Effect of lincRNA PVT1 associated with Enhancer of Zeste homolog 2 (EZH2) and with androgen receptor (AR) on the large-scale gene expression in LNCaP prostate cancer cells

O lincRNA PVT1 (Plasmacytoma Variant Translocation 1) é um RNA longo não codificador de proteínas (ncRNA) descrito como um oncogene sendo superexpresso em vários tipos de cânceres. LincRNA PVT1 está localizado na região genômica 8q24, também conhecida como 'gene desert'. O nível de expressão do lincRNA PVT1 está associado ao aumento do risco de câncer de próstata (PCa) e está correlacionado com os níveis de expressão do receptor de andrógeno (AR). No entanto, o mecanismo do envolvimento do lincRNA PVT1 com o AR no desenvolvimento de câncer de próstata ainda não está bem esclarecido. Aqui, nós testamos a hipótese que a formação do complexo AR-EZH2-PVT1 participa na regulação da expressão gênica em câncer de próstata, nas células LNCaP. A imunoprecipitação de ribonucleoproteínas seguida de PCR quantitativo (RIP-qPCR) revelou que o lincRNA PVT1 está associado fisicamente ao AR (12% do input) e à metiltransferase EZH2, proteína componente do complexo repressor Polycomb 2 (36% do input) sob condições suplementadas com andrógeno (+R1881). O lincRNA PVT1 também está associado fisicamente ao AR (10% de input) e à EZH2 (42% de input) em condições de privação de andrógeno (-R1881). Assim, a associação física entre lincRNA PVT1, AR e EZH2 é independente do hormônio andrógeno. Usando uma abordagem de estudo em larga-escala de perda e ganho de função, nossos resultados mostraram que o silenciamento do lincRNA PVT1 em células LNCaP na presença de andrógeno restaura a expressão parcialmente, totalmente ou causa superexpressão de 160 genes que tiveram a expressão inibida por andrógeno. Entre esses genes, destacamos genes envolvidos na regulação da diferenciação celular, em componentes da junção célula-célula, na inibição da migração e invasão celular e no desencadeamento da via apoptótica. Imunoprecipitação da cromatina seguida de PCR quantitativo (ChIP-qPCR), em cultura de células LNCaP suplementada com andrógeno sob silenciamento do lincRNA PVT1, mostrou aumento significativo na ocupação pela marca de histona ativadora H3K27Ac do promotor do gene NOV, um dos genes que tiveram sua expressão aumentada com o silenciamento de PVT1. O ChIP-qPCR também mostrou, após o silenciamento do lincRNA PVT1, um aumento significativo da marca H3K27me3 na região enhancer do gene NOV, uma característica de enhancers poised (prontos para ativação). Em conclusão, nós fornecemos a primeira evidência experimental para um mecanismo de ação do oncogene lincRNA PVT1 em células de câncer de próstata e demonstramos que sua ação inibidora da expressão afeta genes alvo que facilitam a proliferação e migração de células do câncer de próstata, sugerindo que o lincRNA PVT1 é um novo agente no complexo mecanismo de repressão transcricional envolvendo um RNA silenciador, o receptor de andrógeno (AR) e o potenciador de Zeste homólogo 2 (EZH2) no remodelamento da cromatina em células LNCaP

Long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) is an oncogene known to be overexpressed in various types of cancer. PVT1 lincRNA is located in the wellknown cancer-related genomic region 8q24, also known as 'gene desert. PVT1 lincRNA level of expression is associated with increased prostate cancer (PCa) risk and is correlated with androgen receptor (AR) expression levels. However, the mechanism of PVT1 and AR involvement in the development of prostate cancer is still unclear. Here, we tested the hypothesis that formation of the complex AR-EZH2-PVT1 participates in the regulation of gene expression in prostate cancer, in LNCaP cells. Ribonucleoprotein immunoprecipitation followed by quantitative PCR (RIP-qPCR) revealed that PVT1 lincRNA binds both the AR (12 % of PVT1 input) and the methyltransferase EZH2 from the Polycomb repressive complex 2 (36 % of input) under androgen-supplemented conditions (+R1881). PVT1 also binds both AR (10 % of input) and EZH2 (42 % of input) under androgen-deprived conditions (-R1881). Thus, PVT1 binding to AR and EZH2 is independent of the androgen hormone. Using a large-scale loss and gain of function approach, our results show that PVT1 knockdown (KD) in LNCaP in the presence of androgen restores the expression partially, fully or causes overexpression of 160 genes that are inhibited by androgen. Among these genes, we highlight genes involved in regulation of cell differentiation, in components of cell-cell junction, in inhibition of cell migration and invasion and in triggering of the apoptotic pathway. Chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) with LNCaP cells in androgen-supplemented cultures under PVT1 lincRNA knockdown showed a significant increase in occupancy by the histone activation mark H3K27Ac of the promoter region of the NOV gene, one of the genes that had an increased expression upon PVT1 silencing. ChIPqPCR also showed a significant increase upon PVT1 lincRNA silencing of the H3K27me3 histone mark in the enhancer region of the NOV gene, a distinct feature of poised enhancers. In conclusion, we provide first experimental evidence for a mechanism of action of PVT1 lincRNA oncogene in prostate cancer cells, and show that its inhibitory action affects targetgenes that facilitate proliferation and migration of prostate cancer cells, thus suggesting PVT1 lincRNA as a novel lncRNA player in the complex mechanism of transcriptional repression involving a silencer RNA, the androgen receptor (AR) and the Enhancer of zeste homolog 2 (EZH2) in chromatin remodeling in LNCaP cells

Tumor ovárico de células de Leydig en una niña de 7 años que se presentó como pubarquia precoz con elevación de 17 hidroxiprogesterona / Ovarian tumor of Leydig cells in a 7-year-old girl who presented as precocious pubarche with elevation of 17 hydroxyprogesterone

Clinical case: a girl of 7 ½ years who consulted for early pubarche without thelark, with a percentile size of 75 for a genetic target size in the 10th percentile, overweight with a 90th percentile BMI, and normal blood pressure. The biochemical study showed high levels of androgens: testosterone: 7.2 ng/dL, androstenedione of 5.1 ng / ml, 17OHP: 15 ng / dL with low normal DHEAS (0.26 ug/ml), Plasma Renin Activity normal low: 0.22 ng/mL/h. Initial imaging study showed a bone age of 10 years 6 months and normal abdominal and pelvic ultrasound. Molecular study showed no pathogenic variants in the CYP21A2 gene (21 Hydroxylase). With a probable diagnosis of non-classical congenital adrenal hyperplasia (HSRNC) and no known mutation, he started treatment with hydrocortisone (12 mg/m2). At 8.7 years, pubertal development begins and braking begins with LHRH analogues, which are administered for 18 months. Despite the treatment, signs of virilization and elevation of androgens (testosterone up to 130 ng/ml) are progressively accentuated, which do not diminish when trying different corticosteroid schemes. MRI of the abdomen and pelvis shows the normal adrenal glands and a solid nodular image of 2.1 x 1.6 cm in the right ovary (Figure 2), later demonstrated with pelvic ultrasound (Figure 2). Right laparoscopic oophorectomy was performed, whose biopsy demonstrated a Leydig cell tumor. One month after surgery, all androgenic levels were normalized, so the gradual suspension of corticosteroids began. Conclusion: Although HSRNC is the most frequent pathological cause of early pubarche, when it is associated with progressive clinical and biochemical hyperandrogenism despite adequate treatment and without pathogenic variants in the CYP21A2 gene, even with high levels of 17OHP, other causes should be considered, specifically, androgen producing tumors.

Caso clínico: niña de 7½ años que consulta por pubarquia precoz sin telarquia, con talla en percentil 75 para una talla objetivo genético en percentil 10, sobrepeso con IMC percentil 90 y presión arterial normal. El estudio bioquímico mostró niveles elevados de andrógenos: testosterona: 7,2 ng/dL, androstenediona de 5,1 ng/ml, 17OHP: 15 ng/dL con DHEAS normal baja (0,26 ug/ml), Actividad de Renina Plasmática normal baja: 0.22 ng/ mL/h. Estudio de imágenes inicial mostró una edad ósea de 10 años 6 meses y ecografía abdominal y pelviana normales. Estudio molecular no mostró variantes patogénicas en el gen CYP21A2 (21 Hidroxilasa). Con diagnosticó probable de hiperplasia suprarrenal congénita no clásica (HSRNC) y sin mutación conocida,inició el tratamiento con hidrocortisona (12 mg/m2). A los 8.7 años comienza desarrollo puberal y se inicia frenación con análogos de LHRH, los cuales se administran por 18 meses. A pesar del tratamiento se acentúan progresivamente los signos de virilización y hayelevación de los andrógenos (testosterona hasta 130 ng/ml), que no disminuyen intentando diferentes esquemas de corticoides. Se realiza RM de abdomen y pelvis que muestra las glándulas suprarrenales normales y una imagen nodular sólida de 2.1 x 1.6 cm en el ovario derecho (Figura 2), demostrada posteriormente con Ecografía pelviana (Figura 2). Se realiza ooforectomía derecha por vía laparoscópica, cuya biopsia demostró un tumor de células de Leydig. Un mes después de la cirugía, se normalizan todos los niveles androgénicos por lo que se inició la suspensión gradual de los corticoides. Conclusión: Aunque la HSRNC es la causa patológica más frecuente de la pubarquia precoz, cuando se asocia con un hiperandrogenismo clínico y bioquímico progresivo a pesar de un tratamiento adecuado y sin variantes patógenicas en el gen CYP21A2, incluso con niveles elevados de 17OHP, otras causas deben ser consideradas, específicamente tumores productores de andrógenos.

Estudios del laboratorio de diversos parámetros orientados a tratar de dilucidar la fisiopatología del síndrome de ovarios poliquísticos (SOP). Parte 2 / Laboratory studies of several parameters aimed to elucidate the pathophysiology of polycystic ovary syndrome. Second part

RESUMEN Diversos estudios bioquímicos adicionales a la evaluación de Testosterona total (TT), biodisponible (Tbio) y libre (TL) han sido realizados a los efectos que pudieran resultar de mayor utilidad para el diagnóstico de patologías concomitantes en el SOP, entre otros. En la hormona anti Mülleriana, cuando la concentración supera a los 3,0 ng/ml existen evidencias de que el 79% de las mismas pueden ser identificadas correctamente como SOP. El Antígeno Prostático Específico (PSA), marcador de singular importancia en pacientes con cáncer de Próstata, con técnicas ultrasensibles ha podido ser detectado en más del 50% en mujeres. En un grupo de pacientes con SOP, los niveles circulantes de PSA fueron significativamente mayores que en las mujeres sin SOP. El Kiss-1 aislado de la placenta y demostrado en otros tejidos, presenta niveles aumentados que correlacionan con la LH, TT, TL y resistencia a la insulina (RI) en adolescentes con SOP versus adolescentes sin SOP, sugiriendo que el Kiss-1 podría estar involucrado en el desarrollo del SOP en estas pacientes. Algunas pacientes con SOP están asociadas a patologías relevantes, de las cuales han sido comunicadas el aumento del BMI, mayor grado de dislipemia, adiposidad central, RI y Síndrome Metabólico (SMe). En las pacientes con un fenotipo clásico (hiperandrogenismo, alteración del ciclo menstrual y ovarios poliquísticos), estas patologías son de mayor frecuencia y severidad que en los otros fenotipos, particularmente aquellos sin hiperandrogenismo. Otras determinaciones como TNFα, interleuquinas, test de tolerancia a la glucosa, ApoB, partículas pequeñas de LDL e Inhibidor del Activador del Plasminógeno-1 han sido comunicados que podrían ser de utilidad para tener mayor sensibilidad en la definición de patología concomitantes en el SOP. Actualmente se ha comenzado a evaluar otros marcadores como el Fetuin-A; Quemerina, Nesfatina-1, Neopterina y Endocannabinoides, cuyos resultados preliminares parecerían ser un aporte importante para evaluar SMe y RI en paciente con SOP y tratar de definir su prevalencia en los distintos fenotipos de esta patología.

ABSTRACT Several biochemical studies in addition to the evaluation of total Testosterone (TT), bioavailable (bioT) and free (FT) have been performed to the effects that could be of greater use for the diagnosis of concomitant pathologies in the PCOS, among others. The anti-Müllerian hormone whose concentration when exceeds 3.0 ng/ml, there is evidence that 79% of these patients can be correctly identified as PCOS. The Prostate-Specific Antigen (PSA), a marker of singular importance in patients with prostate cancer, with ultra-sensitive techniques, has been detected in more than 50% of women. In a group of patients with PCOS, circulating levels of PSA are significantly higher than in women without PCOS. The Kiss-1 isolated from the placenta and demonstrated in other tissues, has increased levels that correlate with LH, TT, TL and insulin resistance (IR) in adolescents with PCOS respect to adolescents without PCOS, suggesting that Kiss-1 could be involved in the development of the PCOS in these patients. In some patients with PCOS, they are associated with relevant pathologies, of which the increase in BMI, higher degree of dyslipidemia, central adiposity, IR and Metabolic Syndrome (MeS) have been reported. Those that show a classic phenotype (hyperandrogenism, alteration of the menstrual cycle and polycystic ovaries) these characteristics are of greater frequency and severity than in the other phenotypes, particularly those without hyperandrogenism. Other determinations such as TNFα, interleukins, glucose tolerance test, ApoB, small particles of LDL and Plasminogen Activator Inhibitor-1 have been reported that could be useful to have greater sensitivity in the definition of concomitant pathology in the PCOS. Currently, other markers such as Fetuin-A, Chemerin, Nesfatin-1 Neopterin and Endocannabinoids have been evaluated. The preliminary results suggest to be an important contribution to define MeS and IR in patient with PCOS and to try to determine its prevalence in the different phenotypes of this pathology.

Are 2D: 4D finger-length ratios an indicator of androgenetic alopecia in males?

Abstract BACKGROUND: Although the pathogenesis of androgenetic alopecia is not completely understood, the roles of genetic susceptibility and androgens are well-known. A lower ratio of the second digit (index finger = 2D) to the fourth digit (ring finger = 4D) length has been hypothesized to reflect prenatal androgen exposure and/or higher sensitivity to androgens. OBJECTIVES: To determine the relationship between the second to fourth digit length ratio and androgenetic alopecia. METHODS: Finger length measurements were made by a digital vernier calliper. Androgenetic alopecia severity was assessed using the Hamilton-Norwood scale. Subjects with an androgenetic alopecia score of grade III or more were included in the study. RESULTS: A total of 189 males with androgenetic alopecia and 171 healthy controls were enrolled in the study. The age range of participants was 19-65 years. The 2D:4D ratios in patients with androgenetic alopecia were significantly lower than those of healthy controls for the right hand; however, no significant difference was found for the left hand. Average 2D:4D ratios in androgenetic alopecia patients were also lower than in controls. No significant relationship was observed between androgenetic alopecia severity and 2D:4D ratios. CONCLUSION: Our data support the anatomical evidence of in utero androgen exposure and/or an individual’s sensitivity to androgens in patients with androgenetic alopecia. Furthermore, the right hand 2D:4D ratio might be an indicator of androgenetic alopecia development.

Morfometria corpórea, características do sêmen, proteínas seminais e testosterona em cervos Cervus unicolor, em cativeiro / Body morphometry, semen characteristics, seminal proteins and testosterone in sambar deer (Cervus unicolor) in captivity

Objetivou-se estudar a morfometria corpórea, as características do sêmen, o perfil proteico do plasma seminal em SDS-PAGE e a concentração sérica de testosterona em cervos-sambar (Cervus unicolor), criados em cativeiro, na estação reprodutiva da primavera. Quatro machos com idades entre 12 e 36 meses foram avaliados em quatro momentos, com intervalos de sete dias, com peso corpóreo (60,5 a 89,0kg), índice de massa corporal (93,07kg/m2 a 126,56kg/m2), volume do ejaculado (0,50±0,35mL a 0,75±0,28mL), motilidade espermática (87,75±4,78% a 90,00±7,07%), defeitos totais (17,25±5,81% a 47,72±17,55%), testosterona sérica (6,43±4,33ng/dL a 166,00±64,48ng/dL) e proteínas do plasma seminal com bandas entre 7,6 e 142kDa. As características dos ejaculados não diferiram (P>0,05) entre as três primeiras colheitas. Houve diferença (P<0,05) para os defeitos espermáticos com elevação na quarta colheita. No plasma seminal de cada cervo, foram identificadas de 16 a 27 bandas de proteínas entre 7,6 e 142kDa. Conclui-se que a qualidade espermática foi satisfatória na primavera. O estresse das contenções sucessivas causou queda da qualidade espermática. A idade influi na concentração sérica de testosterona, a qual foi maior nos cervos aos 36 meses...

The aim of this work was to study the body morphometry, semen characteristics, seminal plasma protein profile in SDS-PAGE and serum testosterone concentration in Sambar Deer (Cervus unicolor), in captivity in the breeding season (spring). Four males aged between 12 and 36 months were assessed in four moments with intervals of seven days with body weight (60.5 to 89.0kg), body mass index (93.07 to 126.56kg/m2), ejaculate volume (0.50±0.35mL to 0.75±0.28mL), sperm motility (87.75±4.78% to 90.00±7.07% ), total defects (17.25±5.81% to 47.72±17.55%), serum testosterone (6.43±4.33 ng/dL to 166.00±64.48ng/dL) and seminal plasma proteins with bands between 7.6 and 142 kDa. The characteristics of ejaculates did not differ (P>0.05) among ejaculates (1st, 2nd and 3rd). There were differences (P<0.05) for sperm defects elevation on the fourth ejaculate. In seminal plasma 16 to 27 protein bands between 7.6 and 142 kDa were identified. In conclusion, sperm quality was satisfactory in the spring and the stress of successive contentions decreased sperm quality. Also, there is influence of age upon serum testosterone concentration which was higher in deer at 36 months...

Hormonas esteroideas foliculares y su relación con los resultados globales del ciclo de estimulación ovárica / Follicular steroid hormones and their relationship to the overall results of ovarian stimulation cycle

Objetivo: Relacionar los niveles de hormonas esteroideas foliculares con el ciclo de estimulación ovárica y sus resultados globales. Métodos: Se incluyeron pacientes < 38 años, con esterilidad de causa masculina, tubárica o desconocida, que recibieron un protocolo largo con agonistas de GnRH y rFSH. Se recogieron las muestras de la primera y segunda aspiración folicular de cada ovario y se realizó un quimioinmunoanálisis de estradiol, progesterona, testosterona y DHEAS. Resultados: Se obtuvieron cifras menores de DHEAS folicular en las pacientes con más días de frenado con agonistas de GnRH (p=0,0003). Cuantos más días de rFSH administrados, mayores fueron los niveles de testosterona y DHEAS folicular (p=0,03; p=0,03). En los resultados globales del ciclo, se obtuvo una correlación negativa entre las cifras de testosterona folicular y el número de complejos puncionados (r= -0,360; p=0,002) y entre la testosterona folicular y el número de embriones de calidad D (r= -0,233; p=0,047). El número de ovocitos maduros fue menor en pacientes con mayores niveles de testosterona folicular (p=0,008). La progesterona folicular fue superior en ovocitos de buena calidad frente a los de calidad no destacable (p=0,006) y muy mala calidad (p=0,04). Conclusiones: Las cifras altas de testosterona folicular se correlacionaron con menor número de complejos puncionados, ovocitos maduros y embriones de calidad D. La buena calidad ovocitaria se asoció a niveles de progesterona folicular superiores.

Objective: To relate the levels of follicular steroid hormones with the ovarian stimulation cycle and its overall results. Method: It was included patients < 38 years old with sterility of male, tubaric or unknown origin who underwent a long protocol with GnRH agonists and rFSH. Samples were obtained from the first and second follicular aspiration of each ovary. A chemiluminescent immunoassay of estradiol, progesterone, testosterone and DHEAS was performed. Results: Figures of follicular DHEAS decreased as the days of treatment with GnRH agonists increased (p=0.0003) and levels of follicular testosterone and DHEAS increased along with the days of treatment with rFSH (p=0.03, p=0.03). In regard to the outcomes of the overall cycle it was found a negative correlation between follicular testosterone levels and the number of punctured complexes (r= -0.360; p=0.002) and between follicular testosterone and the number of D quality embryos (r= -0.233; p=0.047). The number of mature oocytes was lower in patients with higher levels of follicular testosterone (p=0.008). Follicular progesterone was higher in good quality oocytes as compared to those of no remarkable quality (p=0.006) and very poor quality (p=0.04). Conclusions: High levels of follicular testosterone were correlated with a fewer number of punctured complexes, mature oocytes and D quality embryos. Good oocyte quality was associated with higher follicular progesterone levels.

Hiperplasia suprarrenal congénita no-clásica: avances en la detección, diagnóstico, conducta y tratamiento / Nonclassic adrenal hyperplasia: state of the art

Nonclassical adrenal hyperplasia (NC-CAH) is caused by a deficiency in the activity of the 21-hydroxylase enzyme and is the most common autosomal recessive disorder. The clinical features of the disease sre highly variable, and therefore the diagnosis may be overseen. The disorder is characterized by hyperandrogenism of adrenal origin that may become evident during childhood, adolescence or adulthood. The symptoms vary from premature pubarche, mestrual disturbances, hirsutism and virilization to those cases without any clinical evidence of the disease, as described in the cryptic form. The diagnostic approach includes an initial measurement of plasmatic 17OH-progesterone (17OHP) and androgen levels, and an ACTH test in those with elevated baseline 17OHP. The definitive diagnosis of this entity is performed with the documentation of abnormalities in both alleles of the CYP21A2 gene. This paper reviews the clinical, molecular and treatment of patients with NC-CAH.

Polimorfismos en los genes CYP11α y CYP17 y etiología del hiperandrogenismo en pacientes con poliquistosis ovárica / Polymorphism in CYP11alpha and CYP17 genes and the etiology of hyperandrogenism in patients with polycystic ovary syndrome

El síndrome de poliquistosis ovárica (PCOS) es un desorden endocrino-metabólico de naturaleza multifactorial, con una marcada predisposición genética, que afecta al 6% de las mujeres en edad reproductiva. Se caracteriza por la presencia de hiperandrogenismo, oligo-anovulación y ovarios poliquísticos. Entre los genes candidatos se encuentran aquellos que codifican para enzimas que actúan en la síntesis de andrógenos. Dos de los genes candidatos son el CYP17 y el CYP11alfa que codifican para la 17alfa hidroxilasa (P45017alfa) y para el P450scc (colesterol side chain cleavage) respectivamente. Los polimorfismos en estos genes están asociados al desarrollo del fenotipo hiperandrogénico. Nuestro objetivo fue analizar las frecuencias alélicas de los polimorfismos de los dos genes mencionados en población con PCOS, compararla con población normal y analizar la relación de cada variante alélica con el fenotipo hiperandrogénico correspondiente. Se analizaron 65 pacientes y 58 controles sanos en los que se determinaron niveles de testosterona y frecuencia de polimorfismos en los genes mencionados. Se observó una diferencia estadísticamente significativa cuando se asoció el grupo de mayor nivel de androgenemia con la presencia del genotipo A2/A2 del gen CYP17, y se hallaron mayores niveles de andrógenos circulantes en las pacientes con PCOS portadoras del alelo 216- del gen CYP11alfa. Nuestros resultados sugieren que ambos alelos juegan un rol menor en el desarrollo de PCOS y podrían ser considerados como potenciales marcadores de riesgo genético para el desarrollo del fenotipo hiperandrogénico.

The polycystic ovary syndrome (PCOS) is a heterogeneous multifactorial endocrine metabolic disorder with genetic predisposition affecting 6% of women in the reproductive age. This syndrome is characterized by the presence of oligo-anovulation, hyperandrogenism and polycystic ovaries. Several genes have been postulated as responsible for the etiology of this disorder. Among these genes are those encoding the enzymes involved in the ovarian androgen biosynthesis. Two of the candidate genes are the CYP17 and the CYP11alpha, encoding the 17-alpha-hydroxylase (P45017alpha) and the cholesterol side chain cleavage (P450scc) respectively. The polymorphisms of these genes are linked to the development of an hyperandrogenic phenotype. The aim of this work was to analyze the allelic frequencies of such polymorphisms in a cohort of women with PCOS and to compare them with those of healthy women. Furthermore, the correlation between each allelic variant and the corresponding hyperandrogenic phenotype was also assessed. Therefore, 65 patients and 58 age matched healthy controls were analyzed. The serum levels of testosterone and the frequency of each polymorphism were determined. When the PCOS population was analyzed, a significant statistical difference was found when relating the group with the highest androgenemia level with the presence of A2/A2 genotype of CYP 17 gene, and a higher level of circulating androgen was found in PCO women carrying the 216- allele of CYP11alpha gene (that did not reach statistical significance). Our results suggest that both alleles play a minor role in the development of PCOS and could be a genetic risk marker of the hyperandrogenic phenotype.

Fecal androgen levels in common marmoset (Callithrix jacchus) males living in captive family groups

In captive common marmoset groups, the reproductive inhibition observed in subordinate female seems to be a result of olfactory, visual and behavioral cues from the dominant female. However, few studies have examined the relationship among adult males living in the same social group. These studies have shown that reproductive failure among peer males seems to be based on hormonal and behavioral mechanisms. New insights on sexual strategies in primates have been shown using fecal steroids, but so far no information is available for common marmoset males. In the present study, we evaluated the influence of light-dark cycle, age and reproductive condition on the profile of fecal androgens in males living in the same family group. Feces were collected from six fathers and six sons for androgen determination during the light phase of the 24-h cycle for eight days randomly distributed over a 4-week period. Androgen levels were determined by enzyme immunoassay technique. Adult sons showed higher androgen levels (166.97 ± 22.95 ng/g) than fathers (80.69 ± 44.38 ng/g) and juveniles (49.06 ± 23.15 ng/g; P < 0.05). No diurnal variation (P > 0.05) in fecal androgen profile was observed in adults or juveniles. No indication of androgen-mediated social competition between fathers and adult sons was demonstrable. These results provide basic information on fecal androgen profile useful to investigate the socioendocrinology of free-ranging common marmoset males and verify that, in contrast to daughters, the reproductive suppression of sons is not based on physiological inhibition of their gonads.

Changes in the fecal concentrations of cortisol and androgen metabolites in captive male jaguars (Panthera onca) in response to stress

In the present study we determined the efficacy of the measurement of fecal cortisol and androgen metabolite concentrations to monitor adrenal and testicular activity in the jaguar (Panthera onca). Three captive male jaguars were chemically restrained and electroejaculated once or twice within a period of two months. Fecal samples were collected daily for 5 days before and 5 days after the procedure and stored at -20ºC until extraction. Variations in the concentrations of cortisol and androgen metabolites before and after the procedure were determined by solid phase cortisol and testosterone radioimmunoassay and feces dry weight was determined by drying at 37ºC for 24 h under vacuum. On four occasions, fecal cortisol metabolite levels were elevated above baseline (307.8 ± 17.5 ng/g dry feces) in the first fecal sample collected after the procedure (100 to 350 percent above baseline). On one occasion, we did not detect any variation. Mean (± SEM) fecal androgen concentration did not change after chemical restraint and electroejaculation (before: 131.1 ± 26.7, after: 213.7 ± 43.6 ng/g dry feces). These data show that determination of fecal cortisol and androgen metabolites can be very useful for a noninvasive assessment of animal well-being and as a complement to behavioral, physiological, and pathological studies. It can also be useful for the study of the relationship between adrenal activity and reproductive performance in the jaguar.

Estudio in vitro de tres antiandrógenos empleados en la actualidad en enfermedades andrógenodependientes / In vitro assay of three antiandrogens presently used in the treatment of androgen dependent diseases

Objetivo: Analizar el efecto in vitro del acetato de ciproterona, finasteride y flutamida, sobre la enzima 5Ó-reductasa, principal indicador bioquímico responsable del potente efecto andrógenico de la tetosterona, al convertirla en dihidrotestosterona. Material y Métodos: Se midio la actividad de la enzima 5Ó-reductasa en la próstata de ratas de macho adultos, utilizando concentraciones de 20 a 500µM de cada antiandrógeno. Resultados: El análisis estadístico muestra que la flutamina presenta mejor actividad antiandrogénica a medida que se incrementa su concentración, mientras que en el acetato de ciproterona y el finasteride, el efecto antiandrogénico fue menor a diferentes concentraciones (p<0.05); probablemente por una mayor velocidad de disociación de estos compuestos con su receptor

Evaluation of androgens production in acne vulgaris with and without polycystic ovaries: possible role of insulin-like growth factor-1

In this study, hormonal measurements of serum levels of testosterone [T], dehydroepiandrosterone sulfate [DHEA-S], sex hormone binding globulin [SHBG], luteinizing hormone [LH], follicle stimulating hormone [FSH] and insulin like growth factor- I [IGF-I] were performed in 44 women with Acne vulgaris and ten healthy controls. A pelvic ultrasound examination for polycystic ovaries [PCO] was done in all subjects. The grade of hirsutism and menstrual irregularities if present was recorded. A significant increase in serum levels of T, DHEA-S and a significant decrease in serum levels of SHBG were found in acne patients compared with the controls. Patients with mild acne showed no significant changes for all biochemical parameters compared with the controls. Patients with moderate and severe acne showed serum levels of T, DHEA-S and LH were significantly increased, whereas SHBG was significantly decreased. Serum IGF-I level was significantly elevated only in severe acne form comparing with the controls. A significant positive correlation between T and DHEA-S levels was found in severe acne. In 13 patients with PCO, serum IGF-I and T levels and LH/FSH ratio were elevated significantly compared with patients without. Also, a significant positive correlation between IGF-I and DHEA-S level was found in acne patients with PCO

Dosagem dos andrógenos em mulheres hirsutas: valor diagnóstico e resposta à depressäo com dexametasona / Determination of androgens in hirsute females: diagnosis value and response to depression with dexamethasome

Foram estudadas 30 mulheres hirsutas no sentido de avaliar os valores dos andrógenos basais e após teste dinâmico de supressäo com dexametasona. Os andrógenos dosados (RIE) foram testosterona, androstenediona e sulfato de dehidroepiandrosterona. Do ponto de vista clínico, 73% das pacientes apresentavam hirsutismo de início entre os 12 e 20 anos; 93% com hirsutismo de leve a moderado; as alteraçöes menstruais mais freqüentes, a espaniomenorréia (60%) e 50% apresentavam infertilidade primária. 90% das pacientes apresentavam valores de T acima da normalidade, sendo este andrógeno o mais freqüentemente alterado. Apenas duas pacientes apresentaram todos os andrógenos dosados dentro da normalidade; uma paciente com valor somente de DEA-S alterado; uma paciente com o valor somente de A alterado; em 12 pacientes somente o valor de T alterado e 14 com valor de T e A acima da normalidade. Em relaçäo ao teste de supressäo com dexametasona, verificamos que 16 pacientes apresentavam hiperandrogenismo do tipo dependente de ACTH, cinco de origem independente de ACTH e sete do tipo misto. Pudemos também observar que näo houve associaçäo entre os valores mais altos da T basais com quadro clínico de hirsutismo mais severo e/ou a presença de maiores alteraçöes menstruais

Estudio dinámico del androgenismo en la mujer normal / Dynamic study of androgenism in the normal woman

Se describen en este trabajo los hallazgos obtenidos en el estudio dinámico del androgenismo de 10 mujeres normales mediante la realización de la prueba de supresión/estimulación. Los resultados presentados incluyen para cada esteroide investigado, los niveles críticos de aumento o disminución estadísticamente significativos como límites de normalidad; los rangos corregidos de variación sectorial esteroidea normal, y los perfiles dinámicos correspondientes a las distintas contribuciones sectoriales. A este respecto, el cálculo de la participación sectorial porcentual mostró, para dehidroepiandrosterona, androstenediona y testosterona, junto a una constante y significativa actividad del sector autónomo-periférico, que el sector adrenal es importante para DHEA y A4 pero no para T, mientras que los ovarios estimulados contribuyen escasamente al aporte de DHEA; en forma moderada al de androstenediona, y en gran medida al de T. Los datos aportados, finalmente, se consideran esenciales para la interpretación de los estudios dinámicos efectuados con la prueba S/E en mujeres clínicamente hiperandrogénicas

La prueba de supresion/estimulación en el estudio del androgenismo femenino: protocolo, fundamentos y dinámica segun nuestra experiencia de 115 casos / Supression/stimulation test in the study of female bases androgenization: proctocol bases and dynamics according to our experience in 115 cases

De acuerdo a nuestra experiencia, que incluye la realización de 115 pruebas dinámicas de supresión/estimulación en 10 mujeres normales controles y 105 pacientes clínicamente hiperandrogénicas, dicha prueba resulta idónea y útil para la investigación clínica, el díagnóstico y la orientación terapéutica en los hiperandrogenismos femeninos siempre que en su realización se respeten determinadas pautas: efectuarla en la segunda mitad del ciclo, con una supresión propiamente dicha de por lo menos ocho días de duración y lograda con 3mg díarios de dxm, para agregarle al cabo de ese lapso la estimulación ovárica simultánea con 5000 UI de HCG por día durante tres días consecutivos. Los dosajes esteroideos basales y dinámicos se harán en orina y plasma. En base a los resultados obtenidos se calcularán las contribuciones sectoriales, cuyo análisis e interpretación se harán medíante las Tablas confeccionadas con los datos obtenidos por el monitoreo dinámico de los controles normales