A glucocorticoid receptor antagonist reduces sign-tracking behavior in male Japanese quail.

Addiction is characterized as a chronic debilitating disease. One devastating feature of addiction is the susceptibility of relapse (40-60%) after stretches of abstinence. One theory that may account for relapse suggests that drug cues (e.g., paraphernalia) may increase stress hormones, and this may prompt relapse. Repeatedly pairing a neutral cue with a reward is commonly utilized to measure what subjects learn about a cue that is predictive of reward. Research has shown that animals that attend to a cue more than to the reward (sign trackers) may be more vulnerable to drug addiction. Additionally, research has shown that sign tracking is associated with an increase in corticosterone, a primary stress hormone. PT150 is a novel glucocorticoid receptor antagonist that moderates the release of corticosterone. In the current experiment, it was hypothesized that subjects given repeated administration of PT150 would reduce sign tracking compared to subjects given placebo. Time spent (in seconds) near a cue that predicts reward (conditional stimulus) served as a measure of sign tracking, and PT150 or placebo was administered following sign tracking. An independent-samples t test revealed that subjects that received PT150 had reduced time spent near the conditioned stimulus compared to controls. Given the devastating effects of drug addiction, identification of a potential pharmacological intervention in the reduction of relapse would be of great value. Therefore, future research is needed to validate the use of PT150 in reducing behaviors associated with drug addiction. (PsycINFO Database Record

Uso del tratamiento hormonal sustitutivo en España: tendencias en el período 2000-2014 / Utilization of hormone replacement therapy in Spain: Trends in the period 2000-2014

Fundamento y objetivo: El objetivo de este trabajo fue describir la evolución del consumo, la oferta y la prevalencia de uso del tratamiento hormonal sustitutivo (THS) en España durante el período 2000-2014. Métodos: La prevalencia de uso anual bruta en mujeres≥40 años se estimó a partir de datos individuales procedentes de las historias clínicas anonimizadas de la base de datos BIFAP. Para el cálculo del consumo se utilizaron datos agregados de dispensación. El consumo de THS se expresó en dosis diarias definidas dispensadas por cada 1.000 mujeres≥40 años y día (DHD). Resultados: En el año 2000 el consumo de THS fue de 33,12 DHD: estrógenos (19,81 DHD), tibolona (6,88 DHD) y estrógenos asociados a progestágenos (6,44 DHD). En 2014 el consumo de THS fue de 5,32 DHD: estrógenos (1,08 DHD), tibolona (1,61 DHD) y estrógenos combinados con progestágenos (2,62 DHD). Las presentaciones comercializadas se redujeron un 46,9%. La prevalencia de uso de THS en mujeres≥40 años registradas en la base de datos BIFAP ha pasado de un 7,19% (IC 95% 6,97-7,40) en el año 2001 a un 0,21% (IC 95% 0,20-0,22) en 2014. Las mujeres de 40-45 años registraron la mayor prevalencia de uso en 2014: 0,71% (IC 95% 0,66-0,76). Conclusiones: El consumo y la prevalencia de uso de THS ha descendido de manera notable a partir de la publicación de los estudios Women's Health Iniciative, Million Women Study y las medidas reguladoras, siguiendo una tendencia similar a la observada en otros países occidentales (AU)

Background and objective: The objective of the study was to describe the trends of utilization, supply and prevalence of hormone replacement therapy (HRT) in Spain during the period 2000-2014. Methods: Annual prevalence of HRT use including the 95% CI was calculated for women aged≥40 using individual data from the national population-based database BIFAP. Annual and total-period consumptions were expressed in defined daily doses (DDD) per 1,000 inhabitants per day and were obtained from the databases of medications dispensed in community pharmacies and charged through official prescriptions to the Spanish National Health System. Results: In the year 2000, overall HRT consumption was 33.12 DDDs/1000 inhabitans/day: 19.81 for oestrogen only, 6.88 for tibolone and 6.44 for combined oestrogen and progestagen. In 2014 overall HRT consumption was 5.32 DDDs/1000 inhabitans/day: 1.08 for oestrogen only, 1.61 for tibolone and 2.62 for combinations of oestrogen and progestagen. The marketed presentations of HRT decreased by 46.9%. Prevalence of HRT use in women aged≥40 in BIFAP was 7.19% (95% CI 6.97-7.40) in 2001 and 0.21% (95% CI 0.20-0.22) in 2014. Women aged 40-45 registered the highest prevalence of use in 2014: 0.71% (95% CI 0.66-0.76). Conclusions: A sharp decline in the consumption and prevalence of HRT has been observed in Spain since the publication of the Women's Health Initiative and Million Women Study and the regulatory measures taken restricting conditions of use, showing a similar trend to that of other western countries (AU)

Effects of tibolone and its metabolites on prolactin and insulin-like growth factor binding protein-1 expression in human endometrial stromal cells.

The effects of the postmenopausal replacement steroid tibolone and its 3α-, 3ß-OH and Δ-4 tibolone metabolites were evaluated on progesterone receptor-mediated classic decidualization markers insulin-like growth factor binding protein-1 (IGFBP-1) and prolactin expression in human endometrial stromal cells (HESCs). Supernatants of conditioned medium or erxtracted RNA from experimental cell incubations of confluent HESCs were subjected to ELISAs, Western blot analysis and RT/PCR, and results were statisically assesed. Over 21 days, specific ELISAs observed linear increases in secreted IGFBP-1 and prolactin levels elicited by tibolone and its metabolites. Cultured HESCs were refractory to E2 and dexamethasone, whereas tibolone and each metabolite exceeded medroxyprogesterone acetate in significantly elevating IGFBP-1 and prolactin output. Anti-progestins eliminated IGFBP-1 and prolactin induction by tibolone and its metabolites. Immunoblotting and RT/PCR confirmed ELISA results. These observations of IGFBP-1 and prolactin expression: (a) indicate the relevance of cultured HESCs in evaluating the chronic effects of tibolone administration to women; (b) are consistent with PR-mediated endometrial atrophy and protection against endometrial bleeding despite the persistence of circulating ER-binding, but not PR-binding metabolites following tibolone administration to women.

Molecular and structural basis of androgen receptor responses to dihydrotestosterone, medroxyprogesterone acetate and Δ(4)-tibolone.

Medroxyprogesterone acetate (MPA) has widely been used in hormone replacement therapy (HRT), and is associated with an increased risk of breast cancer, possibly due to disruption of androgen receptor (AR) signaling. In contrast, the synthetic HRT Tibolone does not increase breast density, and is rapidly metabolized to estrogenic 3α-OH-tibolone and 3ß-OH-tibolone, and a delta-4 isomer (Δ(4)-TIB) that has both androgenic and progestagenic properties. Here, we show that 5α-dihydrotestosterone (DHT) and Δ(4)-TIB, but not MPA, stabilize AR protein levels, initiate specific AR intramolecular interactions critical for AR transcriptional regulation, and increase proliferation of AR positive MDA-MB-453 breast cancer cells. Structural modeling and molecular dynamic simulation indicate that Δ(4)-TIB induces a more stable AR structure than does DHT, and MPA a less stable one. Microarray expression analyses confirms that the molecular actions of Δ(4)-TIB more closely resembles DHT in breast cancer cells than either ligand does to MPA.

Activated protein C resistance among postmenopausal women using transdermal estrogens: importance of progestogen.

OBJECTIVE: Although the route of estrogen administration is known to be an important determinant of the thrombotic risk among postmenopausal women using hormone therapy, recent data have shown that norpregnane derivatives but not micronized progesterone increase venous thromboembolism risk among transdermal estrogens users. However, the differential effects of progesterone and norpregnanes on hemostasis have not yet been investigated. METHODS: We set up a cross-sectional study among healthy postmenopausal women aged 45 to 70 years. The impact of activated protein C (APC) on endogenous thrombin potential was investigated in the plasma samples of 108 women who did not use any hormone therapy (n = 40) or who were treated with transdermal estrogens combined with micronized progesterone (n = 30) or norpregnane derivatives (n = 38). RESULTS: After exclusion of women with factor V Leiden and/or G20210A prothrombin gene mutations, there was no significant change in APC sensitivity among women who used transdermal estrogens combined with micronized progesterone compared with nonusers. Women using transdermal estrogens combined with norpregnanes were less sensitive to APC than were nonusers (P = 0.003) or users of transdermal estrogens combined with micronized progesterone (P = 0.004). In addition, prothrombin fragment 1 + 2 concentration was higher in users of transdermal estrogens plus norpregnanes than in nonusers (P = 0.004). Other hemostatic parameters did not vary significantly across the different subgroups. CONCLUSIONS: Transdermal estrogens combined with norpregnanes may induce APC resistance and activate blood coagulation. These results provide a biological support to epidemiological data regarding the potential thrombogenic effects of norpregnanes. However, these findings need to be confirmed in a randomized trial.

Regulation of activities of steroid hormone receptors by tibolone and its primary metabolites.

This work was undertaken (i) to study deeply the estrogen, androgen and progestative activities of tibolone and its metabolites (ii) to determine whether tibolone and its metabolites present glucocorticoid or mineralocorticoid activity. For this purpose, we used human cell lines bearing a luciferase gene with a responsive element under the control of human estrogen receptor alpha (ERalpha) or estrogen receptor beta (ERbeta) or androgen receptor (AR) or chimeric Gal4 fusion with progesterone receptor (PR), glucocorticoid receptor (GR) or mineralocorticoid receptor (MR). The major tibolone metabolites, the two hydroxymetabolites, bind and activate ER with a preference for ERalpha. Tibolone and the Delta(4)-tibolone are agonists for AR and PR and surprisingly 3alpha- and 3beta-OH-tibolone are antagonists for them. Moreover we showed for the first time that tibolone and its primary metabolites are GR and MR antagonists with a stronger affinity for MR than for GR. In conclusion, tibolone by these actions on different receptors and by this capacity to transform in different metabolites, has more complex effects than initially supposed.

6beta-Hydroxy-5beta-methyl-20-oxo-19-norpregn-9(10)-en-3beta-yl acetate.

In the title compound, C(23)H(34)O(4), which is an intermediate in the synthesis of pregnane derivatives with a modified skeleton that show potent abortion-inducing activity, the conformation of ring B is close to half-chair due to the presence of both the C=C double bond and the axial 5beta-methyl group. Rings A and C have conformations close to chair, while ring D has a twisted conformation around the bridgehead C-C bond. Molecules are hydrogen bonded via the hydroxyl and acetoxy groups into infinite chains. Quantum-mechanical ab initio Roothan Hartree-Fock calculations show that crystal packing might be responsible for the low values of the angles between rings A and B, and between ring A and rings C and D, as well as for a different steric position of the methyl ketone side chain compared to the geometry of the free molecule.

3beta-OH-tibolone acutely dilates rat muscle arterioles similar to 3alpha-OH-tibolone.

In an earlier study, we focused on the vasoactive effect of 3alpha-OH-tibolone on spontaneously constricted isolatedfemale rat gracilis muscle arterioles. Vasodilator effects (from 10 to 10 M) of 3alpha-OH-tibolone were similar to those of 17beta-estradiol. It was reported that 3beta-OH-tibolone like estradiol altered GABAB activation in neurons through a membrane estrogen receptor, whereas the 3alpha-OH metabolite did not. We therefore hypothesized that the 3beta-OH metabolite may also have a vasodilating effect in our isolated arteriole model. The results indicate that 3beta-OH-tibolone induces a vasodilator effect in small arterioles that is comparable with that of 3alpha-OH-tibolone at the same concentration. This is intriguing because the binding affinity of 3alpha-OH-tibolone to the estrogen receptor is almost twice that of 3beta-OH-tibolone. Other mechanisms may play a role.

The microbial hydroxylation of levonorgestrel.

The microbial transformation of levonorgestrel (1) by Cunningham elegans resulted in the formation of five hydroxylated metabolites, 13-ethyl-10beta, 17beta-dihydroxy-18,19-dinor-17alpha-pregn-4-en-20-yn-3-one(2), 13-ethyl-6beta,17beta-dihydroxy-18,19-dinor-17alpha-pregn-4-en-20-yn-3-one (3) 13-ethyl 6beta, 10beta, 17beta-trihydroxy-18,19-dinor-17alpha-pregn-4-en-20-yn-3-one (4) 13-ethyl-15alpha-17beta-dihydroxy-18,19-dinor-17alpha-pregn-4-en-20-yn-3-one (5) and 13-ethyl-11alpha, 17beta-dihydroxy-18,19-dinor-17alpha-pregn-4en-20-yn-3-one. The fermentation of one with Rhizopus stolonifer, Fusarium lini and Curvularia lunata afforded compound 2 as a major metabolise. These metabolites were structurally characterized on the basis of spectroScopic techniques. Metabolite 6 was identified as a new compound. Compounds 2 2 ad 5 displayed inhibitory activity against the acetylcholinesterase ( AChE, EC. 3.1.1.7) with IC50 values of 79.2 and 24.5 microM, respectively. The metabolites 2 and 5 also showed inhibitory activity against the butyryLcholinesterase ( BChE, E.C 3.1.1.8) with IC50 values ranging between 9.4 and 309.8 microM.

Avaliaçäo da densidade mamográfica em mulheres na menopausa, sob terapia de reposiçäo hormonal convencional e tibolona / Mammographic density evaluation in menopausal womwn on conventional hormonal replacement therapy and tibolone

Objetivou-se analisar as alteraçöes de densidade mamográficas em mulheres na menopausa, submetidas a diferentes esquemas de terapia de reposiçäo hormonal (TRH).Estudaram-se em três grupos de 32 mulheres: G1 controle, G2 usuárias de estrogênios conjugados isolados (0,625 mg/dia) ou associados a medroxiprogesterona (2,5 mg/dia ou 5 mg/12 dias/mês) e G3 usuárias de tibolona (2,5 mg/dia). Critérios de inclusäo: duas mamografias, intervalo de 12 meses, a primeira prévia à TRH. Utilizaram-se dois métodos de classificaçäo de densidade mamográfica: Wolfe (N1, P1, P2, DY) e Colégio Brasileiro de Radiologia (CBR 1, 2, 3,4). Dois radiologistas revisaram os laudos, separadamente, sem o conhecimento dos grupos. Observou-se que as pacientes do G1 e G3 eram mais idosas e com maior tempo de menopausa quando comparadas as do G2(p < 0,05). Relataram mastalgia e aumento do volume mamário, 9 pacientes do G2 e 4 de G3. Näo houve relato no G1 (p < 0,05). Observamos que mulheres com mamas pouco densas (N1, P1 ou CBR 1,2) tinham 3,6 gestaçöes e aquelas com mamas densas (P2, DY ou CBR3,4), 2,3 gestaçöes (p < 0,01). Encontrou-se aumento de densidade mamográfica, segundo os critérios de Wolfe, em 6,2 porcento no G2, 9,4 porcento no G2 e 3,2 porcento no G3 e, segundo os critérios do CBR. em 3,2 porcento, 12,3 porcento e 6,2 porcento, respectivamente. Houve discordância interobservadores, nos laudos inicial e final, em 20,8 porcento e 24 porcento, segundo os critérios de Wolfe e 14,6 porcento e 11,5 porcento, segundo os critérios do CBR, respectivamente. Pela interpretaçäo de Kappa, a concordância interobservadores foi boa e a concordância entre os critérios de Wolfe e do CBR, fraca. A tibolona parece provocar menos sintomas mamários e menores alteraçöes de densidade mamários, quando comparada à TRH convencional

Determination of delta4-3-ketosteroids based on oxime formation by difference circular dichroism spectroscopy.

delta4-3-Ketosteroids exhibit an intensive negative Cotton effect on the circular dichroism (CD) spectra in the wavelength range for the n-pi* electronic transition (270-350 nm). With hydroxylamine hydrochloride, delta4-3-ketosteroid compounds can be transformed into oxime derivatives. Following oxime formation, positive ellipticity with low intensity can be registered in this wavelength range. The quantitative determination of delta4-3-ketosteroids is based on the considerable difference between the ellipticities before and after oxime formation. The difference ellipticity for the six ketosteroids examined (norethisterone, levonorgestrel, levonorgestrel acetate, methyltestosterone, testosterone phenylpropionate, nortestosterone phenylpropionate) varies linearly with the concentration in the interval 6 x 10(-6)-3 x 10(-3) mol/L. The method can be well applied to determination of delta4-3-ketosteroid contamination of norgestimate [(+)-13-ethyl-17-hydroxy-18,19-dinor-17alpha-pregn4-en-20-yn-3-one oxime acetate]; 0.02-10% impurity can be measured.

Norpregnane glycosides from a Thai soft coral, Scleronephthya pallida.

Two new norpregnane glycosides, 19-norpregna-1,3,5(10), 20-tetraen-3-O-alpha-fucopyranoside (3) and 19-norpregna-1,3,5(10), 20-tetraen-3-O-beta-arabinopyranoside (4), were isolated from the soft coral Scleronephthya pallida, along with two known steroids, pregna-1,20-dien-3-one (1) and 19-norpregna-1,3,5(10), 20-tetraen-3-ol (2). 19-Norpregna-1,3,5(10), 20-tetraen-3-O-alpha-fucopyranoside (3) exhibits moderate antimalarial and cytotoxic activities. The chemical structures of 1-4 were elucidated from spectroscopic data.

Lack of estrogenic potential of progesterone- or 19-nor-progesterone-derived progestins as opposed to testosterone or 19-nor-testosterone derivatives on endometrial Ishikawa cells.

Estrogen receptors of human endometrial cancer Ishikawa cells were found to be present in moderate amounts (160-200 fmol/mg protein), and to specifically bind moxestrol (R2858) with a very high affinity characterized by a Kd around 60 pM, when measured under equilibrium conditions. The binding specificity respected a decreasing order as follows: estradiol (E2: 100%) > 4-hydroxy-tamoxifen (4OHTAM: 52.7%) > estriol (E3: 5.7%) > estrone (E1: 2.1%) > TAM (0.2%). The induction of alkaline phosphatase activity (APase) used as an estrogen-specific response, confirmed the intrinsic estrogenicity of progestins derived from 19-nor-testosterone (19NT): norethindrone (NOR), norethynodrel and levonorgestrel, at concentrations ranging from 10(-8) to 10(-6) M. The effect of NOR was partially blocked by the antiestrogen 4OHTAM, which was also partially agonistic in this model, but neither by the antiprogestin mifepristone (RU486) nor by the aromatase inhibitor aminoglutethimide. A simulatory effect was also detected at 10(-7) or 10(-6) M with ethindrone, the testosterone- (T) derived progestin homologous to NOR, and with both androgenic parent-compounds, i.e. T and 19NT themselves. In contrast, progesterone (P) derivatives like medroxyprogesterone acetate (MPA) and chlormadinone acetate (CMA) remained totally inactive, as well as 19-nor-progesterone (19NP) itself or its progestagenic derivatives: ORG 2058 and nomegestrol acetate (NOM). Structure-activity relationships deduced from these studies suggest that it is not the absence of the 19-methyl group which can account for the estrogenic potential of the so-called "19-norprogestins", but rather their steroid structure derived from T in a broad sense (including the 19NT derivatives), as opposed to the non-estrogenic therapeutic progestins derived from P like MPA or CMA, or from 19NP like NOM.

Experiencia clínica con un nuevo anticonceptivo: desogestrel/etinil estradiol / Clinical experience with a new contraceptive: desogestrel/ethynil stradiol

Se estudió el efecto anticonceptivo, los efectos secundarios y las modificaciones en los sangrados menstruales, el peso corporal, la presión arterial y constantes de laboratorio, en dos grupos de mujeres: el primero utilizando la mezcla anticonceptiva de 150 mcg de desogestrel y 30 mcg de etinil estradiol y el segundo la de 150 mcg de levo-norgestrel y 30 mcg de etinil estradiol. Se estudiaron en total 252 mujeres, durante 4.332 ciclos, con un tiempo máximo de consumo de 34 ciclos. Los resultados mostraron un muy favorable efecto del desogestrel/etinil estradiol sobre el peso corporal, que no aumentó sino aun disminuyó con esta mezcla, la cual además produjo menores alteraciones del sangrado menstrual, mínimos e insignificantes efectos secundarios, gran tolerancia y un alto índice de continuidad de 89%

[The effect of low dose gestagens on the hypothalamic-pituitary-ovarian axis]. / Wirkung niedrig dosierter Gestagene auf die hypothalamisch-hypophysär-ovarielle achse.

The contraceptive effect of continuous treatment with low dose progestogens (minipill) has been attributed mainly to alterations of the cervical mucus and the endometrium. This study was undertaken to investigate the effect of low dose progestogens on hypothalamic-pituitary-ovarian function. Plasma concentrations of follicle stimulating hormone (FSH), luteininzing hormone (LH), estradiol-17 beta (E-2) and Progesterone were measured daily during apparently ovulatory menstrual cycles and during treatment cycles with different low dose progestogens. From the results obtained it was concluded that the minipill has a clear-cut effect on the LH/FSH peak at midcycle and on corpus luteum function. A cyclic secretion of E-2 is maintained in the majority of cases. In a few treatment cycles however, follicular maturation was suppressed as indicated by low E-2 concentrations. It was concluded that the minipill exerts a profound effect at the central and ovarian level which contributes to its satisfactory contraceptive efficacy.

PIP: Different doses of the progestagens Lynestrenol and ORG 3236 were administered continuously in small quantities. Radioimmunological assays were made to determine the different effects of these "minidoses" on the function of the hypothalamic-pituitary-ovarian system. This is determined by the levels of follicular stimulating hormone (FSH), luteinizing hormone (LH), estradiol-17beta (E2), and progesterone during ovulation and treatment cycles. Low progestagen doses had a clear-cut effect on the LH/FSH peak in midcycle and on the corpus luteum function. The cyclical secretion of E2 is maintained at normal levels in most cases. A few cases did show follicular maturation as being suppressed, as indicated by low concentrations of E2. The "minipill" was shown to produce a profound effect at the central and ovarian level, which contributes to its efficiency and safety as a contraceptive.