Age-Specific Modulation of Prefrontal Cortex LTP by Glucocorticoid Receptors Following Brief Exposure to HFD

The corticolimbic circuits in general and the medial prefrontal cortex in particular, undergo maturation during juvenility. It is thus expected that environmental challenges in forms of obesogenic diet can exert different effects in juvenile animals compared to adults. Further, the relationship between glucocorticoids and obesity has also been demonstrated in several studies. As a result, glucocorticoid receptor (GR) antagonists are currently being tested as potential anti-obesity agents. In the present study, we examined the effects of short-term exposure to high-fat diet (HFD) on prefrontal long-term potentiation (LTP) in both juvenile and adult rats, and the role of glucocorticoid receptors (GRs) in modulating these effects. We found HFD impaired prefrontal LTP in both juveniles and adults, but the effects of GR modulation were age- and diet-dependent. Specifically, GR antagonist RU-486 reversed the impairment of LTP in juvenile animals following HFD, and had no effect on control-diet animals. In adult animals, RU-486 has no effect on HFD-impaired LTP, but abolished LTP in control-diet animals. Furthermore, impairments in the prefrontal LTP following HFD are involved with an increase in the mPFC GR levels only in the juveniles. Further, we found that in vivo application of GR agonists into adult mPFC rescued HFD-induced impairment in LTP, suggesting that these receptors might represent strategic therapeutic targets to potentially combat obesity and metabolic related disorder.

Ethical and Legal Aspects of Using the Abortion Pill RU-486 in Italy in the Years 2020-2021

In some countries, one of the most difficult bioethical challenges of the SARS-CoV-2 coronavirus pandemic has been a sharp rise in the number of chemical abortions performed in hospitals, outpatient clinics, or at home. Limited access to medical services at public and private healthcare facilities and the development of telemedicine have resulted in the practice of chemical abortion having largely moved from hospitals and clinics to the home. Chemical abortion is a method used only in the early stages of pregnancy, i.e. up to 7-9 weeks. The first stage of abortion involves taking RU-486, a pill containing a preparation called Mifepristone which kills the newly conceived life in the mother’s womb. The second stage of chemical abortion involves the use of a preparation called Misoprostol, which leads to the expulsion of the dead embryo from the woman’s body. The main goal of the article is to analyze the ethical and legal dispute in Italy in the years 2020 and 2021 concerning new ways of using the abortion pill RU-486. In August 2020, the Italian Ministry of Health decided that chemical abortion – performed using the medical preparation RU-486 – should not be practiced at hospital gynecological and obstetric wards, but in day hospitals, without the requirement of hospitalization for women performing such abortions. Analyses carried out in the article show that chemical abortion not only kills the life of a human embryo, but in some cases may also be dangerous to the life and health of the mother. The liberalization of chemical abortion in Italy has led to an increase in the number of unborn children who are aborted, as it facilitates access to this type of abortion and makes it a procedure that is largely self-administered at home. The Italian dispute over home abortion and the RU-486 pill is linked to other serious bioethical debates that are currently taking place in many countries around the world

In-Vitro Effects of Active Vitamin-D3 and Vitamin-A on Human Melanoma (BLM and1205Lu) Cell Growth

Though UV ray in the Sun light is essential for vitamin-D3 formation, yet UV rays can lead to skin cancer. Lack of vitamin-D3 also can lead to cancer. Hence, we decided to study the effect of vitamin-D3 on human melanoma cell models. Our aims were to study the in-vitro effects of vitamin-D3 and vitamin-A on human melanoma (BLM, 1205Lu) cell growth, as vitamins D3 and A are important for a healthy skin. Initially dose-curve (100 nM to 100 μM concentration) study was carried out with vitamin-D3 and vitamin-A (retinoic acid) on BLM cells to determine the optimal concentrations of vitamins-D3 and A for co-incubation with progesterone (50 μM) and RU-486 (50 μM). Supernatants from the treated cells were subjected to Elisarray. Vitamin-D3 and vitamin-A showed a dose-dependent decrease in cell growth. Based on the dose-curve, it was decided to use 25 μM (at 57% cell growth) of vitamin-D3 and 50 μM (at 55% cell growth) of vitamin-A for co-incubation studies. Co-incubation of vitamin-D3 with vitamin-A showed an additive effect on the decrease of BLM cell growth (20%). Similarly co-incubation of vitamin-D3 with progesterone (33% cell growth) and with RU-486 (28% cell growth) as well as co-incubation of vitamin-A with progesterone (31% cell growth) and RU-486 (18% cell growth) showed an additive effect on the decrease of BLM cell growth. Based on the BLM co-incubation studies, we decided to repeat the studies on 1205Lu cells. So, 25 μM (at 34% cell growth) of vitamin-D3 and 50 μM (at 44% cell growth) of vitamin-A were used for co-incubation studies. Co-incubation of vitamin-D3 with vitamin-A showed an additive effect on the decrease of cell growth (22%). Though co-incubation of vitamin-D3 with progesterone did not show any difference in cell growth (51%), yet co-incubation with RU-486 showed a decrease in 1205Lu cell growth (21%). Similarly co-incubation of vitamin-A with progesterone (25% cell growth) and with RU-486 (23% cell growth) showed a synergistic effect on the decrease of 1205Lu cell growth. Conclusion: These studies suggested that combination of vitamins and steroids might be effective in decreasing melanoma cell growth. Hence, various combination of vitamins and steroids will be tested for their effect on melanoma cell growth in vitro in order to develop a combo drug treatment for melanoma.

Mifepristone (Korlym ®) Use Interferes With Accurate Serum Measurement of Sex Steroids

Introduction: Mifepristone (MFP) aka RU-486/Korlym is a synthetic steroid analog originally developed in 1980 as an abortifacient that is now used in the management of Cushing’s syndrome (CS). It is both a progesterone (PG) and glucocorticoid (GC) receptor blocker. CS is often associated with reproductive functional anomalies including hypogonadism, menstrual irregularities and infertility. Due to its mode of action, measurement of serum PG and/or cortisol in patients on MFP are inaccurate indices of systemic PG and/or GC deficiency or excess. However, the potential impact of MFP use on other serum steroid assays has not been widely studied. We report the case of a 55 yr old man on treatment with MFP for adrenal CS found to have striking artefactual changes in serum androgen and estrogen levels. Case Summary: A 55 yr old African American man was referred with poorly controlled type 2 diabetes, hypertension, hyperlipidemia, obesity and untreated hypogonadism. He had a 3.2cm left adrenal incidentaloma associated with adrenal CS and he chose medical management rather than surgery. Upon starting MFP at 300mg QD serum total estrogen (by radio-immunoassay; RIA) and estradiol (by chemiluminescence immunoassay; CIA) were markedly elevated while serum total, free, bioavailable testosterone and dihydrotestosterone were all markedly reduced. His HBA1c, weight and energy levels improved on MFP despite these findings. The serum steroid levels normalized to pre-treatment levels after stopping MFP for ~ 4weeks but the changes recurred after restarting therapy. After MFP dose escalation to 300mg BID the serum steroid levels normalized after stopping MFP for ~ 6 weeks. The artefactual low testosterone levels also occured with measurement by equilibrium dialysis but “normal accurate” results were obtained when measured by liquid chromatography-Tandem mass spectrometry (LC-TMS). He remains on MFP 300mg BID without need for androgen repletion. Discussion: With increased use of MFP for CS, indices for tracking its clinical and biochemical effects assume great importance. There are few reports of the possible effects of MFP on estrogen and testosterone serum assays despite its touted low cross reactivity with sex steroids. Our case suggests that the significance, extent and prevalence of artefactual changes on serum sex steroid assays may be underestimated and under- appreciated. Conclusions: Our case of wide disparities in serum estrogen and androgen measures in a patient on MFP indicates that caution needs to be exercised in the interpretation of such results in patients on current MFP therapy. Our clinical observations suggest depending on the dose that a wash out period of 4–6 weeks is required to ensure accurate measures. Studies to ascertain the prevalence of this artefactual effect are needed and it appears testosterone measurement by LC-TMS obviates the testosterone assay artefact.

TEMPAT ABORSI DI SURABAYA, 08985622115 KLINIK ABORSI KURET SURABAYA, | TEMPAT KURET DI SURABAYA, | KURET AMAN

TEMPAT KURET | BIDAN MELAYANI KURET | BIDAN PRAKTEK SURABAYA | JUAL OBAT ABORSI DI SURABAYA | TEMPAT ABORSI DI TEMPAT ABORSI DI SURABAYA | LOKASI ABORSI DI SURABAYA | KLINIK ABORSI DI SURABAYA | TEMPAT KURET DI SURABAYA | KURET AMAN | PROSES KURET AMAN DI SURABAYA SURABAYA | KLINIK ABORSI KURET SURABAYA | JASA ABORSI DI DAERAH SURABAYA | OBAT MENGGUGURKAN KANDUNGAN DI SURABAYA | OBAT MENGGUGURKAN JANIN DI SURABAYA | JUAL CYTOTEC MURAH DI SURABAYA | MISOPROSTOL DI SURABAYA | MIFEPREX | JUAL GASTRUL DI SURABAYA | RU 486 | MISOPROSTON | SERLEY | HAID TIDAK TERATUR | OBAT TERLAMBAT TELAT BULAN DI SURABAYA | OBAT ABORSI AMPUH | OBAT ABORSI PRODUK PFIZER | OBAT ABORSI STANDART FDA | OBAT ABORSI ORIGINAL ASLI | OBAT PENGGUGUR KANDUNGAN DI SURABAYA | OBAT PENGGUGUR JANIN DI SURABAYA | CARA MENGGUGURKAN KANDUNGAN | PILL TABLET JAMU ABORSI DI SURABAYA | LANCAR BULAN | APOTIK YANG MENJUAL OBAT CYTOTEC DI SURABAYA | APOTIK YANG MENJUAL GASTRUL DI SURABAYA | MISOPROSTOL MIFEPREX SURABAYA | OBAT PIL JAMU TABLET HERBAL TRADISIONAL DI SURABAYA |

Calcium/Calmodulin-Stimulatable Adenylyl cyclases are required for the potentiating effect of acute glucocorticoid exposure in the hippocampal synapse

Experience-dependent synaptic plasticity is important for learning and memory and regulated by the functions of N-methyl-D-aspartate (NMDA) and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate receptors. The stress hormone glucocorticoid, exerts effects on synaptic plasticity through functional modification of these receptors. In this study, we have investigated the mechanisms underlying synaptic potentiation during acute stress and glucocorticoid treatment. We report that stress-induced glucocorticoid increase augments the postsynaptic localisation and phosphorylation of calcium permeable AMPAR, GluA1 and enhances long-term potentiation (LTP) observed 60 minutes following high frequency stimulation. These effects could be reproduced in adrenalectomized rats by the administration of a stress-equivalent dose of corticosterone. The observed response was blocked by the glucocorticoid receptor (GR) antagonist RU-486, thus potentially mediated by the GR. Glucocorticoid treatment further increased expression of hippocampal adenylyl cyclases (ACs), AC1 and AC8, implicating this class of enzymes as mediators of hyper-glucocorticoid effect on synaptic plasticity. In support of this, selective pharmacological AC inhibition with SQ22536 ablated the glucocorticoid modulation of AMPA-dependent neuronal plasticity and enhancement of LTP. These data support a key role for calcium/calmodulin-sensitive adenylyl cyclase enzymes in facilitating the modulatory actions of glucocorticoid hormones on a substrate for cellular mechanism of learning and memory, via AMPA receptor activation.

The Antiasthma Medication Ciclesonide Suppresses Breast Cancer Stem Cells through Inhibition of the Glucocorticoid Receptor Signaling-Dependent YAP Pathway

Ciclesonide is an FDA-approved glucocorticoid used to treat asthma and allergic rhinitis. However, whether it has anticancer and anti-cancer stem cell (CSC) effects is unknown. This study focused on investigating the effect of ciclesonide on breast cancer and CSCs and determining its underlying mechanism. Here, we showed that ciclesonide inhibits breast cancer and CSC formation. Similar glucocorticoids—dexamethasone and prednisone—did not inhibit CSC formation. Ciclesonide-induced glucocorticoid receptor (GR) degradation was dependent on ubiquitination. We showed via GR small interfering RNA (siRNA) that GR plays an important role in CSC formation. We showed via western blot and immunofluorescence assays that ciclesonide reduces the nuclear level of GR. The GR antagonist RU-486 also inhibited CSC formation. Ciclesonide reduced the protein level of the Hippo transducer Yes-associated protein (YAP). GR siRNA induced a decrease in YAP protein expression and inhibited mammosphere formation. The YAP inhibitor verteporfin inhibited CSC formation and transcription of the connective tissue growth factor and cysteine-rich protein 61 genes. The GR/YAP1 pathway regulated breast CSC formation. We showed that the GR/YAP signaling pathway regulates breast CSC formation and revealed a new approach for targeting GR and YAP to inhibit CSC formation.

Cortisol Directly Stimulates Spermatogonial Differentiation, Meiosis, and Spermiogenesis in Zebrafish (Danio rerio) Testicular Explants

Cortisol is the major endocrine factor mediating the inhibitory effects of stress on vertebrate reproduction. It is well known that cortisol affects reproduction by interacting with the hypothalamic–pituitary–gonads axis, leading to downstream inhibitory and stimulatory effects on gonads. However, the mechanisms are not fully understood. In this study, we provide novel data demonstrating the stimulatory effects of cortisol on spermatogenesis using an ex vivo organ culture system. The results revealed that cortisol treatment did not modulate basal androgen production, but it influenced transcript levels of a selected number of genes involved in the zebrafish testicular function ar (androgen receptor), star (steroidogenic acute regulatory), cyp17a1 (17α-hydroxylase/17,20 lyase/17,20 desmolase), cyp11a2 (cytochrome P450, family 11, subfamily A, polypeptide 2), hsd11b2 (11-beta hydroxysteroid dehydrogenase), cyp2k22 (cytochrome P450, family 2, subfamily K, polypeptide 22), fkbp5 (FKBP prolyl isomerase 5), grα (glucocorticoid receptor alpha), and grβ (glucocorticoid receptor beta) in a short-term culture. We also showed that cortisol stimulates spermatogonial proliferation and differentiation in an androgen independent manner as well as promoting meiosis and spermiogenesis by increasing the number of spermatozoa in the testes. Moreover, we demonstrated that concomitant treatment with RU 486, a potent glucocorticoid receptor (Gr) antagonist, did not affect the cortisol effects on spermatogonial differentiation but blocked the induced effects on meiosis and spermiogenesis. Supporting the Gr-mediated effects, RU 486 nullified the cortisol-induced expression of sycp3l (synaptonemal complex protein 3), a marker for the meiotic prophase that encodes a component of the synaptonemal complex. This is consistent with in silico analysis that found 10 putative GREs (glucocorticoid response elements) upstream of the zebrafish sycp3l. Finally, we also showed that grα mRNA is expressed in Sertoli and Leydig cells, but also in several types of germ cells, including spermatogonia and spermatocytes. Altogether, this evidence indicates that cortisol exerts paracrine roles in the zebrafish testicular function and spermatogenesis, highlighting its effects on spermatogonial differentiation, meiosis, and spermiogenesis.