnancy suggests that a shift in steroid receptor expression during late pregnancy does not contribute to the change in TIDA responsiveness to prolactin at this time.

Expression of estrogen receptors-α and -β in primary breast neoplasms and tumors exposed to neoadjuvant hormonal therapy

We studied the content and expression of mRNA for estrogen receptors-alpha and -beta in breast tumors before and after 3-month neoadjuvant hormone therapy with antiestrogen tamoxifen and/or aromatase inhibitors. Expression of estrogen receptors-alpha and -beta was most often detected in ER+PR+ tumors and most significantly decreased in these neoplasms after exemestane therapy. Immunocytochemical and radioligand assays showed that tamoxifen and anastrozole have little effect on the number of estrogen receptors-alpha. The number of progesterone receptors in tumors decreased by the end of anastrozole therapy. Estrogen receptors-beta were immunocytochemically revealed in 50% primary breast tumors. Anastrozole slightly decreased, while tamoxifen increased the incidence of these receptors. Interruption of signaling through estrogen receptors and suppression of estrogen biosynthesis had different effects on the receptor status of neoplasms and distribution of estrogen receptors-alpha and -beta.

Immunohistochemical distribution of oestrogen and progesterone receptors and tissue concentrations of oestrogens in the cervix of non-pregnant cows.

An immunohistochemical study of the expression of oestrogen (ER) and progesterone receptors (PR) in different regions along the longitudinal and vertical axes of the cervix of non-pregnant cows was performed. Animals were separated into two groups depending on the presence or absence of a functional corpus luteum in their ovaries, as indicated by blood progesterone concentrations. The high progesterone group (HP4) had serum progesterone concentrations > 2.0 ng mL(-1) (n = 6) and the low progesterone group (LP4) had serum progesterone concentrations < or = 0.5 ng mL(-1) (n = 4). Significantly higher concentrations of oestrogen were found in the cervical tissue of animals in the LP4 group than those in the HP4 group (473 +/- 53 v.149 +/- 46 pg g(-1) wet weight; P < 0.01). Furthermore, there was a significant effect of tissue layer (epithelium to deep stroma) on the number of ER (P < 0.01) and PR (P < 0.05) immunoreactive nuclei per 1000 cells. For both ER and PR the proportion of cells expressing the receptor increased from epithelium to subepithelial stroma (P < 0.01) and from subepithelium to deep stroma (ER P < 0.05; PR P =0.061). When the number of receptor-positive cells were expressed per mm2 tissue, differences between the subepithelial stroma and the deep stroma became even more marked. In addition, the vaginal part of the cervix had significantly more (P < 0.01) ER and PR immunoreactive nuclei per 1000 cells than the uterine part, but these differences were no longer apparent when a correction was made for cell density. There was no relationship between progesterone status of the animals, nor local tissue oestrogen concentrations and ER or PR immunoreactivity in the cervix of these non-pregnant cows. Instead, a strong relationship between both longitudinal and vertical positioning of tissue in the cervix and expression of both receptor types was shown. In addition, a strong correlation between ER and PR expression in the subepithelial stroma (R = 0.85, P < 0.01) and the deep stroma (R = 0.83 P < 0.01) was evident. In conclusion, these results demonstrate that in studies of steroid hormone receptor expression in the cervix, careful description of sampling site and depth are necessary if the results are to be interpreted meaningfully.

Evidence for progesterone receptors in the human fetoplacental vascular tree.

The presence of progesterone receptors (PR) throughout the human term fetoplacental vascular tree was investigated. By reverse transcription-polymerase chain reaction (RT-PCR), we showed expression of PR mRNAs in stem villi vessels, chorionic arteries and veins, and umbilical arteries and veins. Binding studies and Scatchard analysis revealed a single class of high-affinity binding sites for (3)H-R5020 (promegestone) in cytosolic extracts of all placental vessels, with K(d) values in the range of 2.5-4 nM. High levels of PR were detected in placental vessels compared to other vascular tissues. Thus, maximum binding capacities of stem villi vessels, chorionic arteries and veins, and umbilical arteries and veins were 247 +/- 25, 377 +/- 58, 295 +/- 40, 371 +/- 118, and 672 +/- 144 fmol/mg protein, respectively. Endothelial cell elimination in chorionic arteries did not significantly modify the number of PR. RT-PCR and binding studies also assessed PR expression in cultured placental vascular smooth muscle cells isolated from stem villi vessels. All these data suggested that most of the PR of fetoplacental vessels were from the media. In conclusion, we report here the first evidence of the presence of PR in the muscular layer of human term fetoplacental vessels. This finding, together with the high progesterone concentrations in cord blood, suggests that the interactions between the PR and its ligand may play a role in the physiology and physiopathology of human fetoplacental vascularization.

An ongoing validation of a Tier I screening battery for detecting endocrine-active compounds (EACs).

After previously examining an estrogen receptor agonist (17beta-estradiol), several additional compounds have been evaluated in a Tier I screening battery for detecting endocrine-active compounds (EACs): an estrogen receptor antagonist (ICI-182,780, ICI), an androgen receptor antagonist (flutamide, FLUT), a testosterone biosynthesis inhibitor (ketoconazole, KETO), a 5alpha-reductase inhibitor (finasteride, FIN), and an aromatase inhibitor (anastrozole, ANA). The Tier I battery incorporates two short-term in vivo tests (a 5-day ovariectomized female battery and a 15-day intact male battery) and an in vitro yeast transactivation system (YTS). The Tier I battery is designed to identify compounds that have the potential to act as agonists or antagonists to the estrogen, androgen, progesterone, or dopamine receptors, steroid biosynthesis inhibitors (aromatase, 5alpha-reductase, and testosterone biosynthesis), or compounds that alter thyroid function. ICI administration decreased uterine estrogen and progesterone receptor number in the female battery, increased serum follicle-stimulating hormone (FSH) levels and caused spermatid retention in the male battery, and activated gene transcription in the YTS containing the estrogen receptor. FLUT administration increased uterine stromal cell proliferation in the female battery and decreased weights for all androgen-dependent tissues, induced Leydig cell hyperplasia, and caused hormonal alterations (increased testosterone (T), estradiol (E2), dihydrotestosterone (DHT), luteinizing hormone (LH), and FSH) in the male battery, and competed for binding to the androgen receptor in the YTS competition assay. In the male battery KETO decreased weights for all androgen-dependent tissues, caused hormonal alterations (decreased T and DHT and increased LH and FSH), and induced spermatid retention. FIN decreased seminal vesicle and accessory sex gland (ASG) unit weight and caused hormonal alterations (decreased DHT and increased LH, and PRL) in the male battery. KETO was judged not to affect any of the endpoints in the female battery. ANA decreased ASG unit weight and serum E2 levels in the male battery. Using the responses obtained for all the endpoints in the Tier I battery, a distinct "fingerprint" was produced for each type of endocrine activity against which compounds with unknown activity can be compared. These data demonstrate that the described Tier I battery is useful for identifying EACs.