O treinamento aeróbio de forma crônica está correlacionado com menor grau de inflamação e pode ser considerado um dos principais responsáveis por criar condições favoráveis para o equilíbrio intestinal. A mucosa do intestino consiste em uma monocamada de células epiteliais com função principal de absorção de nutrientes e formação de barreira contra entrada de potenciais antígenos que podem prejudicar sua função. A permeabilidade da barreira intestinal pode ser controlada por diferentes processos. Um aumento de antígenos luminais como lipopolissacarídeo (LPS) pode gerar um aumento na permeabilidade paracelular através de quebra das principais proteínas de função barreira, claudina e ocludina, ativando o sistema imune local, levando a um processo de inflamação e dano do tecido. Essas alterações da função barreira estão ligadas com algumas doenças como a doença inflamatória intestinal e o câncer. Por essa razão, faz-se necessário entender o papel do exercício físico na regulação da integridade da mucosa intestinal e seu efeito terapêutico. Assim, o objetivo deste trabalho é avaliar o efeito do treinamento aeróbio sobre a morfologia e função barreira da mucosa intestinal em ratos wistar. Na primeira etapa, os animais foram divididos em 2 grupos: grupo 1 - controle sedentário (CTRL), grupo 2 - treinado (T) e desenvolvido um treinamento aeróbio durante 8 semanas com uma carga correspondente a 65% da velocidade máxima atingida no teste de exercício físico progressivo até a exaustão, por 60 minutos durante 5 vezes por semana. 9 O treinamento físico promoveu uma maior perda de peso nos animais treinados sem aumento do consumo de ração comparado com animais sedentários. A segunda etapa ocorreu após o período de treinamento para um tratamento com 3 dias de injeção de LPS intraperitoneal. Os animais foram divididos em 4 subgrupos: grupo 1 - controle sedentário (CTRL), grupo 2 - treinado (T), grupo 3 - controle sedentário com LPS (CTRL + LPS) e grupo 4 - treinado com LPS (treinado + LPS). Os animais sedentários com LPS apresentaram uma maior perda de peso e menor consumo de ração além de maior perda de altura das vilosidades intestinais em detrimento ao maior grau de inflamação com aumento também de infiltrados no cólon. Em contrapartida, o treinamento físico apresentou um efeito protetor sobre a perda de peso e as vilosidades intestinais observados nos animais treinados com LPS. Além disso, foi analisado a expressão gênica de algumas interleucinas pró e anti-inflamatórias (IL 1?, IL 6, IL 10 e IL 23) e os genes para claudina 1 e ocludina. O principal aumento foi observado no perfil gênico da IL23 uma citocina pró inflamatória e da claudina 1 nos animais sedentários com LPS em comparação ao controle como uma ação compensatória para reparação da permeabilidade intestinal e função barreira. Juntos, esses resultados apoiam a hipótese de que o treinamento físico é um fator importante para proteção da integridade da mucosa e função barreira intestinal / Chronic aerobic training correlates with low level of inflammation and is considered one of the main factors in creating favorable conditions for intestinal balance. The intestinal mucosa consists of a monolayer of epithelial cells whose the main function is to absorb nutrients and form a barrier against entry of potential antigens that may impair its function. Different processes control the permeability of the intestinal barrier. An increase in luminal antigens such as lipopolysaccharides (LPS) may lead to an increase in paracellular permeability through the breakdown of the main barrier proteins, claudin and occludin, activating the local immune system, leading to a process of inflammation and tissue damage. These changes in barrier function are linked to some pathologies including inflammatory bowel disease and cancer. For this reason, it is necessary to understand the role of physical exercise in regulating the integrity of the intestinal mucosa and its therapeutic effect. Thus, the objective of this study was to evaluate the effect of aerobic training on the morphology and barrier function of the intestinal mucosa in wistar rats. For the first stage of this project, animals were separated into 2 groups: group 1 - sedentary control (CTRL), group 2 - trained (T) which involved 8 weeks of aerobic training, five sessions of one hour, with a load corresponding to 65% of the maximum speed attained in a progressive exercise test to exhaustion. Physical training promoted weight loss in trained animals without increased 11 feed intake compared to sedentary animals. The second stage of this project was performed after the training period and involved 3 days of intraperitoneal LPS injections. The animals were separated into 4 subgroups: group 1 - sedentary control (CTRL), group 2 - trained (T), group 3 - sedentary control with LPS (CTRL + LPS) and group 4 - trained with LPS (trained + LPS). Sedentary LPS animals presented greater weight loss and lower feed intake, as well as greater height loss of intestinal villi, a higher degree of inflammation and also increased colon infiltrates. In contrast, physical training had a protective effect on weight loss and intestinal villi in animals who trained and received LPS. In addition, gene expression of some pro and anti-inflammatory interleukins (IL-1?, IL-6, IL-10 and IL-23) and the genes for claudin 1 and occludin were analyzed. The main increase was shown in the gene profile of IL23 a proinflammatory cytokine and claudin 1 in sedentary with LPS animals as a compensatory action for repair of intestinal permeability and barrier function. Together, these results support the hypothesis that physical training is an important factor for protection of mucosal integrity and intestinal barrier function

O objetivo do presente estudo foi avaliar o desempenho, saúde intestinal e metabolizabilidade de dietas para frangos de corte alimentados com o uso contínuo ou mudança brusca de diferentes rações à base de milho e sorgo moído ou inteiro. No Exp. 1 foi estudada a mudança no tipo de grão com as dietas: M100% (ração à base de milho); S100% (ração à base de sorgo); M:S50% (ração à base de 50% milho e 50% sorgo); PS-M (ração à base de sorgo na fase pré-inicial e milho nas demais fases); PM-S (ração à base de milho na fase pré-inicial e sorgo nas demais fases). No Exp. 2 foi estudada a mudança na forma do grão de sorgo: Sm100% (ração à base de sorgo moído); Si100% (ação à base de sorgo inteiro); PSm-Si (ração à base de sorgo moído na fase pré-inicial e inteiro nas demais fases) e PSi- Sm (ração à base de sorgo inteiro na fase pré-inicial e moído nas demais fases). Os experimentos foram conduzidos em um delineamento em blocos casualizados (blocos no tempo) para avaliar o desempenho e epitélio jejunal (r=8), microbiota intestinal (r=4) e metabolizabilidade das dietas (r=10). As mudanças entre milho e sorgo não alteraram o desempenho, epitélio jejunal nem a metabolizabilidade das rações, mas influenciaram a porcentagem de Clostridium, Weissella, Bacillus e Alkaliphilus no intestino delgado e Lactobacillus e Desulfotomaculum nos cecos. O uso de sorgo inteiro piorou o desempenho aos sete dias. Aos 40 dias, Sm100% e PSm-Si apresentaram desempenhos semelhantes, PSi-Sm apresentou menor ganho de peso, mas melhor conversão alimentar e Si100% apresentou o pior desempenho. Si100% e PSm-Si apresentaram aumento no peso relativo da moela e na metabolizabilidade das rações, assim como diminuição de Clostridium e aumento de bactérias das famílias Actinomycetales e Bacillales no intestino delgado, Si100% resultou em menor porcentagem de Alkaliphilus e Enterococcus que Sm100% nos cecos. Conclui-se que a melhor estratégia de uso do sorgo é a substituição total de milho por sorgo moído ao alojamento com posterior mudança para sorgo inteiro, pois não afeta o desempenho e epitélio jejunal das aves, melhora a metabolizabilidade das rações e potencializa a redução de Clostridium no intestino delgado de frangos de corte. / The aim of this study was to evaluate performance, intestinal health and metabolizability of diets in broilers fed different corn- or sorghum (ground or whole)- based diets, continuously or with abrupt change between the diets. Exp. 1 - Change in the type of grain: C100% (corn-based diet); S100% (sorghum-based diet); C:S50% (50% corn and 50% sorghum-based diet); PC-S (corn-based diet in pre-starter phase and sorghum-based diet in other phases); PS-C (sorghum-based diet in pre-starter phase and corn-based diet in other phases). Exp. 2 - Change in the form of sorghum grain: Gs100% (ground sorghum-based diet); Ws100% (whole sorghum-based diet); PGs-Ws (ground sorghum-based diet in pre-starter phase and whole sorghum-based diet in other phases); PWs-Gs (whole sorghum-based diet in pre-starter phase and ground sorghum-based diet in other phases). Experiments were conducted in a randomized block design for to evaluate performance and jejunal epithelium (r=8), intestinal microbiota (r=4) and metabolizabilty of diets (r=10). The changes between corn and sorghum did not affect performance, jejunal epithelium or metabolizability of the diets, but influenced the genera Clostridium, Weissella, Bacillus and Alkaliphilus in the small intestine, and Lactobacillus and Desulfotomaculum in the caecum. Whole sorghum resulted in decreased performance at seven days of age. At 40 days, Gs100% and PGs-Ws showed similar performance, PWs-Gs showed lower weight gain and the best feed conversion rate, and Ws100% showed the worst performance. Ws100% and PGs-Ws resulted in the biggest gizzard relative weight and the highest diet metabolizability values, as well as the lowest level of Clostridium and highest level of Actinomycetales and Bacillales in the small intestine. Ws100% showed lower level of Alkaliphilus and Enterococcus than Gs100% in the caecum. The best strategy to use sorghum in broilers diets is replacing 100% of corn for ground sorghum since the first day followed by change to whole sorghum, because this diet did not affect performance or jejunal epithelium, improved diet metabolizability values, and reduced Clostridium in the small intestine of broilers.

Com o intuito de verificar a eficácia do açúcar e de edulcorante a base de sacarina sódica e neoesperidina nas dietas de leitões, foram realizados dois experimentos, ambos com 42 leitões recém-desmamados aos 21 dias de idade. Foram oferecidas três dietas: dieta controle (DC), dieta com açúcar (DA) e dieta com edulcorante (DE). Os experimentos foram em blocos causalizados, sendo o primeiro com sete repetições e dois animais por unidade experimental e o segundo com seis repetições, um animal por unidade experimental e arranjo fatorial 3 x 2 + 1 (três dietas, duas épocas de abate: aos 28 e 35 dias de idade e o abate no dia do desmame). No primeiro experimento, os animais permaneceram nas instalações de creche até os 63 dias de idade para avaliação do desempenho, da incidência de diarreia e da viabilidade econômica das dietas. No segundo experimento, os animais foram abatidos para se proceder à pesagem de órgãos, medição do comprimento do intestino delgado e coleta dos segmentos do duodeno e jejuno para realização da morfometria intestinal. Não houve diferenças entre as dietas para o CDR, GDP e CA nos Períodos 1 (21 aos 28 dias), 3 (21 aos 50 dias) e 4 (21 aos 63 dias de idade). No Período 2 (21 aos 35 dias), os animais que consumiram a DE apresentaram maior (P<0,05) CDR e GDP, se comparados aos que consumiram a DA. Nos Períodos 1, 2 e 3 foi observado menor custo da dieta por kg de peso vivo ganho e melhores índices econômico e de custo para os animais que consumiram DE. Ao avaliar a morfometria intestinal e peso de órgãos, não se observou diferença entre as dietas, com exceção para o peso do estômago, que foi maior nos animais que consumiram DCE. A inclusão de edulcorante é eficaz em melhorar o desempenho no Período 2, além de ser viável sob o ponto de vista econômico. / In order to verify the effectiveness of sugar and sweetener containing saccharin and neohesperidin in diets of piglets, two experiments were conducted, both with 42 newborn piglets weaned at 21 days old. Were offered three diets: control diet (DC), diet with sugar (DA) and diet with sweetener (DE). The experiments were in randomized blocks, the first of which had seven replicates and two animals per experimental unit and the second, six replicates and one animal per experimental unit comprising a factorial 3 x 2 + 1 (three diets, two slaughter ages: at 28 and 35 days of age and slaughter on the day of weaning). In the first experiment, the animals remained on the nursery facilities until 63 days of age to evaluate the performance, the incidence of diarrhea and the economic viability of the diets. In the second experiment, the animals were slaughtered to weigh the organs, measure the length of the small intestine and collecting segments of the duodenum and jejunum to determine intestinal morphology. There were no differences between diets for the ADFI, FC and ADG in the Periods 1 (21 to 28 days), 3 (21 to 50 days) and 4 (21 to 63 days old). In the Period 2 (21 to 35 days), animals that consumed the DE had higher (P<0.05) ADFI and ADG, compared to those fed the DA. In Periods 1, 2 and 3 was observed lower cost per kg of the diet of live weight gain and better economic indicators to the animals fed DE. No differences were observed between the diets when evaluating intestinal morphology and organ weights, except for the weight of the stomach, which was higher in animals fed DE. The inclusion of sweetener is effective in improving performance in Period 2, and is viable under the economical point of view.

by Wong Ka-Hing. / "September 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 226-279). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Les particules de dioxyde de titane (TiO2) sont utilisées dans de nombreux secteurs industriels du fait de leurs propriétés physiques et chimiques intéressantes. Depuis une dizaine d’années, elles sont également utilisées sous forme nanoparticulaire car la taille nanométrique leur apporte de nouvelles propriétés, recherchées dans certaines applications industrielles. Elles sont par exemple utilisées comme colorant blanc dans le secteur de la cosmétologie, de la pharmacologie et dans les industries agroalimentaires. Dans ces dernières, l’utilisation de ces particules est autorisée car le TiO2 est un composé insoluble et relativement inerte. Le colorant alimentaire E171, autorisé depuis 1966, est ainsi constitué de particules de TiO2, initialement sous forme micrométrique, mais il s’avère que selon les procédés de fabrication, entre 10 et 43 % (selon les études) de ces particules présentent un diamètre inférieur à 100 nm, i.e. sont sous forme nanométrique. Ce n’est pas un nanomatériau du point de vue de la définition européenne, il n’est donc pas soumis à l’obligation d’étiquetage dans les produits alimentaires. Le E171 est présent dans de nombreux aliments sans que son impact sur la santé humaine, après ingestion, n’ait été clairement documenté. De plus en plus d’études s’intéressent à la toxicité des nanoparticules (NPs) après leur ingestion, mais peu d’entre elles ont été menées avec le E171 à proprement parler. Les études in vivo et in vitro publiées à ce jour démontrent que les NPs de TiO2 sont peu toxiques. Leur absorption intestinale et leur translocation vers le système sanguin puis des organes secondaires est faible. Les principaux effets décrits sont une augmentation des espèces réactives de l’oxygène associées à un stress oxydant, l’induction de marqueurs de l’inflammation, et plus récemment l’induction du stress du réticulum endoplasmique. Des effets sont également rapportés sur différents paramètres de la barrière intestinale, i.e. le microbiote, le mucus, les transporteurs membranaires, les jonctions cellulaires et l’immunité intestinale. Chez certaines personnes, cette barrière est compromise, elles sont donc potentiellement plus sensibles aux micros et nanos-particules contenues dans l’alimentation. Leur épithélium intestinal est enflammé, et à long terme, ces personnes peuvent développer des maladies inflammatoires chroniques de l’intestin et dans les cas les plus graves, des cancers.L’objectif de cette thèse est d’étudier la toxicité du colorant alimentaire E171 et d’approfondir les connaissances relatives à l’impact des NPs de TiO2 sur le système gastro-intestinal. Pour cela, nous avons travaillé avec différents modèles cellulaires d’épithélia intestinaux humain, un modèle d’épithélium jointif composé d’entérocytes Caco-2, un modèle d’épithélium sécrétant une couche de mucus, composé de cellules Caco-2 et HT29-MTX et enfin un modèle d’épithélium bordant les plaques de Peyer, composé des cellules Caco-2(C1) et RajiB. Ces modèles cellulaires ont été exposés de façon aigüe (6 h, 24 h et 48 h) ou chronique (21 jours), au colorant E171 ainsi qu’à deux NPs de TiO2 : A12, qui a la même structure cristalline que le E171 et P25, une NP très documentée dans la littérature. Nos résultats montrent que le E171 et les NPs de TiO2 sont modérément toxiques, ils n’engendrent pas de mortalité cellulaire ni de dommages à L’ADN. Néanmoins, ils provoquent une accumulation d’espèces réactives de l’oxygène intracellulaires et modulent certains marqueurs impliqués dans le stress oxydant, le stress du réticulum endoplasmique et l’inflammation. Ils impactent également la sécrétion et la composition de la couche de mucus, l’expression des transporteurs ABC, qui sont des paramètres impliqués dans la fonction de barrière de l’épithélium intestinal, le rendant possiblement plus vulnérable aux agressions extérieures. / Micro-sized titanium dioxide (TiO2) particles are used for years by industrials for their attractive physical and chemical properties. The use of TiO2 nanoparticles (NPs) is also constantly increasing, because the nanometric size gives new interesting properties to particles which industrials are looking for. In some daily-life products including paints, plastics, paper, medicines and food, micro-sized TiO2 particles are used as a pigment for their opacifying and whitening capacities. The use of TiO2 as a food additive, i.e. E171 in the EU, has been authorized in most countries since the 60ies, without any established acceptable daily intake, because of their low toxicity and intestinal absorption. However, it was recently shown that E171 can contain up to 43% of particles with diameter ranging from 1 to 100 nm, i.e. NPs. Still, E171 is not a nanomaterial as described in the European recommendation of definition because it contains less than 50% of NPs (in number). Food grade TiO2 is present in a wide range of food products while little is known about its toxicological impact to human health. The toxicity of ingested TiO2, either nano- or micro-sized, is increasingly documented, still E171 itself is rarely used in these studies.According to in vivo and in vitro studies, TiO2 particles were proven relatively safe for intestinal cells, no cytotoxicity neither genotoxicity were reported. Nevertheless, particles were often reported to increase reactive oxygen species (ROS) cell content, to impair autophagic processes and modulate gene expression and the content of proteins involved in oxidative stress, endoplasmic reticulum stress and inflammatory response regulation. Interestingly, their reported impact on intestinal cells suggests alteration of almost all the components of the intestinal barrier function, i.e. microbiota, mucus, cell junctions and transporters. This intestinal barrier function is altered in patients suffering from intestinal bowel diseases, these persons are thus possibly more sensitive to mineral particulate in food.The present study aimed at improving knowledge on the toxicity of food-grade TiO2. To this purpose, the impact of E171 was evaluated on in vitro cell models representative of the human intestinal epithelium, i.e. a model of differentiated Caco-2 enterocytes, a model of mucus-secreting epithelium obtained by coculture of Caco-2 and HT29-MTX mucus-secreting cells and a model of the follicle-associated epithelium, which lines Peyer patches, obtained by coculture of Caco-2(C1) and RajiB cells. These cell models were either acutely exposed for 6 h, 24 h and 48 h or chronically exposed for 21 days to E171. In parallel, they were exposed to two model TiO2-NPs, A12 which has the same crystalline structure as E171 and P25, a well-documented TiO2-NPs. Our results show that E171 and TiO2-NPs induced no overt cell mortality but significant oxidative stress, and that they oxidatively damage DNA. They modulate the expression of genes involved in oxidative stress and endoplasmic reticulum stress regulation. They also modulate the expression of genes, as well as the content of proteins from mucus, ABC transporters and inflammatory markers, which are the main players of the intestinal barrier function and presumably increase epithelium sensitivity to xenobiotics. These data suggest that they may be implicated in the development or aggravation of inflammatory bowel diseases.

Le cancer colorectal représente la deuxième cause de mortalité par cancer en France. Dans l’intestin, l’initiation et la progression tumorale sont corrélées à l’accumulation séquentielle de mutations génétiques et épigénétiques au niveau du compartiment de renouvellement de l’épithélium. Ces altérations ont pour conséquence une croissance incontrôlée de l’épithélium et, à terme, la formation de lésions cancéreuses. En tenant compte du nombre croissant de cas de cancers colorectaux, la découverte de facteurs de prédisposition de cette pathologie reste d’un intérêt majeur. Les données de séquençage du génome humain ne suffisant pas à expliquer la prévalence de la maladie à l’échelle de la population, nous nous intéressons aux mécanismes permettant le contrôle de l’expression de certains gènes : les mécanismes épigénétiques. Dans notre équipe nous disposons de modèles animaux génétiquement modifiés nous permettant d’étudier, dans des conditions proches de la pathologie humaine, les phases précoces de l’initiation tumorale. Ces souris, bien que génétiquement identiques, développeront pourtant un nombre de tumeurs intestinales très variable. En comparant les profils moléculaires de souris développant peu ou beaucoup de tumeurs intestinales, nous souhaitons mettre en évidence les facteurs épigénétiques mis en jeu pour expliquer cette différence de susceptibilité à la tumorigenèse. Pour ce faire, nous avons mis en place une stratégie visant à prélever un échantillon intestinal, par chirurgie, sur de jeunes souris qui ont ensuite été suivies jusqu’à l’âge adulte et à l’apparition des tumeurs. Cette stratégie innovante nous a permis de corréler les profils d'expression et de methylation d'ADN d’intestins développant peu ou beaucoup de tumeurs, ouvrant la possibilité de disposer de nouveaux marqueurs prédictifs quant aux chances de développer un cancer. Ces données sont complétées par une étude sur les conséquences de la perte du gène suppresseur de tumeur Apc, un gène couramment muté dans les cancers colorectaux humain. A l’aide de modèles de souris d’invalidation inductible, nous avons déterminé les conséquences de la perte d’Apc sur les profils de méthylation de l'ADN des cellules souches intestinales, et leur capacité à initier une tumeur. L’ensemble de ces différentes projets développés dans le cadre de ma thèse nous ont permis de mieux comprendre les mécanismes cellulaires liés à la prédisposition et à l’initiation tumorale et de proposer des nouvelles stratégies diagnostiques et d’évaluation du risque individuel. / Cancer initiation and progression represent the outcome of the progressive accumulation of genetic and epigenetic alterations. Global changes in the epigenome are now considered as a common hallmark of malignancies. However, most of our present knowledge represents the result of the comparison between fully established malignancies and their surrounding healthy tissue. Such comparison is not informative about the epigenetic contribution to the very early steps of cancer onset. By performing DNA methylation and gene expression profiling of the intestinal epithelium of relevant in vivo models we aim at shedding light on the correlation between the interindividual epigenetic polymorphisms within the population and the relative risk to develop malignancies, and establish the existence of a molecular signature associated with an increased susceptibility to develop intestinal cancer. Our results confirm that a considerable degree in the variability associated to cancer susceptibility cannot be ascribed to major genetic changes and that such heterogeneity seems to correlate with distinct molecular profiles associated to classes of poorly or highly susceptible isogenic animals.We also investigated in vivo the timing at which the remodeling occur at the epigenomic scale by analyzing the alterations in the DNA methylation and gene expression profiles of intestinal stem cells upon the loss of the Apc gene, the most common genetic lesion associated with human colorectal cancer initiation. We found that the loss of function of Apc in the Lgr5-positive intestinal stem cell compartment is rapidly accompanied by a reprogramming of the DNA methylation profiles resulting in altered gene expression and impaired fate determination in those cells. The results show that part of the phenotype resulting from the constitutive activation of the Wnt pathway upon Apc loss is acquired via differential epigenetic regulation of key biological processes controlling the balance between self-renewal and differentiation. By using conditional genetic ex vivo models we found part of these oncogenic effects to be reversible via the modulation of the machinery responsible for de novo methylation of the DNA.Overall, this work confirms that the epigenetic remodeling is an early event in tumorigenesis that might even precede actual cell transformation. The functional impact of our findings on cancer initiation is currently under investigation.

Submitted by Jaqueline Silva (jtas29@gmail.com) on 2014-09-22T17:08:11Z No. of bitstreams: 2 Santos Filho, Edvande-2014-dissertação.pdf: 3465269 bytes, checksum: fd50a85f70b6b596e0c5c4fb5fd18cb9 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2014-09-22T18:45:16Z (GMT) No. of bitstreams: 2 Santos Filho, Edvande-2014-dissertação.pdf: 3465269 bytes, checksum: fd50a85f70b6b596e0c5c4fb5fd18cb9 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-09-22T18:45:16Z (GMT). No. of bitstreams: 2 Santos Filho, Edvande-2014-dissertação.pdf: 3465269 bytes, checksum: fd50a85f70b6b596e0c5c4fb5fd18cb9 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-03-20 / Introduction: Intestinal mucositis is a frequent limiting factor in antineoplastic therapy. There is no truly effective treatment targeted to cure this side effect. Curcuma longa L. has been reported to have antioxidant, antitumor and anti-inflammatory properties. Objective: This study aimed to evaluate the effects of a mucoadhesive formulation with Curcuma longa L. extract (MCL) in mice bearing intestinal mucositis induced by 5-FU. Methods: Swiss male adult mice (35-40g) were randomly allocated into three experimental groups of 5 animals each: control (mucoadhesive formulation 0.6mL/animal, via gavage, from day 1-6); exposed to 5-fluorouracil (5-FU 200mg/kg, i.p., in days 4-6); treated prophylactically and throughout mucositis with FMCL and exposed to 5-FU (MCL 15mg/kg, via gavage, from day 1-6, plus 5-FU 200mg/kg, i.p., in days 4-6). In the 7th day, animals were anesthetized by xylazine-ketamine followed by cervical dislocation. Duodenal samples, 10 cm after pyloric sphincter were removed to perform the essays. The parameters evaluated were: body weight assessment, morphometrics and histo-pathological analysis, apoptosis (p53/Bax, Bcl-2), cell proliferation (Ki-67), myelo-peroxidase (MPO) and malondialdehyde (MDA). Results: 5-FU induced intestinal mucositis characterized by villus shortening, crypt deepening, intense inflammatory infiltration, vacuolization and mucosal edema. Besides, 5-FU induced severe mice body mass reduction, apoptosis in cells of villus and crypts (p<0.001), increase in MPO activity and MDA formation (p<0.05), when compared to the control group. Treatment with MCL attenuated body mass loss, protected intestinal mucosa from villus shortening and crypt deepening, decrease MPO activity and MDA formation (p<0.05). In this group of animals was also observed high expression of the cell proliferation marker Ki-67 in epithelial cells lining of villus and crypts. Conclusion: Our data confirm the therapeutic potential of MCL for the treatment of intestinal mucositis in mice. Further studies are needed to assess this formulation potential for human use. / Introdução: A mucosite intestinal é um dos efeitos adversos limitantes das terapias antineoplásicas. Não existe tratamento realmente efetivo direcionado a cura deste efeito colateral grave. A Curcuma longa L. tem sido proposta como candidata ao tratamento de várias doenças por possuir propriedades antioxidante, antitumoral e anti-inflamatória. Objetivo: Este estudo objetivou avaliar os efeitos da formulação mucoadesiva com extrato de Curcuma longa L. (FMCL) em animais portadores de mucosite intestinal induzida por 5-FU. Métodos: Camundongos Swiss adultos, machos (35-40g) foram aleatoriamente alocados em três grupos experimentais de 5 animais cada: [1] controle (formulação mucoadesiva 0.6mL/animal, via gavagem, do dia 1-6); [2] exposto ao 5-fluorouracil (5-FU 200mg/kg, i.p., dos dias 4-6), [3] tratado profilaticamente e ao curso da mucosite com FMCL e exposto ao 5-FU (FMCL 15mg/kg, via gavagem dos dias 1-6, mais 5-FU 200mg/kg, i.p., dos dias 4-6). No sétimo dia, os animais foram anestesiados por xilazina-ketamina seguido por deslocamento cervical. Amostras de duodeno, 10 cm após o esfíncter pilórico, foram coletadas para realização dos ensaios. Os parâmetros avaliados foram: avaliação de massa corporal, análise morfométrica e histopatológica, apoptose (p53/Bax; Bcl-2), proliferação celular (Ki-67), mieloperoxidase (MPO) e malondialdeído (MDA) Resultados: A administração de 5-FU induziu mucosite intestinal caracterizada por encurtamento das vilosidades, aprofundamento das criptas, intenso infiltrado infla-matório, vacuolização e edema na mucosa. Além disso, o 5-FU induziu grave redução de massa corporal nos camundongos e apoptose nas células das vilosida-des e criptas, quando comparado ao grupo controle (p<0,001). Foi observado ainda aumento na atividade de MPO e formação de MDA, quando comparado ao grupo controle (p<0,05). Por outro lado, o tratamento com a FMCL atenuou a perda de massa corporal dos animais com mucosite. Ademais, protegeu a mucosa intestinal da redução no tamanho das vilosidades e criptas induzidas pelo 5-FU. Neste grupo de animais foi observado ainda aumento na expressão do marcador de proliferação celular Ki-67 nas células epiteliais de revestimento das vilosidades e criptas intestinais. O tratamento também diminuiu significantemente a atividade de MPO e formação de MDA (p<0,05). Conclusão: Os dados confirmam o potencial terapêutico da FMCL no tratamento da mucosite intestinal em camundongos. Estudos adicionais são necessários para que esta formulação possa se tornar uma alternativa segura para uso em seres humanos.

Em nosso estudo pretendemos estudar a via oral para o desenvolvimento de vacina para toxoplasmose utilizando parasitas irradiados por Cobalto-60, como uma alternativa para a prevenção desta importane parasitose. Para tanto, avaliamos o desenvolvimento da imunidade sistêmica e intestinal e a resistência à infecção, em camundongos imunizados por esta via com taquizoítos irradiados a 255Gy, e desafiados com cistos da cepa ME-49. Camundongos isogênicos C57Bl/6j foram imunizados com 107 taquizoitos de T. gondii irradiados a 255Gy em diferentes esquemas e utilizando adjuvantes leite (anti-péptico) e hidróxido de alumínio (anti-ácido). As preparações de taquizoítos irradiados por via oral induziram produção de imunoglobulinas IgG e IgA no soro de camundongos, sendo predominante a subclasse de IgG2a, similar a infecção crônica, determinadas por ELISA. Seu uso com adjuvantes anti-pépticos ou anti-ácidos induziu produção de IgA fecal e menos significativamente de IgG fecal. Existem indícios de indução de tolerância em nível intestinal, com queda da produção de anticorpos e da proliferação celular antígeno dirigida, especialmente quando o leite foi utilizado como adjuvante, em ensaios de produção in vitro de anticorpos ou proliferação estimulada por antígenos de T.gondii, por esplenócitos e linfócitos intestinais. As preparações orais induziram proteção quantitativa ao desafio dos animais imunizados por cepa cistogênica, que foi semelhante a imunização parenteral, quando o hidróxido de alumínio foi usado como adjuvante. Todos estes dados mostram a possibilidade de desenvolvimento de uma vacina oral para toxoplasmose utilizando taquizoítos irradiados, com aplicação prática num futuro próximo para uso em campo, utilizando iscas atrativas para imunizar felinos domésticos e selvagens. / We study the oral route for the development of a vaccine for toxoplasmosis, using parasites irradiated with 60 Cobalt, as an alternative for vaccine development to this worldwide parasitic infection. We evaluated the development of immunity at serum or mucosal levels, and their efficiency in protect the mice against challenge with oral cysts of the ME-49 strain. C57Bl/6j isogenic mice were immunized by oral route with 107 255 Gy irradiated tachyzoites from RH strain, at several protocols using milk as anti-peptic adjuvant and alum hydroxide as antacid. The preparations of irradiated tachyzoites induced production of serum IgG and IgA in immunized mice, as determined by ELISA, with IgG2a as the dominant subclass, similar to chronic infection. Their use with adjuvant allowed the excretion of significant amounts of IgA in stools also IgG, despite a lesser extent. There are suggestion of tolerance induction at mucosal level, with lower antigen induced proliferation and lower in vitro antibody production by spleen and gut lymphocytes, with the latter doses, specially when milk was used as adjuvant. All oral preparations induced some quantitative protection against challenge, which was similar to the parenteral route only isolated alum hydroxide was used as adjuvant. All these data support the possibility of the development of an oral vaccine against toxoplasmosis, using irradiated tachyzoites, which would be possible tool in near future for use in field baits, for immunizing either domestic or wild felids.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-03T19:19:19Z No. of bitstreams: 1 2015 - Marina Jorge de Lemos.pdf: 6290720 bytes, checksum: 466915088ab49af16fcd11435ccc2913 (MD5) / Made available in DSpace on 2017-05-03T19:19:19Z (GMT). No. of bitstreams: 1 2015 - Marina Jorge de Lemos.pdf: 6290720 bytes, checksum: 466915088ab49af16fcd11435ccc2913 (MD5) Previous issue date: 2015-12-08 / CAPES / An experiment was conducted in order to evaluate the intestinal morphology, performance, egg quality and analysis of the financial feasibility of including various additives in feed during the whole production period of Japanese quails. 360 quails were distributed in a completely randomized design in a time split plot scheme (9 to 23 weeks of age and 24 to 39 weeks old) with five treatments and eight repetitions of nine birds each. The used treatments were: control, antibiotic, prebiotic, probiotic and synbiotic. The intestinal morphology was evaluated by the height and width and width/height ratio of the villus, crypt depth and villus/crypt ratio; performance, by the feed intake, egg production, egg weight average, feed conversion (kg / kg and kg / dozen), egg mass and viability; egg quality, by the Haugh unit, yolk index, percentage of egg components, shell thickness measured by the micrometer and the scanning electron microscope (SEM); financial analysis was computed by the internal rate of return, net present value and cost benefit. The inclusion of additives in the feed increased the height and width of the villi, decreased crypt depth and increased villous / crypt ratio compared to the control. The inclusion of the additives reduced the feed intake in the two periods of production of the quails in comparison with the control. Throughout the production period, consumption was lower after inclusion of antibiotic and synbiotic in the feed. Egg production, egg weight average, egg mass and feed conversion (kg / kg and kg / dozen) were improved in the quails fed with additives in the two periods of production. The supply of additives within 24 to 39 weeks of age increased the Haugh unit of the eggs compared to the control. The inclusion of additives in the feed increased the yolk and shell percentage, and improved shell and membrane thickness measured by the micrometer compared to the control in both periods of production. The shell thickness (SEM) and the shell palisade layer were improved after inclusion of antibiotic and synbiotic in the feed within 24 to 39 weeks of age. The financial analysis showed best values in the treatments that used antibiotics and synbiotic. The inclusion of antibiotics, probiotic, prebiotic and synbiotic in the feed improves performance and egg quality throughout the production phase of Japanese quails, and these improvements increase effects when the birds receive the additives in the first productive period. The inclusion of antibiotics and synbiotic is more effective than the inclusion of the other additives in reducing food intake throughout the production period and improving shell thickness during the second production period. Intestinal integrity of Japanese quails improved after the inclusion of the tested additives. The financial analysis proved to be more viable to include antibiotic or synbiotic in the feed of Japanese quails in comparison to the other studied additives. / Foi conduzido um experimento com o objetivo de avaliar a morfologia intestinal, o desempenho, a qualidade dos ovos e an?lise da viabilidade financeira da inclus?o de diferentes aditivos na ra??o durante todo o per?odo produtivo de codornas japonesas. Foram utilizadas 360 codornas distribu?das em um delineamento inteiramente casualizado em esquema de parcela subdividida no tempo (de 9 at? 23 semanas de idade e de 24 at? 39 semanas de idade) com cinco tratamentos e oito repeti??es de nove aves cada. Os tratamentos utilizados foram: controle, antibi?tico, prebi?tico, probi?tico e simbi?tico. A morfologia intestinal foi avaliada atrav?s da altura e largura e propor??o largura/altura das vilosidades, profundidade de cripta e rela??o vilo: cripta; o desempenho pelo consumo de ra??o, produ??o de ovos, peso m?dio dos ovos, convers?o alimentar (kg/kg e kg/d?zia), massa dos ovos e viabilidade; a qualidade de ovos pela unidade Haugh, ?ndice de gema, percentagem dos componentes do ovo, espessura de casca medida pelo micr?metro e pelo microsc?pio eletr?nico de varredura (MEV); a an?lise financeira foi computada pela taxa interna de retorno, valor presente l?quido e benef?cio custo. A inclus?o dos aditivos na ra??o aumentou a altura e largura das vilosidades, diminuiu a profundidade de cripta e aumentou a rela??o vilo:cripta em compara??o com o controle. A inclus?o dos aditivos reduziu o consumo de ra??o nos dois per?odos de produ??o das codornas em compara??o com o controle. Durante todo o per?odo produtivo, o consumo foi menor ap?s inclus?o de antibi?tico e simbi?tico na ra??o. A produ??o de ovos, o peso m?dio dos ovos, a massa de ovos e a convers?o alimentar (kg/kg e kg/d?zia) foram melhores nas codornas alimentadas com os aditivos nos dois per?odos de produ??o. O fornecimento dos aditivos no per?odo de 24 at? 39 semanas de idade aumentou a unidade Haugh dos ovos em rela??o ao controle. A inclus?o dos aditivos na ra??o aumentou a porcentagem de gema e de casca, e melhorou a espessura da casca avaliada pelo micr?metro e das membranas da casca em compara??o com o controle nos dois per?odos de produ??o. A espessura da casca (MEV) e da camada pali?ada da casca foi melhor ap?s inclus?o de antibi?tico e simbi?tico na ra??o no per?odo de 24 at? 39 semanas de idade. A an?lise financeira mostrou os melhores valores nos tratamentos que utilizaram antibi?tico e simbi?tico. A inclus?o de antibi?tico, probi?tico, prebi?tico e simbi?tico na ra??o melhora o desempenho e a qualidade dos ovos durante toda a fase produtiva de codornas japonesas, sendo esse efeito potencializado quando as aves recebem os aditivos no primeiro per?odo produtivo. A inclus?o de antibi?tico e simbi?tico ? mais eficaz que a inclus?o dos outros aditivos na redu??o do consumo de ra??o em todo per?odo produtivo e em melhorar a espessura da casca durante o segundo per?odo produtivo. A integridade intestinal das codornas japonesas melhora ap?s a inclus?o dos aditivos testados. A an?lise financeira demonstrou ser mais vi?vel a inclus?o de antibi?tico ou de simbi?tico na ra??o das codornas japonesas em compara??o com os outros aditivos estudados.

Com o intuito de verificar a eficácia do açúcar e de edulcorante a base de sacarina sódica e neoesperidina nas dietas de leitões, foram realizados dois experimentos, ambos com 42 leitões recém-desmamados aos 21 dias de idade. Foram oferecidas três dietas: dieta controle (DC), dieta com açúcar (DA) e dieta com edulcorante (DE). Os experimentos foram em blocos causalizados, sendo o primeiro com sete repetições e dois animais por unidade experimental e o segundo com seis repetições, um animal por unidade experimental e arranjo fatorial 3 x 2 + 1 (três dietas, duas épocas de abate: aos 28 e 35 dias de idade e o abate no dia do desmame). No primeiro experimento, os animais permaneceram nas instalações de creche até os 63 dias de idade para avaliação do desempenho, da incidência de diarreia e da viabilidade econômica das dietas. No segundo experimento, os animais foram abatidos para se proceder à pesagem de órgãos, medição do comprimento do intestino delgado e coleta dos segmentos do duodeno e jejuno para realização da morfometria intestinal. Não houve diferenças entre as dietas para o CDR, GDP e CA nos Períodos 1 (21 aos 28 dias), 3 (21 aos 50 dias) e 4 (21 aos 63 dias de idade). No Período 2 (21 aos 35 dias), os animais que consumiram a DE apresentaram maior (P&lt;0,05) CDR e GDP, se comparados aos que consumiram a DA. Nos Períodos 1, 2 e 3 foi observado menor custo da dieta por kg de peso vivo ganho e melhores índices econômico e de custo para os animais que consumiram DE. Ao avaliar a morfometria intestinal e peso de órgãos, não se observou diferença entre as dietas, com exceção para o peso do estômago, que foi maior nos animais que consumiram DCE. A inclusão de edulcorante é eficaz em melhorar o desempenho no Período 2, além de ser viável sob o ponto de vista econômico. / In order to verify the effectiveness of sugar and sweetener containing saccharin and neohesperidin in diets of piglets, two experiments were conducted, both with 42 newborn piglets weaned at 21 days old. Were offered three diets: control diet (DC), diet with sugar (DA) and diet with sweetener (DE). The experiments were in randomized blocks, the first of which had seven replicates and two animals per experimental unit and the second, six replicates and one animal per experimental unit comprising a factorial 3 x 2 + 1 (three diets, two slaughter ages: at 28 and 35 days of age and slaughter on the day of weaning). In the first experiment, the animals remained on the nursery facilities until 63 days of age to evaluate the performance, the incidence of diarrhea and the economic viability of the diets. In the second experiment, the animals were slaughtered to weigh the organs, measure the length of the small intestine and collecting segments of the duodenum and jejunum to determine intestinal morphology. There were no differences between diets for the ADFI, FC and ADG in the Periods 1 (21 to 28 days), 3 (21 to 50 days) and 4 (21 to 63 days old). In the Period 2 (21 to 35 days), animals that consumed the DE had higher (P&lt;0.05) ADFI and ADG, compared to those fed the DA. In Periods 1, 2 and 3 was observed lower cost per kg of the diet of live weight gain and better economic indicators to the animals fed DE. No differences were observed between the diets when evaluating intestinal morphology and organ weights, except for the weight of the stomach, which was higher in animals fed DE. The inclusion of sweetener is effective in improving performance in Period 2, and is viable under the economical point of view.