RESUMO
BACKGROUND: The FIFA has implemented an important antidoping programme for the 2014 FIFA World Cup. AIM: To perform the analyses before and during the World Cup with biological monitoring of blood and urine samples. METHODS: All qualified players from the 32 teams participating in the World Cup were tested out-of-competition. During the World Cup, 2-8 players per match were tested. Over 1000 samples were collected in total and analysed in the WADA accredited Laboratory of Lausanne. RESULTS: The quality of the analyses was at the required level as described in the WADA technical documents. The urinary steroid profiles of the players were stable and consistent with previously published papers on football players. During the competition, amphetamine was detected in a sample collected on a player who had a therapeutic use exemption for attention deficit hyperactivity disorder. The blood passport data showed no significant difference in haemoglobin values between out-of-competition and postmatch samples. CONCLUSIONS: Logistical issues linked to biological samples collection, and the overseas shipment during the World Cup did not impair the quality of the analyses, especially when used as the biological passport of football players.
Assuntos
Dopagem Esportivo/prevenção & controle , Futebol/fisiologia , Anfetamina/análise , Androstenodiona/análogos & derivados , Androstenodiona/análise , Análise Química do Sangue/métodos , Brasil , Clembuterol/análise , Glucocorticoides/análise , Humanos , Manejo de Espécimes/métodos , Esteroides/análise , Detecção do Abuso de Substâncias/métodos , Tramadol/análise , Urinálise/métodosRESUMO
Cysticerci and tapeworms from Taenia crassiceps WFU, ORF and Taenia solium synthesize sex-steroid hormones in vitro. Corticosteroids increase the 17ß-estradiol synthesis by T. crassiceps cysticerci. T. crassiceps WFU cysticerci synthesize corticosteroids, mainly 11-deoxycorticosterone (DOC). The aim of this work was to investigate whether classical steroidogenic inhibitors modify the capacity of T. crassiceps WFU cysticerci to synthesize corticosteroids and sex steroid hormones. For this purpose, T. crassiceps WFU cysticerci were obtained from the abdominal cavity of mice, pre-cultured for 24h in DMEM+antibiotics/antimycotics and cultured in the presence of tritiated progesterone ((3)H-P4), androstendione ((3)H-A4), or dehydroepiandrosterone ((3)H-DHEA) plus different doses of the corresponding inhibitors, for different periods. Blanks with the culture media adding the tritiated precursors were simultaneously incubated. At the end of the incubation period, parasites were separated and media extracted with ether. The resulting steroids were separated by thin layer chromatography (TLC). Data were expressed as percent transformation of the tritiated precursors. Results showed that after 2h of exposure of the cysticerci to 100 µM formestane, the (3)H-17ß-estradiol synthesis from tritiated androstenedione was significantly inhibited. The incubation of cysticerci in the presence of (3)H-DHEA and danazol (100 nM) resulted in (3)H-androstenediol accumulation and a significant reduction of the 17ß-estradiol synthesis. The cysticerci (3)H-DOC synthesis was significantly inhibited when the parasites were cultured in the presence of different ketoconazole dosis. The drug treatments did not affect parasite's viability. The results of this study showed that corticosteroid and sex steroid synthesis in T. crassiceps WFU cysticerci can be modified by steroidogenic enzyme inhibitors. As was shown previously by our laboratory and others, parasite survival and development depends on sex steroids, therefore the inhibition of their synthesis is a good starting point exploited in situations where the inhibition of steroidogenesis could help to control the infection for the development of new treatments, or replacement of the usual therapy in resistant parasite infections. We raise the possibility that these drug actions may be beneficially.
Assuntos
Inibidores Enzimáticos/farmacologia , Esteroides/metabolismo , Taenia/efeitos dos fármacos , Taenia/metabolismo , Androstenodiona/análogos & derivados , Androstenodiona/farmacologia , Animais , Cromatografia em Camada Fina , Danazol/farmacologia , Desoxicorticosterona/farmacologia , Estradiol/metabolismo , Cetoconazol/farmacologiaRESUMO
The aim of this study was to synthesize three different D-homoandrostadiene derivatives (2-4) and study their biological activity. We carried out in vivo and in vitro experiments using female cycling mice, which were synchronized for estrus with luteinizing hormone-releasing hormone (LHRH) and injected with the steroidal compounds. It was also determined the binding of these compounds to the progesterone receptors (PR). Since these steroids have a new D-homoandrostandienone skeleton in their molecular structure, it was of interest also to study their binding to the androgen receptors (AR). After LHRH treatment, the mice of the control group showed the presence of 14+/-4 corpus lutea in the ovary whereas the animals treated with steroids 2-4, with RBAs of 100%, exhibited 11+/-7, 12+/-2, and 10+/-4 respectively. As a result of this study, it is evident that these steroids did not inhibit the ovulation in these animals. The uterus of the control group, showed the typical progestational activity with an enlarged endometrial thickness with a secretory activity. However, the endometrium of the mice treated with steroids 2-4 did not show an enlargement of the endometrium and no secretory activity could be detected. This fact indicates that compounds 2-4 had antagonistic activity in this tissue. The overall data show that steroids 2-4 are antagonists of the PR. However, they do not bind to the AR. These results also demonstrate that 2-4 have an antiprogestational activity in vivo, but do not decrease the number of corpus lutea in the ovary of mice treated with LHRH.
Assuntos
Androstadienos/química , Androstadienos/farmacologia , Receptores de Progesterona/antagonistas & inibidores , Antagonistas de Receptores de Andrógenos , Androstadienos/metabolismo , Androstenodiona/análogos & derivados , Androstenodiona/química , Androstenodiona/farmacologia , Animais , Ligação Competitiva , Corpo Lúteo/efeitos dos fármacos , Corpo Lúteo/metabolismo , Endométrio/efeitos dos fármacos , Endométrio/metabolismo , Feminino , Hormônio Liberador de Gonadotropina/farmacologia , Homosteroides/química , Homosteroides/farmacologia , Antagonistas de Hormônios/química , Antagonistas de Hormônios/metabolismo , Antagonistas de Hormônios/farmacologia , Masculino , Camundongos , Mifepristona/química , Mifepristona/metabolismo , Mifepristona/farmacologia , Estrutura Molecular , Ovário/efeitos dos fármacos , Ovário/metabolismo , Fenilacetatos/química , Progesterona/química , Progesterona/metabolismo , Progesterona/farmacologia , Coelhos , Ratos , Receptores Androgênicos/metabolismo , Receptores de Progesterona/metabolismo , Relação Estrutura-Atividade , Útero/efeitos dos fármacos , Útero/metabolismoRESUMO
A simple and novel HPLC-MS method for the simultaneous quantification of testosterone, 11-ketotestosterone, and 11beta-hydroxyandrostenedione in fish serum was developed and validated. Separation was achieved on a C-18 column using a water-acetonitrile mobile-phase with a cycle time of 12 min. Ion detection was performed using ESI positive SIM at [M+H] (m/z 303, 303, 289). The linear ranges (0.2-50 ng/ml), limits of detection (0.1-0.2 ng/ml) and quantification (0.2-0.5 ng/ml) were established. The method was validated by measuring the three androgens in goldfish sera, displaying comparable values to those reported by other analytical techniques (RIA, EIA).
Assuntos
Androstenodiona/análogos & derivados , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Testosterona/análogos & derivados , Testosterona/sangue , Androstenodiona/sangue , Animais , Carpa DouradaRESUMO
The present study was designed to obtain new insights into fish gonadal sex differentiation by comparing the effects of two different masculinizing treatments on some candidate gene expression profiles. Masculinization was induced in rainbow trout, Oncorhynchus mykiss, genetic all-female populations using either an active fish androgen (11betaAnd, 11beta-hydroxyandrostenedione) or an aromatase inhibitor (ATD, 1,4,6-androstatriene-3,17-dione). The expression profiles of 100 candidate genes were obtained by real-time RT-PCR, and 46 profiles displayed a significant differential expression between control populations (males and females) and ATD/11betaAnd-treated populations. These expression profiles were grouped in four temporally correlated expression clusters. Among the common responses shared by the two masculinizing treatments, the inhibition of some early female differentiating genes (cyp19a1, foxl2a, fst, and fshb) appears to be crucial for effective masculinization, suggesting that these genes act together via a short regulation loop to maintain high sex-specific ovarian expression of cyp19a1. This simultaneous down-regulation of female-specific genes could be triggered by some testicular genes, such as dmrt1, nr0b1 (also known as dax1), and pdgfra, which are quickly up-regulated by the two masculinizing treatments. In contrast to 11betaAnd, ATD quickly restored the expression levels of steroidogenesis related genes (cyp11b2.1, cyp11b2.2, hsd3b1, cyp17a, star, and nr5a1) and some Sertoli cell markers (sox9a2 and amh) to the expression levels observed during control testicular differentiation. This demonstrates that these genes are probably not needed for active masculinization and that the inhibition of endogenous estrogen synthesis produces a much more complete and specific testicular pattern of gene expression than that observed following androgen-induced masculinization.
Assuntos
Androgênios/farmacologia , Estrogênios/metabolismo , Oncorhynchus mykiss/fisiologia , Ovário/fisiologia , Diferenciação Sexual/fisiologia , Testículo/fisiologia , Androstatrienos/farmacologia , Androstenodiona/análogos & derivados , Androstenodiona/farmacologia , Animais , Aromatase/genética , Aromatase/metabolismo , Receptor Nuclear Órfão DAX-1 , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Inibidores Enzimáticos/farmacologia , Feminino , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Perfilação da Expressão Gênica , Genótipo , Masculino , Oncorhynchus mykiss/genética , Ovário/efeitos dos fármacos , Fenótipo , Receptores do Ácido Retinoico/genética , Receptores do Ácido Retinoico/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Diferenciação Sexual/genética , Testículo/efeitos dos fármacos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
In male rats, the antidepressant-like effect of fluoxetine (FLX) and desipramine (DMI) in the forced swimming test (FST) is reduced by orchidectomy and partially restored by testosterone (T). It is unknown if this modulation of T is produced by its estrogenic metabolites. The objectives of this study were to evaluate if the aromatase inhibitor, formestane, interferes with the antidepressant-like effect of DMI and FLX in intact male rats, and to analyze if 17beta-estradiol (E2) modifies the FST and interacts with the antidepressants in orchidectomized (Orx) males. Intact males received DMI (1.25-5.0 mg/kg) and FLX (2.5-10 mg/kg) alone or in combination with formestane (17.5 mg/kg). Orx rats received E2 (5, 10, 20 and 40 microg/rat) or the combination of E2 [at sub-threshold (5 microg/rat) and optimal (10 microg/rat) doses] plus sub-effective doses of DMI (2.5 mg/kg) or FLX (10 mg/kg). Serum testosterone and estradiol levels were measured in intact-control and -formestane treated animals as well as in castrated males replaced with various doses of E2. Formestane in intact males lacked of an action in the FST, but cancelled the antidepressant-like effect of DMI and FLX. E2 at the supra-physiological doses of 10 and 20 microg/rat produced antidepressant-like effects. E2 at 5 microg/rat (that re-established the levels of this hormone to physiological levels) and at 10 microg/rat restored the antidepressant-like action of DMI and FLX in Orx rats. It was concluded that estrogens participate in the antidepressant-like effect of DMI and FLX in the FST.
Assuntos
Antidepressivos de Segunda Geração/farmacologia , Antidepressivos Tricíclicos/farmacologia , Desipramina/farmacologia , Estrogênios/fisiologia , Fluoxetina/farmacologia , Androstenodiona/análogos & derivados , Androstenodiona/farmacologia , Animais , Inibidores da Aromatase/farmacologia , Química Encefálica/efeitos dos fármacos , Estradiol/sangue , Estradiol/farmacologia , Feminino , Masculino , Atividade Motora/efeitos dos fármacos , Orquiectomia , Ovariectomia , Ratos , Natação/psicologia , Testosterona/sangueRESUMO
OBJECTIVE: To determine the sensitivity of 11beta-hydroxyandrostenedione (11-OHA4) and delta5-androstenediol (ADIOL) as markers of excessive adrenal androgen production. DESIGN: Prospective study. SETTING: Academic medical centers. PATIENT(S): Thirteen women with untreated 21-hydroxylase-deficient nonclassic adrenal hyperplasia (NCAH) and 18 healthy, eumenorrheic, nonhirsute controls matched for age and body mass index. INTERVENTION(S): All subjects were studied before and after acute adrenal stimulation with 0.25 mg of IV ACTH-(1-24). MAIN OUTCOME MEASURE(S): Basal levels of total testosterone, sex hormone-binding globulin, DHEAS, and free testosterone were measured. Levels of androstenedione (A4), DHEA, 11-OHA4, and ADIOL were determined before (Steroid0) and 60 minutes after (Steroid60) acute ACTH-(1-24) stimulation. RESULT(S): Patients with NCAH had higher median basal levels of DHEAS and total and free testosterone than controls. Patients with NCAH had higher median A4(0), A460, DHEA(0), DHEA60, 11-OHA4(0), ADIOL0, and ADIOL60 levels but similar 11-OHA4(60) levels compared with controls. Among patients with NCAH, 30%, 54%, 15%, and 85% had 11-OHA4(0), ADIOL0, 11-OHA4(60), and ADIOL(60) levels, respectively, above the 95th percentile of controls. CONCLUSION(S): Overall, serum levels of 11-OHA4 did not appear to be a very sensitive marker of excessive adrenal androgen production, at least in patients with NCAH. Although ACTH-stimulated ADIOL levels were elevated in 85% of the patients studied, they did not appear to have any advantage over the measurement of A4 or DHEA levels.
Assuntos
Hiperplasia Suprarrenal Congênita/metabolismo , Androgênios/biossíntese , Androstenodiona/análogos & derivados , Adulto , Androstenodiona/sangue , Biomarcadores/sangue , Índice de Massa Corporal , Estudos de Casos e Controles , Feminino , Humanos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The effect of androstenedione on luteal progesterone production was studied during luteolysis preceding parturition as well as that induced by the antiprogestin RU486 in late pregnant rats. Luteal cells from animals on days 19, 20 or 21 of pregnancy and incubated with 10 microM androstenedione increased progesterone production by 99, 136, and 277%, respectively. The animals receiving androstenedione (10 mg/rat s.c.) on day 19 of pregnancy showed an increase in serum progesterone levels, a decline in luteal 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity and an increase in corpus luteum weight without modifying 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD) activity on day 21 of pregnancy. Androstenedione and testosterone but not dihydrotestosterone were able to prevent the decrease in serum progesterone concentration and corpus luteum weight observed 58 h after treatment with RU486 (2 mg/kg) on day 18 of pregnancy. However, the three androgens studied inhibited the luteal 3 beta-HSD activity but 20 alpha-HSD activity was not affected, when compared with animals receiving RU486 alone. The co-administration of androstenedione with the aromatase inhibitor 4-hydroxyandrostenedione or with the specific antioestrogen ICI 164,384 did not modify the effects induced by androstenedione in RU486-treated rats, indicating that the action of androstenedione on progesterone production and secretion at the time of luteolysis seems to occur through an androgenic mechanism and is not mediated by previous conversion of the androgens to oestrogens. In all experiments the high luteal 20 alpha-HSD activity, that characterizes a luteolytic process, was not modified by androgens. Androstenedione administered to adrenalectomized rats was also able to prevent the decrease in serum progesterone concentration observed in spontaneous or RU486-induced luteolysis. The administration of androstenedione to RU486-treated rats induced a decrease in luteal progesterone content concomitant with an increase in serum progesterone levels. These studies demonstrate that androgens during luteolysis, are able to stimulate luteal progesterone secretion, prevent the loss in corpora lutea weight and enhance the decrease in 3 beta-HSD activity, without affecting the increase in 20 alpha-HSD activity.
Assuntos
Androstenodiona/farmacologia , Corpo Lúteo/metabolismo , Luteólise/metabolismo , Mifepristona/farmacologia , Prenhez , Progesterona/biossíntese , 17-Hidroxiesteroide Desidrogenases/metabolismo , 20-Hidroxiesteroide Desidrogenases/metabolismo , 20-alfa-Hidroxiesteroide Desidrogenase , Adrenalectomia , Androstenodiona/análogos & derivados , Androstenodiona/metabolismo , Animais , Inibidores da Aromatase , Células Cultivadas , Corpo Lúteo/citologia , Corpo Lúteo/efeitos dos fármacos , Di-Hidrotestosterona/farmacologia , Estradiol/análogos & derivados , Estradiol/farmacologia , Feminino , Antagonistas de Hormônios/farmacologia , Luteolíticos/farmacologia , Indutores da Menstruação/farmacologia , Tamanho do Órgão/efeitos dos fármacos , Alcamidas Poli-Insaturadas , Gravidez , Progesterona/sangue , Ratos , Ratos Wistar , Testosterona/farmacologiaRESUMO
Serum cortisol, 17-hydroxyprogesterone and five androgens (androstenedione, 11-hydroxyandrostenedione, dehydroepiandrosterone and its sulphate, and testosterone) were measured by radioimmunoassay at 8:00, 16:00, 20:00 and 24:00 h and again at 8:00 h on the next day in eight healthy female subjects aged 16-40 years in the midfollicular phase of the menstrual cycle. Regression analysis revealed a significant (alpha = 0.05) circadian periodicity, with estimated peaks between 8:00 a.m. (testosterone) and 2:00 p.m. (dehydroepiandrosterone sulphate), and with estimated amplitudes between 76% (11-hydroxyandrostenedione) and 33% (17-hydroxyprogesterone, testosterone and dehydroepiandrosterone sulphate) of the rhythm adjusted mean. The inferential statistical approach illustrated here should contribute significantly to the diagnostic elucidation of adrenocortical disorders.