RESUMO

Escherichia coli é uma bactéria gram-negativa da família das Enterobacteriaceae, inicialmente descrita como uma bactéria comensal, mas que pela aquisição de fatores de virulência tornar-se patogênica. Dentre os fatores de virulência importantes na patogenicidade destacam-se membros da família das Serino-proteases Autotransportadoras de Enterobacteriaceae. (SPATEs). Sat (toxina autotransportadora secretada) tem se destacado como fator de virulência presente nas E. coli patogênicas, sendo a SPATE mais prevalente em E. coli isolada de sepse. No entanto, até o momento, não está claro se Sat poderia conferir alguma vantagem no escape da E. coli do intestino e/ou na sua colonização e manutenção da bactéria em nichos extraintestinais, tais como circulação sanguínea, acarretando a bacteremia que, dependendo das condições do hospedeiro, pode vir a evoluir para um quadro grave de sepse. Desta forma, o objetivo deste trabalho foi investigar se uma cepa de E. coli extraintestinal, portadora do gene sat, isolada de bacteremia (EC071) secreta a toxina durante a infecção de células endoteliais, in vitro. Inicialmente as condições ideais para o cultivo bacteriano e estabelecimento do MOI foram determinadas. Em seguida, células endoteliais da linhagem HUVEC foram infectadas com a cepa EC071. Na análise da cinética de infecção, por Western blotting detectou-se a presença de Sat no sobrenadante das células a partir de 4h de infecção. Contudo, nos ensaios preliminares para avaliação da citotoxicidade que avaliou morfologia e destacamento celular a partir da comparação com células infectadas com a cepa HB101, utilizada como controle, os danos celulares observados não foram significativos. Desta forma, o esclarecimento sobre a ação de Sat sobre o endotélio e manutenção da bactéria dentro do contexto da infecção na corrente sanguínea, dependerá de novos ensaios.

RESUMO

We are far behind the 2025 World Health Organization (WHO) goal of a zero increase in obesity. Close to 360 million people in Latin America and the Caribbean are overweight, with the highest rates observed in the Bahamas, Mexico, and Chile. To achieve relevant progress against the obesity epidemic, scientific research is essential to establish uniform practices in the study of obesity pathophysiology (using pre-clinical and clinical models) that ensure accuracy, reproducibility, and transcendent outcomes. The present review focuses on relevant aspects of white adipose tissue (WAT) expansion, underlying mechanisms of inefficient expandability, and its repercussion in ectopic lipid accumulation in the liver during nutritional abundance. In addition, we highlight the potential role of disrupted circadian rhythm in WAT metabolism. Since genetic factors also play a key role in determining an individual's predisposition to weight gain, we describe the most relevant genes associated with obesity in the Mexican population, underlining that most of them are related to appetite control.

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Several strategies are used by Escherichia coli to evade the host innate immune system in the blood, such as the cleavage of complement system proteins by secreted proteases. Members of the Serine Proteases Autotransporters of Enterobacteriaceae (SPATE) family have been described as presenting proteolytic effects against complement proteins. Among the SPATE-encoding genes sat (secreted autotransporter toxin) has been detected in high frequencies among strains of E. coli isolated from bacteremia. Sat has been characterized for its cytotoxic action, but the possible immunomodulatory effects of Sat have not been investigated. Therefore, this study aimed to evaluate the proteolytic effects of Sat on complement proteins and the role in pathogenesis of BSI caused by extraintestinal E. coli (ExPEC). E. coli EC071 was selected as a Sat-producing ExPEC strain. Whole-genome sequencing showed that sat sequences of EC071 and uropathogenic E. coli CFT073 present 99% identity. EC071 was shown to be resistant to the bactericidal activity of normal human serum (NHS). Purified native Sat was used in proteolytic assays with proteins of the complement system and, except for C1q, all tested substrates were cleaved by Sat in a dose and time-dependent manner. Moreover, E. coli DH5α survived in NHS pre-incubated with Sat. EC071-derivative strains harboring sat knockout and in trans complementations producing either active or non-active Sat were tested in a murine sepsis model. Lethality was reduced by 50% when mice were inoculated with the sat mutant strain. The complemented strain producing active Sat partially restored the effect caused by the wild-type strain. The results presented in this study show that Sat presents immunomodulatory effects by cleaving several proteins of the three complement system pathways. Therefore, Sat plays an important role in the establishment of bloodstream infections and sepsis.

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Increased homocysteine (Hcy) levels have been associated with a higher risk of cardiovascular and neurodegenerative diseases. Passive DNA demethylation has been suggested as one of the mechanisms implicated in the development of these conditions, and most studies have investigated this relationship in older adult populations. Therefore, this study aimed to evaluate the relationship between corporal composition and biochemical and haematological indicators with plasma homocysteine levels and genome-wide methylation (Alu, LINE-1, and SAT2) in a population of healthy young adults (median age, 18 years). We showed that the prevalence of hyperhomocysteinemia was significantly higher in men (18.5%) than in women (6.6%) (P = 0.034). Increased Hcy level was substantially associated with higher levels of body mass index and visceral fat in females, whereas in males, it was significantly associated with reduced red cell distribution width and high-density lipoprotein (HDL) cholesterol (HDL-C) levels and increased low-density lipoprotein/HDL ratio. Hypomethylation of Alu was significantly associated with reduced levels of HDL-C (<40.0 mg dL-1), whereas hypomethylation of LINE-1 and SAT2 was significantly associated with higher levels of skeletal muscle (<39.3%) in males. These results highlight the participation of hormonal factors in regulating Hcy metabolism, primarily in the female population, whereas changes in DNA methylation observed in males might be associated with the consumption of a protein diet with high levels of methionine, independent of increased Hcy levels.

Assuntos

RESUMO

Several strategies are used by Escherichia coli to evade the host innate immune system in the blood, such as the cleavage of complement system proteins by secreted proteases. Members of the Serine Proteases Autotransporters of Enterobacteriaceae (SPATE) family have been described as presenting proteolytic effects against complement proteins. Among the SPATE-encoding genes sat (secreted autotransporter toxin) has been detected in high frequencies among strains of E. coli isolated from bacteremia. Sat has been characterized for its cytotoxic action, but the possible immunomodulatory effects of Sat have not been investigated. Therefore, this study aimed to evaluate the proteolytic effects of Sat on complement proteins and the role in pathogenesis of BSI caused by extraintestinal E. coli (ExPEC). E. coli EC071 was selected as a Sat-producing ExPEC strain. Whole-genome sequencing showed that sat sequences of EC071 and uropathogenic E. coli CFT073 present 99% identity. EC071 was shown to be resistant to the bactericidal activity of normal human serum (NHS). Purified native Sat was used in proteolytic assays with proteins of the complement system and, except for C1q, all tested substrates were cleaved by Sat in a dose and time-dependent manner. Moreover, E. coli DH5α survived in NHS pre-incubated with Sat. EC071-derivative strains harboring sat knockout and in trans complementations producing either active or non-active Sat were tested in a murine sepsis model. Lethality was reduced by 50% when mice were inoculated with the sat mutant strain. The complemented strain producing active Sat partially restored the effect caused by the wild-type strain. The results presented in this study show that Sat presents immunomodulatory effects by cleaving several proteins of the three complement system pathways. Therefore, Sat plays an important role in the establishment of bloodstream infections and sepsis.

RESUMO

Diante das exigências crescentes das agências regulatórias do mundo todo quanto à redução/eliminação de ácidos graxos trans nos alimentos industrializados, bem como da conscientização do consumidor sobre a relação entre alimentação e saúde, o desenvolvimento de alternativas mais saudáveis aos óleos parcialmente hidrogenados e a outras fontes lipídicas com alto grau de saturaçã o se faz necessário. O oleogel, um sistema composto por um óleo preso em uma rede tridimensional formada por um agente estruturante, se apresenta como uma solução promissora. Dentre os diversos agentes estruturantes, as ceras vegetais se destacam por sua excelente capacidade de gelificação de óleos. Contudo, apresentam uma desvantagem sob o aspecto sensorial, pois podem conferir cerosidade e sabor residual desagradável aos alimentos. Com o objetivo de viabilizar o uso das ceras como agentes estruturantes em oleogéis face ao seu excelente desempenho tecnológico, este projeto propõe o estudo e a aplicação de oleogéis à base de óleo de soja (SBO) estruturado com ceras de farelo de arroz (RBW) a 2 e 4 % (m/m) ou carnaúba (CBW) a 3 e 6% (m/m), isoladamente. As matérias-primas foram caracterizadas e o comportamento de gelificação de cada cera foi avaliado por análises de textura por penetração de cone, estabilidade à perda de óleo por centrifugação, energia coesiva por parâmetro de solubilidade de Hansen (HSP) e comportamento de cristalização e fusão por calorimetria exploratória diferencial (DSC). Os resultados mostraram que ambas as ceras são capazes de formar oleogéis estruturalmente estáveis, contudo, o oleogel com 2% de RBW apresentou maior firmeza a 20 °C (190,4 gf/cm2) do que o oleogel com 6% de CBW a 5 °C (186,1 gf/cm2). Ao final de 5 dias, a capacidade de retenção de óleo do oleogel preparado com RBW foi de 100% às concentrações de 2 e 4% (m/m), contra 61 e 99,3% do oleogel elaborado com CBW às concentrações de 3 e 6% (m/m), respectivamente. Esses resultados podem ser explicados pela diferença entre as energias coesivas, ou seja, do grau de interação molecular entre o solvente e o soluto de cada oleogel. De acordo com os resultados de distância, que prevê se o gel formado será forte, fraco ou se não haverá formação de gel, o soluto CBW apresentou menor interação com o óleo (3,3 MPa1/2) do que o soluto RBW (3,7 MPa1/2). Os oleogéis foram aplicados como ingredientes em diferentes formulações de cream cheese, que foram analisados quanto a diferentes parâmetros de textura e esses resultados foram comparados a uma referência comercial. Nenhuma das amostras produzidas obteve resultados de textura estatisticamente iguais aos do cream cheese comercial (CC), o que pode ser explicado pelas diferenças de formulação e processamento dos produtos. Face aos resultados para textura e estabilidade à perda de óleo dos oleogéis de RBW, este agente estruturante apresenta ria maior potencial de aplicação, porém o oleogel CBW6 obteve alta capacidade de retenção de óleo (99,3%) e quando aplicado na formulação de cream cheese (CCBW6) apresentou resultados de firmeza e espalhabilidade mais próximos da amostra de referência, feita com gordura do leite (CMF)

Given the growing demands of regulatory agencies around the world regarding the reduction/elimination of trans fatty acids in processed foods, as well as consumer awareness about the relationship between food and health, the development of healthier alternatives to partially hydrogenated oils and others lipid sources with a high degree of saturation are necessary. Oleogel, a system composed of an oil trapped in a three-dimensional network formed by a structuring agent, presents itself as a promising solution. Among the various structuring agents, vegetable waxes stand out for their excellent oil gelling capacity. However, they have a sensory disadvantage, as they can give waxy and unpleasant aftertaste to foods. Aiming at enabling the use of waxes as structuring agents in oleogels in view of their excellent technological performance, this study proposes the evaluation and application of oleogels based on soybean oil (SBO) structured with rice bran wax (RBW) at 2 and 4% (m/m) or carnauba (CBW) at 3 and 6% (m/m). The raw materials were characterized and the gelling behavior of each wax was evaluated by analysis of texture by cone penetration, stability to oil loss by centrifugation, cohesive energy by Hansen solubility parameter (HSP) and crystallization and melting behavior. by differential scanning calorimetry (DSC). The results showed that both waxes are able to form structurally stable oleogels, however, oleogel with 2% RBW showed greater firmness at 20 °C (190.4 gf/cm2) than oleogel with 6% CBW at 5° C (186.1 gf/cm2). At the end of 5 days, the oil retention capacity of oleogel prepared with RBW was 100% at concentrations of 2 and 4% (m/m), against 61 and 99.3% of oleogel prepared with CBW at concentrations of 3 and 6% (m/m), respectively. These results can be explained by the difference between the cohesive energies, that is, the degree of molecular interaction between the solvent and the solute of each oleogel. According to the distance results, which predicts if the formed gel will be strong, weak or if there will be no gel formation, the CBW solute showed less interaction with the oil (3.3 MPa1/2) than the RBW solute (3 ,7 MPa1/2). Oleogels were applied as ingredients in different cream cheese formulations, which were analyzed for different texture parameters and these results were compared to a commercial reference. None of the samples produced had texture results statistically equal to those of commercial cream cheese (CC), which can be explained by the differences in formulation and processing of the products. Given the results for texture and oil binding capacity of RBW oleogels, this structuring agent would present greater application potential, but CBW6 oleogel obtained high oil biding capacity (99.3%) and when applied in cream cheese formulation (CCBW6) showed firmness and spreadability results closer to the reference sample, made with milk fat (CMF)

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As gorduras ricas em ácidos graxos saturados e/ou trans são ingredientes que brindam propriedades nos alimentos como textura, fusão e estabilidade. Porém, a relação entre seu consumo e enfermidades cardiovasculares, força a busca de novas alternativas. Uma alternativa potencial são os oleogéis, pois são um material plástico e com melhor balanço lipídico, do ponto de vista nutricional. Os oleogéis são sistemas coloidais onde uma fase orgânica líquida, representada por um óleo, é imobilizada por uma rede sólida tridimensional formada por agentes estruturantes. Atualmente, tem sido estudados oleogéis conformados com variados agentes estruturantes, com o intuito de encontrar a melhor opção. Além disso, alguns estudos indicam que o uso combinado de dois ou mais agentes estruturantes pode resultar no aumento de seu potencial de estruturação de óleos ou na possibilidade de customizar as propriedades adequadas para produção alimentar. Assim, o objetivo deste estudo é avaliar a interação entre os componentes de oleogéis preparados com óleo de girassol alto oleico (HOSO), como base lipídica, e combinações de cera de candelilla (CW) e monoestearato de sorbitana (SMS), como agentes estruturantes. A avaliação dos oleogéis por meio dos diagramas de fases e a consistência, mostrou que há uma sinergia entre os componentes, na concentração de 8 e 4% de agentes estruturantes e 79-89% de cera de candelilla. Estes oleogéis mistos tiveram uma faixa de fusão maior do que os oleogéis não mistos. Sugerindo a construição de cristais mais estáveis, que demoraram mais tempo em fundir. A capacidade de retenção de óleo se mostrou adequada (99%) nas amostras contendo mais de 1% de agentes estruturantes e com mais de 40% de Cera de Candelilla. Nas imagens obtidas por PLM, foi observada uma variedade de tamanho de cristais, sugerindo que cada componente do oleogel formou um tipo de cristal. Com a elaboração de bigéis, foi notado que, para obter um produto satisfatoriamente espalhável ou para observar o aumento de sua consistência em comparação com o oleogel isoladamente, é necessária uma concentração maior do que 4% de agentes estruturantes no oleogel e/ou a adição de menos de 20% de água na formulação total. A estabilidade dos bigéis foi beneficiada pela goma xantana e pela mistura de 2:1 de goma xantana e goma guar na concentração de 0,3%. Dessa forma, o desenvolvimento de alimentos com teores reduzidos de gorduras saturadas e ao mesmo tempo livres de gorduras trans se faz possível com a formulação de oleogéis, o que poderá contribuir para melhorar a qualidade nutricional dos alimentos industrializados oferecidos à população

Fats rich in saturated and / or trans fatty acids provide properties in food such as texture, fusion and stability. But, since those fats increase cardiovascular disease risk, alternatives have been searched. A potential alternative is oleogels, as they are a plastic material with better lipid balance from a nutritional point of view. Oleogels are colloidal systems where a liquid organic phase, represented by an oil, is immobilized by a three-dimensional solid network formed by structuring agents. Currently, oleogels conformed with various structuring agents have been studied in order to find the best option. In addition, some studies indicate that the combined use of two or more structuring agents may result in increased oil structuring potential or the possibility of customizing the properties suitable for food production. Thus, the aim of this study is to evaluate the interaction between the components of oleogels prepared with high oleic sunflower oil (HOSO) as a lipid base and combinations of candelilla wax (CW) and sorbitan monostearate (SMS) as structuring agents. The evaluation of oleogels by phase diagrams and consistency (Yield value) showed that there is a synergy between the components, in the concentration of 8 and 4% of structuring agents and 79-89% of candelilla wax. These mixed oleogels had a higher melting range than unmixed oleogels. Suggesting the construction of more stable crystals, which took longer to fuse. Oil binding capacity was adequate (99%) in samples containing more than 1% of structuring agents and over 40% of candelilla wax. In the PLM images, a variety of crystal sizes were observed, suggesting that each oleogel component formed a crystal type. With the elaboration of bigels, it was noted that in order to obtain a satisfactorily spreadable product or to observe the increase of its consistency compared to the oleogel alone, a concentration greater than 4% of structuring agents in the oleogel and / or the addition of less than 20% water in the total formulation. The stability of the bigels was benefited by xanthan gum and the 2: 1 mixture of xanthan gum and guar gum at a concentration of 0.3%.Thus, the development of foods with reduced levels of saturated fat and at the same time free of trans fats is possible with the formulation of oleogels, which may contribute to improve the nutritional quality of processed foods offered to the population

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Angiogenesis in inflammation are hallmarks for adipose tissue expansion in obesity. The role of angiopoietin/Tie-2 system in adipose tissue expansion and immune cell recruitment is unclear. We studied the effect of sleeve gastrectomy and the influence of FTO rs9930506 polymorphism on Tie-2, angiopoietin-1 and angiopoietin-2 expression in morbid obesity. Fifteen morbidly obese subjects (4 men and 11 women) aged 24-55 years were followed-up 3 and 6 months after sleeve gastrectomy. Serum sTie-2, angiopoietin-1, angiopoietin-2, and hypoxia-inducible factor-1α concentrations were determined by ELISA. Tie-2 and its ligands in visceral and subcutaneous adipose tissue were localized by immunohistochemistry. Tie-2 expression was measured by flow cytometry in circulating monocytes and infiltrated macrophages. Comparisons before and after sleeve gastrectomy were carried out using ANOVA for repeated measures. rs9930506FTO genotyping was performed by PCR-RFLP. Circulating sTie-2 and angiopoietin-2 were higher before sleeve gastrectomy. Tie-2 and angiopoietin-2 mRNA levels were higher in subcutaneous adipose tissue than visceral and both decreased after surgery. Monocytes and infiltrated macrophages showed a pro-inflammatory phenotype, with increased Tie-2 expression that decreased 3 and 6 months after sleeve gastrectomy. Baseline sTie-2 correlated inversely with adiponectin levels. At baseline the rs9930506FTO AG ó GG genotypes carriers had more 34 kg than genotype carriers of rs9930506 AA. Weight and body mass index decreased at 6 months. We found that angiopoietin/Tie-2 system is mainly expressed in subcutaneous adipose tissue, contributing to expandability, fat accumulation, and monocytes attachment in obesity. Bariatric surgery favorably modifies the pro-angiogenic profile, allowed a reduced angiogenic expression in the circulation and adipose tissue.

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Over a decade ago, foot-and-mouth disease (FMD) re-emerged in Southern Africa specifically in beef exporting countries that had successfully maintained disease-free areas in the past. FMD virus (FMDV) serotype SAT2 has been responsible for a majority of these outbreaks. Epidemiological studies have revealed the importance of the African buffalo as the major wildlife FMD reservoir in the region. We used phylogeographic analysis to study dynamics of FMD transmission between buffalo and domestic cattle at the interface of the major wildlife protected areas in the region currently encompassing two largest Transfrontier conservation areas: Kavango-Zambezi (KAZA) and Great Limpopo (GL). Results of this study showed restricted local occurrence of each FMDV SAT2 topotypes I, II, and III, with occasional virus migration from KAZA to GL. Origins of outbreaks in livestock are frequently attributed to wild buffalo, but our results suggest that transmission from cattle to buffalo also occurs. We used coalescent Bayesian skyline analysis to study the genetic variation of the virus in cattle and buffalo, and discussed the association of these genetic changes in the virus and relevant epidemiological events that occurred in this area. Our results show that the genetic diversity of FMDV SAT2 has decreased in buffalo and cattle population during the last decade. This study contributes to understand the major dynamics of transmission and genetic variation of FMDV SAT2 in Southern Africa, which will could ultimately help in designing efficient strategies for the control of FMD at a local and regional level.