Submitted by Clebson Anjos (clebson.leandro54@gmail.com) on 2016-03-04T20:35:17Z No. of bitstreams: 1 arquivototal.pdf: 1623954 bytes, checksum: 8a3abb877583d97539e60ac1388438ee (MD5) / Made available in DSpace on 2016-03-04T20:35:18Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1623954 bytes, checksum: 8a3abb877583d97539e60ac1388438ee (MD5) Previous issue date: 2015-02-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The study of proteolysis, lipolysis volatiles profile of caprine Coalho cheese to which were added probiotic bacteria during storage. The cheeses were named: QS, control sample of Coalho cheese prepared with a starter culture of Lactococcus lactis, subsp. lactis, and Lactococcus lactis, subsp. cremoris; QLA, Coalho cheese with culture of Lactobacillus acidophilus; QLP, Coalho cheese sample formulated with Lactobacillus paracasei; QB, Coalho cheese with Bifidobacterium lactis; and QC, Coalho cheese with co-culture containing the 3 microorganisms in associated. The cheeses were analyzed after 1, 7, 14, 21 and 28 days of storage at the temperature of 10 °C ± 2 °C. The probiotic cell counts in cheese was superior to 6.5 log CFU g⁻¹ of cheese in the first day of storage, and surpassed 7 log CFU g⁻¹ of cheese in the 28th day of storage. Based on comparative studies carried out, it must be highlighted that the use of the various probiotic lactic cultures affects the lipid, protein, and aromatic profiles of caprine Coalho cheese during storage. It was verified that, when they were added to the Coalho cheese samples, the lactic cultures, regardless of type, presented lipolytic action (p < 0.01) during storage, causing a reduction in long chain fatty acids and an increase in short chain fatty acids. The acids released have direct influence in particular compounds of the aroma of the product, especially the characteristic caprine aroma, with an increase in caprylic, caproic and capric acids. It was observed that the addition of the Bifidobacterium lactis culture and the co-culture of probiotics had a positive effect on the fatty acid profile, with a higher production of volatiles, resulting in more concentrations during the CLA storage period (0.74 and 0.54 g/100 g of fatty acids), optimal values for the PUFA/SFA relationships (0.23 and 0.065) and DFA (50.23 and 45.08), associated with a reduction in the atherogenicity index (2.15 and 2.38) and in the trombosidade index (1.69 and 1.82). Twenty-five volatile compounds were identified in caprine Coalho cheese: six alcohols, four hydrocarbons, four terpenes, three acids, three ketones, three aldehydes and two esters. In the five types of cheese samples analyzed, one was able to observe the production of esters during storage and the reduction of aldehydes. The fruity aroma of esters is desirable, since it lessens undesirable aromas which are found in some compounds such as acids and aldehydes. The use of a combination of probiotic cultures (co-culture) also influenced (p < 0,01) the proteolysis of caprine Coalho cheese, providing a higher percentage of soluble protein, a higher proteolytic depth index and a greater release of fatty acids in the first day after processing.The probiotic caprine Coalho cheese samples presented a proteolysis rate which was at its highest after seven days of storage, and their fractions of αs₂ and k-casein were the ones with the highest degree of hydrolysis during storage. Among the samples prepared using isolated probiotic bacteria, it must be emphasized that B. lactis presented a more intense proteolytic action, showing a high percentage of soluble protein and a greater degradation of the αs₂ casein during storage. Probiotic cheeses produced using goat milk have excellent characteristics in terms of nutritional and aromatic attributes; highlighted the B. lactis and prodizidos adding the co-culture of probiotics. / O estudo da proteólise, lipólise e perfil de voláteis de queijo de coalho caprino ao qual foram adicionadas bactérias probióticas durante o armazenamento. Os queijos foram denominados de: QS, queijo de coalho controle elaborado com a cultura “starter” de Lactococcus lactis subsp. lactis e Lactococcus lactis subsp. cremoris; QLA, queijo de coalho com cultura de Lactobacillus acidophilus; QLP, queijo de coalho elaborado com Lactobacillus paracasei; QB, queijo de coalho com Bifidobacterium lactis; e QC, queijo de coalho com co-cultura, contendo os 3 micro-organismos em associado. Os queijos foram analisados após 1, 7, 14, 21 e 28 dias de armazenamento a 10 °C ± 2 °C. A contagem das células probióticas nos queijos foi superior a 6,5 log UFC g-1 de queijo no 1° dia de armazenamento e 7 log UFC g-1 de queijo, no 28° dia. Com base nos estudos comparativos realizados, ressalta-se que o uso das diferentes culturas láticas probióticas afetou o perfil lipídico, proteico e aromático do queijo de coalho caprino, durante o armazenamento. Verificou-se que as culturas láticas, independente do tipo, quando adicionadas aos queijos de coalho, apresentaram ação lipolítica (p < 0,01) durante o armazenamento, provocando a redução dos ácidos graxos de cadeia longa e aumento dos ácidos graxos de cadeia curta. Os ácidos liberados influenciaram diretamente nos compostos do aroma peculiar do produto, principalmente, o aroma característico de caprinos, com aumento dos ácidos caprílico, caproico e cáprico. Observou-se efeito positivo da adição da cultura de Bifidobacterium lactis e da co-cultura de probióticos no perfil de ácidos graxos, e uma maior produção de voláteis, que resultou em maiores concentrações durante o período de estocagem do CLA (0,74 e 0,54 g/100g de ácidos graxos), e ótimos valores das relações AGPI/AGS (0,23 e 0,065) e DFA (50,23 e 45,08), associados à redução dos índices de aterogenicidade (2,15 e 2,38) e trombosidade (1,69 e 1,82). Vinte e cinco compostos voláteis foram identificados no queijo de coalho caprino: sendo seis alcoóis, quatro hidrocarbonetos, quatro terpenos, três ácidos, três cetonas, três aldeídos e dois ésteres. Nos cinco tipos de queijos de coalho caprino avaliados, observou-se produção de ésteres ao longo do armazenamento e redução de aldeídos. O aroma frutado dos ésteres é almejável, uma vez que suaviza aromas indesejáveis e característicos de alguns compostos como ácidos e aldeídos. A utilização da combinação das culturas probióticas (co-cultura) também influenciou (p < 7 0,01) a proteólise do queijo de coalho caprino, proporcionando maior teor de proteína solúvel, maior índice proteolítico de profundidade e maior liberação de aminoácidos no 1° dia após o processamento. Os queijos de coalho caprinos probióticos apresentaram índice de proteólise em extensão máxima após sete dias de estocagem, sendo que suas frações de αs2 e k-caseína foram as que obtiveram maior grau de hidrólise ao longo do armazenamento. Dentre os queijos elaborados com bactérias probióticas isoladas, enfatiza-se que a B. lactis exibiu ação proteolítica mais intensa, proporcionando elevado teor de proteína solúvel e maior degradação da fração de αs2 caseína ao longo do armazenamento. Os queijos probióticos fabricados a partir de leite de cabra possuem excelentes características em relação aos atributos nutricionais e aromáticos, em destaque os prodizidos adicionando Bifidobacterium lactis e a co-cultura de probióticos.

abstract: A mule deer herd exists on the northern rim of the Grand Canyon, located on the North Kaibab Plateau. Historical references to this indigenous mule deer herd presented reports of periodic population irruption and collapse. Partially funded by the Arizona Game and Fish Department and the Arizona Deer Association, examination of herd nutritional and metabolic status from the Fall 2005 - Spring 2008 was completed at the request of AzGFD and ADA. Habitat analysis included forage micro-histological, protein, and caloric content plus whole blood and plasma assays gauging herd metabolic response. Modelling was completed using best management practices wildlife energy demand calculations and principal component analysis. Forage quality analysis and modelling suggest a sufficient amount of nitrogen (N) available (DPI) to the deer for protein synthesis. Energy analysis (MEI) of forage suggest caloric deficiencies are widely prevalent on the north Kaibab plateau. Principal component analysis integrates forage and metabolic results providing a linear regression model describing the dynamics of forage utilization, energy availability, and forage nitrogen supply with metabolic demand and response of the mule deer herd. Most of the plasma and blood metabolic indicators suggest baseline values for the North Kaibab mule deer. Albumin values are in agreement with albumin values for mule deer in the Southwest. I suggest that the agreed values become a standard for mule deer in the Southwestern U.S. As excess dietary N is converted to a caloric resource, a continual state of under-nutrition exists for the deer upon entering the N. Kaibab winter range. The population is exceeding the nutritional resource plane that the winter habitat provides. Management recommendations include implementation of multiple small-scale habitat rehabilitation efforts over time, including invasive juniper (Juniperous osteosperma) and piñon (Pinus edulis) management, prescribed burning to control big sage (Artemesia tridentata) populations, and reseeding treated areas with a seed mix of native shrubs, grasses and forbs. I recommended that the population size of the North Kaibab deer herd is maintained at the current size with natural selection controlling growth, or the population be artificially reduced through increased hunting opportunities. / Dissertation/Thesis / Ph.D. Environmental Design and Planning 2014

ABSTRACTThe prevalence of diabetes is increasing dramatically, incurring a major health and socioeconomic burden. Type 2 diabetes (T2D), the most prevalent form of diabetes, results from a variable combination of insulin resistance and insulin deficiency, secondary to pancreatic β-cell failure. These defects are caused by a complex interplay between genetic and environmental/ lifestyle factors. Among the latter, poor dietary quality is a crucial driver of T2D development. Although adopting healthy dietary habits is considered as a mainstay for T2D prevention, what constitutes a healthy diet remains controversial. Epidemiological studies examining the association of dietary fat quality with T2D incidence have yielded equivocal results and may suffer from confounding. On the other hand, randomized trials assessing the impact of dietary fat saturation on glucose homeostasis have major methodological shortcomings, precluding reliable conclusions. In order to elucidate this question, we compared the effects of palm oil vs olive oil on glucose homeostasis and other relevant metabolic parameters, in a mouse model of high-fat diet-induced obesity. The saturated fatty acid-rich palm oil is the most abundantly used oil worldwide. Olive oil is a staple food of the Mediterranean diet, rich in monounsaturated fatty acids and widely regarded as healthful. In this model, palm oil was not more harmful than olive oil with regard to glucose/insulin homeostasis. However, palm oil was associated with increased visceral adiposity and triglyceridemia compared to olive oil. Circulating and tissue free fatty acid (FFA) concentration and composition are determined by dietary factors, as well as genetic and metabolic factors. There is accumulating evidence indicating that increased FFA levels and/or an unbalanced FFA composition with excess palmitate, induce β-cell dysfunction and apoptosis (lipotoxicity). To characterize the mechanisms underlying lipotoxicity, we combined RNA-sequencing with proteomics of β-cells exposed to palmitate, the most prevalent SFA in humans. This cross-omics study showed that palmitate altered lipid and amino-acid metabolism, and affected amplifying pathways of insulin secretion and exocytosis. Furthermore, palmitate induced stress pathways, including mitochondrial dysfunction, oxidative stress and endoplasmic reticulum (ER) stress. ER stress is triggered when protein folding demand exceeds ER folding capacity. This response aims to restore ER homeostasis but if unresolved, it can become deleterious. Islets from T2D patients display signs of ER stress, pointing to a potentially pathogenic role of the latter.Monogenic and neonatal diabetes are rare forms of diabetes caused by single gene mutations. These forms are of particular interest, as they can serve as ‘human knockout’ models of diabetes. Recent evidence shows that there is overlap in the genetic basis of monogenic diabetes and T2D, suggesting that they may be part of a pathologic continuum. To explore the role of ER stress in diabetes pathogenesis, we studied two different genetic syndromes involving neonatal or early-onset diabetes, caused by mutations in genes related to ER function (DNAJC3 and YIPF5). Using in vitro knockdown models, we showed that ER stress elicited by impaired chaperone function (DNAJC3) or by impaired ER-to-Golgi protein transport (YIFP5) causes β-cell apoptosis. Altogether, our findings support that lipotoxic and genetic ER stress contribute to diabetes pathogenesis. Preventing or modulating ER stress thus holds anti-diabetic therapeutic potential. Future research should focus on defining optimal strategies to restore a balanced FFA profile and enhance ER function, aiming to prevent ER-stress induced β-cell failure. RésuméLa prévalence du diabète progresse constamment, posant un défi sanitaire et socioéconomique majeur. Le diabète de type 2 (DT2), la forme la plus courante de diabète, résulte de la résistance à l’insuline, en association avec un déficit insulinique dû à la défaillance des cellules β pancréatiques. Ces anomalies découlent d’une interaction complexe entre des facteurs génétiques et des facteurs liés au mode de vie. Parmi ces derniers, la qualité du régime alimentaire est un facteur crucial pour le développement du DT2. Bien que le suivi d’un régime alimentaire sain est considéré comme le pilier pour la prévention du DT2, ce qui constitue un régime sain demeure un sujet de controverse.Les études épidémiologiques examinant l’association entre la qualité de la graisse alimentaire et l’incidence du DT2 ont donné des résultats équivoques, affectés éventuellement par des facteurs confondants. En outre, les études randomisées évaluant l’impact du degré de saturation de la graisse alimentaire sur l’homéostasie du glucose comportent des limitations méthodologiques majeures. Afin d’élucider cette question, on a comparé les effets de l’huile de palme aux effets de l’huile d’olive sur l’homéostasie du glucose et d’autres paramètres métaboliques pertinents. Dans ce but, on a utilisé un modèle murin d’obésité induite par un régime riche en graisse. L’huile de palme est riche en acides gras saturés et elle est l’huile la plus utilisée globalement. L’huile d’olive est un aliment phare du régime Méditerranéen, riche en acides gras monoinsaturés et généralement reconnu comme un aliment sain. Dans notre modèle murin, la consommation d’huile de palme n’était pas plus néfaste que celle de l’huile d’olive sur l’homéostasie du glucose, la sensibilité à l’insuline et l’insulinosécrétion. Par contre, l’huile de palme était associée à une adiposité viscérale et une triglycéridémie plus élevée comparée à l’huile d’olive.La concentration et la composition des acides gras libres (AGL) sont déterminées par des facteurs alimentaires, génétiques et métaboliques. Des données abondantes démontrent que la présence des niveaux élevés d’AGL et/ou d’une composition déséquilibrée d’AGL induit la dysfonction et l’apoptose des cellules β (lipotoxicité). Pour caractériser les mécanismes sous-jacents de la lipotoxicité, on a combiné un séquençage ARN à une étude protéomique des cellules β exposées au palmitate, l’AGL saturé le plus courant chez l’homme. Cette étude conjointe a montré que le palmitate altère le métabolisme des lipides et des acides aminés, les voies d’amplification de la sécrétion d’insuline et l’exocytose. Le palmitate induit également des voies de stress cellulaires, telles que la dysfonction mitochondriale, le stress oxydatif et le stress du réticulum endoplasmique (RE). Le stress du RE est activé quand les besoins en sécrétion protéique dépassent les capacités de l’organite. Cette réponse a pour but de rétablir l’homéostasie du RE mais si le stress reste non résolu, ceci peut s’avérer délétère. Des îlots des patients avec un DT2 montrent des signes de stress du RE, évoquant un rôle potentiellement pathogénique de ce dernier.Le diabète monogénique et néonatal sont des formes rares de diabète causées par des mutations d’un seul gène. Ces formes sont particulièrement intéressantes sur le plan physiopathologique car elles représentent des ‘knockout’ humains. Des données récentes montrent que la base génétique du diabète monogénique n’est pas complètement distincte de celle du diabète de type 2 et les deux entités pourraient faire partie d’un continuum. Afin d’explorer le rôle du stress du RE dans la pathogénèse du diabète, on a étudié deux syndromes génétiques entraînant un diabète néonatal ou à début très précoce. Ces syndromes sont causés par des mutations dans des gènes impliqués dans la fonction du RE (DNAJC3 et YIPF5). En silençant ces gènes in vitro, on a montré que le stress du RE, déclenché soit par une dysfonction des chaperones (DNAJC3), soit par un retard du trafic de protéines du RE vers le Golgi (YIPF5), induit l’apoptose des cellules β.Ces résultats suggèrent que le stress du RE génétique et lipotoxique contribuent à la pathogénèse du diabète. La prévention ou modulation du stress du RE présente donc un potentiel thérapeutique anti-diabétique. Des études futures pourraient permettre de définir des stratégies optimales pour rétablir un profil d’AGL équilibré ou renforcer la fonction du RE, en vue de prévenir la défaillance des cellules β. / Doctorat en Sciences médicales (Médecine) / info:eu-repo/semantics/nonPublished

Detta arbete undersöker förhållandet mellan kvalitetsparametrar i frityrolja, såsom halten fria fettsyror (FFA), anisidinvärdet (AV) och %TPM (Total Polar Materials) samt Agtronvärdet hos köttbullar efter fritering. Kvalitetsparametrar hos frityrolja, såsom primära och sekundära oxidationsprodukter, kan mätas för att utvärdera oljans kvalitet då nivåerna ofta ökar i samband med oljans användning. Primära oxidationsprodukter, såsom peroxider, kan mätas med hjälp av ett så kallat peroxidvärde (PV) och sekundära oxidationsprodukter, såsom aldehyder, kan mätas med hjälp av anisidinvärdet. Analys av %TPM och FFA är exempel på andra metoder för att mäta kvaliteten hos en olja då polariteten ökar i takt med oljans användning. Halten FFA i oljan undersöktes med hjälp av titrering med natriumhydroxid och %TPM mättes med hjälp av mätinstrumentet Testo 270. Agtronvärdet hos köttbullarna mättes med hjälp av instrumentet Agtron E30FP, som använder IR-teknologi för att bestämma färgen på en produkt. Resultaten visade att halten FFA, AV och %TPM i frityroljan ökade med tiden under samtliga fyra produktionsdagar. Hypotesen att produkten bör vara mörkare, det vill säga att Agtronvärdet bör vara lägre vid produktionens slut jämfört med början stödjs av resultaten från två av fyra produktionsdagar. Slutsatsen drogs att det fanns skillnader i oljans sammansättning, med hänsyn till FFA, AV och %TPM vid produktionsdagens slut jämfört med start. Däremot kunde ingen tydlig koppling ses mellan dessa förändringar och en minskning av Agtronvärdet hos köttbullarna över tiden. Ytterligare studier behövs således för att kunna dra en slutsats kring sambandet mellan kvalitetsparametrar i frityrolja och färgen på det friterade livsmedlet. / This study examines the relationship between quality parameters of frying oil, such as Free fatty acids (FFA), the Anisidine Value (AV) and Total Polar Materials (%TPM), as well as the Agtron Value of meatballs after deep frying. Quality parameters in frying oil, such as primary and secondary oxidation products, measure the quality of the frying oil since levels increase with the use of the oil. Primary oxidation products, such as peroxides, can be measured by the Peroxide Value (PV) and secondary oxidation products, such as aldehydes, can be measured by the Anisidine Value. Analysis of %TPM and FFA are other methods to measure the quality of an oil, since the polarity increases as the oil is used for frying. The instrument Testo 270 was used to measure %TPM in the oil and FFA was determined by titration with sodium hydroxide. The Agtron E30FP instrument was used to determine the Agtron Values of the meatballs, wich used IR technology to determine the color of the product. Results showed that the FFA, AV and %TPM of the oil increased over time in all four production days. It was expected that the product should be darker, that is, the Agtron Value should be lower at the end of production compared to the beginning, but this was only seen in two out of four production days. It was concluded that there are differences in the composition of the oil, with regard to FFA, AV and %TPM at the end of the production day compared to the beginning of the day, but no clear connection was seen between these changes and a decrease in the Agtron Value of the meatballs over time. Further studies are needed to be able to draw a conclusion about the relationship between quality parameters of frying oil and the color of the fried food.

L’athérosclérose est étroitement liée au diabète de type 2. De fortes concentrations plasmatiques en acides gras libres (AGL) et en insuline sont des caractéristiques retrouvées chez les patients souffrant de ces deux pathologies. Les AGL, présents dans notre alimentation, font partie de l’environnement auquel les cellules sont exposées. Leurs effets dépendent de leur nature, les acides gras saturés (AGS) étant néfastes et les acides gras monoinsaturés (AGMI) plus protecteurs. Ils ont donc des effets variés sur les cellules musculaires lisses vasculaires (CMLV) impliquées dans la pathogénèse de l’athérosclérose. Ainsi, l’objectif principal de ce projet de maîtrise était d’évaluer l’impact de deux combinaisons d’AGL sur la viabilité des CMLV, en condition hyperinsulinémique ou non. Les deux combinaisons renfermaient les mêmes AGL mais en proportions différentes, l’une étant plus riche en AGS et l’autre en AGMI. Nos résultats ont montré que les combinaisons d’AGL ont un effet pro-apoptotique principalement dû aux AGS. L’acide oléique présent dans les combinaisons atténue cependant cet effet. Il diminue même plus fortement l’apoptose des CMLV lorsqu’associé à un AGS que lorsqu’utilisé seul. Cet impact est significatif uniquement dans certaines proportions de ces AGL et est plus efficace en présence d’insuline. Ces résultats mettent en lumière la présence d’une compétition entre mécanismes anti- et pro-apoptotiques en fonction des proportions d’AGS versus AGMI et de l’insulinémie chez les CMLV. Ils soulignent également l’importance de la présence des AGMI dans les diètes riches en AGS et pourraient être utiles pour l’élaboration de nouvelles diètes adaptées aux patients athérosclérotiques et diabétiques. / Atherosclerosis is closely linked to type 2 diabetes. High plasmatic concentrations of free fatty acids (FFA) and insulin are common features in patients suffering from both diseases. FFA, present in our diet, are part of the environment to which body cells are exposed. Their effects are dependent of their nature, being harmful for saturated fatty acids (SFA) and more protective for monounsaturated fatty acids (MUFA). They can have therefore various effects on vascular smooth muscle cells (VSMC) implicated throughout the development of atherosclerosis. Thus, this study aimed to assess the impact of two FFA combinations on VSMC viability, whether or not in a hyperinsulinemic condition. Both combinations contained the same FFA but in different proportions, one being richer in SFA and the other in MUFA. Our results showed that FFA combinations have a pro-apoptotic impact, mainly due to SFA. However, the presence of oleic acid in the combinations attenuated this effect. Furthermore, oleic acid had the capacity to reduce more strongly VSMC apoptosis when combined with a SFA than when used alone, although only under specific FFA ratios. This impact is even more effective in presence of insulin. These results highlight the presence of a competition between anti- and pro-apoptotic mechanisms dependent of FFA ratios (SFA vs. MUFA) and insulinemia to which are exposed VSMC. They also underline the importance of the presence of MUFA such as oleic acid in diets rich in SFA and could be useful for the development of new diets adapted to atherosclerotic and diabetic patients.

A reposição de glicogênio pós treino foi investigada usando uma mistura de ambos, glicose e glicose-frutose, como repositores, em atletas corredores de longa distância. Dez corredores competitivos (30±4,7 anos) foram avaliados para composição corporal e consumo alimentar antes do exercício de 90 minutos em esteira elétrica a 70% V02máx. Amostras de sangue venoso foram coletadas antes e após o término do exercício, quando eles receberam 1g de carboidrato.kg-1 de uma mistura de ambos, glicose-frutose (CHO1) e glicose (CHO2), e outra com sucralose ou placebo (controle). Amostras de sangue foram retiradas 30, 60, 90 e 120 minutos após o consumo dos suplementos e usadas para determinação de glicose e de ácidos graxos plasmáticos, por reações colorimétricas. Não foram observadas diferenças significativas na composição corporal e na hidratação durante o exercício. Os tratamentos CHO1 e CHO2 produziram elevação na glicemia, em 30 minutos, comparados ao grupo placebo. Entretanto, em 90 min, somente no tratamento CHO1, uma queda na glicemia foi observada (CHO2>CHO1). Os ácidos graxos livres aumentaram comparativamente após o exercício (CHO1=CHO2=placebo) e, diferente do placebo, em 90 minutos, reduziram nos tratamentos CHO1 e CHO2. Ambos os repositores atuaram similarmente nas concentrações de ácidos graxos livres, mas o declínio da glicose plasmática (remoção da glicose) ocorreu primeiramente com o tratamento glicose-frutose do que com o glicose. Assim, a mistura glicose-frutose parece promover uma reposição de glicogênio mais rápida após exercícios de longa duração. / The post training glycogen replacement was investigated using either glucose or glucose-fructose mixture as repositors in long-distance athletes. Ten males (30±4,7yrs) voluntary competitive long distance runners were assessed for body composition and dietary intake prior to a 90 min 70% vo2 max exercise in treadmill. Blood samples were collected before and after the exercise when they received orally 1g of CHO.kg-1 either as maltodextrin-glucose-fructose (CHO1) or maltodextrin - glucose (CHO2) mixtures, having sucralose or placebo (control). Blood samples were drawn 30,60, 90 and 120 minutes after supplement intake and used for plasma glucose and free-fatty acids determinations by colorimetric reactions. There were no significant changes on body composition, mass or hydration, during the treadmill exercise. CHO1 and CHO2 loading increased the 30 min blood glucose both significatively above the placebo. However at 90 min only CHO1 decreased significantily (CHO2>CHO1). The FFA increased comparatively (CHO1=CHO2=placebo) after exercise and differently from placebo CHO1 and CHO2 decreased (similarly) up to 90 min. Both energetic repositors acted similarly on FFA levels but the plasma glucose decreasing (glucose removal) occurred earlier with glucose-fructose loading rather than glucose alone. Thus glucose-fructose mixtures seems to promote faster glycogen repletion after long lasting aerobic exercises.

In der vorliegenden Arbeit wurde eine Beeinflussung der Aktivität der humanen neutrophilen Elastase (EC 3.4.21.37) durch wässrige und Dichlormethan-Extrakte von 15 Basidiomyceten festgestellt. Durch aktivitätsgeleitete Fraktionierung (mehrfache SC, GC-MS) der Dichlormethan-Extrakte von Heterobasidion annosum (Fr.) Bref. und Lactarius deterrimus Grög. wurden Fraktionen freier langkettiger Fettsäuren als ein wirksames Prinzip der Elastase-Hemmung und auch der Kollagenase-Hemmung (Clostridium histolyticum Kollagenase, EC 3.4.24.3) isoliert und identifiziert. Das Screening von 17 freien langkettigen Fettsäuren zeigte, dass einfach ungesättigte Fettsäuren eine stärkere Hemmung der Elastase-Aktivität bewirkten als ihre gesättigten bzw. mehrfach ungesättigten Homologa: Ölsäure (C18:1 cis-9): IC50 5µM; Stearin-(C18:0), Linolsäure (C18:2 cis-9,12): IC50 10µM; alpha- (C18:3 cis-9,12,15), gamma-Linolensäure (C18:3 cis-6,9,12): IC50 15µM. Inhibitorisch am stärksten wirksam war Erucasäur! e (C22:1 cis-13): IC50 450nM. Für Kollagenase wurde hingegen gezeigt, dass die gesättigten Fettsäuren eine erheblich stärkere Hemmaktivität als ihre ungesättigten Homologa aufwiesen. Aktivste Verbindungen waren Palmitin- (C16:0), Heptadecan- (C17:0), Stearin- und Nonadecansäure (C19:0) mit IC50-Werten von 20-45µM. Die Untersuchung von 9 ausgewählten Fettsäuren bezüglich der Hemmung der Aktivität der MMP-9 (EC 3.4.24.35) zeigte als aktivste Verbindungen Palmitolein- (16:1 cis-9), alpha- und gamma-Linolensäure. Die wirksamen Konzentrationen (250µM) lagen jedoch sehr hoch. Zytotoxizitätsuntersuchungen (ECV-304) der Extrakte von H. annosum und L. deterrimus sowie der freien Fettsäuren schlossen sich ebenso wie Untersuchungen zur Proteaseaktivität der Zelllinien ECV-304, MCF-7 und MDA-MB 231 an. Die Proteaseaktivität der Zellen nahm in der Reihenfolge ECV-304 < MCF-7 < MDA-MB 231 zu. Die einzig untersuchte Fettsäure gamma-Linolensäure zeigte keine reproduzierbare Beeinflussung d! er Proteaseaktivität. / In the present paper it was established that the activity of humane neutrophil elastase (EC 3.4.21.37) is affected by aqueous and dichloromethane extracts of 15 basidiomycetes. Bioassay-guided fractionation (repeated CC, GC-MS) of dichloromethane extracts of Heterobasidion annosum (Fr.) Bref. and Lactarius deterrimus Grög. led to isolation and identification of fractions of free fatty acids as one active principle of elastase inhibition as well as collagenase inhibition (Clostridium histolyticum collagenase, EC 3.4.24.3). By testing 17 free fatty acids for elastase inhibition it was shown that the inhibition rate of unsaturated acids was much higher than the rate of the saturated ones: oleic acid (C18:1 cis-9): IC50 5µM; stearic acid (C18:0), linoleic acid (C18:2 cis-9,12): IC50 10µM; linolenic acid (C18:3 cis-9,12,15), gamma-linolenic acid (C18:3 cis-6,9,12): IC50 15µM. The highly active erucic acid with an IC50 value of 450nM is remarkable. As a result for collagenase we can assume that the saturated fatty acids were more potent than the unsaturated ones. Palmitic acid (C16:0), heptadecanoic acid (C17:0), stearic acid, and nonadecanoic acid (C19:0) were the most potent fatty acids with IC50 values of 20-45µM. 9 selected fatty acids were investigated for their ability to inhibit the activity of MMP-9 (EC 3.4.24.35). Palmitoleic acid (16:1 cis-9), linolenic acid, and gamma-linolenic acid were the most potent fatty acids but their inhibiting concentrations were very high (250µM). Investigation of cytotoxicity of the extracts of H. annosum, L. deterrimus, and free fatty acids as well as investigation of protease activity of ECV-304, MCF-7, and MDA-MB 231 cells followed. Protease activity of cells increased in the following manner: ECV-304 < MCF-7 < MDA-MB 231. The only investigated fatty acid gamma-linolenic acid did not influence protease activity reproducibly.

A reposição de glicogênio pós treino foi investigada usando uma mistura de ambos, glicose e glicose-frutose, como repositores, em atletas corredores de longa distância. Dez corredores competitivos (30±4,7 anos) foram avaliados para composição corporal e consumo alimentar antes do exercício de 90 minutos em esteira elétrica a 70% V02máx. Amostras de sangue venoso foram coletadas antes e após o término do exercício, quando eles receberam 1g de carboidrato.kg-1 de uma mistura de ambos, glicose-frutose (CHO1) e glicose (CHO2), e outra com sucralose ou placebo (controle). Amostras de sangue foram retiradas 30, 60, 90 e 120 minutos após o consumo dos suplementos e usadas para determinação de glicose e de ácidos graxos plasmáticos, por reações colorimétricas. Não foram observadas diferenças significativas na composição corporal e na hidratação durante o exercício. Os tratamentos CHO1 e CHO2 produziram elevação na glicemia, em 30 minutos, comparados ao grupo placebo. Entretanto, em 90 min, somente no tratamento CHO1, uma queda na glicemia foi observada (CHO2>CHO1). Os ácidos graxos livres aumentaram comparativamente após o exercício (CHO1=CHO2=placebo) e, diferente do placebo, em 90 minutos, reduziram nos tratamentos CHO1 e CHO2. Ambos os repositores atuaram similarmente nas concentrações de ácidos graxos livres, mas o declínio da glicose plasmática (remoção da glicose) ocorreu primeiramente com o tratamento glicose-frutose do que com o glicose. Assim, a mistura glicose-frutose parece promover uma reposição de glicogênio mais rápida após exercícios de longa duração. / The post training glycogen replacement was investigated using either glucose or glucose-fructose mixture as repositors in long-distance athletes. Ten males (30±4,7yrs) voluntary competitive long distance runners were assessed for body composition and dietary intake prior to a 90 min 70% vo2 max exercise in treadmill. Blood samples were collected before and after the exercise when they received orally 1g of CHO.kg-1 either as maltodextrin-glucose-fructose (CHO1) or maltodextrin - glucose (CHO2) mixtures, having sucralose or placebo (control). Blood samples were drawn 30,60, 90 and 120 minutes after supplement intake and used for plasma glucose and free-fatty acids determinations by colorimetric reactions. There were no significant changes on body composition, mass or hydration, during the treadmill exercise. CHO1 and CHO2 loading increased the 30 min blood glucose both significatively above the placebo. However at 90 min only CHO1 decreased significantily (CHO2>CHO1). The FFA increased comparatively (CHO1=CHO2=placebo) after exercise and differently from placebo CHO1 and CHO2 decreased (similarly) up to 90 min. Both energetic repositors acted similarly on FFA levels but the plasma glucose decreasing (glucose removal) occurred earlier with glucose-fructose loading rather than glucose alone. Thus glucose-fructose mixtures seems to promote faster glycogen repletion after long lasting aerobic exercises.

Submitted by Marilene Donadel (marilene.donadel@unioeste.br) on 2017-08-11T14:43:57Z No. of bitstreams: 2 Andriele Magarinos Focking 2017.pdf: 2481446 bytes, checksum: 763f767d058face92edaaa394dc5b780 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-08-11T14:43:57Z (GMT). No. of bitstreams: 2 Andriele Magarinos Focking 2017.pdf: 2481446 bytes, checksum: 763f767d058face92edaaa394dc5b780 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-03-09 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The utilization of biodiesel as a substitute for diesel oil is one of the alternatives that aims to minimize the utilization of non-renewable fuels (petroleum) in the world energy matrix. Worldwide, biodiesel production occurs through the transesterification process. In Brazil, the most commonly used raw material for this purpose is soybean oil. A new way that has been promising for the biodiesel production is the Hidroesterification process. With that, the present work a imed to evaluate the soy oil enzymatic hydrolysis, using the comercial enzyme Lipozyme TL IM as the catalyst of the reaction. Was performed hydrolysis test of soy oil in closed and batch system, based on two experimental plannings (Placket-Burmann ad DCCR), to evaluate the effects of the variables involved in the process(pH; temperature; molar fraction water/oil; stirrings peed of the batch reactor and mass fraction enzyme/substrate). Since that, was perfomed kinetic studies with different conditions of molar fraction water/oil and different dosages of catalyst, as well as the utilization of a tip ultrasonic and a chemical agent to evaluateit’s effects on the kinetic of the reaction. To describe the kinetic of enzymatic hydrolysis of soy oil, was used a mathematical model based on the Michaelis-Menten kinetic. The results of experimental planningsindicatedthattheaciditylevelswerefavoredwhenthetemperatureof 52,2ºC, pH 6.1, stirring speed 150 rpm and molar fraction water/oil 37,7:1, getting 82,4% in the value of acidity level. The kinetic studies showed that the utilization of the probe ultrasonic did not accelerated the velocity of the enzymatic hydrolysis reaction. The kinetic involving the utilization of the chemical agent suffered a high decrease in the velocity of reaction, showing that this would be a inhibitor of enzymatic activity of Lipozyme TL IM. By the mathematical model, was verified that the model described well the experimental datas, as well made able the visualization of others components concentration (triacylglycerols, diacylglycerols, monoacylglycerols, glycerol, free fatty acids and water). It is believed that the values of the constants of the reaction velocity obtained in the model has more physical meaning and could be used to conducting the simulations in anothers conditions. / A utilização do biodiesel como substituto do óleo diesel é uma das alternativas que visa minimizar a utilização de combustíveis não renováveis (petróleo) na matriz energética mundial.Mundialmente, a produção de biodiesel ocorre pelo processo de transesterificação. No Brasil, a matéria-prima mais utilizada para esta finalidade é o óleo de soja. Uma nova rota que se mostra promissora para a produção de biodiesel é o processo de hidroesterificação. Desta maneira, o presente trabalho objetivou avaliar a hidrólise enzimática do óleo de soja, utilizando a enzima comercial Lipozyme TL IM como catalisador da reação. Foram realizados ensaios de hidrólise do óleo de soja em sistema fechado e batelada, baseados em dois planejamentos experimentais (Placket-Burmann e DCCR), visando avaliar os efeitos das variáveis envolvidas no processo (pH; temperatura; razão molar água/óleo; velocidade de agitação no reator batelada e razão mássica enzima/substrato). A partir disto, foram realizados estudos cinéticos com diferentes condições de razão molar água/óleo e diferentes dosagens de catalisador, bem como a utilização de ultrassom de ponta e um agente químico visando avaliar seus efeitos na cinética da reação. Para descrever a cinética da hidrólise enzimática do óleo de soja foi utilizado um modelo matemático baseado na cinética de Michaelis-Menten. Os resultados dos planejamentos experimentais indicaram que o índice de acidez foi favorecido quando temperatura de 52,2ºC, pH 6,1, velocidade de agitação de 150 rpm e razão molar água/óleo 37,7:1, obtendo 82,4% no valor do índice de acidez. Os estudos cinéticos demonstraram que a utilização do ultrassom de sonda não acelerou a velocidade da reação de hidrólise 13 enzimática. A cinética envolvendo a utilização de agente químico sofreu um alto decaimento na velocidade da reação, demonstrando que este seria um inibidor da atividade enzimática da Lipozyme TL IM. Pela modelagem matemática verificou-se que o modelo descreveu bem os dados experimentais, bem como possibilitou a visualização da concentração dos demais componentes (triacilgliceróis, diacilgliceróis, monoacilgliceróis, glicerol,ácidos graxos livreeágua). Acredita-se que os valores das constantes da velocidade de reação obtidos no modelo tenham maior significado físico e possam ser utilizados para realização de simulações em outras condições.

Chronic hyperlipidemia (lipotoxicity) and hyperglycemia (glucotoxicity) have recently been shown to induce Endoplasmic Reticulum (ER) stress, which may contribute to pancreatic beta-cell dysfunction in type 2 diabetes. This thesis examined the involvement of ER stress in beta-cell lipotoxicity and glucotoxicity. Although chronic treatment with saturated free fatty acids (FFA) in vitro induced ER stress, altering ER stress by increasing or knocking-down GRP78 chaperone expression had no effect on apoptosis induction. Conversely, overexpression of ER chaperones rescued the reduction in proinsulin protein levels caused by chronic exposure to high glucose, although it had no effect on the decreased insulin mRNA levels and proinsulin translation rate. Thus, ER stress is likely not the main mechanism involved in saturated FFA-induced beta-cell apoptosis in vitro, but it may contribute to glucotoxic effects on proinsulin levels. These findings have increased our understanding of the link between ER stress and beta-cell dysfunction in type 2 diabetes.