Obesity is a major health problem in the Western world. Mean birth weight has increased during the last 25 years. One explanation is that the proportion of large for gestational age (LGA) infants has increased. Such infants risk developing obesity, cardiovascular disease and diabetes later in life. Despite the risk of neonatal hypoglycemia, their postnatal metabolic adaptation has not been investigated. Our data, obtained with stable isotope labeled compounds, demonstrate that newborn LGA infants have increased lipolysis and decreased insulin sensitivity. After administration of glucagon, the plasma levels of glucose and the rate of glucose production increased. The simultaneous increase in insulin correlated with the decrease in lipolysis, indicating an antilipolytic effect of insulin in these infants. We also demonstrated an intergenerational effect of being born LGA, since women born LGA, were at higher risk of giving birth to LGA infants than women not born LGA. Further, the LGA infants formed three subgroups: born long only, born heavy only, and born both long and heavy. Infants born LGA of women with high birth weight or adult obesity were at higher risk of being LGA concerning weight alone, predisposing to overweight and obesity at childbearing age. In addition we found that pregnant women with gestational diabetes were at increased risk of giving birth to infants that were heavy alone. This could explain the risk of both perinatal complications and later metabolic disease in infants of this group of women. To identify determinants of fetal growth, 20 pregnant women with a wide range of fetal weights were investigated at 36 weeks of gestation. Maternal fat mass was strongly associated with insulin resistance. Insulin resistance was related to glucose production, which correlated positively with fetal size. The variation in resting energy expenditure, which was closely related to fetal weight, was largely explained by BMI, insulin resistance, and glucose production. Lipolysis was not rate limiting for fetal growth in this group of women. Consequently, high maternal glucose production due to a high fat mass may result in excessive fetal growth.

Erst kürzlich wurde der Lipolyse-stimulierte Lipoproteinrezeptor (LSR, engl. lipolysis-stimulated lipoprotein receptor) als der zelluläre Oberflächenrezeptor von CDT und Iota-Toxin, zweier Vertreter der Iota-Toxin-Familie der clostridialen Aktin-ADP-ribosylierenden Toxine, identifiziert. In dieser Arbeit sollte geprüft werden, ob CST, ein weiterer Vertreter der Iota-Toxin-Familie, ebenfalls LSR für den Zelleintritt nutzt. Zunächst wurden die Toxinkomponenten CSTa und CSTb erstmals rekombinant hergestellt. Dazu wurden die für CSTa und CSTb codierenden Genabschnitte mittels PCR amplifiziert und anschließend in einen Expressionsvektor kloniert. Als Expressionsvektor wurde in dieser Arbeit der pHis1522-Vektor verwendet. Zur Amplifizierung wurden die Plasmide in E. coli transformiert und anschließend aufgereinigt. Die Proteinexpression erfolgte in B. megaterium, weil dieses Bakterium sich bereits zur Expression anderer clostridialer Toxine bewährt hatte. Zur Aufreinigung der 6xHis-getaggten Proteine wurde die Nickel-Affinitätschromatographie eingesetzt. Als nächstes wurde gezeigt, dass die rekombinant hergestellten Toxinkomponenten CSTa und CSTb biologisch aktiv waren. Dazu wurden CaCo2-Zellen mit CST behandelt und anschließend die Morphologie der Zellen untersucht. CaCo2-Zellen, die mit CSTa und CSTb behandelt wurden, wiesen Vergiftungserscheinungen wie eine typische Zellabrundung auf. Mit dem „Aktin-Nach-ADP-Ribosylierungs-Assay“ und der fluoreszenzmikroskopischen Untersuchung von TRITC-Phalloidin-gefärbtem Aktin wurde gezeigt, dass das rekombinant hergestellte CST Aktin-ADP-ribosylierende Eigenschaften besaß. Nachdem gezeigt war, dass rekombinant hergestelltes CST sich wie ein biologisch aktives, binäres Aktin-ADP-ribosylierendes Toxin verhält, konnte mithilfe der Vergiftung von H1-HeLa(+LSR)-Zellen und nativen H1-HeLa-Zellen, die kein LSR exprimierten, nachgewiesen werden, dass die Wirkung des Toxins LSR-abhängig ist. FACS-Analysen und Kolokalisationsstudien mit Alexa488-gefärbtem CSTb und Antikörper-gefärbtem LSR erbrachten zusätzlich den Beweis, dass CSTb auf der Zelloberfläche an LSR bindet und bei der Aufnahme in die Zellen mit LSR in endozytischen Vesikeln kolokalisiert. Die Ergebnisse dieser Arbeit zeigen, dass das C. spiroforme Toxin (CST) ebenfalls LSR als Rezeptor für den Zelleintritt verwendet.

Diss. (sammanfattning). Uppsala : Sveriges lantbruksuniv. / Härtill 4 uppsatser.

Diss. (sammanfattning) Umeå : Umeå universitet, 2006. / Härtill 4 uppsatser.

The objective of the study was to analyze the relationship of psychrotrophic lipolytic bacteria in bulk samples of cow?s raw milk and their metabolic activity based on the content of free fatty acids and identifying the factors that may influence their levels. The contents of free fatty acids (FFA) and counts of total bacteria, psychrotrophic lipolytic bacteria (PLiBC) and somatic cells were determined in 150 samples of cow?s bulk raw milk on 20 farms with three different milking technologies in South Bohemia during 2008?10. FFA were determined using an extraction-titration method. Within the compared technologies, the highest mean values of FFA (38,8 mmol?kg-1; P<0,001) and PLiBC (696 CFU?ml?1) were observed on farms with pipeline milking in stalls. The lowest mean FFA level (15,4 mmol?kg-1) was determined on farms with an automatic milking system. Medium values were determined on farms with parlour milking. From the aspect of the inhibition of an increase in psychrotrophic bacteria, and mainly in psychrotrophic lipolytic bacteria in conditions of cold storage of raw milk the temperature of 4°C seems optimum as it markedly inhibits the growth of mesophilic and psychrotrophic bacteria and at the same time the increase in the values of free fatty acids is slower at this temperature compared to the temperatures of 6,5 and 10°C.

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.

No description available.

O objetivo deste estudo foi avaliar os efeitos do ácido linoléico conjugado (CLA), especificamente do isômero trans-10, cis-12 em culturas de explante de tecido adiposo de suínos, em crescimento. / The objective of this study was to evaluate the effects of conjugated linoleic acid (CLA), specifically the tnas-10, cis-12 isomer on cultures of adipose tissue explants from growing pigs.

A crescente prevalência de diabetes mellitus (DM) em nível mundial, a sua incidência cada vez maior em crianças e adolescentes e resultados prévios obtidos em nosso laboratório (piora do quadro diabético em ratos na puberdade) nos levaram a investigar o metabolismo de glicose em tecido adiposo branco na puberdade em ratos controle (CO) e intolerantes à glicose (STZ). Parâmetros de desenvolvimento corporal e maturação sexual foram estudados entre a 5ª e a 10ª semanas de vida dos animais, identificando-se a 6ª e 7ª semanas como importante fase da puberdade. Desta forma, os estudos metabólicos foram realizados neste período. Os animais CO apresentaram franca resistência à insulina no período da puberdade, com prejuízos no metabolismo de glicose em adipócitos isolados. A injeção de estreptozotocina no quinto dia de vida produziu um quadro de intolerância à glicose em nossos animais, resultando em atraso no início do estirão de crescimento e desenvolvimento sexual. A resistência fisiológica à insulina da puberdade não produziu piora da resposta à insulina no tecido adiposo branco. / The growth in the prevalence of diabetes mellitus (DM), a larger incidence of type 1 and 2 DM in children and adolescents and the previous results of our laboratory (worsening of the metabolic picture in diabetic rats during puberty) motivate us to study the glucose metabolism in the puberty in order to assess the possible influence of the physiological insulin resistance characteristic of this period on the course of the diabetic status. Five-day old male Wistar rats were divided into two experimental groups: control group (CO) and streptozotocin group (STZ). Body development and sexual maturation were weekly evaluated between the 5th and 10th weeks of life. We identify the 6th and 7th weeks as an important early phase of the puberty. The insulin resistance and a slight degree of glucose intolerance were determined in non-diabetic rats. On the other hand, STZ group showed insulin resistance and it was impaired in the course of puberty suggesting that the developing of this resistance during the puberty presents a different characteristic of that seen in normal rats.

Este trabalho avaliou os efeitos de diferentes períodos de armazenagem (0, 3 e 6 dias) e temperaturas (4º e 10ºC) do leite de cabra sobre o produto pasteurizado. Foram avaliadas as alterações microbiológicas e físico-químicas do leite caprino cru e a influência nas características microbiológicas, físico-químicas e sensoriais do produto pasteurizado durante o armazenamento por até 6 dias. Utilizou-se a pasteurização lenta (65ºC/30 min.) e as amostras foram mantidas a 10ºC por 0, 3 e 6 dias. Foram realizadas contagens de microrganismos mesófilos, psicrotróficos totais, lipolíticos, proteolíticos, coliformes totais e fecais. As populações de microrganismos tiveram maior desenvolvimento na temperatura de 10ºC do que a 4º C. A população de microrganismos mesófilos e psicrotróficos atingiu 106 UFC/mL em todos os experimentos realizados a 10ºC no terceiro dia de estocagem. A 4ºC, este valor foi ultrapassado no sexto dia. A pasteurização foi eficiente na redução microbiana, sendo que os microrganismos lipolíticos, proteolíticos e psicrotróficos não foram detectados durante a armazenagem do produto. Os microrganismos mesófilos também foram reduzidos. Contudo alterações decorrentes do período de estocagem crua influenciaram as características físico químicas e sensoriais na estocagem após a pasteurização. A acidez do leite cru manteve-se aceitável em todos os experimentos realizados a 4ºC. No leite estocado a 10ºC, a acidez manteve-se aceitável apenas no terceiro dia após a coleta em 2 dos 3 experimentos. Foi observado decréscimo no teor de gordura do leite cru durante o armazenamento em ambas as temperaturas e o aumento gradativo da quantidade de ácidos graxos livres (AGL). Após a pasteurização, os valores de acidez e teor de gordura permaneceram estáveis, enquanto a quantidade de AGL continuou aumentando significativamente. Para a estocagem do leite cru a 10ºC a pasteurização só foi eficiente para manter a qualidade sensorial em níveis aceitáveis quando o leite cru foi pasteurizado no mesmo dia da ordenha e do resfriamento, enquanto na estocagem a 4ºC, o leite cru estocado por 6 dias foi rejeitado apenas no 3º dia após a pasteurização. Na análise sensorial, o leite estocado a 10ºC teve como principal causa de rejeição o odor e sabor azedos, enquanto a 4ºC, a rejeição deu-se principalmente pelo sabor cáprico. Concluiu-se que o leite de cabra cru poderia ser mantido por até 3 dias a 4ºC, para o produto pasteurizado ter uma vida útil de 6 dias e que a temperatura 10ºC não deve ser utilizada como instrumento na conservação do leite cru. / Different periods of storage (0, 3 and 6 days) and temperatures (4 and 10ºC) effects of raw goat milk on pasteurized product were evaluated. Microbiological and physical-chemical alterations in raw and pasteurized goat milk stored for up six days were evaluated. Pasteurization (65ºC/30 minutes) was used and samples were kept at 10ºC for 0, 3 and 6 days. Counting of mesophiles, total, lipolytic and proteolytic psichotrophic bacteria, total and fecal coliforms were determined. The microorganisms populations had higher growth at 10ºC than 4ºC and samples didn´t resist for 6 days in this condition. Pasteurization was efficient to reduce the number of microorganisms. Lipolytic, proteolytic and total psicrotrophic bacteria weren´t detected after pasteurization. Mesophiles and psichrotophic microrganisms populations reached 106 cfu/mL in all experiments at 10ºC at third day of storage. At 4ºC, this population was exceeded at sixth day. Pasteurization was efficient to reduce the number of microorganisms. Lipolytic, proteolytic and total psicrotrophic bacteria were not detected during pasteurized product storage. Mesophiles microorganisms also reduced. Alterations in raw milk during storage influenced physicalchemical characteristics after pasteurization. Acidity in raw milk remained acceptable in all of experiments at 4ºC. In stored raw milk at 10ºC, acidity remained acceptable only up to third day after milking in 2 of 3 experiments. It was observed decrease in raw milk fat content during storage in both temperatures and gradual increase of free fatty acids (FFA). After heat treatment, the values of acidity and fat content remained stable, while FFA continued increasing significantly. For raw milk storage at 10ºC pasteurization was efficient to keep sensorial quality in acceptable levels when raw milk was pasteurized in the same day of milking and cooling, while in storage at 4ºC, raw milk stored per 6 days was rejected only in third day after pasteurization. In sensorial analysis, stored milk at 10ºC had as main cause of rejection the acid odor and flavor, while at 4ºC, the rejection was given mainly for the capric flavor. It was concluded that raw goat milk can be kept for up to 3 days at 4ºC before the heat treatment for pasteurized product have six days of shelf-life and that the temperature 10ºC does not have to be used as an instrument of raw milk conservation.