The study includes separate papers, which are all linked by their emphasis on the effect of different diets on the impact of the development of the small intestine of the young ostrich. This abstract is intended to provide readers with a broad overview of the outcomes of the study. Part 1: Yolk utilisation and the development of the small intestines Chapter 2 deals with the composition of egg yolk as it is absorbed by starved ostrich Struthio camelus L. ) chicks from one to seven days post-hatching and for ostrich (Struthio camelus L.) chicks from one to sixteen days post-hatching on a pre-starter broiler diet. Chapter 3 provides information on the intestinal enzymes of ostrich (Struthio camelus L. ) chicks from one to sixteen days post-hatching on a pre-starter broiler diet. The effect of growth on enzyme activity immediately post-hatching up to sixteen days of age was studied. Chapter 4 details a histological and morphological study of the gastrointestinal tract of ostrich (Struthio camelus L.) chicks from two to sixteen days post-hatching on a pre-starter broiler diet. The effect of growth on histological and morphological changes in the various parts of the small intestines was examined. Part 2: Influence on various pre-starter diets on growth and the development of the intestinal tract Chapter 5 discussed a growth and digestibility study of ostrich (Struthio camelus L.) chicks on eight different pre-starter diets. The performance, growth and physiological development of the chicks on high and low variations of different nutritional components were examined. Chapter 6 provides information on certain intestinal enzymes of ostrich (Struthio camelus L.) chicks on the eight different pre-starter diets. Chapter 7 dealt with the histological and morphological changes of the gastrointestinal tract of ostrich (Struthio camelus L.) chicks as a result of high and low variations of different nutritional components in prestarter diets. Implications The first part of this study revealed that although it appeared that yolk content was absorbed faster in starved ostrich chicks, intake of external feed had a positive influence on the absorption of fat from the yolk. The changes in the fatty acid composition of the yolk fat content that were observed may indicate that ostrich chicks have the ability to withdraw certain fatty acid components, especially unsaturated fatty acids. The digestive tract of ostrich chicks was compared with that of broiler chicks and differences observed in amylase and lipase activity between the birds in this trial and in poultry could be due to genetic differences between species. It could also be due to the genetic homogeneity of the poultry chicks that were used in the trials, whereas ostrich chicks have not been subjected to the same extent of advanced breeding improvement to date. Results on histological and morphological growth of the small intestines of the ostrich chick indicate increased absorption with an increase in age. The second part of this study revealed that ostrich chicks performed differently on diets formulated with different nutritional components. It appears that a high fat and low sugar content in the diet is advantageous in ostrich pre-starter diets. Although there were a few mortalities during the trials, there is no clear indication as to why the chicks died, as the causes of mortalities were not investigated. Differences in composition of the eight pre-starter diets did not seem to have an influence on the protein content or enzyme activity of the small intestines of ostrich chicks. An anti-nutritional factor within certain feedstuffs may, however, have had an influence on enzyme activity. Similar results were obtained for the histological and morphological parameters which were measured in chicks fed the different diets. There was no single diet that promoted intestine development, although it seemed as if the small intestines of the chicks fed the high protein diet were under higher digestive stress than those of the chicks fed the other diets. / Thesis (PhD)--University of Pretoria, 2011. / Anatomy and Physiology / Unrestricted

This study investigated the effects of chronic stress on mucin expression in the GIT of mice. Sixteen ICR male mice were randomly divided into treated and control animals. Restraint stress was applied to the treated group for 21 days. Body weight (BW) changes and feed consumed (FC) were regularly recorded. After 21 days mice were euthanized and blood, GIT mucosa, and GIT sections were collected. Serum corticosterone levels were determined. The GIT sections were fixed, paraffin embedded, sectioned, and stained with AB-PAS to visualize goblet cells. Mucin expression was determined using qPCR. As expected stressed mice showed higher corticosterone than the control animals. BW decreased in the stressed group. Histology showed a decrease in goblet cells in stressed mice. Muc1 and Muc4 in the colon and Muc3 in the duodenum were upregulated in stressed animals. These findings suggest that chronic stress may affect mucin expression in the GIT.

Estimates for 2017 indicate that 20% of cancers globally are gastrointestinal tract (GIT) cancers, with oesophageal cancer being the 8th most common cancer. Oesophageal squamous cell carcinoma (OSCC) occurs in the upper to mid oesophagus and is present at high incidence in developing countries including South Africa. There are no early symptoms, resulting in late diagnosis and poor prognosis. In this study, tumour and blood DNA was obtained from 35 OSCC patients and subjected to whole genome sequencing (WGS). Bioinformatics analysis pipelines were designed to identify the possibility of novel viral insertions, investigating Human Endogenous Retroviruses (HERV's) insertions alongside the presence of somatic mutations in patient samples. The aims being to identify integration of any foreign DNA, to investigate if there is any linkage between HERV insertion and somatic mutations, and to identify any somatic mutations of potential interest in the OSCC cohort. The novel virus investigations however, proved to be inconclusive and there appeared to be no link between HERV insertions and somatic mutations present in the patients. Very significantly, it was determined that numerous somatic mutations were present in the MUC3A gene of the patient cohort, an interesting observation as no such previous association with OSCC has been recorded. MUC3A is a membrane-bound glycoprotein component of mucous gels, and its aberrant expression has been correlated with invasion and metastasis in a variety of other cancers. However, due to the complexity of the particular gene sequence and the known inconsistencies of variant calling performed on complex data sets, these mutations should be viewed with extreme caution as they are likely to be false positives. Analysis of RNA-seq data showed a 4.6 log2 fold increase in MUC3A expression in the tumour samples of these OSCC patients, with a P-adjusted value of 7.05e-06, suggesting highly significant differential gene expression. Functional enrichment analysis further showed that MUC3A was significantly associated with one of the top 5 gene ontologies (extracellular matrix structural constituent) for molecular function ontology class together with a number of collagen (COL) and MMP genes known to play a role in oncogenic progression and membrane stiffness. GSEA and KEGG analysis indicated predominantly chemokine/cytokine pro-inflammatory enriched pathways. Immunohistochemistry staining showed 10 out of 13 of the samples had no detectable levels of MUC3A protein, suggesting that the production of a non-functional truncated protein may lead to the upregulation of MUC3A expression that could possibly play a role in downstream pro-oncogenic signalling.

No description available.

O objetivo do presente trabalho foi avaliar o efeito da adição de inulina (I-X1), concentrado proteico de soro de leite (WC-X2) e/ou isolado proteico de soro de leite (WI-X3) em sorvete simbiótico de açaí (Euterpe oleracea Mart.) sobre a viabilidade do probiótico L. rhamnosus GG e a sua sobrevivência frente às condições encontradas no trato gastrintestinal simuladas in vitro, bem como sobre as características tecnológicas e a aceitabilidade sensorial dos produtos resultantes, ao longo de seu armazenamento a -18°C por até 112 dias. Adicionalmente, os sorvetes de açaí foram caracterizados quanto à sua composição centesimal, ácidos graxos insaturados e potencial antioxidante. Por último, objetivou-se a otimização do sorvete simbiótico de açaí a partir dos parâmetros de dureza instrumental e fração de derretimento. Para este fim, foi empregado um delineamento experimental para misturas simples (centroid simplex) incluindo um ponto axial, utilizando diferentes proporções dos fatores X1, X2 e X3. No total, 9 formulações foram avaliadas por um período de até 112 dias de armazenamento congelado. As populações de L. rhamnosus GG permaneceram estáveis e entre 8 e 9 log ufc/g para todas as formulações e ao longo de todo o período de armazenamento. Entretanto, houve redução em, pelo menos, 5 ciclos logaritmos da população do probiótico, após 6 horas de simulação gastrintestinal em todos os períodos estudados, com uma recuperação de L. rhamnosus GG próxima a 4 log ufc/g. Apenas ao 7&#186 dia de armazenamento, a resistência in vitro do probiótico nas formulações contendo inulina, WC e WI superou em até 0,93 log ufc/g a formulação controle. A composição de ácidos graxos revelou que o ácido oleico esteve em maior quantidade nas formulações (teor médio 46,47%), seguido do ácido palmitoleico (teor médio 42,12%). Os sorvetes apresentaram atividade antioxidante na faixa de 302,03 a 505,41 &#181mol TE/g. A maior atividade antioxidante foi observada para a formulação controle, seguida das formulações WC (4% de WC) e WI (4% de WI). As formulações WC (4% WC) e WI (4% WI) apresentaram os maiores valores de dureza, revelando que os fatores WC e WI exerceram forte influência na dureza dos produtos (p<0,05). As menores taxas de derretimento foram observadas nas formulações WI (4% de WI) e WC-WI (2% de WC; 2% de WI). Exceto para a formulação I-WC-WI (p<0,05), a presença de inulina nas demais formulações não alterou de forma significativa a velocidade de derretimento quando comprada à amostra controle. As formulações I-WI (2% de inulina; 2% de WI) e I-WC-WI (1,33% de inulina; 1,33% WC; 1,33 WI), apresentaram os maiores valores de overrun (37,95% e 39,18%, respectivamente). Os escores de aceitabilidade sensorial variaram de 5,83 a 7,63. Apenas as formulações WI (4% WI) e WC-WI (2% WC; 2% WI) aumentaram seus escores médios de aceitabilidade durante os 84 dias de armazenamento (p<0,05). De acordo com os modelos obtidos, WC foi o fator que mais contribuiu tanto para o aumento de dureza como para a velocidade de derretimento das formulações. A partir da otimização simultânea, considerando as respostas avaliadas (textura e fração de derretimento), é desejável que a formulação de sorvete simbiótico açaí seja produzida contendo 1,33% de inulina, 1,33% de WC e 1,33% de WI. Os resultados obtidos sugerem que a utilização dos três ingredientes (inulina, WC e WI) pode ser vantajosa no desenvolvimento de um sorvete probiótico de açaí, uma vez que a presença desses ingredientes contribuiu com o valor nutricional das formulações e também melhorou as características tecnológicas de overrun, derretimento e textura. / The aimed of the present study was to evaluate the effect of inulin (I-X1), whey protein concentrate (WC-X2) and whey protein isolate (WI-X3) addition on the viability and in vitro survival of L. rhamnosus GG through in vitro simulated gastrointestinal tract (GIT), technological properties and sensory acceptance of symbiotic açai ice cream. Additionally, açai ice cream were characterized for their chemical composition, unsaturated fatty acids and antioxidant activity. Finally, we aimed to optimized the symbiotic açai ice cream using parameters of hardness and melting rate. For this purpose, an experimental design was used for simple mixture (simplex centroid) including an axial point, using different proportions of X1, X2 and X3, nine formulations of synbiotic açai ice cream were studied during frozen storage up to 112 days. A reduction of at least 5 log cycles of L. rhamnosus GG was observed in all formulations after 6 hours of the in vitro assays for all storage period evaluated. However, L. rhamnosus GG recovery was close to 4 log cfu/g for all formulations studied in evaluated periods. Nevertheless, on the 7th day of storage, the probiotic in vitro resistance in formulations supplemented with inulin, WC, and WI exceeded up to 0.93 log cfu/g the control formulation. The fatty acids composition showed that oleic acid was the highest fatty acid in synbiotic açai ice cream (mean content 46.47%), followed by palmitoleic acid (mean content 42.12%). The antioxidant activity of ice creams ranged from 302.03 to 505.41 µmol TE/g. Formulation C showed the highest antioxidant activity, followed by the formulations WC (WC 4%) and WI (WI 4%). Formulations WC (4% of WC) and WI (4% of WI) showed the highest hardness values, revealing that WC and WI exerted a strong influence on the product firmness. The lowest melting rate values were observed in formulation WI (4% of WI) and WC-WI (2% of WC, 2% of WI). However, the presence of inulin and WC showed strong influence on the melting rate of the ice creams, resulting in higher melting down of these products. I-WI and I-WC-WI formulations presented the highest overrun (37.95 and 39.18%, respectively). The sensory acceptability scores ranged from 5.83 to 7.63. WI and WC-WI formulations increased their mean acceptability scores significantly during 84 days of storage, whereas no significant differences were observed in acceptability throughout storage for the other formulations. According to the obtained models, WC was the factor that most contributed to increase both hardness and melting rate. From the simultaneous optimization, considering the responses evaluated (hardness and melting rate), it is desirable a symbiotic acai ice cream containing 1.33% of inulin, 1.33% of WC and 1.33% of WI. The results suggest that the use of the three ingredients (inulin, WC and WI) may be advantageous in the development of a synbiotic açai ice cream, since the presence of these ingredients contributed to the nutritional value of the formulations and also improved the technological characteristics of overrun, melting rate and texture.

A distrofia muscular de Duchenne (DMD) é uma doença neuromuscular fatal, a mais comum das distrofias musculares, causada pela ausência da proteína distrofina, componente importante do complexo glicoproteínadistrofina, que envolve as células musculares lisas, cardíacas e esqueléticas, estabilizando a membrana dessas células durante contrações e relaxamentos. Por ser uma doença genética ligada ao cromossomo X, acomete indivíduos do sexo masculino, sendo que casos raros de meninas afetadas também são relatados. Indivíduos afetados pela DMD apresentam alterações significantes na composição corpórea, comparados com a população normal. Cães da raça golden retriever que possuem distrofia muscular (GRMD) representam o melhor modelo animal para ensaios terapêuticos na busca do tratamento da DMD, sendo esses animais fundamentais para o estudo do desenvolvimento dessa anomalia, devido ao fato de apresentarem genes e sintomas clínicos homólogos aos dos pacientes humanos. A similaridade entre as duas doenças é surpreendente, ambas são caracterizadas por precoce miopatia degenerativa, com progressiva necrose, fagocitose, deposição de cálcio e fibrose endomisial e perimisial. O foco principal deste trabalho foi esclarecer dúvidas quanto ao comportamento da musculatura lisa dos animais distróficos e portadores da GRMD, para se conhecer os efeitos da doença nesse tipo de tecido. Para avaliação da musculatura lisa uterina foram utilizados quatro úteros de cadelas golden retriever saudáveis, cinco úteros de portadoras do gene e quatro úteros de animais afetados pela GRMD. Analisando as fotomicrografias uterinas dos grupos, podemos notar que não há diferenças morfológicas através dos estudos histológicos convencionais. Porém, quando analisamos as fotomicrografias polarizadas , nota-se nas cadelas saudáveis exclusivamente colágeno do tipo III, de coloração esverdeada. Quanto às portadoras do gene da GRMD, observa-se colágeno predominantemente vermelho, do tipo I, e pouco em verde, tipo III. Fotomicrografias das fêmeas afetadas pela GRMD encontraram colágeno tipo III na parte interna do endométrio, miométrio e perimétrio, e colágeno tipo I em epitélio vascular, porém ambos em menor quantidade. É possível visualizar colágeno entre as fibras musculares através da luz polarizada. Para a avaliação da digestibilidade dos nutrientes em cães GRMD e sua capacidade absortiva, foram utilizados nove cães GRMD, sete fêmeas portadoras e dez animais saudáveis; todos receberam a mesma dieta adicionada de oxido crômico. O período experimental constou de um intervalo de 10 dias, sendo os cinco primeiros dias de adaptação e os cinco restantes de coleta de amostras. Os coeficientes de digestibilidade aparente (CDAs) obtidos pelos grupos estudados foram comparados e analisados: os resultados indicaram que os cães com GRMD avaliados não apresentaram redução do aproveitamento nutricional. Novas pesquisas que avaliem o gasto energético e composição corpórea destes animais são necessárias para melhor compreensão dos mecanismos envolvidos no emagrecimento e perda de peso que apresentam. Para a avaliação do tempo de trânsito gastrintestinal (TTGI) em cães golden retriever saudáveis, portadores e afetados pela distrofia muscular, foram utilizados 18 cães divididos em três grupos animais saudáveis, portadores e afetados pela GRMD, com seis cães cada. Os animais foram acondicionados em canis individuais e receberam a mesma dieta; esferas de polietileno (BIPS) impregnadas com bário foram fornecidas durante a alimentação. Cada animal recebeu 10 marcadores de tamanho grande (5 mm de diâmetro) e 30 marcadores de tamanho pequeno (1.5mm de diâmetro). Os animais foram radiografados nos momentos antes (basal) da 1ª refeição, para confirmação da ausência de conteúdo alimentar e fecal no trato gastrintestinal, e após a alimentação, em intervalos de duas horas, até que os marcadores atingissem o cólon. Resultados apontaram que os cães GRMD avaliados apresentam TTGI mais rápido quando comparados aos demais grupos. Este trabalho comprovou que a mm. lisa de cães distróficos é, de diversas maneiras, afetada pela doença, apresentando tanto alterações morfológicas como morfofuncionais. Muitos estudos nessa área são necessários para melhor compreensão dos mecanismos da enfermidade, bem como maneiras de intervenção para melhoria da qualidade de vida dos indivíduos afetados. Cães GRMD(s) são o melhor modelo animal para estudos pré-clínicos da DMD, assim esses resultados são muito relevantes para aprimoramento e melhora de manejo em biotérios que abrigam esses animais e para extrapolação de dados na própria medicina / Duchenne muscular dystrophy (DMD) is a fatal neuromuscular disease, it is the most common muscular dystrophy caused by absence of dystrophin protein, an important component of the dystrophin-glycoprotein complex, involving cell membranes of smooth, cardiac and skeletal cells, stabilizing these cells during contractions and relaxations. Because it is a genetic disorder linked to the X chromosome, it affects males, with rare cases of girls affected also reported. Individuals affected by DMD show significant changes in body composition, compared with the normal population. Golden retriever dogs who have muscular dystrophy (GRMD) represent the best animal model for therapeutic trials in the treatment of DMD,; these animals are fundamental to the study of the development of this anomaly, due to the fact that they show clinical symptoms and genes homologous to the human patients. The similarity between the two diseases is staggering; both are characterized by early degenerative myopathy with progressive necrosis, phagocytosis, calcium deposition and endomysial and perimysial fibrosis. The main focus of this thesis was to answer questions about the behavior of smooth muscles of dystrophic animals and carriers of GRMD, to know the effects of the disease in this tissue type. For evaluation of uterine smooth muscle we utilized four uteri of golden retriever healthy bitches, five uteri of gene carriers and four uteri of animals affected by GRMD. Analyzing the uterine photomicrographs of the groups we note that there are no morphological differences in conventional histological studies. However, when we analyze the polarized photomicrographs, we note that in healthy bitches we can see only collagen type III of greenish color. As regards the carriers of the GRMD gene, there is predominantly red collagen , type I, and very little green, type III. Photomicrographs of females affected by GRMD found type III collagen inside the endometrium, myometrium and perimetrium and collagen type I in vascular epithelium, but both in a very little quantity. It is possible to visualize collagen between muscle fibers by polarized light. For the assessment of nutrient digestibility in dogs GRMD and its absorptive capacity, we used nine GRMD, seven carriers females and ten healthy animals and all of them received the same diet with added chromic oxide. The experimental period consisted of an interval of 10 days, with the first five days to adaptation and the last ones to sample collection. The apparent digestibility coefficients obtained by the groups were compared and analyzed; the results showed that dogs with GRMD evaluated showed no reduction in energy utilization. The experimental period consisted of an interval of 10 days, with the first five days to adaptation and the last of them to sample collection. The apparent digestibility coefficients obtained by the groups were compared and analyzed; the results showed that dogs with GRMD evaluated showed no reduction in nutritional absorption. New research to assess energy expenditure and body composition in these animals are needed to a better understanding of the mechanisms involving reduction and weight loss they present. For the assessment of gastrointestinal transit time (TTGI) in golden retriever dogs healthy carriers and affected by GRMD 18 dogs divided into three groups were used - healthy animals, carriers and affected by GRMD, with six dogs each. The animals were placed in individual cages and were fed the same diet; polyethylene spheres (BIPS) impregnated with barium were supplied during feeding. Each animal received 10 markers large size (5mm diameter) and 30 markers small size (1.5mm diameter). Radiographs were taken in the moments before (baseline) the 1 st meal, to confirm the absence of food and fecal content in the gastrointestinal tract and after feeding at two-hour intervals until markers reach the colon. Results indicated that GRMD dogs evaluated showed TTGI faster when compared to the other groups. This thesis has shown that the smooth muscles of dystrophic dogs are, in many ways, affected by the disease, with both morphological and morphofunctional alterations; many studies in this area are needed so we can better understand the mechanisms of disease, as well as ways of intervention to improve the quality of life of affected individuals. GRMD dogs are the best animal model for preclinical studies of DMD, so these results are very relevant to enhancement and improvement of management in bioteries these animals have

O objetivo do presente trabalho foi avaliar o efeito da adição de inulina (I-X1), concentrado proteico de soro de leite (WC-X2) e/ou isolado proteico de soro de leite (WI-X3) em sorvete simbiótico de açaí (Euterpe oleracea Mart.) sobre a viabilidade do probiótico L. rhamnosus GG e a sua sobrevivência frente às condições encontradas no trato gastrintestinal simuladas in vitro, bem como sobre as características tecnológicas e a aceitabilidade sensorial dos produtos resultantes, ao longo de seu armazenamento a -18°C por até 112 dias. Adicionalmente, os sorvetes de açaí foram caracterizados quanto à sua composição centesimal, ácidos graxos insaturados e potencial antioxidante. Por último, objetivou-se a otimização do sorvete simbiótico de açaí a partir dos parâmetros de dureza instrumental e fração de derretimento. Para este fim, foi empregado um delineamento experimental para misturas simples (centroid simplex) incluindo um ponto axial, utilizando diferentes proporções dos fatores X1, X2 e X3. No total, 9 formulações foram avaliadas por um período de até 112 dias de armazenamento congelado. As populações de L. rhamnosus GG permaneceram estáveis e entre 8 e 9 log ufc/g para todas as formulações e ao longo de todo o período de armazenamento. Entretanto, houve redução em, pelo menos, 5 ciclos logaritmos da população do probiótico, após 6 horas de simulação gastrintestinal em todos os períodos estudados, com uma recuperação de L. rhamnosus GG próxima a 4 log ufc/g. Apenas ao 7&#186 dia de armazenamento, a resistência in vitro do probiótico nas formulações contendo inulina, WC e WI superou em até 0,93 log ufc/g a formulação controle. A composição de ácidos graxos revelou que o ácido oleico esteve em maior quantidade nas formulações (teor médio 46,47%), seguido do ácido palmitoleico (teor médio 42,12%). Os sorvetes apresentaram atividade antioxidante na faixa de 302,03 a 505,41 &#181mol TE/g. A maior atividade antioxidante foi observada para a formulação controle, seguida das formulações WC (4% de WC) e WI (4% de WI). As formulações WC (4% WC) e WI (4% WI) apresentaram os maiores valores de dureza, revelando que os fatores WC e WI exerceram forte influência na dureza dos produtos (p<0,05). As menores taxas de derretimento foram observadas nas formulações WI (4% de WI) e WC-WI (2% de WC; 2% de WI). Exceto para a formulação I-WC-WI (p<0,05), a presença de inulina nas demais formulações não alterou de forma significativa a velocidade de derretimento quando comprada à amostra controle. As formulações I-WI (2% de inulina; 2% de WI) e I-WC-WI (1,33% de inulina; 1,33% WC; 1,33 WI), apresentaram os maiores valores de overrun (37,95% e 39,18%, respectivamente). Os escores de aceitabilidade sensorial variaram de 5,83 a 7,63. Apenas as formulações WI (4% WI) e WC-WI (2% WC; 2% WI) aumentaram seus escores médios de aceitabilidade durante os 84 dias de armazenamento (p<0,05). De acordo com os modelos obtidos, WC foi o fator que mais contribuiu tanto para o aumento de dureza como para a velocidade de derretimento das formulações. A partir da otimização simultânea, considerando as respostas avaliadas (textura e fração de derretimento), é desejável que a formulação de sorvete simbiótico açaí seja produzida contendo 1,33% de inulina, 1,33% de WC e 1,33% de WI. Os resultados obtidos sugerem que a utilização dos três ingredientes (inulina, WC e WI) pode ser vantajosa no desenvolvimento de um sorvete probiótico de açaí, uma vez que a presença desses ingredientes contribuiu com o valor nutricional das formulações e também melhorou as características tecnológicas de overrun, derretimento e textura. / The aimed of the present study was to evaluate the effect of inulin (I-X1), whey protein concentrate (WC-X2) and whey protein isolate (WI-X3) addition on the viability and in vitro survival of L. rhamnosus GG through in vitro simulated gastrointestinal tract (GIT), technological properties and sensory acceptance of symbiotic açai ice cream. Additionally, açai ice cream were characterized for their chemical composition, unsaturated fatty acids and antioxidant activity. Finally, we aimed to optimized the symbiotic açai ice cream using parameters of hardness and melting rate. For this purpose, an experimental design was used for simple mixture (simplex centroid) including an axial point, using different proportions of X1, X2 and X3, nine formulations of synbiotic açai ice cream were studied during frozen storage up to 112 days. A reduction of at least 5 log cycles of L. rhamnosus GG was observed in all formulations after 6 hours of the in vitro assays for all storage period evaluated. However, L. rhamnosus GG recovery was close to 4 log cfu/g for all formulations studied in evaluated periods. Nevertheless, on the 7th day of storage, the probiotic in vitro resistance in formulations supplemented with inulin, WC, and WI exceeded up to 0.93 log cfu/g the control formulation. The fatty acids composition showed that oleic acid was the highest fatty acid in synbiotic açai ice cream (mean content 46.47%), followed by palmitoleic acid (mean content 42.12%). The antioxidant activity of ice creams ranged from 302.03 to 505.41 µmol TE/g. Formulation C showed the highest antioxidant activity, followed by the formulations WC (WC 4%) and WI (WI 4%). Formulations WC (4% of WC) and WI (4% of WI) showed the highest hardness values, revealing that WC and WI exerted a strong influence on the product firmness. The lowest melting rate values were observed in formulation WI (4% of WI) and WC-WI (2% of WC, 2% of WI). However, the presence of inulin and WC showed strong influence on the melting rate of the ice creams, resulting in higher melting down of these products. I-WI and I-WC-WI formulations presented the highest overrun (37.95 and 39.18%, respectively). The sensory acceptability scores ranged from 5.83 to 7.63. WI and WC-WI formulations increased their mean acceptability scores significantly during 84 days of storage, whereas no significant differences were observed in acceptability throughout storage for the other formulations. According to the obtained models, WC was the factor that most contributed to increase both hardness and melting rate. From the simultaneous optimization, considering the responses evaluated (hardness and melting rate), it is desirable a symbiotic acai ice cream containing 1.33% of inulin, 1.33% of WC and 1.33% of WI. The results suggest that the use of the three ingredients (inulin, WC and WI) may be advantageous in the development of a synbiotic açai ice cream, since the presence of these ingredients contributed to the nutritional value of the formulations and also improved the technological characteristics of overrun, melting rate and texture.

O comprometimento do trato gastrintestinal pela endometriose em 40 pacientes com endometriose pélvica foi avaliado pelo método da COPE. A coorte estudada compreendeu 21 pacientes com sinais e sintomas gastrintestinais e 19 pacientes sem sinais e sintomas gastrintestinais, visando a estabelecer: 1. associações e correlações entre os parâmetros clínicos que sinalizam a presença de endometriose e as localizações das lesões em cada segmento do trato gastrintestinal; 2. correlação entre o estadiamento da endometriose (ASRM, 1996) e o comprometimento gastrintestinal e 3. correlação entre a classificação histológica da endometriose e o comprometimento gastrintestinal. Através da COPE, o diagnóstico e as correlações entre sinais e sintomas ginecológicos, ultra-sonografia vaginal, classificação da ASRM e histologia, com as características distributivas da doença no trato gastrintestinal, demonstraram que: 1. A idade foi significativamente mais elevada nas pacientes com sinais e sintomas no TGI; 2. A detecção de lesões no TGI ocorreu em 70% das pacientes; 3. A dismenorréia em intensidade severa ou incapacitante e dispareunia em intensidade severa correlacionaram-se com a endometriose ginecológica e a endometriose do trato gastrintestinal na presença de sinais e sintomas no TGI; 4. Os sinais e sintomas gastrintestinais correlacionados com a endometriose ginecológica e do TGI, foram o puxo e o tenesmo cíclico, a dor em cólica cíclica, a obstipação cíclica, a diarréia cíclica, a dor pélvica acíclica, as fezes afiladas e o sangramento intestinal cíclico; 5. A endometriose que provoca sinais e sintomas no TGI está localizada no segmento retossigmóide e/ou no íleo. A dispareunia e a dismenorréia apontam para o acometimento do íleo. A infertilidade sinaliza para a endometriose no apêndice; 6. Nas correlações do toque vaginal com os achados da COPE, o aumento anexial correlaciona-se com retossigmóide e íleo, o espessamento ou nódulo ligamentar, correlaciona-se com o reto e a presença de nódulo no fundo de saco ou lesão no septo reto-vaginal, sinaliza para o acometimento do cólon sigmóide; 7. A COPE aplicada a pacientes com suspeita de endometriose no segmento retossigmóide levantada pela ultra-sonografia transvaginal mostra que a doença se estende em associação significante ao reto, ao cólon sigmóide, cólon ascendente e íleo; 8. A COPE demonstrou que a endometriose acomete mais freqüentemente o íleo, o apêndice, o segmento retossigmoideano e o cólon ascendente na existência ou não de sinais e sintomas do TGI; 9. Os padrões histológicos distribuíram-se igualmente pelas lesões endometrióticas do TGI exceto nas lesões do mesentério; 10. O estádio IV da ASRM, 1996, é fator de risco significativo para o acometimento do reto, sigmóide e íleo. / The importance of the laparoscopic procedure padronization to the run in the peritoneal organs was established in 40 patients with pelvic endometriosis through the observation of its efficient in the characterization of the gastrointestinal tract endometriosis distribution and in the establishment of the meaningful correlations among the most important aspects of the gynecological clinic, vaginal ultrasonagraphic exam, ASRM and histology with gastrointestinal signs and symptoms. The run in the peritoneal organs in the endometriosis has permitted the diagnosis of the endometriosis disease real extension if compared to the competing methods previously described in the pertinent literature

This thesis describes investigations into the genetic basis and pathophysiology of three distinct inherited diseases in humans, two of which are strongly associated to the function of the ectodomain sheddase enzyme ADAM17. The first of these is a novel inherited syndrome of neonatal onset inflammatory skin and bowel disease, which is associated in a consanguineous family with homozygous loss-offunction mutations in ADAM17. This thesis describes investigations of the expression and function of ADAM17 – and downstream proteins it regulates – in an individual affected by this disease. This is accompanied by genetic investigations into other individuals suspected of suffering from the same syndrome. The second investigated disease is Tylosis with Oesophageal Cancer (TOC), an inherited cutaneous disease which represents the only known syndrome of familial oesophageal cancer susceptibility. This disease was associated to dominantly inherited mutations in the Rhomboid protein iRHOM2. This work describes investigations of immortalised keratinocyte cell lines and tissues derived from TOC-affected individuals, and illustrates that the pathogenesis of TOC is characterised by increased iRHOM2-dependent activation and activity of ADAM17, and upregulation of the shedding of ADAM17 substrates, particularly in the EGFR ligand family, accompanied by increased desmosome turnover and transglutaminase 1 activity. This pattern of upregulation results in attendant increases in growth factor signalling, proliferation and motility in TOC keratinocytes, dependent on ADAM17. The third focus of this thesis is a life-threatening inherited gastrointestinal disease (accompanied by severe extraintestinal complications) whose symptoms correspond to Cryptogenic Multifocal Ulcerative Stenosing Enteritis. This work describes the identification of mutations in cytosolic phospholipase A2-α (cPLA2α) – an enzyme responsible for arachidonic acid production, the first step in the eicosanoid synthesis pathway – as associated with this condition in a single affected family. The expression and function of cPLA2α in this disease was investigated, using platelet aggregation stimulated by a downstream product of cPLA2α (Thromboxane A2) as a model.

Mucins are heavily glycosylated epithelial proteins. Under or overexpression of mucins may lead to several different types of dysfunctions. Mucins are under investigation as possible diagnostic markers for malignancies and other disease. This study examined the over/under expression of Mucin-3 in the presence of IP6. Three groups of six mice were fed varying levels of phytate (inositol 6-phosphate, IP6) for six days before being subjected to carbon dioxide asphyxiation. Sections of the duodenum, jejunum, ileum, stomach, and colon were collected, lysed, and the tissue proteins were prepared and analyzed by Western Blot to determine Mucin-3 expression. The hypothesis was that the presence of phytate in the digestive tract would modulate the expression of Mucin 3. Results showed that on average, Mucin 3 expression in untreated tissue was higher in areas of the duodenum and jejunum and lower in the ileum and stomach. Presence of IP6 decreased expression of Mucin 3 in the stomach by one percent in the 1g/kg phytate group while increasing in the 2g/kg phytate group by one percent. The expression of Mucin 3 in the duodenum increased fifty-five and thirty-nine percent in the 1g/kg and 2g/kg phytate groups, respectively. Jejunum Mucin 3 expression was increased by ninety percent in both 1g/kg and 2g/kg phytate groups. The ileum 0g/kg and 2g/kg phytate groups had the same expression of Mucin 3, but the 1g/kg group had a thirty-six percent decrease.