Thesis advisor: Joseph Quinn / The obesity epidemic is highly prevalent, growing and costly to the American public. The burden of obesity-related healthcare and productivity costs fall on employers, the government and the public at large, making it a cause of concern for all. Processed foods have received much of the blame for rising obesity rates in the United States, creating an "organic revolution." However, fast food giants still dominate the market. Changes such as mandated calorie labeling on menus and fat taxes have been attempted, but obesity rates are still growing. Policy action to successfully reduce and prevent obesity must target food environments, influence individual behavior, and support health services. / Thesis (BA) — Boston College, 2018. / Submitted to: Boston College. College of Arts and Sciences. / Discipline: Departmental Honors. / Discipline: Economics.

A domesticação dos equinos e a sua utilização para esportes fez com que a nutrição destes animais passasse por mudanças significativas. O alto requerimento energético pode ser suprido com forragem conservada e grandes quantidades de amido. Este manejo, porém, predispõe os equinos a problemas digestivos e metabólicos, podendo levar a cólica, endotoxemia e laminite. Outra forma de atingir o requerimento energético é a adição de gordura na dieta, já que óleos vegetais e animais podem conter até três vezes mais energia que grãos em igual volume. Sabe-se que óleos de diferentes fontes diferem quanto a propriedades relacionadas aos seus componentes ativos; óleos podem conter componentes como ω-3, ω-6, γ-orizanol em diferentes quantidades e estes compostos podem influenciar o metabolismo. O presente estudo teve por objetivo avaliar o efeito de óleos de diferentes origens e composição sobre o metabolismo bioquímico, oxidativo e função de células polimorfonucleares de equinos. Durante 15 semanas foram utilizados cinco equinos machos, mestiços, que receberam ou óleo mineral, ou óleo de soja, ou óleo de arroz, ou óleo de linhaça, ou óleo de peixe, em delineamento experimental quadrado latino 5x5. Cada período experimental foi composto por três semanas. Foram avaliados perfil bioquímico, metabolismo oxidativo e \"burst\"-respiratório de granulócitos. Houve efeito do tratamento sobre os teores de triglicérides, fração HDL de colesterol e hemoglobina, contagem de eritrócitos, hematócrito e na intensidade de produção de EROs por granulócitos estimulados com Sa-PI. Das variáveis do perfil bioquímico, pode-se observar que a adição dos óleos de soja, arroz, linhaça e peixe na dieta reduziu os triglicérides plasmáticos, a inclusão de óleos em até 5% da MS nas dietas não representou grande desafio oxidativo, apenas as concentrações de hemoglobina foram menores no tratamento com óleo de soja. O óleo de linhaça aumentou o teor de ácido linolênico plasmático e teve efeito sobre a produção de EROs in vitro por granulócitos dos equinos. / Sports use made significant changes in horses nutrition necessary. The high energy requirement is supplied with conserved forage, and large amounts of starch. This management, however, predisposes horses to digestive and metabolic disorders, leading to colic endotoxemia and laminitis. Another way to achieve the energy requirement is adding fat to the diet, as vegetable and animal oils may contain up to three times more energy than grains in equal volume. It is known that oils from different sources differ in their active components related properties, thay may contain ω-3, ω-6, γ-oryzanol, in different quantities and these compounds can influence metabolism. The aim of this study was to evaluate the effect of different oils from different origins and composition on the maintenance of the horses biochemical and oxidative metabolism and on polymorphonuclear cells function. A Latin square design was made with five crossbreed horses fed during three weeks with the five treatments: mineral (control), soy, rice, linseed and fish oil. Biochemical profiles, oxidative metabolism respiratory-burst of granulocytes were evaluated. Treatment effect was found on the triglycerides, HDL cholesterol and hemoglobin levels, erythrocyte count, hematocrit and intensity of ROS production by granulocytes stimulated with Sa-PI. Conclusions: The addition of oil to the diet reduced plasma triglycerides, fat diets didnt offer major oxidative challenge and hemoglobin showed to be a more sensitive oxidative marker. Linseed oil increases the plasma levels of linolenic acid and had effects on the production of in vitro ROS by granulocytes of horses.

Quatro potros machos, sem raça definida, com peso médio de 197,25 Kg, foram utilizados em um delineamento experimental Quadrado Latino 4X4 (quatro animais e quatro períodos), com arranjo fatorial do tipo 2X2, para tratamentos, onde os fatores foram: processamento do feno de alfafa (em cubos X em ramas); com adição ou não de óleo refinado de soja, (com óleo X sem óleo), adicionado no cocho, sobre o concentrado em dietas contendo concentrado comercial na forma de peletes para : avaliar a digestibilidade aparente total da matéria seca, matéria orgânica, proteína bruta, extrato etéreo, fibra em detergente neutro, fibra em detergente ácido e a viabilidade econômica das dietas estudadas. A adição de óleo refinado de soja afetou positivamente a digestibilidade aparente total da MS (66,18%), (p< 0,0093), MO (70,66%), (p< 0,0111), para a dieta com a alfafa na forma de cubos; EE (90,52%), (p< 0,0001) e FDN (53,24%), (0,0257) da dieta com feno de alfafa na forma de ramas. O processamento da alfafa interferiu positivamente sobre a digestibilidade aparente total da PB (80,02%), (p< 0,0048), para a dieta com feno na forma de cubos; FDN (47,92%), (p< 0,0147) e FDA (43,40%), (p< 0,0237), para a dieta com feno na forma de ramas. Independentemente do processamento sobre o feno de alfafa, a adição de óleo refinado de soja aumentou de maneira significativa e econômica a digestibilidade da fração EE. Feno de alfafa na forma de ramas, com adição de óleo, seria a opção mais econômica para utilização na dieta de potros devido ao custo de mercado ser mais baixo. A adição de óleo refinado de soja na dieta composta por : concentrado comercial, em uma base volumosa de feno de alfafa, aumentou a densidade calórica (NDT ou ED (predita)) das dietas estudadas, não deprimindo a digestibilidade aparente total dos macros nutrientes orgânicos e de maneira economicamente viável, para as condições desse experimento. / Four male weanlings, without defined breed, were used in this study, with middleweight 197, 25 Kg in an experimental Latin-square design for four (four animals and four periods), with factorial arrangement of the type 2 X 2 for treatments, where the factors were: processing of the alfalfa hay (cubes X long-stem) and addition or not of refined soybean oil top dressed (with oil X without oil). Diets containing commercial concentrate in the pellets form aimed: to evaluate the total apparent digestibility of the crude matter, organic matter, crude protein, fat, neutral detergent fiber, acid detergent fiber and the economical aspects. The addition of refined soybean oil, affected positively the total apparent digestibility of the CM (66,18%), (p < 0, 0093), OM (70,66%), (p <0, 0111), for cubes fat diet; EE (90,52%), (p <0, 0001) and NDF (53,24%), (p <0, 0257), for long-stem hay more fat diet. The processing in the alfalfa hay interfered positively on the total apparent digestibility of CP (80,02%), (p < 0, 0048), for cube diet; NDF (47,92%), (p <0, 0147) and ADF (43,40%), (p <0, 0237) for long-stem diet. Independently of the processing on the alfalfa hay, the addition of refined soybean oil increased, significant and economical way the total apparent digestibility of the fat fraction. Alfalfa hay in the long-stem forms, with soybean oil addition, would be the most economical option for use in the diet of colts due to the cost of lower market. The addition of refined oil of soy in the composed diet for: commercial concentrate, in a forage base of alfalfa hay, increased the caloric density (TDN or DE (predict)) in the studied diets, not depressing the total apparent digestibility of the organic nutritious macros and in way economically viable, for the conditions of that experiment.

A seleção de animais mais produtivos e a melhoria da qualidade do leite são os principais objetivos dos caprinocultores. Desta forma, o presente estudo, sob a hipótese de que o estresse pode intensificar o processo de apoptose durante a lactação, propõe estudar o efeito do estresse (via administração de ACTH) em animais estressados pontualmente (durante toda a lactação) e submetidos a estresse prolongado (durante três dias seguidos), sobre os níveis de cortisol, quantidade e qualidade do leite produzido e taxa de apoptose das células mamárias. Durante o experimento foram utilizadas 12 cabras primíparas da raça Saanen, subdivididas em 2 grupos e submetidas a aplicação de ACTH/Placebo bimensalmente. Ao longo de todo experimento foram realizadas coletas de sangue pontuais e durante os desafios, além de coletas de leite para mensurar os componentes e a Contagem de Células Somáticas (CCS). Não houve diferenças entre os grupos na produção leiteira e dos componentes do leite (proteína, gordura, lactose e CCS). Nos dias de desafio, animais que receberam ACTH obtiveram picos de produção de cortisol, diferentemente das fêmeas que receberam Placebo. Portanto conclui-se que mesmo produzindo cortisol, as fêmeas não obtiveram queda na qualidade tampouco na quantidade de leite produzido, o estresse de curta duração não traz prejuízos produtivos ao animal. / The selection of productive animals and milk quality improvement are the main goals of goat breeders. According to the hypothesis that stress can intensify the apoptosis process during lactation, the purpose of this research is to study stress effect through ACTH administration in animals stressed during lactation and submitted to a three day stress period. Twelve first-rate pregnancy Saanen goats were subdivided in two groups and submitted to ACTH and Placebo treatment once every two months. During the experiment, blood and milk samples were collected to measure Somatic Cell Count (SCC), protein, fat and lactose. Cortisol levels, quantity and quality of produced milk and mammary cells apoptosis rate were analyzed. As a result, there were no differences in milk production, protein, fat, lactose and SCC levels between both groups. On the other hand, animals who received ACTH obtained cortisol peaks, differently than those who received Placebo. Despite cortisol production, the quality and quantity of produced milk did not changed. Concluding, a short stress period does not impact on the goat milk production.

Este trabalho examina o papel dos alimentos industrializados no acesso às gorduras. Ele analisa os determinantes socioeconâmicos da quantidade e do tipo de gordura disponível para consumo pelas famílias paulistanas, presentes nos alimentos industrializados. Essa análise envolveu uma amostra de 576 famílias no município de São Paulo, cujos dados foram coletados no período de 2008 a dezembro de 20 10, selecionada após alguns critérios excludentes. A fonte dos dados é a Pesquisa de Orçamentos Familiares da FIPE, que está em andamento desde 2008, atualizada através da ponderação do Índice de Custo de Vida da FIPE. Baseado em métodos analíticos de estatística descritiva (média, desvio-padrão, mediana), testes t de student, e métodos econométricos de regressão múltipla, alguns resultados foram encontrados. Os fatores associados aos gastos com alimentos industrializados foram renda, tamanho da família, idade e grau de instrução do chefe. Em relação à gordura total identificou-se como fatores associados renda, tamanho da família, idade do chefe, a presença de crianças na família e a proporção do gasto com alimentação com derivados de leite e derivados de carne. / This work examines the paper of industrialized foods in the access to the fats. It analyzes the socioeconomics determinants of the amount and the type of available fat for consumption for the paulistians families, gifts in industrialized foods. This analysis involved a sample of 576 families in the city of São Paulo, whose given had been collected in the period of 2008 the December of 2010, selected some criteria after exculpatory. The source of the data is the POF (Research of Familiar Budgets) of the FIPE, that is in progress since 2008, brought up to date through the balance of the Index of Cost of living of the FIPE. Based in analytical methods of descriptive statistics (average, shunting line-standard, medium), tests \'t\' of student, and econometrical methods of multiple regression, some results had been found. The factors associated with spending on processed foods were income, family size, age and level of education of the head of family. In relation to total fat was identified as factors associated with income, family size, age of the head of family, the presence of children in the family and the proportion spent on food with dairy and meat products.

糖尿病是一种呈现流行趋势的代谢紊乱综合症，现如今，全球大约有3.46亿糖尿病患者， 这庞大的数字给各国的公共健康安全支出带来了严重的财政负担。 其中，二型糖尿病（T2DM）占90%。其特点是周围组织的胰岛素抵抗以及后期损伤的胰岛β细胞的功能。在饮食后，小肠会分泌两种肠促胰岛素，葡萄糖依赖性促胰岛素多肽（GIP）和胰高血糖素样肽-1（GLP-1）。两种多肽的主要功能是促进餐后胰岛细胞中胰岛素的分泌，另外他们还可以通过其自身的G蛋白偶联受体，GIPR和GLP-1R发挥其他作用，如葡萄糖依赖性的刺激胰岛素的生成，刺激胰岛β细胞的增殖，抑制细胞的凋亡等。这些功能也使肠促胰岛素成为糖尿病治疗的一种手段，比如Exendin-4和DPP4抑制剂。 然而，除了在胰岛中的作用，肠促胰岛还可能和脂质代谢相关，其中GIP和脂质代谢的报导研究的更加深入。在肥胖的状态下，血液中GIP含量高于正常水平；GIPR基因敲除老鼠和GIPR的抑制剂喂养的小鼠可以抵抗高脂饮食诱导的肥胖和2型糖尿病；GIP还可以直接调节脂肪细胞的脂肪生成和脂解。这些数据表明GIP在肥胖和糖尿病的发生过程中可能存在促进作用,这使得GIP治疗药物的开发需要谨慎的对待。 / 为了进一步研究GIP在脂肪细胞中发挥的生物学效应，在本研究中，我们利用腺病毒介导技术通过在脂肪细胞中过表达GIPR来增加GIP的活性，然后检查GIP在脂肪细胞中所起的作用。实验结果表明,GIP可以通过cAMP-PKA信号通路迅速并且长期的刺激脂肪细胞的炎症反应，增强IKKβ-NFκB信号通路和增加炎症基因的表达。更深入的机制研究表明，JNK 信号通路也参与GIP诱导的炎症反应，抑制JNK通路可以大部分恢复GIP增加的炎症因子的表达和IKKβ的磷酸化水平。由于长期的炎症反应，脂肪细胞的胰岛素信号通路受到GIP的损伤，在GIPR过表达的脂肪细胞中，胰岛素刺激的AKT磷酸化水平和葡萄糖吸收能力都被GIP降低，葡萄糖转运蛋白4（Glut-4）的表达水平也同时减少。因此，本研究结果表明GIP可能在肥胖的发展过程中，通过诱导脂肪细胞的炎症反应来损伤胰岛素敏感性而最终导致2型糖尿病的发生。 / Diabetes mellitus is a type of metabolic syndrome that has prevailed all over the world with the development of economic and over-nutrient lifestyle. It is estimated to 346 million diabetes patients in the worldwide most recently. The huge population put a major burden on the cost of public health care to all the countries. Among the types of diabetes, type 2 diabetes (T2DM) makes up 90% of recorded cases. The characteristics of T2DM are insulin resistance of peripheral tissues and impaired pancreatic cell function and mass. Two major incretins GIP (glucose-dependent insulinotropic peptide) and GLP-1 (glucagon-like peptide 1) are secreted from gut in response to food ingestion. The prominent role of GIP and GLP-1 is to stimulate glucose-dependent insulin release in pancreatic β cell. In addition, they both exert multiple biological effects via their relative G-protein coupled receptors, GIPR and GLP-1R, including glucose-stimulated insulin production, cell proliferation and anti-apoptosis in pancreatic β cells. The beneficent effects of incretins potentiate them as targets for the treatment of diabetes. GLP-1 analog, exendin-4 and DDP4 (dipeptidyl peptidase-4) inhibitors (to prevent GIP and GLP-1 from degradation) have been already used in clinical research. However, in addition to their effects on pancreatic β cell, both peptides are also related to lipid metabolism. The role of GIP has been studied more extensively. In obese state, the circulating level of GIP is elevated. GIPR knockout (KO) mice are resistant to high fat diet (HFD) induced obesity, a similar phenotype is found in GIPR antagonist administrated HFD-mice. Moreover, GIP also directly promotes lipogenesis and lipolysis in adipocytes. The rising evidence suggests a potential role of GIP in adipocyte biology and lipid metabolism, which diminishes the enthusiasm of GIP as a candidate therapeutic reagent for T2DM. / In order to further understand the biological effects of GIP in adipocytes, here, we over-expressed GIPR in 3T3-L1 CAR adipocytes via adenovirus-mediated gene transfer technology to enhance the activity of GIP. The results demonstrate that GIP impairs the physiological functions of adipocytes as a consequence of increasing the production of inflammatory cytokines, chemokines, and phosphorylation of IkB kinase (IKK) β through activation of the cyclic AMP-protein kinase A (cAMP-PKA) pathway. Activation of Jun N-terminal Kinase (JNK) pathway is also observed in GIP-induced inflammatory responses in adipocytes. An inhibitor of JNK blocks GIP-stimulated secretion of inflammatory cytokines and chemokines, as well as phosphorylation of IKKβ. The chronic inflammatory response eventually impairs insulin signaling in adipocytes, as demonstrated by reduction of protein kinase B (PKB/AKT) phosphorylation. The subsequently physiological analysis also indicates that GIP inhibits insulin-stimulated glucose uptake, and gene expression analysis reveals a decrease of glucose transporter 4 (Glut-4) in the meanwhile. The results suggest that GIP may be one of stimuli attributable to obesity induced insulin resistance via induction of adipocyte inflammation. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Nie, Yaohui. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 95-111). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgements --- p.v / INTRODUCTION --- p.1 / Chapter Part 1 --- Obesity and Type 2 diabetes --- p.1 / Chapter 1.1 --- Introduction to diabetes --- p.1 / Chapter 1.1.2 --- Physiology of adipocyte --- p.4 / Chapter 1.1.3 --- Mechanism of obesity induced diabetes --- p.10 / Chapter Part 2 --- Incretins and T2DM --- p.12 / Chapter 2.1 --- History of incretins --- p.12 / Chapter 2.2 --- Physiological actions of incretins --- p.14 / Chapter 2.3 --- Molecular mechanism of incretin actions in pancreas --- p.16 / Chapter 2.4 --- Incretins and T2DM --- p.19 / Chapter Part 3 --- Incretins and lipid metabolism --- p.23 / Objective --- p.26 / Methods and materials --- p.28 / Chapter 1 --- Cell culture --- p.28 / Chapter 1.1 --- 3T3-L1 culture and differentiation --- p.28 / Chapter 1.2 --- 3T3-L1 CAR culture and differentiation --- p.29 / Chapter 2 --- Cloning and recombinant adenovirus construction --- p.30 / Chapter 2.1 --- Plasmid construct --- p.30 / Chapter 2.2 --- Construct of recombinant adenoviruses --- p.30 / Chapter 2.3 --- Generation and infection of the adenoviruses --- p.31 / Chapter 3 --- Physiological and morphological assays --- p.32 / Chapter 3.1 --- Lipolysis assay --- p.32 / Chapter 3.2 --- TUNEL assay --- p.32 / Chapter 3.3 --- Glucose uptake --- p.33 / Chapter 3.4 --- Glut-4 localization --- p.33 / Chapter 4 --- Gene expression analysis --- p.35 / Chapter 4.1 --- Quantitative real-time PCR --- p.35 / Chapter 4.2 --- Immunoblot analysis --- p.35 / Chapter 4.3 --- ELISA assay --- p.36 / Chapter 5 --- Isolation of primary adipocytes --- p.37 / Results --- p.38 / Chapter Part 1 --- Role of GIP in 3T3-L1 cells --- p.38 / Chapter 1.1 --- Differentiation of 3T3-L1 adipocytes --- p.38 / Chapter 1.2 --- GIP slightly stimulates phosphorylation of p-CREB and lipolysis in 3T3-L1 cells. --- p.40 / Chapter 1.3 --- Analysis of gene expression in GIP-treated adipocytes --- p.42 / Chapter 1.4 --- Discussion --- p.44 / Chapter Part 2 --- Role of GIP in GIPR over-expressing 3T3-L1 CAR adipocytes --- p.46 / Chapter 2.1 --- Differentiation of 3T3-L1 CAR adipocytes --- p.46 / Chapter 2.2 --- Functional tests in GIPR over-expressing 3T3-L1 CAR adipocytes. --- p.48 / Chapter 2.3 --- Effect of GIP on cell viability --- p.50 / Chapter 2.4 --- Analysis of gene expression in GIP-treated adipocytes --- p.52 / Chapter 2.5 --- GIP activates inflammatory responses in GIPR over-expressing adipocytes --- p.54 / Chapter 2.6 --- Inhibition of IKKb pathway restores GIP-induced inflammatory responses --- p.56 / Chapter 2.7 --- Effects of GIP on adipocytes are partially dependent on the cAMP-PKA pathway --- p.58 / Chapter 2.8 --- Activation of cAMP-PKA pathway induces adipocyte inflammation. --- p.60 / Chapter 2.9 --- cAMP-Epac pathway is not involved in GIP-induced inflammation --- p.62 / Chapter 2.10 --- GIP stimulates cell stress activated kinases --- p.64 / Chapter 2.11 --- JNK partially mediates GIP-induced adipocyte inflammation --- p.65 / Chapter 2.12 --- Inhibition of JNK pathway partially restores GIP-induced inflammatory responses --- p.67 / Chapter 2.13 --- GIP impairs insulin signaling in GIPR over-expressing 3T3-L1 CAR adipocytes via inducing inflammatory response --- p.69 / Chapter 2.14 --- GIP enhances basal glucose uptake but impairs insulin stimulated glucose uptake in 3T3-L1 CAR GIPR over-expressing adipocytes --- p.71 / Chapter 2.15 --- Discussion --- p.73 / Chapter Part 3 --- Role of GIP in primary adipocytes --- p.78 / Chapter 3.1 --- GIPR expression level in primary adipocytes --- p.78 / Chapter 3.2 --- Analysis of gene expression in primary adipocytes after GIP treatment --- p.80 / Chapter 3.3 --- Discussion --- p.81 / SUMMARY --- p.82 / Chapter Future investigation --- p.83 / Chapter Appendix 1: --- Abbreviations --- p.86 / Chapter Appendix 2: --- Protocols --- p.90 / Preparation of competent cells --- p.90 / Outlines of recombinant adenovirus preparation --- p.91 / Virus titering (TCID50) --- p.92 / Primers for real-time PCR --- p.93 / Chapter Publications and Scientfic activities --- p.94 / Thesis related publication: --- p.94 / Other pubiliations: --- p.94 / Scientific activities: --- p.94 / References --- p.95

This thesis presents the results of zooarchaeological investigations into diet in Neolithic central Europe. The aim of these investigations was to gain a better understanding of animal carcass processing, particularly dietary decisions made concerning intensity of exploitation of meat and fat resources. The primary focus was the Linearbandkeramik (LBK) culture, a sedentary community of farmers dating from c. 5500-4900BC in central Europe suspected to be the first society to utilise milk and its products in this region. The adoption of dairying, currently under scrutiny by the NeoMilk project, would have increased the availability of fat on settlements, and could have affected the way in which people utilised primary animal products. Using in-depth zooarchaeological analysis of butchery, fracture and fragmentation, this thesis presents a snapshot of Neolithic meat and fat exploitation. Patterns of butchery and heat exposure suggest differential cooking practices between sites, with a possible focus on nutrient retention at some, contrasted with a cultural preference for roasting at others. Intensive processing of bone fats, namely bone grease, was not detected at any site and it is likely that the domesticated LBK diet rendered this practice unnecessary to subsistence. Bone marrow was a much more commonly exploited resource, but variation was considerable between sites. It is possible that the intensification of dairying had a significant effect on the utilisation of bone marrow. Sites with the most evidence for milk use, detected through lipid residue and osteoarchaeological evidence, show less intensive exploitation of bone marrow than those with little or no evidence of dairying. This thesis therefore presents evidence of zooarchaeologically detectable dietary decisions being made in the face of adoption of new foodstuffs.

O foco principal do projeto foi estudar e avaliar os parâmetros da obtenção de Biodiesel tendo como matéria prima o resíduo gorduroso (OGR) obtido de gordura animal, particularmente a de frango. Os objetivos do projeto vão ao encontro de questões intimamente ligadas a área ambiental e socioeconômica que cercam a produção do biodiesel. Para isso, tornou-se necessário seguir as etapas do trabalho proposto e assim, determinar a viabilidade do projeto. Foi necessário obter os resíduos gordurosos de frango para posterior caracterização e fazer também uma avaliação dos parâmetros convencionais de análise de biodiesel, traçando um estudo comparativo entre a obtenção do biodiesel com reatores convencionais e de ultrassom. Após obtenção da matéria prima, foi feito um tratamento térmico da gordura bruta e posterior separação do resíduo sólido por filtração. O resíduo sólido foi descartado e o filtrado desidratado por aquecimento e adição de cloreto de cálcio. Após obtenção do filtrado (óleo) colocou-se, em um balão de fundo chato de 1000 mL contendo três bocas, 200 mL do óleo obtido e adicionou-se 50 mL do acilante com Na°(catalisador). Realizou-se a reação a 45º C em um reator de vidro (500 mL) acoplado ao balão contendo três bocas e munido de condensador, termômetro e agitador mecânico. A agitação foi constante por 30 minutos. Após a reação ter se completado, obteve-se uma emulsão onde se encontrava biodiesel, glicerina, catalisador e excesso de álcool. Para separar os mesmos do nosso principal produto, adicionou-se água destilada que solubilizou o catalisador, se misturou com o álcool juntando-se à glicerina, formando por sua vez uma solução heterogênea com duas fases que posteriormente foi separado por decantação por funil de bromo. Por fim têm-se o biodiesel propriamente dito. O biodiesel obtido foi caracterizado por cromatografia gasosa utilizando o cromatógrafo CG CP3800 - Varian, e o método D_CARBÔNICA, onde obteve-se o percentual de ácidos graxos. As demais e convencionais análises foram realizadas com base nas normas especificadas pela Agência Nacional do Petróleo (ANP): determinação do índice de acidez pelo método de titulação potenciométrica (NBR 14448); determinação de Índice de Iodo (EN 14111); determinação de água, Método de Titulação Karl Fischer (EN ISO 12937); determinação do Ponto de Fulgor (NBR 14598); e determinação da viscosidade cinemática e cálculo da viscosidade dinâmica (NBR 10441). / The main focus of the project was to study and evaluate the parameters of obtaining Biodiesel having as raw material the greasy residue (OGR) obtained from animal fat, especially chicken. The project objectives are going to meet with issues closely linked to the environmental and socio-economic surrounding the production of biodiesel. For this, it became necessary to follow the steps of the proposed work and thus determine the feasibility of the project. It was necessary to get the greasy chicken waste for further characterization and also make an assessment of conventional parameters for the analysis of biodiesel, making a comparative study between getting the biodiesel reactor and conventional ultrasound. After obtaining the raw material a heat treatment of crude and fat was made subsequent separation of the solid residue by filtration. The solid residue was discarded and the filtrate dried by heating and addition of calcium chloride. After obtaining the filtrate (oil) put himself in a flat bottom flask containing 1000 mL with three necked, 200 mL of the oil obtained and added to 50 mL of acylating with Naº (catalyst). We carried out the reaction at 45 ° C in a glass reactor (500 mL) connected to the bag containing three mouths and fitted with condenser, thermometer and mechanical stirrer. The agitation was constant for 30 minutes. After the reaction have been completed, we obtained an emulsion in which it was biodiesel, glycerin, catalyst and excess alcohol. To separate them from our main product, was added distilled water that solubilized the catalyst, was mixed with alcohol adding to the glycerine, in turn forming a heterogeneous solution with two phases which subsequently was separated by decanting funnel bromine. Finally have the biodiesel itself. The biodiesel was characterized by gas chromatography using GC CP3800 - Varian, and the method D_CARBÔNICA where we obtained the percentage of fatty acids. The other, conventional tests were conducted under the rules specified in the National Petroleum Agency: determination of acid value by the method of potentiometric titration (NBR 14 448); determination of iodine value (EN 14 111); determination of water - Karl Fischer Titration Method (EN ISO 12 937); determination of Flash Point (NBR 14 598); determination of cinematic viscosity and calculation of dynamic viscosity (NBR 10441).

O aumento da adiposidade corpórea pode gerar diversos mediadores inflamatórios com capacidade de influenciar a proliferação e a diferenciação hematopoética e, consequentemente, a complexa regulação da hematopoese. Por isso, propusemo-nos, neste trabalho, avaliar a influência do aumento da adiposidade corpórea sobre a proliferação e a diferenciação de células hematopoéticas, bem como sua capacidade em sintetizar citocinas. Ratos Wistar, machos foram alimentados com uma dieta rica em lipídios durante 14 semanas. Após esse período foram avaliados hemograma, mielograma, perfil lipídico, concentrações séricas de leptina, insulina e adiponectina. Citômetria de fluxo foi utilizada para avaliação da porcentagem de células CD34+/CD133+, bem como o ciclo celular de células medulares. Células medulares foram utilizadas para avaliar a atividade proliferativa in vitro e a capacidade de diferenciação, in vitro, na presença de IL-3, EPO, GM-CSF e G-CSF. Animais, alimentados com dieta hiperlipídica, apresentaram maiores concentrações de leptina circulante, com aumento de gordura corporal, aumento da concetração de proteína C reativa, colesterol total, LDL, VLDL e triacilglicerol. O hemograma apresentou neutrofilia absoluta e a medula óssea apresentou-se hipercelular com aumento do número de granulócitos maduros e da população celular CD133-/CD34+. Os resultados dos testes in vitro demonstraram aumento da capacidade de síntese de IL-3 e aumento de G-CSF, com aumento do potencial proliferativo, também evidenciado pelo maior número de células medulares na fase S/G2/M, bem como o aumento da diferenciação granulocítica. Esses resultados sugerem que a leucocitose e neutrofilia observadas em situações de aumento da adiposidade corpórea são decorrentes de uma complexa modulação do sistema hematopoético. / The body fat increase can generate various inflammatory mediators, that are capable to influence the proliferation and differentiation of hematopoietic cells and consequently modulate the complex regulation of the hematopoiesis. In this study we have proposed to evaluate the effect of increase body fat on the proliferation and differentiation of hematopoietic cells, as well as its ability to synthesize cytokines. Male Wistar rats were subjected to a high fat diet during a period of 14 weeks. After that period were evaluated hemogram, mielogram, lipid profile and the serum concentrations of leptin, insulin and adiponectin. Flow cytometry was used to evaluate the percentage of CD34+/CD133+, as well as the cell cycle of bone marrow cells. Bone marrow cells were used to perform the proliferation and differentiation capacity in vitro in the presence of IL-3, EPO, GM-CSF and G-CSF. Animals fed high-fat diet had higher concentrations of circulating leptin with increase body fat, and increase of C-reactive protein, total cholesterol, LDL, VLDL and triacylglycerol concentrations. The hemogram showed absolute neutrophilia and a hypercellular bone marrow with increase of granulocytic mature population and CD133-/CD34+ cells. The results in vitro, showed an increase of IL-3 and G-CSF production, and higher proliferative potential with an increase in S/G2/M bone marrow cell cycle phases, as well as an increase of the granulocytic differentiation. The results suggest that leukocytosis and neutrophilia observed in this model of body fat increase are in fact a result of a complex modulation of the hematopoietic system.

Cholecystokinin (CCK) secreting enteroendocrine (EEC) I cells which distribute in gastrointestinal tract play an important role in lipid sensing, digestion and fatty acids uptake. Although a lot of research has been performed, the whole mechanism of fat sensing and fatty acid uptake and hormone expression in the CCK cells is still unclear. Global analysis to characterise the CCK cells is essential. CCK cells have an indistinct morphology, a diffuse distribution and a small percentage of population in the small intestine. However, the generation of genetic fluorescence tagged animal model facilitates the study of these cells. In this thesis, single cell dissociation methods and RT-PCR methodologies for detecting nutrient sensing receptors and fatty acids transporters were established and optimised. Expression of mRNAs for fat sensing GPCRs were detected in mouse duodenal epithelium. Expression of FATP family and CD36 in CCK cells and enterocytes was studied by RT-PCR. FATP2, FATP4 and CD36 mRNA were found in both CCK cells and enterocytes. Cell culture methodologies enabling the study of function (calcium imaging and FACS analysis) were established and optimised by checking the cell viability as a criterion. The methodology combining the immunochemistry and FACS analysis to study the hormone was established but requires further optimisation.