L'exposition in utero au diabète maternel aboutit à des changements cardiovasculaires permanents en lien avec des altérations de la fonction vasculaire. En effet, alors que le rat exposé in utero à l'hyperglycémie maternelle modérée (DMO) développe une hypertension artérielle à 6 mois, une altération spécifique de la vasodilatation induite par la prostacycline dans l'aorte est présente dès l’âge de 3 mois. La prostacycline jouant un rôle clé dans le contrôle du tonus vasculaire dans les artères de résistance, nous avons émis l'hypothèse que la programmation foetale du tonus vasculaire suite à l'hyperglycémie maternelle pouvait jouer sur la régulation de la pression artérielle chez les DMO. Notre étude montre que la relaxation induite par la prostacycline est diminuée dans les artères mésentériques aussi bien à 3 mois qu’à 18 mois; de plus, une dysfonction endothéliale tardive associée à une augmentation de l'activité contractile des cellules musculaires lisses (tonus myogénique) est observée à 18 mois. Le tonus vasculaire est étroitement lié au remodelage vasculaire ; aussi nous avons étudié l'effet du diabète maternel sur le remodelage vasculaire après altération chronique de la pression ou du flux. Chez les DMO âgés, l'hypertension n’induit aucun remodelage de l'aorte, ni des artères mésentériques.Chez les jeunes DMO, la réduction du flux carotidien gauche (ligature) induit un remodelage constrictif ; la carotide droite subissant, quant à elle, un remodelage expansif en réponse à l’augmentation de flux. Par contre, la ligature des artères mésentériques n’entraîne aucun remodelage en réponse à la diminution de flux chez les DMO, alors que le remodelage expansif suite à l’augmentation de flux est maintenu. Ces résultats mettent en évidence une programmation foetale du système vasculaire en réponse au diabète maternel,qui pourrait être impliquée dans le développement de l'hypertension à plus long terme. / Cardiovascular alterations including hypertension are lifelong consequences of in utero exposure to maternal diabetes. One possibility was that hypertension could be caused by alterations of vascular function. Preliminary studies demonstrated that rat exposed in utero to moderated maternal hyperglycemia (DMO) developed hypertension around 6 months of age, associated with a specific alteration of prostacyclin-induced vasodilation in aorta. Because prostacyclin plays a key role in the control of vascular tone in resistance arteries, we aimed to determine whether maternal hyperglycaemia could affect this parameter in offspring. We hypothesized that fetal programming of vascular tone could impact regulation of arterial pressure in DMO. We found that prostacyclin induced relaxation was reduced in resistance arteries both at 3- and 18-month of age ; endothelium-mediated relaxation was reduced in 18-monthold DMO although an increase of contractile activity of smooth muscle cells (myogenic tone) was measured at the same age. Secondly, because vascular tone is closely linked to vascular remodeling, we studied the effect of maternal diabetes on vascular remodeling after chronic alteration of pressure or flow. In old DMO, hypertension did not induce inward remodeling neither in aorta nor in mesenteric arteries. In young DMO, the reduction of left carotid flow (induced by ligation) induced an inward remodeling ; the right carotid underwent an increased flow involving outward remodeling. Conversely, in mesenteric arteries, after 1 or 3 weeks of ligation, DMO did not exhibitin ward remodeling in response to low-flow although outward remodeling in high-flow arteries is maintained.These results evidenced a fetal programming of vascular system in adult rats exposed in utero to maternal diabetes, which may be involved in the development of hypertension.

The prenatal environment has been shown to have lasting effects on cardiovascular health. In the present study, pregnant rats were fed a 0.7% NaCl normal salt (NS) diet or an 8% NaCl high salt (HS) diet throughout pregnancy. Adult offspring were fitted with radiotelemetry probes to continuously measure blood pressure and heart rate. Rats were placed in restraining cages to test for a programmed acute stress hyperresponsiveness. Offspring were challenged with HS diet for one week to determine if blood pressure salt sensitivity had been programmed by the prenatal HS diet. Animals were killed following resting and acute stress conditions, after which brains and blood were collected for in situ hybridization for corticotropin releasing hormone (CRH) and radioimmunoassay for corticosterone. In order to determine the contribution of gene expression to differences seen, total brain RNA was analyzed with microarray. Rats were injected with a ganglionic blocker and an adrenergic receptor antagonist before restraint to examine the autonomic component of the stress response. High salt offspring of either sex did not have basal hypertension. Female HS offspring had an increased pressor and tachycardic response to acute stress compared with NS females. There were no differences between male NS and HS offspring during acute stress. Salt sensitivity was not induced during high salt challenge. According to the microarray, 11 genes were upregulated and 10 were downregulated in adult brains, while 17 were upregulated and 17 were downregulated in pup brains. These data indicate that there are long-term changes due to HS diet. CRH levels were higher in the paraventricular nucleus (PVN) of female HS offspring than in female NS offspring during both basal and stressed conditions, though no differences were seen in CRH expression levels of male offspring. Autonomic blockade completely abolished the enhanced tachycardic response seen in female HS offspring. However, a difference in NS and HS blood pressures remained. Thus, female offspring of mothers fed an 8% NaCl diet have alterations in cardiovascular control, indicated by an enhanced tachycardic response as adults, due to changes in the autonomic nervous system, and enhanced pressor response to stress mediated by unknown mechanisms.

Seminal plasma is commonly known to serve as a transport medium for sperm as it moves through the female reproductive tract for fertilization, however, more recent evidence demonstrates seminal plasma induces an expansive inflammatory response in the uterus. Murine models have found this inflammatory response is important for clearing pathogens and poor-quality sperm, eliciting the secretion of cytokines, chemokines, and embryokines that aid in embryo attachment and growth, placental angiogenesis, and blunting maternal immunity to the embryo. However, there is minimal research on the impacts of seminal plasma uterine priming in bovine, and more specifically embryo growth, uterine blood flow, offspring growth and metabolism, and production efficiency. There is significant evidence that malnutrition and environmental stress during gestation alters uterine blood flow resulting in poor placental efficiency and poor fetal growth and development which persists postnatally. Animal production is vital in providing high-quality protein for human consumption but recent challenges of public misconception, consumer preferences, high input costs, and environmental impacts threaten the security of these production systems. Growth efficiency is imperative for improving economic and environmental sustainability, and in turn ensuring the longevity of animal production systems. Knowing the impact of seminal plasma on the uterus, and its potential role in placental efficiency and subsequent offspring growth and metabolic function, and the negative impacts these can have on economic and environmental sustainability drive the need to better understand seminal plasma uterine priming in bovine.

Ionizing radiation exposure during pregnancy raises concerns of potentially harmful effects for both the mother and the unborn child. Fetal programming involves permanent changes in offspring phenotype due to stress experienced in-utero. This phenomenon has been well characterized in cardiometabolic disorders such as hypertension. The effects of prenatal ionizing radiation exposure on offspring cardiovascular endpoints following birth were studied in a mouse model. Pregnant wildtype C57Bl/6J mice were irradiated on day 15 of pregnancy with whole-body 137Cs gamma radiation at nominal doses of 5, 10, 50, 100, 300 or 1000 mGy. Post-natal measurements of offspring weight and blood pressure were completed. In female pups, blood pressure was significantly increased at 300 mGy and heart rate significantly decreased at 1000 mGy. Female pups were growth restricted over the study period at 50, 100 and 1000 mGy. Growth restriction in male pups was only observed at the highest dose of 1000 mGy. Unintended effects on the study measures caused by transportation of pregnant mothers to the irradiation facility were most evident in male offspring with increased blood pressure and heart rate and decreased body size. These unintended effects caused by transportation may have been attenuated with the 10 mGy in-utero exposure. Overall, these results suggest that prenatal radiation effects in mice are both dose- and gender-dependent, with even fairly low doses demonstrating (potentially adaptive) effects. There is a need for further study to better characterize the mechanism of this response. / Thesis / Master of Science (MSc)

Breast cancer is a persistent public health problem. Interesting hypothesis suggests that its risk can be modulated in early life periods, a phenomenon known as fetal programming. In this context, most fetal programming studies focus on maternal influence, due to the greater interaction between mother and fetus in both fetal and lactation periods. However, recent studies show that paternal preconception diet has also a major role in the offspring\'s susceptibility to metabolic chronic non-communicable diseases. Therefore, this direct doctoral project aimed to assess whether the paternal consumption of different high fat diets during the development period of the reproductive system of male rats increased the susceptibility of female offspring to mammary carcinogenesis. In addition we sought to evaluate which mechanisms could be involved in this process. We used male rats of the Sprague-Dawley strain (n = 20/group) that consumed high fat diet with 60% of calories from lipids from lard (LB group) or corn oil (CB group), or AIN-93G control diet (CO group) for nine weeks, during development and sexual maturation periods. These rats were mated with females who consumed only commercial diet in 1:1 ratio. Their 50 days old offspring were subjected to mammary carcinogenesis model using 7,12-dimethylbenz[a]anthracene (50mg/kg). Paternal consumption of high fat diet of animal or plant source had opposite effects, with the paternal consumption of diet with high content of saturated fatty acids (LB) increasing and consumption of diet with high content of n-6 polyunsaturated fatty acids (CB) reducing the risk of breast cancer development in female offspring. These effects were due to changes in the expression of 89 miRNAs in the father\'s sperm and 23 miRNAs in the offspring\'s mammary gland, with overlapping of three miRNAs (miR-1897-5p, miR- 219-1-3p and miR-376a #) that were altered in both tissues. Additionally, female offspring of males fed diets with high content of saturated fatty acids showed less differentiation of the mammary gland, higher levels of cell proliferation, lower levels of apoptosis and altered expression of keys proteins that regulate important cellular functions, such as epithelial to mesenchymal transition. Finally, these females had also altered lipid profile of the fat pad similar to their male parent as well as epigenetic changes that may be related to the etiology of breast cancer. Thus, we conclude that the high-fat preconception paternal diet programmed the susceptibility of female offspring to mammary carcinogenesis, but this effect was dependent on the type of fatty acid consumed and the observed effects possibly results from changes in miRNA expression profile. / O câncer de mama é um persistente problema de saúde pública. Hipótese intrigante sugere que a suscetibilidade à doença pode ser modulada em períodos precoces da vida, fenômeno conhecido como programação fetal. Nesse sentido, a maior parte dos estudos de programação fetal refere-se à influência materna, dada a intensa interação existente entre mãe e feto tanto no período fetal, quanto na lactação. Entretanto, estudos recentes mostram que a dieta paterna pré-concepcional também tem um papel de grande importância na suscetibilidade da prole à uma série de doenças crônicas não-transmissíveis de origem metabólica. Portanto, o presente projeto de doutorado direto teve como objetivo avaliar se o consumo paterno de diferentes dietas hiperlipídicas, durante o período de desenvolvimento do sistema reprodutivo de ratos machos, aumentaria a suscetibilidade da prole feminina à carcinogênese mamária. Adicionalmente buscou-se avaliar quais mecanismos poderiam estar envolvidos nesse processo. Utilizaram-se ratos machos da linhagem Sprague-Dawley (n=20/grupo) que consumiram dieta hiperlipídica com 60% de calorias provenientes de lipídeos de banha (grupo LB) ou óleo de milho (grupo CB), ou dieta controle AIN-93G (grupo CO), por nove semanas, durante os períodos de desenvolvimento e maturação sexual. Esses ratos foram acasalados com fêmeas, que consumiram apenas dieta comercial, na proporção 1:1. Sua prole de 50 dias foi submetida ao modelo de carcinogênese mamária com o uso de 7,12-dimetil-benza[a]antraceno (50mg/kg). O consumo paterno de dietas hiperlipídicas de origem animal ou vegetal conferiram efeitos opostos, com o consumo de dieta com alto teor de ácidos graxos saturados (LB) aumentando e o consumo de dieta com alto teor de ácidos graxos poli-insaturados n-6 (CB) diminuindo o risco de desenvolvimento de câncer de mama na prole feminina. Esses efeitos foram associados à alteração da expressão de 89 miRNAS no espermatozoide dos pais e 23 miRNAs na glândula mamária da prole, com sobreposição de 3 miRNAs (miR-1897-5p, miR-219-1-3p e miR-376a#) que estavam alterados em ambos tecidos. Adicionalmente, a prole feminina de machos que consumiram dieta com alto teor de ácidos graxos saturados apresentou menor diferenciação da glândula mamária, maior nível de proliferação celular, menor nível de apoptose e alteração da expressão de proteínas chaves da regulação celular, como na transição epitélio-mesenquimal. Finalmente, essas fêmeas também apresentaram perfil lipídico alterado semelhante à do seu progenitor masculino, bem como modificações epigenéticas que podem estar relacionadas à etiologia do câncer de mama. Assim, concluímos que a dieta paterna hiperlipídica pré-concepcional programou a suscetibilidade da prole feminina à carcinogênese mamária, porém esse efeito é dependente do tipo de ácido graxo consumido e os efeitos observados possivelmente decorrem de alterações no perfil de expressão de miRNAs.

O câncer de mama é o segundo tipo de neoplasia mais prevalente no mundo e o mais comum entre as mulheres. É descrito que o padrão de consumo alimentar materno e paterno está relacionado à suscetibilidade da prole ao desenvolvimento de doenças crônicas não transmissíveis, inclusive o câncer. A amora-preta é uma das frutas com maior conteúdo antioxidante e seus compostos bioativos possuem atividade antioxidante, anticarcinogênica e anti-inflamatória. Sendo assim, o presente trabalho propõe avaliar os efeitos do consumo materno e/ou paterno de extrato de amora-preta (Rubus spp.) na suscetibilidade da prole feminina ao desenvolvimento de neoplasias mamárias quimicamente induzidas. Para tanto, camundongos da linhagem C57BL/6 foram divididos aleatoriamente em 4 grupos: pai amora (PA), mãe amora (MA), pai e mãe amora (PMA) e controle (CTRL). Os pais receberam extrato de amora-preta logo após o desmame durante 8 semanas e as mães receberam o extrato durante a gestação e lactação. O extrato de amora-preta foi administrado na água de beber (0.84g de antocianinas/L) ad libitum. Os pais tratados com extrato de amora apresentaram redução na atividade enzimática da superóxido dismutase (SOD) e da catalase (CAT) no testículo (p<0.05 e p<0.001, respectivamente), aumento na capacidade antioxidante plasmática, na porcentagem de espermatozoides normais e na produção diária de espermatozóides em relação ao grupo controle (p<0.001 para todos). Além disso, os grupos PA, MA e PMA apresentaram aumento na taxa de prenhez (p<0.05) e redução da mortalidade perinatal (p<0.01, p<0.05 e p<0.001, respectivamente). Em relação à prole feminina não submetida à carcinogênese foi observada redução na capacidade antioxidante plasmática nos grupos PA (p<0.001) e MA (p<0.01), enquanto o grupo PMA apresentou aumento nesse parâmetro (p<0.001). No desenvolvimento da glândula mamária, houve aumento do desenvolvimento epitelial nos grupos PA, MA e PMA (p<0.001 para todos), de diferenciação nos grupos MA e PMA (p<0.01 para ambos) e da taxa de apoptose nos grupos MA e PMA (p<0.05), além de redução no número de TEBs nos grupos PA, MA e PMA (p<0.01, p<0.001 e p<0.001, respectivamente). Não foram observadas alterações significativas nas filhas submetidas à indução química da carcinogênese mamária por DMBA. Assim, é possível concluir que apesar de ter alterado o desenvolvimento da glândula mamária, o consumo materno e/ou paterno de extrato de amora-preta não foi capaz de impactar sobre a suscetibilidade da prole feminina à carcinogênese mamária quimicamente induzida. / Breast cancer is the second most prevalent type of cancer in the world and the most common among women. It is known that maternal and paternal food intake pattern are related to offspring susceptibility to non-communicable diseases, including cancer. Blackberry is one of the fruits with high antioxidant content and its compounds have antioxidant, anticarcinogenic and anti-inflammatory properties. So, the aim of the present study was evaluate the effects of maternal and/or paternal blackberry extract consumption on female offspring susceptibility to chemically-induced breast carcinogenesis. Thus, C57BL/6 mice were divided into 4 groups: father blackberry (FB), mother blackberry (MB), father and mother blackberry (FMB) and control (CTRL). Fathers received blackberry extract from weaning during 8 weeks and the mothers were treated during gestation and lactation. Blackberry extract was given in the drink water (0.84g anthocyanins/L) ad libitum. Fathers treated with blackberry had a reduction on superoxide dismutase (SOD) and catalase (CAT) activities in the testis (p<0.05 and p<0.001, respectivelly), an increase on plasmatic antioxidant capacity, percentage of normal sperm and daily sperm production in relation to control group (p<0.001 for all comparisons). Moreover, FB, MB and FMB groups had an increase of pregnancy rate (p<0.05) and a decrease of perinatal mortality (p<0.01, p<0.05 and p<0.001, respectively). Female offspring had a reduction of plasmatic antioxidant capacitity in FB (p<0.001) and MB (p<0.01) groups, while FMB group showed an increase in this parameter (p<0.001). On mammary gland development, it was observed higher epithelial development in FB, MB and FMB groups (p<0.001 for all comparisons), increased differentiation in MB and FMB groups (p<0.01 for both) and higher apoptosis rate in MB and FMB groups (p<0.05 for both), besides decreased TEBs number in FB, MB and FMB groups (p<0.01, p<0.001 and p<0.001, respectively). It was not found significant differences in the female offspring submitted to chemically-induced breast carcinogenesis. So, it is possible to conclude that in spite of maternal and/or paternal blackberry extract consumption changed the mammary gland development, it was not able to change the female offspring susceptibility to chemically-induced breast carcinogenesis.

Os objetivos deste estudo foram: 1- Avaliar o efeito da suplementação proteica, no terço final da gestação de vacas nelore, sobre a programação fetal e o desempenho de suas crias, submetidas à diferentes ritmos de crescimento até os 318 dias de vida. 2- Avaliar economicamente as diferentes estratégias de suplementação adotadas. Na fase I foram utilizadas 411 vacas da raça Nelore, no início do terço final da gestação, com idade média de 7,6 ± 2,1 anos, peso corporal médio de 429 ± 49 kg e escore de condição corporal (ECC) de 2,84 ± 0,44 (escala de 1 a 5). As vacas foram distribuídas em dois tratamentos conforme o ECC, peso corporal, idade e touro pai da cria. Os tratamentos foram: 1- grupo Vaca- Suplementada (VS) (n = 212) e 2- grupo Vaca-Controle (VC) (n = 199). Os animais do tratamento VS receberam diariamente 0,56 kg de um suplemento mineral proteico composto por 0,50 kg de farelo de soja e 0,06 kg de suplemento mineral, enquanto os animais do tratamento VC receberam somente o suplemento mineral. Na fase II, que corresponde ao período no qual a suplementação foi aplicada às crias, as vacas da primeira fase, juntamente com suas crias, foram redistribuídas, a partir do segundo mês de idade das crias em dois tratamentos. Os tratamentos foram suplementação em creep-feeding, oferecida ad libitum, representando o tratamento Bezerro-Suplementado (BS) (n = 201) e suplementação somente com sal mineral, representando o tratamento Bezerro-Controle (BC) (n = 188). Durante a fase II os animais foram pesados quatro vezes, sendo a primeira aos 108, a segunda aos 149, a terceira aos 182, e a quarta aos 208 dias de idade média dos grupos. Após a quarta pesagem os bezerros foram desmamados e encaminhados a um pasto onde permaneceram por 25 dias até o início da fase III. O ganho médio diário (GMD) foi calculado pela diferença entre duas pesagens consecutivas. Na fase III, que corresponde ao início do período de recria e da estação de inverno, e que teve duração de 85 dias, os bezerros (as) foram reagrupados em dois tratamentos. Em um dos tratamentos os animais foram alimentados em sistema de confinamento, originando o tratamento Recria-Suplementado (RS) (n = 209). No outro tratamento os animais foram mantidos em pastagem formada predominantemente por Panicum maximum cv. Colonhão ou Brachiaria brizantha cv. Marandu, formando o tratamento Recria-Controle (RC) (n = 180). Machos e fêmeas foram mantidos separados tanto no tratamento RS como no RC. As pesagens ocorreram com 16 horas de jejum hídrico e de sólidos, e foram realizadas aos 233, 261, 289 e 318 dias de idade média dos grupos. A análise econômica foi feita com base no cálculo da margem bruta, obtida pela subtração dos custos diferenciais da receita total. Foi observado efeito (P<0,05) da suplementação materna nas três primeiras pesagens da fase II, sendo que o grupo controle apresentou maiores pesos. No entanto, o peso aos 208 dias, momento do desmame, não sofreu (P>0,05) influência da suplementação materna. Foi observado efeito (P<0,05) da suplementação das crias em creep-feeding sobre o peso aos 208 dias, sendo que os animais suplementados foram mais pesados (187 x 180 kg). Quanto ao efeito de sexo das crias, os machos foram (P<0,05) mais pesados, tiveram maior (P<0,05) GMD e ganho de peso total (GTC) do que as fêmeas em todas as pesagens da fase II. Ao avaliar o efeito da suplementação materna sobre o GMD das crias na fase II, não foram (P>0,05) verificadas diferenças entre os tratamentos. Ao analisar o efeito da suplementação das crias em creep-feeding sobre o GMD, e GTC, verificou-se que os animais suplementados obtiveram resultados superiores (P<0,05). O efeito da suplementação materna sobre o peso das crias no decorrer da fase III, não foi diferente (P>0,05) entre os tratamentos. A suplementação das crias em creep-feeding foi efetiva em manter a diferença de peso obtida ao desmame, por toda a fase III, pois os animais suplementados foram mais pesados (P<0,05) ao final desta. Observou-se efeito de interação (P<0,05) entre manejo alimentar na fase III e sexo, sendo que, tanto para os animais alimentados em confinamento quanto para os mantidos à pasto, os machos foram mais pesados que as fêmeas. O GMD, ganho de peso médio diário da fase de recria (GMDR), ganho de peso total da fase de recria (GTR), e ganho total (GT) das crias até os 318 dias de vida não foram afetados (P>0,05) pela suplementação pré-parto materna. Ao avaliar o efeito da suplementação em creep-feeding sobre as variáveis de ganho de peso da fase III, foi verificado que os animais suplementados perderam menos peso (P<0,05) com o estresse do desmame do que os animais do grupo controle, e obtiveram maiores GT (P<0,05) até os 318 dias de idade. No entanto o GTR foi maior (P<0,05) para o tratamento controle. Observou-se efeito de interação (P<0,05) entre manejo alimentar na fase III e sexo, sendo que, tanto para os animais alimentados em sistema de confinamento quanto para os animais mantidos à pasto, os machos tiveram melhores ganhos de peso do que as fêmeas. A margem bruta foi maior para os tratamentos VC-BC-RS (R$ 40.014,48) e VC-BC-RC (R$ 38.266,03), seguido pelos tratamentos VS-BC-RC (R$ 35.574,47) e VC-BS-RS (R$ 35.501,23). A menor margem bruta foi obtida pelo tratamento VS-BS-RC (32.452,03). Os custos com suplementos, hora/máquinas e demais custos diferenciais nos tratamentos onde os animais foram suplementados impactaram significativamente sobre a margem bruta. Os animais dos tratamentos VS-BS-RS, VS-BS-RC, VS-BC-RS, VS-BC-RC, VC-BS-RS, VC-BS- RC e VC-BC-RC deveriam ter um ganho adicional de 29,08; 32,62; 20,61; 19,15; 19,47; 20,01; e 7,54 kg respectivamente, para que a margem bruta desses tratamentos fosse igual à do tratamento VC-BC-RS. A suplementação proteica de vacas Nelore no terço final da gestação não teve efeito sobre o peso e ganho de peso das crias. A suplementação em creep-feeding proporcionou maior peso ao desmame para animais suplementados, no entanto não programou o animal para obter maior desempenho nas fases seguintes. A recria de animais em sistema de confinamento foi eficiente em proporcionar maior peso aos 318 dias e maiores GMD. Os investimentos em suplementação pré-parto e em suplementação pré-desmame não foram eficientes em proporcionar maiores margens brutas, sugerindo que o melhor desempenho produtivo não necessariamente implica em melhor desempenho econômico. / The primary objective of the study was to evaluate the effect of protein supplementation of cows in the final third of pregnancy on fetal programming, as measured by the performance of their offspring submitted to growth rates up to 318 days old. The secondary objective, was to economically evaluate the different strategies of supplementation. In phase I we used 411 Nellore pregnant cows at the beginning of last trimester of gestation, with a mean age of 7.6 ± 2.1 years, mean body weight of 429 ± 49 kg and body condition score (BCS) of 2.84 ± 0.44 (range from1 to 5). Cows were assigned to two treatments according to BCS, body weight, age and father of calves . The treatments consisted of Supplemented-Cow group (SCO) (n = 212) and Control-Cow group (CCO) (n = 199). The cows SCO received daily 0.56 kg of protein supplement composed of 0.50 kg of soybean meal and 0.06 kg of minerals. The cows CCO received only mineral supplement. In phase II, which correspond to the period in which supplementation was offered to calves. Half of cows of each group from the first phase, with their offspring (2 months old), were redistributed in two treatments. 1- creep-feeding, offered ad libitum, Supplemented-Calf (SCA) (n = 201). 2 - Only mineral supplementation, Control-Calf (CCA) (n = 188). During phase II the animals were weighed four times, at 108, 149, 182 208 (weaning time) days old. After weaning they were sent to a pasture and remained for 25 days until the start of phase III. In phase III, which corresponds to winter season, lasted 85 days, the calves were regrouped into two groups. In group I, the calves were kept in feed lot, Supplemented-Growth group (SGG) (n = 209). In group II, the calves were kept in pasture composed predominantly of Panicum maximum or Brachiaria brizantha, Control-Growth group (CGG) group (n = 180). In this phase males and females were kept separated and finished when claves reached 318 days old. The economic analysis was based on the calculation of gross margin, obtained by subtracting the cost differential of total revenue. Effect was observed (P <0.05) of maternal supplementation during the first three weighings of phase II, the control group had higher weights. However, the weight at 208 days, was not (P> 0.05) influenced by maternal supplementation. Effect was observed (P<0.05) supplementing in the creep feeding at weaning time affected the body weight , (P <0.05), calves supplemented were heavier. The effect of sex of calves, bull calves were heavier, had higher (P <0.05) average daily gain (ADG) and total weight gain (TWG) than females in all weighings of phase II. The maternal supplementation on the offspring did not affect (P> 0.05) ADG in phase II between treatments. The effect of supplementation of calves in creep-feeding on daily gain (DG) and total weight gain (TWG) was higher for supplemented calves (P <0.05). The effect of maternal supplementation on weight of calves during phase III, did not showed differences (P> 0.05) between treatments. Supplementation of calves on creepfeeding was effective in keeping the weight difference obtained at weaning time for all phase III. The ADG and TWG of offspring at 318 days old were not affected (P> 0.05) by pre-partum maternal supplementation. Calves that were supplemented on creep-feeding in the phase III lost less weight (P <0.05) than control group. Interaction effect was observed (P <0.05) between feeding management in phase III and sex, both the animals fed in fed lot as well as for animals kept at pasture, males had better weight gains than females. Gross margin was higher for treatments CCO-CCA-SGG (R$ 40,014.48) and CCO-CCA-CGG (R$ 38,266.03), followed by treatments SC-CCA-CGG (R$ 35,574.47) and CCO-SCA-SGG (R$ 35,501.23). The lower gross margin was obtained by treatment SCO-SCA-CGG (32,452.03). The costs of acquiring supplements, machine works and other differential costs of treatments impacted significantly on the gross margin. The animals fed SCO-SCA-SGG, SCO-SCA-CGG, SCO-CCA-SGG, SCO-CCA-CGG, SCO-SCA-SGG, CCO-SCA-CGG CCO-CCA-CGG should have an additional gain of 29.08, 32.62, 20.61, 19.15, 19.47, 20.01, and 7.54 kg respectively, so that the gross margin of these treatments was equal to CCO-CCA-SGG treatment. The protein supplementation of cows in the last trimester of gestation had no effect on weight and weight gain of offspring. Thus, creep-feeding provided greater weaning weight, but the calve were not programmed for greater performance in subsequent phases. Calves in feedlot was effective in providing greater weight to 318 days and higher ADG. Investments in pre-partum supplementation and supplementation pre-weaning were not effective in providing higher gross margins, suggesting that better performance does not necessarily imply better economic performance.

Resumo: Este estudo avaliou o efeito dos níveis de energia (NDT) e proteína bruta (PB) consumidos por vacas de corte durante a gestação sobre o peso dos fetos aos quatro (PF4) e oito meses (PF8) e dos bezerros ao nascimento (PN); sobre o peso pós-natal aos 60 (PV60), 100 (PV100), 180 (PV180) e 205 (PV205) dias de vida; sobre o peso, a idade, o ganho médio diário (GMD), a área de olho de lombo (AOL), o marmoreio e a espessura de gordura (EG) ao abate. Foi conduzida uma revisão sistemática da literatura e uma metanálise (MA) a partir de duas bases de dados eletrônicas (Scopus e Web of Science). A MA para efeitos randomizados foi realizada para cada resultado com as médias do grupo controle e tratado, conduzida com os níveis de NDT e PB em relação às exigências das vacas (NRC, 1996). Foram avaliadas 46 publicações, correspondentes a 50 estudos, 135 ensaios e 9.379 animais. Não houve efeito do consumo de PB e NDT pelas vacas sobre o PF4. Quando as vacas consumiram PB e NDT em níveis iguais às suas exigências, no primeiro (1TRI) e segundo trimestre (2TRI) da gestação, houve redução no PF8 (DM= 2,236kg; P<0,001). O PN reduziu nos bezerros filhos de vacas que consumiram PB igual ou acima das suas exigências no terceiro trimestre da gestação (3TRI) (DM= -2,954kg; P<0,001 e DM= -0,453kg; P=0,045, respectivamente) ou 140% das exigências de NDT (DM= -2,709; P = 0,001). Quando o consumo de NDT foi igual à recomendação do NRC (1996), observou-se aumento de 2,029kg no PN dos bezerros (P<0,001). O PV60 foi inferior quando as vacas consumiram PB (DM= -1,923kg; P<0,001) e NDT (DM= -1,676kg; P<0,001) acima de suas exigências no 3TRI. O PV205 foi 3,122kg inferior (P<0,001) quando as vacas consumiram até 120% de NDT no 2TRI e 3TRI. Os novilhos cujas mães consumiram PB e NDT acima de suas demandas, no 3TRI, foram abatidos 5,5 dias antes (P= 0,015) que os demais. O GMD, o marmoreio e a EG foram superiores em vacas que consumiram NDT e PB acima das suas exigências no 3TRI (P<0,05). Em síntese, os níveis de PB e NDT, consumidos pelas vacas, acima das recomendações do NRC, estão negativamente relacionados com o desenvolvimento dos fetos e com o PN, sendo que esse efeito permanece até os 205 dias de idade. Por outro lado, na fase de terminação o excesso de PB e NDT passa a contribuir com o GMD, o marmoreio e a EG da carcaça das progênies. / Abstract: This study evaluated the effect of total digestible nutrients (TDN) and crude protein (CP) levels, consumed by cows during gestation, on fetal and postnatal development and on carcass traits. We conducted a systematic review of the literature and a metanalysis (MA) on two electronic databases (Scopus and Web of Science). Inclusion criteria were complete studies using pregnant beef cows receiving different diets that analysed fetus weight at four (FW4) and eight months (FW8), and birth weight of calves (BW); postnatal weight at 60 days (PW60), 100 days (PW100), 180 days (PW180) and 205 days (PW205) of life; and on weight, age, mean daily gain (ADG), loin eye area (LEA), marbling and fat thickness (FT) at slaughter. Random effect MA were conducted for each indicator separately with the mean of control and treated group. The analysis was conducted with the TDN and CP levels in relation to cows’ requirements (NRC, 1996). We evaluated 46 publications, reporting 50 studies, 135 trials and 9,379 animals. The CP and TDN level consumed by pregnant cows showed no change on FW4. Cows that consumed CP and TDN in a level equal to their requirements, in the first (1TRI) and second trimester (2TRI) of gestation, showed a reduction in FW8 (DM= 2,236kg; P<0.001). The BW reduced in cows that consumed CP equal to or above their requirements in the third trimester of gestation (3TRI) (DM= -2.954kg, P<0.001 and DM= -0,453kg, P=0.045, respectively), or 140% of TDN requirements (DM= -2.709, P=0.001). The calves of cows that consumed TDN equal to NRC (1996) requirements, increased the BW in 2,029kg (P<0.001). The PW60 was lower when cows consumed CP (DM= -1.923kg; P<0.001) and TDN (DM= -1.676kg; P<0.001) above their requirements in the 3TRI. The PW205 was lower (DM= 3.12kg; P<0.001) when cows consumed 120% of TDN in the 2TRI and 3TRI. Steers whose mothers consumed CP and TDN above their requirements in the 3TRI were slaughtered 5.5 days earlier (P= 0.015) than the others. The ADG, marbling and FT were higher when cows consumed TDN and CP above the NRC (1996) requirements in the 3TRI (P<0.05). In summary, the CP and TDN levels, consumed by cows, above the NRC recommendations, are negatively associated to fetal development and birth calf weight, and this effect remains until 205 days of age. Alternatively, in the feeder phase, CP and TDN in excess contribute to average daily gain, marbling and fat thickness of the progeny.

Breast cancer is a persistent public health problem. Interesting hypothesis suggests that its risk can be modulated in early life periods, a phenomenon known as fetal programming. In this context, most fetal programming studies focus on maternal influence, due to the greater interaction between mother and fetus in both fetal and lactation periods. However, recent studies show that paternal preconception diet has also a major role in the offspring\'s susceptibility to metabolic chronic non-communicable diseases. Therefore, this direct doctoral project aimed to assess whether the paternal consumption of different high fat diets during the development period of the reproductive system of male rats increased the susceptibility of female offspring to mammary carcinogenesis. In addition we sought to evaluate which mechanisms could be involved in this process. We used male rats of the Sprague-Dawley strain (n = 20/group) that consumed high fat diet with 60% of calories from lipids from lard (LB group) or corn oil (CB group), or AIN-93G control diet (CO group) for nine weeks, during development and sexual maturation periods. These rats were mated with females who consumed only commercial diet in 1:1 ratio. Their 50 days old offspring were subjected to mammary carcinogenesis model using 7,12-dimethylbenz[a]anthracene (50mg/kg). Paternal consumption of high fat diet of animal or plant source had opposite effects, with the paternal consumption of diet with high content of saturated fatty acids (LB) increasing and consumption of diet with high content of n-6 polyunsaturated fatty acids (CB) reducing the risk of breast cancer development in female offspring. These effects were due to changes in the expression of 89 miRNAs in the father\'s sperm and 23 miRNAs in the offspring\'s mammary gland, with overlapping of three miRNAs (miR-1897-5p, miR- 219-1-3p and miR-376a #) that were altered in both tissues. Additionally, female offspring of males fed diets with high content of saturated fatty acids showed less differentiation of the mammary gland, higher levels of cell proliferation, lower levels of apoptosis and altered expression of keys proteins that regulate important cellular functions, such as epithelial to mesenchymal transition. Finally, these females had also altered lipid profile of the fat pad similar to their male parent as well as epigenetic changes that may be related to the etiology of breast cancer. Thus, we conclude that the high-fat preconception paternal diet programmed the susceptibility of female offspring to mammary carcinogenesis, but this effect was dependent on the type of fatty acid consumed and the observed effects possibly results from changes in miRNA expression profile. / O câncer de mama é um persistente problema de saúde pública. Hipótese intrigante sugere que a suscetibilidade à doença pode ser modulada em períodos precoces da vida, fenômeno conhecido como programação fetal. Nesse sentido, a maior parte dos estudos de programação fetal refere-se à influência materna, dada a intensa interação existente entre mãe e feto tanto no período fetal, quanto na lactação. Entretanto, estudos recentes mostram que a dieta paterna pré-concepcional também tem um papel de grande importância na suscetibilidade da prole à uma série de doenças crônicas não-transmissíveis de origem metabólica. Portanto, o presente projeto de doutorado direto teve como objetivo avaliar se o consumo paterno de diferentes dietas hiperlipídicas, durante o período de desenvolvimento do sistema reprodutivo de ratos machos, aumentaria a suscetibilidade da prole feminina à carcinogênese mamária. Adicionalmente buscou-se avaliar quais mecanismos poderiam estar envolvidos nesse processo. Utilizaram-se ratos machos da linhagem Sprague-Dawley (n=20/grupo) que consumiram dieta hiperlipídica com 60% de calorias provenientes de lipídeos de banha (grupo LB) ou óleo de milho (grupo CB), ou dieta controle AIN-93G (grupo CO), por nove semanas, durante os períodos de desenvolvimento e maturação sexual. Esses ratos foram acasalados com fêmeas, que consumiram apenas dieta comercial, na proporção 1:1. Sua prole de 50 dias foi submetida ao modelo de carcinogênese mamária com o uso de 7,12-dimetil-benza[a]antraceno (50mg/kg). O consumo paterno de dietas hiperlipídicas de origem animal ou vegetal conferiram efeitos opostos, com o consumo de dieta com alto teor de ácidos graxos saturados (LB) aumentando e o consumo de dieta com alto teor de ácidos graxos poli-insaturados n-6 (CB) diminuindo o risco de desenvolvimento de câncer de mama na prole feminina. Esses efeitos foram associados à alteração da expressão de 89 miRNAS no espermatozoide dos pais e 23 miRNAs na glândula mamária da prole, com sobreposição de 3 miRNAs (miR-1897-5p, miR-219-1-3p e miR-376a#) que estavam alterados em ambos tecidos. Adicionalmente, a prole feminina de machos que consumiram dieta com alto teor de ácidos graxos saturados apresentou menor diferenciação da glândula mamária, maior nível de proliferação celular, menor nível de apoptose e alteração da expressão de proteínas chaves da regulação celular, como na transição epitélio-mesenquimal. Finalmente, essas fêmeas também apresentaram perfil lipídico alterado semelhante à do seu progenitor masculino, bem como modificações epigenéticas que podem estar relacionadas à etiologia do câncer de mama. Assim, concluímos que a dieta paterna hiperlipídica pré-concepcional programou a suscetibilidade da prole feminina à carcinogênese mamária, porém esse efeito é dependente do tipo de ácido graxo consumido e os efeitos observados possivelmente decorrem de alterações no perfil de expressão de miRNAs.

Puerto Rico has the highest prevalence of type 2 diabetes, low birth-weight, and the second highest prevalence of preterm-birth in all the U.S. and its non-incorporated territories. These conditions are related. Birth-weight at both ends of the spectrum and preterm-birth are associated with an increased risk for developing type 2 diabetes and immune-inflammatory dysregulations. Maternal psychosocial stressors during pregnancy have also been recognized as potential risk factors for type 2 diabetes, and have been consistently associated with preterm-birth and low birth-weight across populations. Current evidence points toward epigenetic fetal metabolic-programming as the mechanism that underlies the increased risk for the previously mentioned morbidities. However, the particular psychosocial stressors that may contribute to the high prevalence of low birth-weight and preterm-birth in the population of Puerto Rico have not been well studied. The present study assesses the relationships between particular psychosocial stressors, socioeconomic status, food insecurity, and birth outcomes. The results of this study show that low-risk pregnancy women were more likely to have babies with a higher ponderal index if they were exposed to stressors during gestation months 5, 6, and 7, or if exposed to "relationship stress" at any time during pregnancy. Women exposed to "financial difficulties" at any time during pregnancy were more likely to deliver babies at an earlier gestational age. Differences in birth outcomes between the exposed and non-exposed women were independent of maternal anthropometric measurements, maternal age at birth, number of previous births, and sex of the baby. Significant differences in birth outcomes were found between categories of father's self-identified and identified by others ethnicity, but sample size within categories was small. Although mothers with children at home had higher levels of food insecurity, and the level of food insecurity was correlated with higher levels of stress, no birth outcome measure was associated with food insecurity. Some results are atypical in comparison with other populations, and therefore these findings may contribute to the understanding of population differences in the relationship between maternal stress during pregnancy and birth outcomes. The relatively small sample size and strict exclusion criteria of this study may limit the generalizability of the findings. Epidemiological similarities between Puerto Rico and other populations, and the possibility of a higher ponderal index increasing the risk for type 2 diabetes in the population of Puerto Rico need to be examined in future research.