Zebrafish ambra1b knockout reveals a novel role for Ambra1 in primordial germ cells survival, sex differentiation and reproduction

BACKGROUND: AMBRA1 is an intrinsically disordered protein, working as a scaffold molecule to coordinate, by protein-protein interaction, many cellular processes, including autophagy, mitophagy, apoptosis and cell cycle progression. The zebrafish genome contains two ambra1 paralogous genes (a and b), both involved in development and expressed at high levels in the gonads. Characterization of the zebrafish paralogous genes mutant lines generated by CRISPR/Cas9 approach showed that ambra1b knockout leads to an all-male population. RESULTS: We demonstrated that the silencing of the ambra1b gene determines a reduction of primordial germ cells (PGCs), a condition that, in the zebrafish, leads to the development of all-male progeny. PGC reduction was confirmed by knockdown experiments and rescued by injection of ambra1b and human AMBRA1 mRNAs, but not ambra1a mRNA. Moreover, PGC loss was not rescued by injection with human AMBRA1 mRNA mutated in the CUL4-DDB1 binding region, thus suggesting that interaction with this complex is involved in PGC protection from loss. Results from zebrafish embryos injected with murine Stat3 mRNA and stat3 morpholino suggest that Ambra1b could indirectly regulate this protein through CUL4-DDB1 interaction. According to this, Ambra1+/- mice showed a reduced Stat3 expression in the ovary together with a low number of antral follicles and an increase of atretic follicles, indicating a function of Ambra1 in the ovary of mammals as well. Moreover, in agreement with the high expression of these genes in the testis and ovary, we found significant impairment of the reproductive process and pathological alterations, including tumors, mainly limited to the gonads. CONCLUSIONS: By exploiting ambra1a and ambra1b knockout zebrafish lines, we prove the sub-functionalization between the two paralogous zebrafish genes and uncover a novel function of Ambra1 in the protection from excessive PGC loss, which seems to require binding with the CUL4-DDB1 complex. Both genes seem to play a role in the regulation of reproductive physiology.

The ethics of nursing care for transgender people / A ética do cuidado de enfermagem às pessoas transgênero / La ética de la atención de enfermería a las personas transgénero

ABSTRACT Objectives: to discuss ethical aspects in nursing care for transgender people. Methods: reflective study based on the dilemmas that emerges in nursing care for transgender people. The report was structured around the four bioethical principles. Results: health care for trans people is complex, transversal to many devices and specialties and longitudinal in time, that is why it requires coordinated action. There is an ethical framework in which the nursing care must be observed in the care of this group. Final Considerations: the nurse as a health worker can assume several general lines in the care of transgender patients. So, complementary training should be provided not only to professionals, but also to students of nursing and other health sciences.

RESUMO Objetivos: discutir aspectos éticos na assistência de enfermagem às pessoas transgênero. Métodos: estudo reflexivo a partir dos dilemas que surgem no cuidado de enfermagem às pessoas transgênero. O relato foi estruturado em torno dos quatro princípios bioéticos. Resultados: a atenção à saúde de pessoas trans é complexa, transversal a muitos dispositivos e especialidades e longitudinal no tempo, por isso requer ação coordenada. Existe um referencial ético no qual se enquadram os cuidados de enfermagem que devem ser observados no atendimento a esse grupo. Considerações Finais: o enfermeiro como agente de saúde pode assumir diversas linhas gerais no atendimento a pacientes transgênero. Para tal, deve ser proporcionada formação complementar não só aos profissionais, mas também aos estudantes de enfermagem e outras ciências da saúde.

RESUMEN Objetivos: debatir sobre aspectos éticos en la atención de enfermería a personas transgénero. Métodos: estudio reflexivo fundamentado sobre los dilemas que se plantean en los cuidados de enfermería a personas transgénero. El relato se ha estructurado en torno a los cuatro principios bioéticos. Resultados: la atención sanitaria a las personas trans es compleja, transversal a muchos dispositivos y especialidades y longitudinal en el tiempo por lo que precisa de la actuación coordinada. Existe un marco ético en el que se encuadran los cuidados de enfermería que se precisan en la atención a este colectivo. Consideraciones Finales: la enfermera como agente de salud puede asumir diversas líneas generales en la atención a pacientes transgénero. Para ello, se debe brindar formación adicional no solo a los profesionales, también a los estudiantes de enfermería y de las demás ciencias de la salud.

Paciente masculino con cariotipo 46 XX negativo para el gen SRY y sin ambigüedad genital: reporte de un caso / Male patient 46, XX SRY -negative and unambiguous genitalia: A case report

En la mayoría de los casos, la diferenciación sexual masculina ocurre con la participación del gen SRY. Sin embargo, se pueden presentar otros genotipos excepcionales, como en el caso que se presenta en este reporte. Se trata de un paciente adulto de sexo masculino atendido en el Servicio de Paternidades del Instituto de Genética de la Universidad Nacional de Colombia. Se le hicieron los análisis del gen de la amelogenina y de repeticiones cortas en tándem (Short Tandem Repeat, STR) específicas para el gen SRY con estuches comerciales de identificación humana, así como los de cariotipo convencional e hibridación in situ fluorescente del SRY, y el estudio de microdeleciones del cromosoma Y mediante reacción en cadena de la polimerasa (PCR). Se le hizo la evaluación clínica y se le brindó asesoramiento genético. El paciente no presentaba ambigüedad genital, su cariotipo era 46 XX, y el perfil molecular era negativo para el gen SRY y positivo para el ZFY. Se le diagnosticó un trastorno de diferenciación sexual 46 XX testicular no sindrómico, una rara condición genética. Solo el 20 % de los pacientes con este diagnóstico son negativos para SRY y exhiben perfiles moleculares diversos. La información disponible parece indicar que el ZFY está relacionado con la diferenciación sexual masculina, aún en ausencia del gen SRY.

In most cases, male sexual differentiation occurs with SRY gene mediation. However, exceptional genotypes have been identified, as shown in this paper. This was a male adult patient seen at the Servicio de Paternidades, Instituto de Genética, Universidad Nacional de Colombia. The following procedures were carried out: Amelogenin gene and short tandem repeat analyses using human identification commercial kits, conventional karyotype, SRY fluorescent in situ hybridization, PCR analysis for Y chromosome microdeletions, clinical evaluation, and genetic counseling. We present an adult male with unambiguous genitalia, karyotype 46,XX, and an SRY negative and ZFY positive molecular profile. The diagnosis of nonsyndromic 46,XX testicular disorder of sex development (DSD) -a rare genetic condition- was established. Only 20 % of similarly diagnosed patients are SRY negative and exhibit diverse molecular profiles. Until now, available evidence seems to indicate that, even in the absence of SRY, the ZFY factor is involved in male sexual differentiation.

Complete Androgen Insensitivity Syndrome and Literature Review / Síndrome da insensibilidade androgênica completa e revisão de literatura

BACKGROUNG: Complete Androgen Insensitivity Syndrome (CAIS) has been reported since 1923, but in 1953 it became known as "testicular feminization". It is a rare recessive genetic disorder linked to the X chromosome that results in different mutations in the androgen receptor. The main clinical presentation in childhood is the presence of bilateral inguinal hernia in phenotypically female subjects. Incidence of androgen insensitivity syndrome in phenotypically females with inguinal hernia is estimated in 0.8% to 2.4%. This is a case report of complete androgen insensitivity syndrome and literature review of preoperative diagnostic methods. CASE SUMMARY: We present a 3 years and 6 months old child with female phenotype, born in São Paulo, Brazil which was diagnosed intraoperatively with complete androgen insensitivity syndrome, during inguinal hernia repair and present potential diagnostic alternatives that we consider viable options in order to avoid this kind of surprise during surgery. CONCLUSION: Investigation of CAIS should be standard in pre-pubertal girls with bilateral inguinal hernia, genetic techniques involving X chromatin or Y chromosome tests present the best choices.

INTRODUÇÃO: A síndrome da insensibilidade androgênica completa (SIAC) é relatada desde 1923, mas foi em 1953 que ficou conhecida como "feminilização testicular". É uma doença genética recessiva rara, ligada ao cromossomo X, causando diversas mutações no receptor de androgênio. A principal apresentação clínica na infância é a presença de hérnia inguinal bilateral em indivíduos fenotipicamente femininos com uma incidência estimada de 0,8% a 2,4%. Apresentamos um caso de insensibilidade androgênica completa, com revisão de literatura dos métodos diagnósticos pré operatórios. Relato do Caso: Apresentamos uma criança de 3 anos e 6 meses de idade com fenótipo feminino, nascida em São Paulo, Brasil diagnosticada com síndrome da insensibilidade androgênica completa, durante a cirurgia de herniorrafia inguinal bilateral e apresentamos potenciais alternativas diagnósticas a fim de evitar esse tipo de surpresa durante a cirurgia. CONCLUSÃO: Em meninas pré-puberes, portadoras de hérnia inguinal bilateral, a pesquisa de SIAC se faz necessária, técnicas genéticas que utilizam a pesquisa da cromatina X ou do cromossomo Y seriam as melhores escolhas.

Dimorfismo sexual da vértebra áxis em uma coleção osteológica brasileira / Sex dimorphism by the axis vertebra in a Brazilian osteological collection

O objetivo desse estudo foi verificar o dimorfismo sexual por meio de medidas lineares (Comprimento do corpo e processo odontóide, Comprimento do processo odontóide, Comprimento do forame vertebral, Largura do forame vertebral, Distância dos processos transversos, Distância do processo transverso esquerdo ao processo espinhoso, Distância do processo transverso direito ao processo espinhoso) da segunda vertebra cervical denominada de áxis. Para tanto foram analisadas 181 áxis, sendo 103 masculinas e 78 femininas, na faixa de idade de 22 a 85 anos, pertencentes ao "Biobanco Tomográfico e Osteológico Prof. Eduardo Daruge da FOP/UNICAMP". Tais medidas foram feitas pelo paquímetro digital marca Stainless ­ hardned ® 150 mm Mauá ­ São Paulo, Brasil, após a calibração inter e intraoperador realizada por meio do teste de correlação intraclasse, teve como resultado o valor de 0,98 considerado excelente. Verificou-se que todas as medidas estudadas são dimórficas e foi possível estabelecer um novo modelo de regressão logística, a partir dos dados obtidos junto às ossadas. Concluiu-se que o modelo de regressão logística gerado ­ Germano Sexo = [- 22.7 + (0.16 × Comprimento do corpo) + (0.31 × Comprimento do forame vertebral) + (0.28 × Distância dos processos transversos)] ­ possui 72,4 % de acerto.

The objective of this study was to verify the sexual dimorphism through linear measurements (length of the body and dentinoid process, length of the dentinoid process, length of the vertebral foramen, width of the vertebral foramen, distance of the transverse processes, distance from the left transverse process to the spinal process , Distance from the right transverse process to the spinous process) of the second cervical vertebra known as the Áxis. For this purpose, 181 Axioms were analyzed, of which 103 were male and 78 were female, in the age range of 22 to 85 years, belonging to the "Tomographic and Osteological Biobank Prof. Eduardo Daruge of FOP / UNICAMP ". These measurements were made by the stainless - hardned ® digital pachymetro 150 mm Mauá - São Paulo, Brazil, after the inter and intra - operator calibration performed through the intraclass correlation test, resulting in a value of 0.98 considered excellent. It was verified that all the measures studied are dimorphic and it was possible to establish a new model of logistic regression, based on the data obtained from the bones. It was concluded that the logistic regression model generated Germano Gender = [- 22.7 + (0.16 × Body length) + (0.31 × Vertebral foramen length) + (0.28 × Distance of transverse processes), with 72.4% of hit.

Sexual dimorphism in Ozopherus muricatus Pascoe, 1872 (Coleoptera: Curculionidae) / Dimorfismo sexual em Ozopherus muricatus Pascoe, 1872 (Coleoptera: Curculionidae)

The weevil Ozopherus muricatus Pascoe, 1872 (Coleoptera: Curculionidae) is already considered a pest in Brazil for açaí palm (Euterpe oleracea Mart.) plantations. Its presence has become more frequent and caused significant reduction in the production of açaí fruit. Studies on the biology of this insect pest are still scarce, hindering its management. For studies on pheromones, for instance, it is essential to correctly identify the sex of the insect. The objective of this study was to evaluate the existence of sexual dimorphism in pupae and adults of O. muricatus in order to allow the identification of males and females. Larvae, pupae and adults were collected from açaí plantations and kept in biochemical oxygen demand (BOD) type incubation chambers. Adults and pupae were examined under a stereomicroscope, and dimorphism was determined through observation of external morphological characteristics. In addition, adults were dissected in order to observe genitalia and confirm the sex of specimens. In the pupae, sexual dimorphism was characterized by the presence of two protuberances in the terminal ventral region of the abdomen of females, which were absent in males. In adults, sexual dimorphism was also observed, based on the final abdominal segments. In males, it was possible to see all eight abdominal tergites, while in females tergite VIII is covered by tergite VII. Thus, the sexual dimorphism present in the morphological characteristics of O. muricatus allows the distinction between males and females of this species.(AU)

O gorgulho Ozopherus muricatus Pascoe, 1872 (Coleoptera: Curculionidae) já é considerado uma praga em plantações de açaí (Euterpe oleracea Mart.) no Brasil. Sua presença tem se tornado mais frequente e causado redução significativa na produção de frutos. Estudos sobre a biologia dessa praga ainda são escassos, dificultando o seu manejo. Para estudos com feromônios, por exemplo, é essencial a correta identificação do sexo do inseto. O objetivo deste estudo foi avaliar a existência de dimorfismo sexual em pupas e em adultos de O. muricatus, a fim de permitir a identificação de machos e fêmeas. Larvas, pupas e adultos foram coletados de plantações de açaí e mantidos em câmaras incubadoras para demanda bioquímica de oxigênio (DBO). Adultos e pupas foram examinados sob microscópio estereoscópico, e o dimorfismo foi determinado por meio da observação de características morfológicas externas. Adicionalmente, adultos foram dissecados para a observação da genitália e confirmação do sexo dos espécimes. Nas pupas, o dimorfismo sexual foi caracterizado pela presença de duas protuberâncias na região terminal ventral do abdome das fêmeas, as quais foram ausentes nos machos. Nos adultos, também se observou dimorfismo sexual, com base nos segmentos finais do abdome. Nos machos, é possível visualizar todos os oito tergitos abdominais, enquanto nas fêmeas o tergito VIII é encoberto pelo tergito VII. Assim, o dimorfismo sexual presente nas características morfológicas de O. muricatus permite distinção entre machos e fêmeas dessa espécie.(AU)

Nanos2 in the male reproductive system: Progress in studies / 中华男科学杂志

Nanos2, a member of the Nanos2 gene family, is a specific gene in male germ cells and encodes an evolutionarily conserved RNA binding protein expressed in male primordial germ cells (PGCs) during the embryonic period as well as in the spermatogonial stem cells (SSCs) of the testis. In the embryonic period, Nanos2 promotes the development of male PGCs and inhibits them from meiosis. In the process of spermatogenesis, Nanos2 suppresses the differentiation of SSCs in the testis and maintains the stability of the SSC pool. The knockout of Nanos2 may cause the disappearance of germ cells and sterility in male mice while its overexpression in the testis may lead to accumulation of SSCs in seminiferous tubules. Besides, Nanos2 is involved in the degradation of specific RNAs and possibly associated with some diseases of the male reproductive system. This review focuses on the recent progress in the studies of Nanos2 in the male reproductive system.

Transcriptome analysis of female and male flower buds of Idesia polycarpa Maxim. var. vestita Diels

Background: Idesia polycarpa Maxim. var. vestita Diels, a dioecious plant, is widely used for biodiesel due to the high oil content of its fruits. However, it is hard to distinguish its sex in the seedling stage, which makes breeding and production problematic as only the female tree can produce fruits, and the mechanisms underlying sex determination and differentiation remain unknown due to the lack of available genomic and transcriptomic information. To begin addressing this issue, we performed the transcriptome analysis of its female and male flower. Results: 28,668,977 and 22,227,992 clean reads were obtained from the female and male cDNA libraries, respectively. After quality checks and de novo assembly, a total of 84,213 unigenes with an average length of 1179 bp were generated and 65,972 unigenes (78.34%) could be matched in at least one of the NR, NT, Swiss-Prot, COG, KEGG and GO databases. Functional annotation of the unigenes uncovered diverse biological functions and processes, including reproduction and developmental process, which may play roles in sex determination and differentiation. The Kyoto Encyclopedia of Genes and Genomes pathway analysis showed many unigenes annotated as metabolic pathways, biosynthesis of secondary metabolites pathways, plant­ pathogen interaction, and plant hormone signal transduction. Moreover, 29,953 simple sequence repeats were identified using the microsatellite software. Conclusion: This work provides the first detailed transcriptome analysis of female and male flower of I. polycarpa and lays foundations for future studies on the molecular mechanisms underlying flower bud development of I. polycarpa.

Utilidad clínica de la medición de la hormona antimülleriana en niños / Clinical utility of anti-mullerian hormone measurement in children

Resumen: la hormona antimülleriana, inicialmente denominada sustancia inhibitoria mülleriana, es una glicoproteína homodimérica de 12,5 kDa, que pertenece a la familia del factor de crecimiento transformante beta (TGF-ß) y desempeña un papel crucial en la diferenciación sexual masculina al favorecer la regresión de los conductos de Müller. Dado que su producción en el varón es principalmente por las células de Sertoli inmaduras, en las últimas décadas ha crecido su utilidad más allá de la evaluación de la función ovárica y tratamientos de fertilidad en las mujeres, lo que ha permitido evaluar en el varón la función testicular y los estados de hipogonadismo, trastornos de la diferenciación sexual, pubertad patológica, criptorquidia, entre otras condiciones clínicas revisadas en este manuscrito. Además, esta revisión describe el rol fisiológico de la hormona antimülleriana en los testículos prepuberales y las pruebas de laboratorio disponibles para su medición. (AU)

Abstract: The antimullerian hormone, initially referred as mullerian inhibitory substance, is a 12.5 kDa homodimeric glycoprotein, belonging to the transforming growth factor beta (TGF-ß) family that playing a crucial role in male sexual differentiation by favoring regression of the Mullerian ducts. Since their production in the male is mainly by the immature Sertoli cells, in the last decades its usefulness has growth beyond the evaluation of the ovarian function and female fertility treatments, which has allowed evaluating the testicular function in male and affections such as hypogonadism, disorders of sexual differentiation, pathological puberty, cryptorchidism and others clinical conditions reviewed in this manuscript. In addition, this review describes the physiological role of the antimüllerian hormone in the prepubertal testes and the laboratory tests available for its measurement. (AU)

The analysis of gene mutation and diagnosis and treatment of 5α-reductase 2 deficiency in a child / 临床儿科杂志

Objective To explore the clinical feature and gene mutation in steroid 5α-reductase 2 deficiency (SRD5A2). Method The clinical data of SRD5A2 in a child with vulva abnormality as the first manifestation was retrospectively analyzed. Results This was a 29-month-old child, whose social gender was female. The level of her basic luteinizing hormone (LH) was 0.07 mIU/mL, and follicle-stimulating hormone was (FSH) 0.39 mIU/mL. The baseline levels of testosterone (T), dihydrotestosterone (DHT), 17-hydroxyprogesterone (17-OHP) and androstendione (A2) were 0.06 ng/mL, 19.67 pg/mL, 1.20 ng/mL, and 0.07 ng/mL respectively. Those levels were 3.65 ng/mL, 68.25 pg/mL, 51.72 ng/mL, and 14.70 ng/mL respectively after Human chorionic gonadotropin (HCG) stimulation. The levels of her anti-mullerian hormone (AMH) was 22.97 ng/mL, and inhibin B (INH-B) was 274.4 pg/mL. The uterus and ovaries were not detected by Pelvic ultrasound and MRI. The chromosome showed 46, XY. Sex determination (SRY) gene detection showed normal. Androgen receptor (AR) gene detection showed negative. There was pathogenic mutation of 5α-reductase 2 (SRD5A2) gene in peripheral blood of the child and her parents. The penis grows 2 cm after 4 months of treatment with 2.5% DHT gel. Conclusion SRD5A2 is diagnosed mainly based on the increase of T/DHT after HCG stimulation experiment and it can be confirmed by detection of pathogenic SRD5A2 mutation.

The correlation between phenotype and genotype of 5α-reductase 2 deficiency in 5 children / 临床儿科杂志

Objective To explore the correlation between phenotype and genotype of 5α-reductase 2 deficiency. Methods The clinical data of five children with 5α-reductase 2 deficiency were retrospectively analyzed and the relation between their clinical phenotype and genotype were analyzed. Results All of these five children presented small penis and testicular hypoplasia, three of whom had ones similar to the clitoris appearance. The testosterone/dihydrotestosterone (T/DHT) ratio was 10.26-64.99 after human chorionic gonadotropin (hCG) stimulation. Gene detection showed one case had c.680G>A homozygous mutation and the others were compound heterozygous mutations. The mutations were mainly missense mutations, followed by deletion, duplication and nonsense mutations. Conclusion The 5α-reductase 2 deficiency has different degrees of abnormal genital development. Genetic testing contributed to the diagnosis of this disease.

Disgenesia gonadal mixta como forma de presentación de un desorden de la diferenciación sexual de causa cromosómica / Mixed gonadal dysgenesis as a form of presentation of sex differentiation disorder of chromosomal origin

La diferenciación sexual es un proceso genéticamente determinado y controlado, que puede ser alterado por diferentes tipos de mutaciones genéticas, o por el efecto de hormonas u otros disruptores ambientales que actúan sobre el embrión, resultando en la formación de genitales externos que no se corresponden con el sexo genético y con los genitales internos. La disgenesia gonadal mixta clasifica en los desórdenes de la diferenciación sexual de causa cromosómica. Se describe un paciente de un año de edad que es atendido en el Hospital Pediátrico “Juan Manuel Márquez”, por presentar genitales externos atípicos. El diagnóstico del paciente fue de disgenesia gonadal mixta, y se realizó con los complementarios siguientes: ecografía ginecológica, estudios hormonales y cariotipo. El tratamiento instaurado inicialmente, fue quirúrgico en dos tiempos operatorios, y el seguimiento hormonal hasta la pubertad (14 años), cuando se inició terapia de reemplazo hormonal según lo establecido por la edad de la paciente.

Sex differentiation is a genetically determined and controlled process that may be altered by various types of genetic mutations or by the effect of hormones or other environmental disruptors acting upon the embryo. The result is the formation of external genitalia that does not match with the genetic sex and the internal genitalia. Mixed gonadal dysgenesis is classified into the sexual differentiation disorders of chromosomal cause. Here is a one-year old child, who was seen at “Juan Manuel Marquez” pediatric hospital since he presented with atypical external genitalia. The diagnosis was mixed gonadal dysgenesis, based on supplementary tests like gynecological echography, hormone studies and karyotype. The initial treatment was surgical in two surgical times, and the hormonal follow-up lasted till puberty (14 years) when the hormone replacement therapy started according to the indications for the patient's age.

Hiperplasia suprarrenal congénita: origen de trastornos del desarrollo y diferenciación sexual / Adrenal hyperplasia congenital: beginning of developmental and sexual differentiation disorders

La hiperplasia suprarrenal congénita es una patología de origen genético que se desarrolla por un déficit enzimático secundario a alteraciones en la síntesis de proteínas. Dicho déficit perturba las vías del metabolismo suprarrenal, específicamente las que ocurren en la corteza de la glándula, es decir, el metabolismo glucocorticoide, mineralocorticoide y de hormonas sexuales. Estas variaciones moleculares traducidas a la clínica se expresan en una variedad de presentaciones posibles, cada una dependiente de la enzima que se encuentre afectada, desde alteraciones electrolíticas, metabólicas, renales o del tracto gastrointestinal, pero hay una que es comúnmente encontrada, trastornos de desarrollo y diferenciación sexual. Dentro de las formas más severas de hiperplasia suprarrenal congénita se observan alteraciones de los caracteres sexuales externos que básicamente traducen en virilización de los genitales durante el desarrollo gestacional, así como también pubertad precoz, amenorreas, infertilidad, entre otros...

Adrenal hyperplasia congenital is a genetic disease that develops from an enzyme deficiency secondary to alterations in protein synthesis. This deficit disturbs the pathways of adrenal metabolism, specifically occurring in the cortex of the gland, such as, the glucocorticoid, mineralocorticoid and the sex hormones metabolism. These molecular variations translated to the clinics are expressed in a variety of possible presentations, each depending of the enzyme that is affected, since electrolyte, metabolic, renal abnormalities or and gastrointestinal tract disturbances, but one of these is commonly for all presentation, the development and sexual differentiation disorders. Among the most severe forms of adrenal hyperplasia congenital alterations of the external sexual characters that basically translate into virilization of the genitals duringgestational development, as well as precocious puberty, amenorrhea, infertility observed, among other...

Effectiveness of the aromatase (P450 Arom) inhibitors Letrozole and Exemestane for masculinization of red tilapia (Oreochromis spp.) / Efectividad de los inhibidores de la aromatasa (aromP450) Letrozol y Exemestano para la masculinización de tilapia roja (Oreochromis spp.) / Efetividade dos inibidores da aromatase (aromP450) Letrozol e Exemestano para a masculinização da tilápia vermelha (Oreochromis spp)

Background: cytochrome P450 aromatase enzyme (aromP450) is a key enzyme in the conversion of androgens into estrogens, a crucial step in the control of sexual differentiation in fish. Objective: the aim of this study was to evaluate the efficiency of two aromatase inhibitors (AIs) as an alternative method for the masculinization of red tilapia (Oreochromis spp.). Methods: Letrozole (LT) and Exemestane (EM) aromatase inhibitors were used at two experimental doses (25 and 100 mg/Kg). Five days post-hatching larvae (5 dph) were fed the inhibitors for 30 days (35 dph). The control treatments consisted of 17α metil testosterone (MT) at a concentration of 60 mg/ Kg (positive control) and food with 300 ml/Kg ethanol (negative control). On 60 dph, gonadal extraction of fishes was performed for histological processing and staining with hematoxylin-eosin for analysis. Results: there were no significant differences (p>0.05) between any compound and implemented doses with the controls in terms of larval survival. Percentage of male fish increased for LT, EM, and MT (positive control), which showed significant differences (p<0.05) with the negative control. The dose analysis showed significant differences (p<0.05) for 100 mg/Kg dose and positive control with 25 mg/Kg dose and negative control; there were also differences between 25 mg/Kg dose and negative control. Conclusions: our results suggest that oral administration of third generation AIs (Type I or Type II) is effective for increasing the proportion of males without differences between inhibitor types. There is also a direct effect of the dose on male proportion. Suppression of aromatase activity allows guiding sexual differentiation towards final testicular development.

Antecedentes: la citocromo P450 aromatasa (aromP450) es una enzima clave en la conversión de andrógenos a estrógenos; un paso crucial en el control de la diferenciación sexual en peces. Objetivo: evaluar la eficiencia de dos inhibidores de la aromatasa (IAs) como método alternativo para la masculinización de tilapia roja (Oreochromis spp.). Métodos: se evaluaron los inhibidores de la aromatasa, letrozol (LT) y exemestano (EM), a dos dosis experimentales de 25 y 100 mg/Kg. Larvas de 5 días post eclosión (dpe) fueron alimentadas durante 30 días (35 dpe) con los tratamientos. Como testigos se implementaron 17α metil testosterona (MT) a una concentración de 60 mg/Kg (control positivo), y alimento con etanol 300 ml/Kg (control negativo). El día 60 dpe se realizó extracción gonadal de los peces para procesamiento histológico y tinción con hematoxilina-eosina para su análisis. Resultados: no hubo diferencias significativas (p>0,05) entre ninguno de los tratamientos y los controles para la variable sobrevivencia larvaria. La proporción de machos aumentó con la implementación de LT, EM y MT (control positivo), los cuales presentaron diferencias significativas (p<0,05) con el control negativo. Hubo diferencias significativas (p<0,05) entre la dosis de 100 mg/Kg y el control positivo con la dosis de 25 mg/Kg y el control negativo; también se presentaron diferencias entre las dosis de 25 mg/kg y el control negativo. Conclusiones: nuestros resultados sugieren que el suministro vía oral de IAs de tercera generación, tipo I o tipo II, es eficiente en aumentar la proporción de machos en los grupos tratados, sin diferencias entre el tipo de inhibidor, pero con un efecto directo de la dosis sobre la proporción de machos. La supresión de la actividad de la aromatasa, permitió orientar la diferenciación sexual hacia el desarrollo testicular.

Antecedentes: a enzima aromatase citocromo P450 (aromP450) é uma enzima chave na conversão de andrógenos em estrógenos, um passo fundamental no controle da diferenciação sexual em peixes. Objetivo: avaliar a eficiência de dois inibidores da aromatase (AIs), como um método alternativo para a masculinização da tilápia vermelha (Oreochromis spp.). Métodos: avaliaram-se os inibidores da aromatase, letrozol (LT) e exemestano (EM), em duas doses experimentais de 25 e 100 mg/Kg. Larvas 5 dias pós-eclosão (dpe) foram alimentadas durante 30 dias (35 dpe) com os tratamentos. Como testemunho, controle positivo, se implementaram 17α metil testosterona (MT) a uma concentração de 60 mg/Kg e como controle negativo alimento com etanol 300 ml/Kg. No dia 60 dpe extraíram-se as gônadas dos peixes para processamento histológico e coloração com hematoxilina-eosina para sua analise. Resultados: não houve diferenças significativas (p>0,05) entre os tratamentos e os controles para a variável sobrevivência larvária. A percentagem de machos foi aumentada com a utilização de LT, EM e MT (controle positivo), com diferenças significativas (p<0,05) comparados com o controle negativo. A análise da dose mostrou diferenças significativas (p<0,05) entre a dose de 100 mg/Kg e o controle positivo com a dose de 25 mg/Kg e o controle negativo; também apresentaram diferenças entre a dose de 25 mg/kg e o controle negativo. Conclusões: nossos resultados sugerem que o fornecimento oral de IAs de terça geração, tipo I ou II, são eficientes para aumentar a proporção de machos nos grupos tratados, sem diferenças entre o tipo de inibidor, porem com efeito direto da dose sobre a proporção de machos. A supressão da atividade da aromatase permitiu orientar a diferenciação sexual até um desenvolvimento testicular final.

Validation of a physical anthropology methodology using mandibles for gender estimation in a Brazilian population

Validation studies of physical anthropology methods in the different population groups are extremely important, especially in cases in which the population variations may cause problems in the identification of a native individual by the application of norms developed for different communities. OBJECTIVE: This study aimed to estimate the gender of skeletons by application of the method of Oliveira, et al. (1995), previously used in a population sample from Northeast Brazil. MATERIAL AND METHODS: The accuracy of this method was assessed for a population from Southeast Brazil and validated by statistical tests. The method used two mandibular measurements, namely the bigonial distance and the mandibular ramus height. The sample was composed of 66 skulls and the method was applied by two examiners. The results were statistically analyzed by the paired t test, logistic discriminant analysis and logistic regression. RESULTS: The results demonstrated that the application of the method of Oliveira, et al. (1995) in this population achieved very different outcomes between genders, with 100% for females and only 11% for males, which may be explained by ethnic differences. However, statistical adjustment of measurement data for the population analyzed allowed accuracy of 76.47% for males and 78.13% for females, with the creation of a new discriminant formula. CONCLUSION: It was concluded that methods involving physical anthropology present high rate of accuracy for human identification, easy application, low cost and simplicity; however, the methodologies must be validated for the different populations due to differences in ethnic patterns, which are directly related to the phenotypic aspects. In this specific case, the method of Oliveira, et al. (1995) presented good accuracy and may be used for gender estimation in Brazil in two geographic regions, namely ...

Hiperplasia suprarrenal congénita: reporte de un caso clínico / Congenital adrenal hyperplasia: case report

INTRODUCCIÓN: La Hiperplasia suprarrenal congénita (HSC) es una enfermedad autosómica recesiva cuya principal causa es la deficiencia de 21-hidroxilasa, quien participa en la síntesis del cortisol y aldosterona. Se describen dos formas de HSC, una clásica y otra no clásica, siendo la primera el objetivo de análisis a lo largo del caso clínico. Sus manifestaciones clínicas varían en gravedad, dependiendo del nivel de deficiencia hormonal. Dentro de la clásica se describe la forma perdedora de sal, cuyas consecuencias son el exceso de andrógenos e insuficiencia de cortisol y mineral o corticoides. Así esta se puede manifestar como un trastorno de la diferenciación sexual (virilización de los genitales externos si el feto es femenino) e insuficiencia suprarrenal. Para su diagnóstico se consideran los antecedentes familiares, manifestaciones clínicas, medición de los niveles de 17-hidroxiprogesterona y la detección de la alteración genética. PRESENTACIÓN DEL CASO: Paciente con antecedentes familiares de hermano con HSC, nace con un trastorno de la diferenciación sexual y es dado de alta con sexo legal masculino. Después de 3 meses desarrolla una insuficiencia suprarrenal, diagnosticándose HSC forma clásica perdedora de sal y por cariotipo se determina sexo femenino. DISCUSIÓN: Los pilares del manejo de la HSC son el consejo genético en las familias con riesgo, el tratamiento antenatal con dexametasona, terapia postnatal con glucocorticoides y el tratamiento quirúrgico de las alteraciones de los genitales externos, junto con las nuevas investigaciones en base a terapia genética y el uso de células madre, requiriendo de este modo la HSC una vista integral.

INTRODUCTION: Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease whose main cause is the deficiency of 21-hydroxylase, an enzyme involved in the synthesis of cortisol and aldosterone. There are two forms of CAH, a classical and nonclassical form, being the first objective of analysis in the clinical case. Its clinical manifestations vary in severity, depending on the level of hormone deficiency. Within the classic is described the salt-wasting form, whose consequences are androgen excess and insufficiency of cortisol and mineral o corticoids. So this may manifest as a sex differentiation disorder (virilization of the external genitalia if the fetus is female) and adrenal insufficiency. For diagnosis are considered the family history, clinical manifestations, measuring 17-hydroxyprogesterone levels and detection of genetic alteration. CASE REPORT: Patient with a family history of a brother with HSC brother, born with a disorder of sexual differentiation and is discharged with legal male sex. After three months develops adrenal insufficiency and was diagnosed with classical HSC salt-wasting form and determined female karyotype. DISCUSSION: The Pillars of the HSC are handling genetic counseling in families at risk, prenatal treatment with dexamethasone, postnatal glucocorticoid therapy and surgical treatment of disorders of the external genitalia, along with new research based therapy gene and the use of stem cells, requiring this way an integral view of HSC.

Sexual difference, identification and object choice in individuals with sex differentiation disorders / Diferença sexual, identificação e escolha de objeto em pacientes com distúrbios da diferenciação sexual / Différence sexuelle, identification et choix d'objet parmi des patients porteur de troubles de différenciation sexuelle / Diferencia sexual, identificación e elección de objeto en pacientes con anomalías de la diferenciación sexual

OBJECTIVE: To investigate how sexual identity is structured and also to investigate the relationship between sexual identity, choice of sex object and sexual difference. METHOD: Semi-structured interviews were held with seven adult patients who were born with sex differentiation disorders: Two had 5-alpha-reductase type-2 deficiency and five had congenital adrenal hyperplasia. CONCLUSIONS: Sex is trauma. Neither male nor female nor any other gender identification implies the choice of sex object, genders of partners or sexual practices.

OBJETIVO: Interrogar como se estrutura a identidade sexual. Investigar as relações entre identidade sexual, escolha de objeto sexual e diferença sexual. MÉTODO: Aplicamos entrevista semi-estruturada em pacientes com distúrbios da diferenciação sexual: 2 com deficiência da 5 alfa redutase tipo 2 e 5 com Hiperplasia Adrenal Congênita. CONCLUSÕES: Sexo é trauma. Identificação masculina, feminina ou outras não implicam no sexo do objeto de escolha, gênero dos parceiros ou práticas sexuais.

OBJECTIF: Ce travail a pour but d'analyser la façon comment l'identité sexuelle se structure, ainsi que d'investiguer les rapports entre l'identité sexuelle, le choix de l'objet sexuel et la différence sexuelle. MÉTHODE: Entrevue semi-dirigée de patients porteur de troubles de différenciation sexuelle: Deux patients porteurs du déficit 5-alpha réductase type 2 et cinq patients porteurs d'hyperplasie congénitale des surrénales. CONCLUSION: Le sexe est un traumatisme. L'identification masculine, féminine ou autre n'implique pas le sexe de l'objet du choix, le genre des partenaires ou les pratiques sexuels.

OBJETIVO: Interrogar como se estructura la identidad sexual. Investigar las relaciones entre identidad sexual, elección de objeto sexual y diferencia sexual. MÉTODO: Fueron aplicadas entrevistas semiestructuradas en pacientes con trastornos en la diferenciación sexual: 2 con deficiencias de la 5 Alpha Redutase tipo 2 y 5 con Hiperplasia Adrenal Congénita. CONCLUSIÓN: Sexo es trauma. Identificación masculina, femenina u otras no implican en el sexo del objeto de elección, el género de los compañeros (partenaires) o prácticas sexuales.

Effect of a non steroidal tamoxifen on the gonad and sex differentiation in Nile tilapia, Oreochromis niloticus.

The effect of different doses of anti-estrogenic chemical was studied in a cichlid Oreochromis niloticus on mortality, growth, sexual development and sex differentiation. Percent mortality (5, 10 and 10) appeared to be dose-dependent. All groups of tamoxifen (TA) treated fishes exhibited significant growth as compared to the controlled fishes. Immature fishes treated with TA at 200 mg l-1, showed lower gonado-somatic-index (GSI) (male 0.137; female 0.118) than control group (male 0.138; female 0.122) of fishes. Ovarian and testicular differentiation was adjudged by the appearance of the ovarian cavity (ovary) and the efferent (sperm) duct (testis), respectively. Sex and sexual maturity of individual fishes were determined using standard histological method. Tamoxifen treatment to the fry (yolk sac absorbed stage) of Oreochromis niloticus in the dose of 200 mg l-1 for 60 days produced 90% males and 1% intersex population. Results suggested masculinizing action of tamoxifen reversing the sex of genetic female tilapia.

Disorders of Sex Development

The birth of a new baby is one of the most dramatic events in a family, and the first question is usually "is it a boy or a girl?" The newborn infant with ambiguous external genitalia often comes as a surprise for the doctors as well as the parents and is sometimes described as an endocrine emergency situation presenting a problem of sex assignment. The nomenclature such as 'intersex', 'hermaphrodite', and 'pseudohermaphrodite' is out of date as well as confusing, and many urologists are concerned that these confusing terms could be perceived to be pejorative by some affected families. In response to concerns regarding outdated and controversial terms, the Chicago Consensus held in 2005 recommended new terminology based on the umbrella term disorders of sex differentiation (DSDs). The term DSD has a comprehensive definition including any problem noted at birth in which the genitalia are atypical in relation to the chromosomes or gonads. The karyotype is used as a prefix defining the classification of DSD. DSDs are rare and complex. The optimal management of patients with DSD must be individualized and multidisciplinary, considering all aspects, including psychological care and full disclosure of alternatives relating to surgery type and timing. Although further studies are necessary to confirm guidelines and recommendations fitting for the individual patients with DSD, this article is an attempt to provide a balanced perspective for new taxonomy, clinical evaluation, and medical, surgical, and psychological management of DSD.

Distúrbio da diferenciação sexual testicular XX: relato de caso / XX testicular disorder of sex differentiation: case report

The 46 XX, testicular sex differentiation disorder, or XX male syndrome, is a rare condition detected by cytogenetics, in which testicular development occurs in the absence of the Y chromosome. It occurs in 1:20,000 to 25,000 male newborns and represents 2% of cases of male infertility. About 90% of individuals present with normal phenotype at birth and are generally diagnosed after puberty for hypoganadism, gynecomastia, and/or infertility. The authors present the report of an XX male with complete masculinization and infertility.

O distúrbio da diferenciação sexual testicular 46,XX, ou síndrome do homem XX, é uma condição rara, na qual o desenvolvimento testicular ocorre na ausência do cromossomo Y detectado citogeneticamente. Incide em 1:20.000 a 25.000 recém-nascidos do sexo masculino e representa 2% dos casos de infertilidade masculina. Cerca de 90% dos indivíduos apresentam fenótipo normal ao nascimento e são, geralmente, diagnosticados após a puberdade por hipogonadismo, ginecomastia e/ou infertilidade. Os autores apresentam o relato de um homem XX com masculinização completa e infertilidade.