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Abstract Background: The production of biofuels has caused an increase in the prices of agricultural commodities. Thus, the ecological footprint, social inclusion and profitability of production systems have encouraged the use of agro-industrial products as an alternative in ruminant feeds. Objective: To evaluate carcass yields, non-carcass components, and the economic viability of including licuri oil in diet of Santa Ines ewes. Methods: A total of 32 Santa Ines ewes (multiparous, non-lactating, 2-4 years old, and 36.7±0.87 kg of body weight-BW) were allotted to a randomized block design with four treatments (diets containing 0, 2, 4 or 5% licuri oil) with eight replicates per treatment, and confined for 77 days. Results: The incremental inclusion of licuri oil promoted a quadratic response on slaughter BW, carcass and true yields, chest widths, heart fat, pancreas, omental fat, and ribeye area (p<0.05). Carcass, heart and loin weight, shank yield, fat thickness and loin fat were reduced (p<0.05); while cooling losses, left half-carcass weight, saw blade yield, and proportion of meat in the loin increased (p<0.05) with increasing dietary levels of licuri oil. The control diet resulted in the best gross revenue (USD$609.39); however, there was a loss (USD$50.96) regarding economic performance indicators. Conclusion: The use of up to 2% licuri oil in the diet increased carcass yield of discard ewes.

Resumen Antecedentes: La producción de biocombustibles ha provocado un aumento en los precios de los productos agrícolas básicos. Por lo tanto, la huella ecológica, la inclusión social y la rentabilidad de los sistemas productivos han incentivado la busqueda de productos agroindustriales como alternativa para la alimentación de rumiantes. Objetivo: Evaluar el rendimiento en canal, los componentes ajenos a la canal, y la viabilidad económica de incluir aceite de licuri en dietas para ovejas de raza Santa Ines. Métodos: Un total de 32 ovejas Santa Ines (multíparas, no lactantes, de 2 a 4 años y de 36,7±0,87 kg de peso corporal-PC) fueron asignadas a un diseño de bloques al azar con cuatro tratamientos (dieta basal adicionada con 0, 2, 4 o 5% de aceite de licuri) con ocho repeticiones por tratamiento, y confinadas por 77 días. Resultados: Los niveles incrementales de aceite de licuri tuvieron un efecto cuadrático sobre el PC al sacrificio, el rendimiento real y de la canal, el ancho del pecho, la grasa del corazón, el páncreas, la grasa del epiplón y el área del ojo del lomo (p<0,05). Se redujo (p<0,05) el peso de la canal, el corazón y el lomo, el rendimiento del jarrete, el grosor de la grasa y la grasa del lomo; y aumentaron las pérdidas por enfriamiento, el peso de media canal restante, el rendimiento de la hoja de sierra y la proporción de carne en el lomo (p<0,05) con niveles incrementales de aceite de licuri. La dieta control resultó en el mejor ingreso bruto (USD$609,39); sin embargo, hubo una pérdida (USD$50,96) en los indicadores de desempeño económico. Conclusión: El uso de hasta un 2% de aceite de licuri en la dieta incrementa el rendimiento en canal de las ovejas de descarte.

Resumo Antecedentes: A produção de biocombustíveis tem causado um aumento nos preços das commodities agrícolas. Assim, a pegada ecológica, a inclusão social e a rentabilidade do sistema de produção têm incentivado o uso de produtos agroindustriais como alternativa alimentar para ruminantes. Objetivo: Avaliar os rendimentos de carcaça, componentes não carcaça e a viabilidade econômica da inclusão do óleo de licuri em dietas para ovelhas Santa Inês. Métodos: Um total de 32 ovelhas Santa Inês (multíparas, não lactantes, 2-4 anos e 36,7±0,87 kg de peso corporal-PC) foram distribuídas em delineamento em blocos casualizados com 4 tratamentos (dietas contendo óleo de licuri a 0, 2, 4 e 5% de matéria seca) com 8 repetições por tratamento e confinadas por 77 dias. Resultados: A inclusão de óleo de licuri promoveu efeito quadrático no PC de abate, rendimentos de carcaça e verdadeiros, largura de peito, gordura do coração, pâncreas, gordura omental e área de olho de lombo (p<0,05). Os pesos de carcaça, coração e lombo, rendimento de pernil, espessura de gordura e gordura do lombo foram reduzidos (p<0,05), e as perdas por resfriamento, peso de meia carcaça esquerda, rendimento de lâmina de serra e proporção de carne no lombo aumentaram (p<0,05) com um aumento nos níveis de óleo de licuri. A dieta controle resultou na melhor receita bruta (USD$609,39); no entanto, houve uma perda (USD$50,96) em relação aos indicadores de desempenho econômico. Conclusão: O uso de até 2% de óleo de licuri na dieta proporcionou aumento nos rendimentos de carcaça de ovelhas de descarte.

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We investigated whether the recombinant leptin (1, 10, 100 ng/mL) influences the meiotic maturation of goat oocytes, whether the MAPK and JAK2/STAT3 pathways mediate the effects of leptin during in-vitro maturation, and whether leptin differently affects the abundance of mRNAs relevant to leptin signal transduction and apoptosis in oocytes and cumulus cells. The addition of leptin to the maturation medium positively affected the number of oocytes that completed nuclear maturation. Nuclear oocyte maturation stimulated by leptin was significantly impaired when we added the specific inhibitors of MAPK (U0126) and JAK2/STAT3 (AG490) to the maturation medium. The addition of leptin (10 ng/mL) during maturation did not affect the expression of AMPKα1, PPARα, Caspase 3, and BCL2 genes in oocytes or cumulus cells. The PPARγ and BAX mRNA abundances were significantly reduced in cumulus cells in the leptin group compared to the control group. Our results demonstrate that supplementation of the in-vitro maturation medium with leptin significantly improves nuclear maturation and reveal the important role of the MAPK and JAK2/STAT3 signaling pathways in establishing the leptin-mediated nuclear maturation of goat oocytes. Moreover, leptin treatment affects PPARγ and BAX gene expression in cumulus cells.

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In the present study, the cytotoxic effects of a 1,3-thiazolium-5-thiolate derivative of a mesoionic compound, MIH 2.4Bl, were assessed in the MCF-7 breast cancer cell line. The cytotoxic effects of MIH 2.4Bl were determined using a crystal violet assay. Using a dose-response curve, the IC50 value of MIH 2.4Bl was determined to be 45.8±0.8 µM. Additionally, the effects of MIH 2.4Bl on mitochondrial respiration were characterized using oxygen consumption rate analysis. Treating MCF-7 cells with increasing concentrations of MIH 2.4Bl resulted in a significant reduction in all mitochondrial respiratory parameters compared with the control cells, indicative of an overall decrease in mitochondrial membrane potential. The induction of autophagy by MIH 2.4Bl was also examined by measuring changes in the expression of protein markers of autophagy. As shown by western blot analysis, treatment of MCF-7 cells with MIH 2.4Bl resulted in increased protein expression levels of Beclin-1 and ATG5, as well as an increase in the microtubule-associated protein 1A/1B light chain 3B (LC3B)-II to LC3B-I ratio compared with the control cells. Microarray analysis of changes in gene expression following MIH 2.4Bl treatment demonstrated 3,659 genes exhibited a fold-change ≥2. Among these genes, 779 were up-regulated, and 2,880 were down-regulated in cells treated with MIH 2.4Bl compared with the control cells. Based on the identity of the transcripts and fold-change of expression, six genes were selected for verification by reverse transcription-quantitative (RT-q)PCR; activating transcription factor 3, acidic repeat-containing protein, heparin-binding EGF-like growth factor, regulator of G-protein signaling 2, Dickkopf WNT signaling pathway inhibitor 1 and adhesion molecule with Ig like domain 2. The results of RT-qPCR analysis of RNA isolated from control and MIH 2.4Bl treated cells were consistent with the expression changes identified by microarray analysis. Together, these results suggest that MIH 2.4Bl may be a promising candidate for treating breast cancer and warrants further in vitro and in vivo investigation.

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In this work, we report the synthesis of a new 1,3-thiazolium-5-thiolate derivative of a mesoionic compound (MIH 2.4Bl) and the characterization of its selective cytotoxicity on a panel of breast cancer cells lines. The cytotoxic effect of MIH 2.4Bl on breast cancer cell lines was determined by XTT and crystal violet assays, flow cytometry analysis, electron microscopy characterization, and terminal deoxynucleotidyl transferase (TdT) deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) apoptosis assays. As determined using XTT cell growth and survival assays, MIH 2.4Bl exhibited growth inhibition activity on most breast cancer cell lines tested, compared with normal human mammary epithelial cells. Three breast cancer cell lines (MCF-7, T-47D, and ZR-75-1) showed a more potent sensitivity index to growth inhibition by MIH 2.4Bl than the other breast cancer cell lines. Interestingly, these 3 cell lines were derived from tumors of Luminal A origin and have ER (estrogen receptor), PR (progesterone receptor), and HER2 (human epidermal growth factor receptor 2) positive expression. Additional analysis of cytotoxicity mediated by MIH 2.4Bl was performed using the MCF-7 cell line. MCF-7 cells displayed both time- and dose-dependent decreases in cell growth and survival, with a maximum cytotoxic effect observed at 72 and 96 hours. The MCF-7 cells were also characterized for cell cycle changes upon treatment with MIH 2.4Bl. Using flow cytometry analysis of cell cycle distribution, a treatment-dependent effect was observed; treatment of cells with MIH 2.4Bl increased the G2/M population to 34.2% compared with 0.1% in untreated (control) cells. Ultrastructural analysis of MFC-7 cells treated with MIH 2.4Bl at 2 different concentrations (37.5 and 75 µM) was performed by transmission electron microscopy. Cells treated with 37.5 µM MIH 2.4Bl showed morphologic changes beginning at 6 hours after treatment, while cells treated with 75 µM showed changes beginning at 3 hours after treatment. These changes were characterized by an alteration of nuclear morphology and mitochondrial degeneration consistent with apoptotic cell death. Results of a TUNEL assay performed on cells treated for 96 hours with MIH 2.4Bl supported the observation of apoptosis. Together, these results suggest that MIH 2.4Bl is a promising candidate for treating breast cancer and support further in vitro and in vivo investigation.

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The aims of the study were to characterize leptin and it is receptor (LEPR) proteins immunoexpression in ovine ovaries and to evaluate the effects of leptin on development of secondary follicles cultured in vitro. The ovaries were collected and fixed for immunohistochemical analysis. Additional pairs of ovaries were collected and secondary follicles were isolated and cultured, for 18 days, in α-MEM+ alone or supplemented with 10 or 25 ng/mL of leptin. The antrum formation and fully grown oocytes rates were higher in 25 ng/mL leptin than all treatments. GSH levels and mitochondrial activity were higher in 10 or 25 ng/mL leptin than α-MEM+. 25 ng/mL leptin showed a higher percentage of MII than the α-MEM+. In conclusion, leptin and its receptor are expressed in ovine ovaries and 25 ng/mL leptin promoted higher in vitro maturation rates by improving follicular development, GSH levels and mitochondrial activity of ovine oocytes compared to control medium.

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Mammary tumor is the most frequent among the tumors that affect canine females, with relevant importance in veterinary medicine. The objective of this study was to determine the image characteristics of mammary tumors in female dogs, and compare different ultrasonographic techniques for neoplastic evaluation. During the experiment, 30 bitches with presence of nodular lesion in the mammary gland were used. Initially females were submitted to clinical and laboratory evaluations, and subsequent to the ultrasound examination of the tumor mass, as well as abdominal ultrasound and thoracic x-ray for the metastasis investigation. Quantitative analysis by histogram of the gray levels and categorization of the tumor masses by the BI-RADS system were performed. Later, the bitches were submitted to surgical resection of the tumors, where samples of the neoplastic tissue were collected for histopathological analysis. Carcinoma in mixed tumor showed a higher rate (33.3%), and the malignancy degree of epidermal tumors were classified in grade 1 (n=9), grade 2 (n=12) and grade 3 (n=3). Malignancy degree showed positive correlation with BI-RADS (r=0.55; P<0.05) and with the parameter echotexture - histogram base width (r=0.42, P<0.05). BI-RADS graduation also showed a positive correlation with the echotexture parameters (standard deviation of average echogenicity r=0.66, P<0.05 and base width r=0.55, P<0.05). It was concluded that the BI-RADS method in combination with the echotexture of tumors, can be used to evaluate mammary tumors in dogs and establish the planning of treatment.(AU)

Entre os tumores que acometem cadelas a neoplasia mamária é a de maior incidência, apresentando relevante importância na medicina veterinária. Este trabalho teve o objetivo de determinar as características ultrassonográficas de neoplasias mamárias em cadelas, e comparar diferentes técnicas ultrassonográficas de avaliação tumoral. Durante a realização deste experimento, foram utilizadas 30 cadelas com presença de lesão nodular em glândula mamária. Inicialmente as fêmeas foram submetidas a avaliações clínicas e laboratoriais, e subsequentemente à realização de exame ultrassonográfico da massa tumoral, assim como ultrassom abdominal e raio x torácico para a pesquisa de metástase. Foram realizadas a análise quantitativa por histograma dos níveis de cinza e categorização das massas tumorais pelo sistema BI-RADS. Posteriormente as cadelas foram submetidas à ressecção cirúrgica dos tumores, onde foram coletadas amostras do tecido neoplásico para análise histopatológica. O carcinoma em tumor misto foi o tipo tumoral de maior incidência (33.3%), e a graduação de malignidade dos tumores epiteliais foram classificadas em grau 1 (n=9), grau 2 (n=12) e grau 3 (n=3). A graduação demonstrou correlação positiva com a categorização BI-RADS (r=0,55; P<0,05) e com o parâmetro de ecotextura - largura de base do histograma (r=0,42; P<0,05). A graduação BI-RADS também demonstrou uma correlação positiva com os parâmetros de ecotextura (desvio padrão da média da ecogenicidade r=0,66; P<0,05 e largura de base r=0,55; P<0,05). Conclui-se que o método de categorização BI-RADS, assim como os parâmetros de ecotextura dos tumores, podem ser usados para avaliação de neoplasia mamária em cadelas e assim auxiliar no planejamento de tratamento de cada caso.(AU)

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Mammary tumor is the most frequent among the tumors that affect canine females, with relevant importance in veterinary medicine. The objective of this study was to determine the image characteristics of mammary tumors in female dogs, and compare different ultrasonographic techniques for neoplastic evaluation. During the experiment, 30 bitches with presence of nodular lesion in the mammary gland were used. Initially females were submitted to clinical and laboratory evaluations, and subsequent to the ultrasound examination of the tumor mass, as well as abdominal ultrasound and thoracic x-ray for the metastasis investigation. Quantitative analysis by histogram of the gray levels and categorization of the tumor masses by the BI-RADS system were performed. Later, the bitches were submitted to surgical resection of the tumors, where samples of the neoplastic tissue were collected for histopathological analysis. Carcinoma in mixed tumor showed a higher rate (33.3%), and the malignancy degree of epidermal tumors were classified in grade 1 (n=9), grade 2 (n=12) and grade 3 (n=3). Malignancy degree showed positive correlation with BI-RADS (r=0.55; P<0.05) and with the parameter echotexture - histogram base width (r=0.42, P<0.05). BI-RADS graduation also showed a positive correlation with the echotexture parameters (standard deviation of average echogenicity r=0.66, P<0.05 and base width r=0.55, P<0.05). It was concluded that the BI-RADS method in combination with the echotexture of tumors, can be used to evaluate mammary tumors in dogs and establish the planning of treatment.(AU)

Entre os tumores que acometem cadelas a neoplasia mamária é a de maior incidência, apresentando relevante importância na medicina veterinária. Este trabalho teve o objetivo de determinar as características ultrassonográficas de neoplasias mamárias em cadelas, e comparar diferentes técnicas ultrassonográficas de avaliação tumoral. Durante a realização deste experimento, foram utilizadas 30 cadelas com presença de lesão nodular em glândula mamária. Inicialmente as fêmeas foram submetidas a avaliações clínicas e laboratoriais, e subsequentemente à realização de exame ultrassonográfico da massa tumoral, assim como ultrassom abdominal e raio x torácico para a pesquisa de metástase. Foram realizadas a análise quantitativa por histograma dos níveis de cinza e categorização das massas tumorais pelo sistema BI-RADS. Posteriormente as cadelas foram submetidas à ressecção cirúrgica dos tumores, onde foram coletadas amostras do tecido neoplásico para análise histopatológica. O carcinoma em tumor misto foi o tipo tumoral de maior incidência (33.3%), e a graduação de malignidade dos tumores epiteliais foram classificadas em grau 1 (n=9), grau 2 (n=12) e grau 3 (n=3). A graduação demonstrou correlação positiva com a categorização BI-RADS (r=0,55; P<0,05) e com o parâmetro de ecotextura - largura de base do histograma (r=0,42; P<0,05). A graduação BI-RADS também demonstrou uma correlação positiva com os parâmetros de ecotextura (desvio padrão da média da ecogenicidade r=0,66; P<0,05 e largura de base r=0,55; P<0,05). Conclui-se que o método de categorização BI-RADS, assim como os parâmetros de ecotextura dos tumores, podem ser usados para avaliação de neoplasia mamária em cadelas e assim auxiliar no planejamento de tratamento de cada caso.(AU)

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Recent studies have shown that adiponectin, an adipokine predominantly produced by adipose tissue, regulates several reproductive processes. However, the mechanisms of action of adiponectin on the maturation of goat oocytes remain to be determined. The aim of this study was to investigate whether (a) adiponectin influences the meiotic maturation of goat oocytes; (b) MAPK MEK 1/2 mediates the effects of adiponectin; and 3) adiponectin differentially affects mRNA relative abundance of genes relevant for adiponectin signal transduction in goat oocytes. The addition of adiponectin (5 µg/ml) during the maturation of goat oocytes resulted in a higher percentage of successful nuclear maturation compared to those of the group without adiponectin (p < 0.05). Adiponectin-stimulated nuclear oocyte maturation was significantly impaired by a mitogen-activated protein kinase MEK 1/2 inhibitor, U0126 (p < 0.05). There was no evidence of any adiponectin-induced difference in the relative transcript abundances of AdipoR1, AdipoR2, AMPKα1, AMPKα2, PPARα and PPARγ genes. In conclusion, these results indicate that adiponectin has a positive effect on the meiotic maturation of goat oocytes through the MAPK MEK 1/2 pathway. Furthermore, the adiponectin does not affect the relative abundance of genes relevant for adiponectin signal transduction in goat oocytes.

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Adiponectin is an adipokine secreted primarily by adipocytes and is involved in the control of male and female reproductive functions. Circulating levels of adiponectin are inversely correlated with body fat mass, and its biological effects are predominantly mediated through two receptors, AdipoR1 and AdipoR2. The aim of the present study was to verify the expression of the adiponectin system (adiponectin and its receptors, AdipoR1 and AdipoR2) in goat ovary using qPCR and immunohistochemistry analyses and further investigate the in vitro effects of recombinant adiponectin (5 µg/mL and 10 µg/mL) on goat oocyte nuclear maturation. We demonstrated that the mRNA and proteins of the adiponectin system are present in goat ovary. Gene and protein expression of AdipoR1 and AdipoR2 was detected in follicular cells (oocyte, cumulus, granulosa and theca) of small and large antral follicles, while adiponectin mRNA was not detected in oocytes from small and large follicles or in large follicle cumulus cells. Finally, addition of various concentrations of adiponectin in maturation medium affected the number of oocytes that reached metaphase II. In conclusion, in the present study, we detected expression of adiponectin and its receptors AdipoR1 and AdipoR2 in goat ovarian follicles. Furthermore, we demonstrated that recombinant adiponectin increases nuclear maturation of goat oocytes in vitro.

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Objetivou-se avaliar a expressão do mRNA para o gene do fator de crescimento IGF-2 em oócitos e células do cumulus de ovelhas em diferentes estágios do desenvolvimento folicular. Os folículos classificados morfologicamente como antrais (terciários e pré-ovulatórios) foram aspirados manualmente para obtenção dos oócitos e células do cumulus. Os folículos pré-antrais (secundários) foram extraídos do córtex ovariano, por microdissecção, e os oócitos retirados. Nos dois grupos, os oócitos foram desnudados e agrupados em pools de dez células cada (Grupo A, n=10; Grupo B, n=10) e dez amostras com grupos de células do cumulus (Grupo A1, n=10, B1, n=10). O mRNA foi extraído e convertido em cDNA utilizando a técnica da RT-PCR, utilizando Oligo DT randômico para o mRNA. A análise da expressão confirmou a expressão gênica para IGF-2 nos grupos de oócitos e células do cumulus. Houve um aumento da expressão relativa do mRNA para IGF-2 nos grupos de oócitos durante a fase mais tardia do desenvolvimento folicular e as diferenças foram consideradas significantes (p<0,05). Não houve variação significante da expressão de IGF2 entre os grupos de células do cumulus. Conclui-se que o fator de crescimento IGF-2 tem níveis mais elevados de expressão em oócitos ovinos, na segunda fase do desenvolvimento folicular, mas expressão semelhante em células do cumulus durante as fases estudadas do desenvolvimento folicular.(AU)

The aim of this study was to analyze the mRNA expression of IGF-2 growth factor in oocytes and cumulus cells from native sheep follicles at different stages of follicular development. The classified morphologically as antral follicles (tertiary preovulatory) were aspirated manually to obtain the oocyte and the cumulus cells. The preantral follicles (secondary) were extracted from the ovarian cortex by microdissection, and oocytes were removed. In both groups, oocytes were denuded and grouped into pools of ten cells each (Group A, n=10, Group B, n=10) and ten samples with groups of cumulus cells (Group A1, n=10; B1, n=10). The mRNA was extracted and converted to cDNA using the RT-PCR technique. The expression analysis confirmed the expression of IGF-2 gene for groups of oocyte and the cumulus cells. There was an increase in the relative expression of mRNA for IGF-2 for groups of oocytes during the later stage of follicular development and differences were considered significant (p<0.05). There was no significant variation in the expression of IGF2 between groups of cumulus cells. It is concluded that the growth factor IGF-2 has higher levels of expression in sheep oocytes in the second stage of follicular development in the conditions adopted and similar expression in cumulus cells during various stages of follicular development.(AU)

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Objetivou-se avaliar a expressão do mRNA para o gene do fator de crescimento IGF-2 em oócitos e células do cumulus de ovelhas em diferentes estágios do desenvolvimento folicular. Os folículos classificados morfologicamente como antrais (terciários e pré-ovulatórios) foram aspirados manualmente para obtenção dos oócitos e células do cumulus. Os folículos pré-antrais (secundários) foram extraídos do córtex ovariano, por microdissecção, e os oócitos retirados. Nos dois grupos, os oócitos foram desnudados e agrupados em "pools" de dez células cada (Grupo A, n=10; Grupo B, n=10) e dez amostras com grupos de células do cumulus (Grupo A1, n=10, B1, n=10). O mRNA foi extraído e convertido em cDNA utilizando a técnica da RT-PCR, utilizando Oligo DT randômico para o mRNA. A análise da expressão confirmou a expressão gênica para IGF-2 nos grupos de oócitos e células do cumulus. Houve um aumento da expressão relativa do mRNA para IGF-2 nos grupos de oócitos durante a fase mais tardia do desenvolvimento folicular e as diferenças foram consideradas significantes (p<0,05). Não houve variação significante da expressão de IGF2 entre os grupos de células do cumulus. Conclui-se que o fator de crescimento IGF-2 tem níveis mais elevados de expressão em oócitos ovinos, na segunda fase do desenvolvimento folicular, mas expressão semelhante em células do cumulus durante as fases estudadas do desenvolvimento folicular.(AU)

The aim of this study was to analyze the mRNA expression of IGF-2 growth factor in oocytes and cumulus cells from native sheep follicles at different stages of follicular development. The classified morphologically as antral follicles (tertiary preovulatory) were aspirated manually to obtain the oocyte and the cumulus cells. The preantral follicles (secondary) were extracted from the ovarian cortex by microdissection, and oocytes were removed. In both groups, oocytes were denuded and grouped into "pools" of ten cells each (Group A, n=10, Group B, n=10) and ten samples with groups of cumulus cells (Group A1, n=10; B1, n=10). The mRNA was extracted and converted to cDNA using the RT-PCR technique. The expression analysis confirmed the expression of IGF-2 gene for groups of oocyte and the cumulus cells. There was an increase in the relative expression of mRNA for IGF-2 for groups of oocytes during the later stage of follicular development and differences were considered significant (p<0.05). There was no significant variation in the expression of IGF2 between groups of cumulus cells. It is concluded that the growth factor IGF-2 has higher levels of expression in sheep oocytes in the second stage of follicular development in the conditions adopted and similar expression in cumulus cells during various stages of follicular development.(AU)

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The effects of deslorelin acetate use in inducing ovulation need to be clarified to improve the results of equine embryo transfer. The mRNA abundance for angiogenic factors and LH receptor (LHR) in corpus luteum (CL) was studied in mares with natural (control group [CG]) and induced ovulation with deslorelin acetate (treatment group [TG]; follicles: ≥ 35 mm). Transrectal ultrasonography was used to verify the ovulation day, and on Days 4, 8, and 12 after ovulation (Day 0), CL samples were obtained through ultrasound-guided biopsy. The messenger RNA expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and LHR genes were analyzed by real-time polymerase chain reaction. A positive correlation was observed between VEGF and LHR (P < 0.00001, r = 0.78), and it was possible to detect higher LHR expression in the TG than in the CG on Day 4 (P < 0.05). Moreover, this expression was higher on Days 4 and 8 than on Day 12 in the TG. Basic fibroblast growth factor was also expressed in luteal tissue on all days for both groups; however, these differences were not significant. In conclusion, deslorelin acetate was effective for the induction of ovulation in mares, resulting in higher expression of LHR, especially on the fourth day after ovulation. In addition, VEGF expression was influenced by induced ovulation, with a lower level on Day 12, which is expected in nonpregnant mares.

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O Programa de Pós-Graduação em Biotecnologia (PPGB) da Renorbio, em nível apenas de doutorado, tem por missão formar pessoal qualificado para o exercício da pesquisa e do magistério superior, assim como promover pesquisa articulada em Biotecnologia incentivando a interação, intercâmbio e transferência de tecnologias para o setor produtivo, depósito de patentes e fortalecer a interação e intercâmbio Universidade-Empresa. O PPGB agrega 36 diferentes Instituições da Região Nordeste e Estado do Espirito Santo, entre universidades e centros de pesquisa, sendo 12 nucleadoras (instituições que podem emitir diplomas). O PPGB possui 4 áreas de conhecimento da biotecnologia: agropecuária, industrial, recursos naturais e saúde. Na área de reprodução animal, vários projetos vem sendo desenvolvidos, inclusive com depósito de patente.

The Graduate Program in Biotechnology (PPGB) ­RENORBIO, in the doctoral level only, has the mission of training students to the research and university teaching and promote articulated research in Biotechnology encouraging interaction, exchange and transfer of technologies to the productive sector, patent deposit and strengthen interaction and exchanges University-Industry. The PPGB adds 36 different institutions (universities and research centers) in the Northeastern Region and Espirito Santo State, 12 of them are institutions that may issue diplomas. Four knowledge areas of biotechnology are developed: Agricultural, industrial, natural resources and health. In the animal reproduction area, several projects have been developed, including patent deposit

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O Programa de Pós-Graduação em Biotecnologia (PPGB) da Renorbio, em nível apenas de doutorado, tem por missão formar pessoal qualificado para o exercício da pesquisa e do magistério superior, assim como promover pesquisa articulada em Biotecnologia incentivando a interação, intercâmbio e transferência de tecnologias para o setor produtivo, depósito de patentes e fortalecer a interação e intercâmbio Universidade-Empresa. O PPGB agrega 36 diferentes Instituições da Região Nordeste e Estado do Espirito Santo, entre universidades e centros de pesquisa, sendo 12 nucleadoras (instituições que podem emitir diplomas). O PPGB possui 4 áreas de conhecimento da biotecnologia: agropecuária, industrial, recursos naturais e saúde. Na área de reprodução animal, vários projetos vem sendo desenvolvidos, inclusive com depósito de patente.(AU)

The Graduate Program in Biotechnology (PPGB) RENORBIO, in the doctoral level only, has the mission of training students to the research and university teaching and promote articulated research in Biotechnology encouraging interaction, exchange and transfer of technologies to the productive sector, patent deposit and strengthen interaction and exchanges University-Industry. The PPGB adds 36 different institutions (universities and research centers) in the Northeastern Region and Espirito Santo State, 12 of them are institutions that may issue diplomas. Four knowledge areas of biotechnology are developed: Agricultural, industrial, natural resources and health. In the animal reproduction area, several projects have been developed, including patent deposit.(AU)

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This study was undertaken with the aim to evaluate the p53 expression, applying the immunohistochemical technique to malignant primary mammary neoplasms histopathologically diagnosed in female dogs, and to investigate exon 8 of the Tp53 suppressor gene for mutation types by means of PCR-RFLP pattern of bands. Nineteen healthy mammary glands were used as a control group (group 1). Samples from 18 cases diagnosed with malignant mammary tumors (group 2), and the contralateral mammary glands (group 3) were collected during the UFRPE Veterinary Hospital routine. The tumors were diagnosed by histopathology and subdivided into grades of malignity. The streptavidin-biotin peroxidase method was used to analyze the immunohistochemical expression of p53, evaluated according to the location and intensity of stain. Expression of p53 protein was not observed in the samples of group 1. On the contrary, it was observed in all malignant tumors; the protein p53 was localized either only in the nucleus or in the nucleus and in the cytoplasm, in samples of group 2. In group 3, expression of p53 protein was observed in the tumors (2 samples) and in normal mammary tissues (4 samples). For the molecular analyses, genomic DNA was extracted and submitted to PCR-RFLP with the following endonuclease enzymes: AluI, BsoBI, DdeI and SmaI. The band pattern showed polymorphism between groups, but not between histological variants of tumors. This polymorphism detected mutations in the fragment studied - exon 8 of Tp53 - which could account for changes in nucleotides, located in the restriction sites of the endonuclease enzymes. In conclusion, the immunoexpression of p53 had no relationship with histological subtype or malignity grade, but it can be related to the presence of mammary tumors in female dogs. The PCR-RFLP technique can be an important tool for the study of mammary carcinogenesis in bitches because the polymorphism obtained may allow early diagnosis in tissues of mammary glands.(AU)

Este estudo foi realizado com o objetivo de avaliar a expressão da proteína p53, pela técnica de imuno-histoquímica, em neoplasmas mamários malignos em cadelas, além de investigar mutações no éxon 8 do gene supressor Tp53 por meio do padrão de bandas obtidas por PCR-RFLP. Dezenove mamas de cadelas saudáveis foram usadas como controle (Grupo 1). Amostras de 18 casos de tumores malignos (Grupo 2) e suas glândulas mamárias contralaterais (Grupo 3) foram obtidas na rotina do Hospital Veterinário da UFRPE. Os tumores foram identificados histologicamente e classificados em graus de malignidade. O método da estreptoavidina-biotina peroxidase foi utilizado para a análise da expressão de p53 por imuno-histoquímica, de acordo com a localização e intensidade da coloração. A expressão da proteína p53 não foi observada nas amostras do Grupo 1, mas foi encontrada em todas as amostras de tumores malignos (Grupo 2) seja só no núcleo, ou também no citoplasma. No Grupo 3, a expressão foi observada em quatro amostras normais e em duas que apresentavam tumor. Para a análise molecular, o DNA genômico foi extraído e submetido à PCR-RFLP com as seguintes endonucleases: AluI, BsoBI, DdeI e SmaI. O padrão de bandas foi polimórfico entre os grupos, mas não entre as variantes tumorais. Esse polimorfismo detectou mutações no fragmento estudado - éxon 8 do gene Tp53 - que podem resultar em alterações nos nucleotídeos, localizados nos sítios de restrição das enzimas. Esses achados levam a conclusão de que a imunoexpressão da p53 não tem relação com o subtipo histológico ou grau de malignidade do tumor, mas sim com a presença dos tumores no tecido mamário de cadelas. A PCR-RFLP pode ser usada como importante ferramenta para o estudo da carcinogênese mamária na cadela, possibilitando gerar diagnósticos precoces através do polimorfismo obtido com endonucleases de restrição pré-selecionadas.(AU)

Asunto(s)

RESUMEN

This study was undertaken with the aim to evaluate the p53 expression, applying the immunohistochemical technique to malignant primary mammary neoplasms histopathologically diagnosed in female dogs, and to investigate exon 8 of the Tp53 suppressor gene for mutation types by means of PCR-RFLP pattern of bands. Nineteen healthy mammary glands were used as a control group (group 1). Samples from 18 cases diagnosed with malignant mammary tumors (group 2), and the contralateral mammary glands (group 3) were collected during the UFRPE Veterinary Hospital routine. The tumors were diagnosed by histopathology and subdivided into grades of malignity. The streptavidin-biotin peroxidase method was used to analyze the immunohistochemical expression of p53, evaluated according to the location and intensity of stain. Expression of p53 protein was not observed in the samples of group 1. On the contrary, it was observed in all malignant tumors; the protein p53 was localized either only in the nucleus or in the nucleus and in the cytoplasm, in samples of group 2. In group 3, expression of p53 protein was observed in the tumors (2 samples) and in normal mammary tissues (4 samples). For the molecular analyses, genomic DNA was extracted and submitted to PCR-RFLP with the following endonuclease enzymes: AluI, BsoBI, DdeI and SmaI. The band pattern showed polymorphism between groups, but not between histological variants of tumors. This polymorphism detected mutations in the fragment studied - exon 8 of Tp53 - which could account for changes in nucleotides, located in the restriction sites of the endonuclease enzymes. In conclusion, the immunoexpression of p53 had no relationship with histological subtype or malignity grade, but it can be related to the presence of mammary tumors in female dogs. The PCR-RFLP technique can be an important tool for the study of mammary carcinogenesis in bitches because the polymorphism obtained may allow early diagnosis in tissues of mammary glands.

Este estudo foi realizado com o objetivo de avaliar a expressão da proteína p53, pela técnica de imuno-histoquímica, em neoplasmas mamários malignos em cadelas, além de investigar mutações no éxon 8 do gene supressor Tp53 por meio do padrão de bandas obtidas por PCR-RFLP. Dezenove mamas de cadelas saudáveis foram usadas como controle (Grupo 1). Amostras de 18 casos de tumores malignos (Grupo 2) e suas glândulas mamárias contralaterais (Grupo 3) foram obtidas na rotina do Hospital Veterinário da UFRPE. Os tumores foram identificados histologicamente e classificados em graus de malignidade. O método da estreptoavidina-biotina peroxidase foi utilizado para a análise da expressão de p53 por imuno-histoquímica, de acordo com a localização e intensidade da coloração. A expressão da proteína p53 não foi observada nas amostras do Grupo 1, mas foi encontrada em todas as amostras de tumores malignos (Grupo 2) seja só no núcleo, ou também no citoplasma. No Grupo 3, a expressão foi observada em quatro amostras normais e em duas que apresentavam tumor. Para a análise molecular, o DNA genômico foi extraído e submetido à PCR-RFLP com as seguintes endonucleases: AluI, BsoBI, DdeI e SmaI. O padrão de bandas foi polimórfico entre os grupos, mas não entre as variantes tumorais. Esse polimorfismo detectou mutações no fragmento estudado - éxon 8 do gene Tp53 - que podem resultar em alterações nos nucleotídeos, localizados nos sítios de restrição das enzimas. Esses achados levam a conclusão de que a imunoexpressão da p53 não tem relação com o subtipo histológico ou grau de malignidade do tumor, mas sim com a presença dos tumores no tecido mamário de cadelas. A PCR-RFLP pode ser usada como importante ferramenta para o estudo da carcinogênese mamária na cadela, possibilitando gerar diagnósticos precoces através do polimorfismo obtido com endonucleases de restrição pré-selecionadas.

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Background: Understanding ovarian folliculogenesis is critical to the study of fertility and in the development of fertility techniques as well as contraception. Mares and women share striking similarities in ovarian folliculogenesis, and in insulin resistance and obesity syndromes. The effects of insulin resistance and obesity on follicular development and the surrounding endocrinology and genes in mares may shed light on the causes and effects of metabolic and reproductive disorders such as polycystic ovarian syndrome (PCOS) in women using an appropriate research model. Review: Studies in laboratory animals (e.g. mice and rats) have demonstrated that, in general, these animals are not good research models to study ovarian function in women because of the remarkable physiological differences in ovarian folliculogenesis and luteogenesis/luteolysis. Therefore, there is an urgent need for the development of in vivo (whole animal) research models using species (e.g. mare and cow) that have a similar physiological ovarian function to the woman. The use of such models will allow for an understanding of the causes and effects of different pathological reproductive processes involved in diseases. Several studies from our group and others have shown that nowadays the mare seems to be one, if not the best, animal model to study ovarian function in women. In addition, the recent elucidation of the equine genome has provided evidence of the high gene conservation and similar chromosomal order of this species to humans, reinforcing the importance of this species for comparative studies with humans. Conclusions: The use of farm animal models is also relevant for agricultural and biomedical research because this might help to improve reproductive efficiency and health in animals and humans, as well as the quality of products (e.g. oocytes, embryos, etc.). This review will focus on the potential use of the mare as a model to study the effects of obesity and insulin resistance syndromes on ovarian function in women.

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Editorial

RESUMEN

A população de raças caprinas naturalizadas e Sem Raça Definida (SRD) do Nordeste do Brasil é pouco caracterizada, apesar da importância que se tem dado aos estudos de recursos genéticos em animais domésticos. Com o objetivo de estudar o polimorfismo do gene da αS1-caseína em DNA genômico de cabras Moxotó e SRD provenientes do Semiárido do Nordeste brasileiro, por meio da técnica de PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Lenght Polymorfism), foram genotipadas 215 cabras pertencentes aos grupos genéticos Moxotó e SRD, provenientes dos estados de Pernambuco, Paraíba, Rio Grande do Norte e Ceará. Com base na frequência alélica das raças estudadas, não se observou diferença significativa (p> 0,05) entre as populações de cada Estado e entre animais Moxotó e SRD. Diante da maior frequência observada do alelo B (forte) do gene da αS1-caseína nos animais estudados, admite-se a possibilidade de que fenotipicamente esses animais venham a exibir a característica de uma forte produção de proteínas, característica importante para o leite destinado à produção de queijos, favorecendo a caprinocultura da região.

The population of native and mixed-breeds (MB) goats from the Northeast of Brazil is little characterized, despite the importance given to studies of genetic resources in domestic animals. With the objective of studying the polymorphism of the αS1-casein gene in genomic DNA of Moxotó and mixed-breed goats from the semi-arid of Brazilian Northeast, by the PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Lenght Polymorfism) technique, 215 Moxotó and mixed-breeed goats, which came from the Brazilian states of Pernambuco, Paraíba, Rio Grande do Norte and Ceará, were used. Based on the allelic frequency from the breeds studied, there was no significant difference (p> 0.05) among the populations of each studied state and between Moxotó and mixedbreed animals. In face of the detection of a highest presence of allele B (strong) from the αS1-casein gene in the animals studied, it is admitted the possibility that, phenotypically, these animals present the characteristic of a strong production of proteins, an important feature for the milk destined to cheese production, favoring the goat raising in the region.

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Desenvolveu-se este estudo para avaliar os efeitos das condições reprodutivas (multíparas ou nulíparas) e climáticas (período seco ou chuvoso) sobre o número e qualidade de embriões colhidos de cabras da raça Boer superovuladas. Cinquenta fêmeas caprinas, sendo 33 multíparas e 17 nulíparas, foram submetidas à sincronização do estro com o auxílio de CIDR®, durante onze dias, e superovuladas com 250UI de FSH-p. Observaram-se as fêmeas quanto ao estro e realização de cobertura a intervalos de doze horas, após a remoção dos CIDR®. Os embriões foram colhidos seis dias após a última cobertura, por via transcervical. Não houve diferença significativa (P>0,05) entre o número médio de estruturas e embriões viáveis recuperados das multíparas ou nulíparas. O número médio de estruturas e embriões viáveis recuperados de doadoras multíparas não diferiu significativamente (P>0,05) entre os períodos seco e chuvoso. O número médio de estruturas e de embriões viáveis, classificados como G1, recuperados de doadoras nulíparas foi maior (P<0,05) no período chuvoso que no período seco. Conclui-se que a condição climática, neste experimento, não teve efeito sobre a produção embrionária de fêmeas multíparas, entretanto as nulíparas foram mais sensíveis ao período seco, apresentando redução no número e qualidade dos embriões recuperados.

This study was conducted to evaluate the effect of the reproductive (multiparous or nulliparous) and climatic (dry or rainy period) conditions on the number and quality of embryos collected from Boer does. Fifty does, being 33 multiparous and 17 nulliparous, were synchronized for oestrus with CIDR® devices for a period of 11 days and superovulated with 250UI of FSH-p. The animals were observed for oestrous behaviour at 12 h intervals, after CIDR® withdrawal. Does were mated and six days later flushed transcervically to recover the embryos. There was no significant difference (P>0.05) among the average number of structures and viable embryos recovered from the multiparous and nulliparous females. The average number of structures and viable embryos recovered from multiparous does had no significant difference (P>0.05) between dry and rainy periods. The average number of structures and viable embryos classified as G1 recovered from nulliparous was lower (P<0.05) in the dry period than in the rainy period. It could be concluded that the climatic condition did not affect the embryonic production of multiparous females, however the nulliparous were more sensitive to the dry period, presenting reduction in the number and quality of the collected embryos.

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