Vitrified sheep isolated secondary follicles are able to grow and form antrum after a short period of in vitro culture.

The risk of reintroducing malignant cells after ovarian graft into patients following post-cancer treatment is an obstacle for clinical applications (autotransplantation). In this context, in vitro follicle culture would be an alternative to transplantation in order to minimize such risks. Therefore, the aim of this study was to compare the development of secondary follicles after vitrification in isolated form (without stroma) with vitrification in in situ form (within fragments of ovarian tissue). Follicles were first isolated from ovarian fragments from mixed-breed ewes and then vitrified; these comprised the Follicle-Vitrification group (Follicle-Vit), or fragments of ovarian tissue were first vitrified, followed by isolation of the follicles, resulting in the Tissue-Vitrification group (Tissue-Vit). Control and vitrified groups were submitted to in vitro culture (6 days) and follicular morphology, viability, antrum formation, follicle and oocyte diameter, growth rate, ultrastructural characteristics and cell proliferation were evaluated. The percentages of morphologically normal follicles and antrum formation were similar among groups. Follicular viability and oocyte diameter were similar between Follicle-Vit and Tissue-Vit. The follicular diameter and growth rate of Follicle-Vit were similar to the Control, while those of Tissue-Vit were significantly lower compared to the Control. Both vitrified groups had an augmented rate of granulosa cellular proliferation compared to Control. Secondary follicles can be successfully vitrified before or after isolation from the ovarian tissue without impairing their ability to survive and grow during in vitro culture.

Activin-A promotes the development of goat isolated secondary follicles in vitro.

The role of activin-A in follicular development and on the mRNA expression levels of different genes in goat secondary follicles was evaluated. Goat secondary follicles (≥ 150 µm) were cultured for 18 days under control conditions or with the addition of either 50 or 100 ng/ml activin-A (Experiment 1). The mRNA levels for the genes that code for activin-A, ActR-IA, ActR-IB, ActR-IIA, ActR-IIB, follicle stimulating hormone receptor (FSH-R) and P450 aromatase were measured in each condition (Experiment 2). We observed that after 6 days of culture, the antrum formation rate was higher in cultures with added activin-A than in the cultured control (P < 0.05). The addition of 50 ng/ml activin-A increased the follicular growth rate in the final third of the culture (days 12-18), resulting in a higher percentage of meiosis resumption (P < 0.05). On day 6, the addition of activin-A (50 ng/ml) increased the levels of ActR-IA mRNA compared with the cultured control (P < 0.05). After 18 days, the addition of 50 ng/ml activin-A significantly increased the levels of its own mRNA compared with the non-cultured control. Moreover, this treatment reduced the mRNA levels of P450 aromatase in comparison with the cultured control (P < 0.05). Higher levels of P450 aromatase mRNA were found for both activin-A treatments compared with the non-cultured control (P < 0.05). No difference in estradiol levels was detected among any of the tested treatments. In conclusion, the addition of activin-A to culture medium stimulated early antrum formation as well as an increase in the daily follicular growth rate and the percentage of meiosis resumption.

Effect of medium composition on the in vitro culture of bovine pre-antral follicles: morphology and viability do not guarantee functionality.

Summary This study investigated the effect of three different culture media (α minimum essential medium (α-MEM), McCoy or TCM199 during the in vitro culture (IVC) of bovine isolated pre-antral follicles. Pre-antral follicles greater than 150 µm in size were isolated and cultured for 0 (control), 8 or 16 days in one of the abovementioned culture media. Follicles were evaluated for survival, growth and antrum formation at days 8 and 16. The results showed that TCM199 was the most suitable medium to preserve follicular viability and ultrastructure, resulting in the highest rates of antrum formation. In conclusion, TCM199 promotes the in vitro development of isolated pre-antral follicles without hampering follicular functionality by sustaining in vitro growth and antrum formation.

Responses of plasma adipokines to high intensity interval training: systematic review / Respostas das adipocinas plasmáticas ao treino intervalado de alta intensidade: revisão sistemática / Respuestas de las adipocinas plasmáticas al entrenamiento por intervalos de alta intensidad: revisión sistemática

ABSTRACT Introduction Obesity is one of the major diseases of modern times. However, the explanation for its pathophysiology is recent and has not yet been fully elucidated. White adipose tissue synthesizes and secretes adipokines that affect several pathologies related to obesity. Excessive growth of this tissue results in increased levels of pro-inflammatory adipokines and a consequent decrease in anti-inflammatory adipokines. Nevertheless, most studies use moderate intensity training, limiting the understanding of high intensity interval training in these proteins. Objective To verify the latest information on the effects of HIIT in improving the profile of circulating adipokines. Methods A search was performed on the databases PUBMED, Lilacs, HighWire, BVS and the Cochrane Database of Systematic Reviews, with the following keywords: HIIT adipokines, HIIT leptin, HIIT adiponectin. Eleven studies were selected, published in English and Portuguese between 2013 and 2017. Results HIIT proved to be effective in increasing adiponectin in the adolescent population and in Olympic athletes, but this depended on a good prescription parameter and exercise intensity. However, maximum or supramaximal intensities were superior to low and moderate intensities. In turn, leptin presented a significant decrease in response to HIIT due to the reduction of adipose tissue, demonstrating a directly proportional relation. Other adipokines, such as omentin-1 and interleukin-10, also responded positively to HIIT, resulting in improved anti-inflammatory status. Conclusion HIIT proved to be an efficient method to reduce inflammation due to obesity, as well as inducing an improvement in sports performance. However, the effects depend on training volume, intensity and prescription method. Level of evidence I; Therapeutic study-Investigating the results of treatment.

RESUMO Introdução A obesidade é uma das principais doenças dos tempos modernos. Entretanto, a explicação da sua fisiopatologia é recente e ainda não foi totalmente esclarecida. O tecido adiposo branco sintetiza e secreta adipocinas que acometem diversas patologias relacionadas à obesidade. O aumento excessivo desse tecido resulta no aumento dos níveis de adipocinas pró-inflamatórias e na consequente diminuição de adipocinas anti-inflamatórias. Entretanto, a maioria dos estudos utiliza o treinamento de intensidade moderada, limitando o entendimento do treinamento intervalado de alta intensidade nessas proteínas. Objetivo Verificar as mais recentes informações sobre os efeitos do HIIT na melhoria do perfil das adipocinas circulantes. Métodos Foi realizada uma pesquisa nos bancos de dados PUBMED, Scielo, Lilacs, HighWire, BVS e Cochrane Database of Systematic Reviews com as seguintes palavras chaves: HIIT adipokines, HIIT leptin, HIIT adiponectin. Onze estudos foram selecionados, publicados em inglês e em português, entre os anos de 2013 e 2017. Resultados O HIIT mostrou-se eficiente para aumentar a adiponectina na população adolescente e em atletas olímpicos, mas isso depende de um bom parâmetro de prescrição e da intensidade do exercício. Entretanto, as intensidades máximas ou supramáximas se mostraram superiores às intensidades baixas e moderadas. Por sua vez, a leptina apresentou significativa diminuição em resposta ao HIIT devido à redução do tecido adiposo, demonstrando uma relação diretamente proporcional. Outras adipocinas, como a omentina-1 e a Iiterleucina-10, também responderam de forma positiva ao HIIT, resultando em um melhor estado anti-inflamatório. Conclusão O HIIT demonstrou ser um método eficiente para diminuir a inflamação decorrente da obesidade, assim como induzir uma melhora no rendimento esportivo. Entretanto, os efeitos dependem do volume de treino, intensidade e método de prescrição. Nível de evidência I; Estudo terapêutico-Investigação dos resultados do tratamento.

RESUMEN Introducción La obesidad es una de las principales enfermedades de los tiempos modernos. Entretanto, la explicación de su fisiopatología es reciente y aún no se ha dilucidado completamente. El tejido adiposo blanco sintetiza y secreta adipocinas que afectan diversas patologías relacionadas a la obesidad. El aumento excesivo de este tejido resulta en el aumento de los niveles de adipocinas proinflamatorias y la consiguiente disminución de las adipocinas antiinflamatorias. Entretanto, la mayoría de los estudios usa el entrenamiento de intensidad moderada, limitando el entendimiento del entrenamiento por intervalos de alta intensidad en estas proteínas. Objetivo Verificar las más recientes informaciones sobre los efectos de HIIT en la mejora del perfil de las adipocinas circulantes. Métodos Se realizó una búsqueda en los bancos de datos PUBMED, Lilacs, HighWire, BVS y Cochrane Database of Systematic Reviews con las siguientes palabras llave: HIIT adipokines, HIIT leptin, HIIT adiponectin. Se seleccionaron 11 estudios, publicados en inglés y portugués entre 2013 y 2017. Resultados El HIIT se mostró eficiente para aumentar la adiponectina en la población adolescente y en atletas olímpicos, pero eso depende de un buen parámetro de prescripción e intensidad del ejercicio. Entretanto, las intensidades máximas o supramáximas se mostraron superiores a las intensidades bajas y moderadas. A su vez, la leptina present ó disminución significativa en respuesta al HIIT debido a la reducción del tejido adiposo, demostrando una relación directamente proporcional. Otras adipocinas, como omentina-1 e interleucina-10, también respondieron positivamente al HIIT, resultando en un mejor estado antiinflamatorio. Conclusión El HIIT demostró ser un método eficiente para disminuir la inflamación proveniente de la obesidad, así como inducir una mejora en el rendimiento deportivo. Entretanto, los efectos dependen del volumen de entrenamiento, la intensidad y el método de prescripción. Nivel de evidencia I, Estudio terapéutico - Investigación de los resultados del tratamiento.

Expression and localization of Aquaporin 3 (AQP3) in folliculogenesis of ewes.

The mRNA expression and localization of Aquaporin 3 (AQP3) were investigated in the ovarian follicles of ewes at different stages of development (primordial, primary, secondary, small, and large antral). The gene expression was quantified by qPCR, while the protein identification and localization were determined by Western blot and immunohistochemistry, respectively. Analysis revealed that AQP3 mRNA was detected only in the antral follicles, whereas the protein expression was detected in the oocyte and granulosa cells in all stages of follicular development. The latter observation suggests that the presence of AQP3 in follicles of all categories, especially in the antral follicles, provides novel insights on the mechanisms that regulate the flow of water between cells during the formation of antral follicles in sheep.

Effects of GDF-9 and FSH on mRNA expression for FSH-R, GDF-9 and BMPs in in vitro cultured goat preantral follicles

The aim of the present study was to determine the role of GDF-9 and/or FSH on the growth and mRNA expression for FSH-R, GDF-9, and BMPs in goat secondary follicles after culture in vitro. Goat secondary follicles (~200µm) were isolated and cultured for six days in minimum essential medium (MEM) supplemented with GDF-9 (200 ng/mL), FSH (50 ng/mL) or both. At the beginning and end of culture, the follicular diameter was evaluated and compared. The levels of mRNA for GDF-9, FSH-R and BMPs -2, -4, -6, -7 and -15 in cultured follicles were quantified by real time PCR. The results showed that a significant increase of follicle diameter after six days when compared to day 0, but the presence of GDF-9 and FSH did not influence the follicular growth in comparison with those cultured in MEM. Real time PCR showed that GDF-9 down-regulated the levels of mRNA for BMPs -2 and -15, while FSH either alone or in combination with GDF-9 did not affect the expression of GDF-9, FSH-R and BMPs. In conclusion, GDF-9 reduced the expression of BMP-2 and -15 in caprine preantral follicles after their culture, but FSH either alone or in association with GDF-9 did not control the expression of GDF-9, FSH-R and BMPs.