München, Techn. Universiẗat, Diss., 2001.

Tierärztl. Hochsch., Diss., 2003--Hannover.

Les patients atteints de MICI (maladie de Crohn et recto-colite hémorragique) présentent souvent des lésions des cellules de l'épithélium intestinal. La rémission de ces maladies nécessite à la fois un arrêt de l'inflammation et une migration des entérocytes pour réparer les dommages épithéliaux. Cette migration cellulaire appelée restitution intestinale requiert des adhérences cellule-MEC et cellule-cellule réalisées par les complexes protéiques associés aux intégrines et cadhérines. Le but de cette thèse a été d'étudier l'impact deSaccharomyces boulardii (Sb) sur la réparation de l'épithélium intestinal lésé. Nous avons montré que le surnageant de Sb contenait des facteurs modulant la restitution de l'épithélium intestinal in vivo et in vitro sans affecter la prolifération des cellules épithéliales. Ces effets motogéniques du surnageant de Sb s'exercent via la modulation des molécules d'adhérence. En effet, le surnageant de Sb augmente l'affinité de l'intégrine α2β1 pour son ligand le collagène de type I mais entre en compétition avec les intégrine αvβ5, pour inhiber l'adhérence des entérocytes sur la vitronectine. Ces modifications de l'adhérence avec la matrice extracellulaire entraînent une régulation des voies de signalisation émanant des intégrines et une réorganisation des plaquesd'adhérence. Ces évènements vont accroître la migration des entérocytes. De plus, nos résultats préliminaires portant sur Sb et l'adhérence cellule-cellule durant la restitution intestinale ont montré une implication de la E-cadhérine dans la migration induite par Sb. / Intestinal epithelial cell damage is frequently seen in IBD patients with ulcerative colitis or Crohn's disease. The remission of these diseases requires both the cessation of inflammation and the migration of enterocytes torepair the damaged epithelium. Adhesions with the ECM and the adjacent cells using complex of proteins associated with integrins and cadherins are necessary for this cell migration called intestinal restitution. Theaim of this thesis was to study the effect of S.boulardii on the resealing of a wound in intestinal epithelia. First of all, we demonstrated that the supernatant of S.boulardii contains factors that modulate intestinal epithelial cell restitution both in vitro and in vivo without affecting cell proliferation. We showed that the motogenic factors of S.boulardii act by modulating adhesion molecules. Indeed, the supernatant of S.boulardii increase the the affinity between 21 and it ligand the collagen type I, but also compete with integrin v5 to block theadhesion of enterocytes on vitronectin. These modifications of adhesion on extracellular matrix lead to aregulation of signaling pathway mediated by integrins, and a reorganization of focal adhesions. These eventscontribute to an increase of the migration of enterocytes. Add to this, our preliminaries results on S.boulardiiand cell-cell adhesion during intestinal restitution show an involvement of E-cadherin in the migrationS.boulardii-induced. With this work, we have demonstrated that heat-sensitive motogenic factors secreted by S.boulardii can enhance intestinal restitution with a dynamic regulation of adhesion between integrin and the ECM.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-08-29T13:41:36Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_giana_carla_gaboardi.pdf: 846790 bytes, checksum: 476ac8accf8427c699effbc54192a57d (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-08-29T19:50:59Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_giana_carla_gaboardi.pdf: 846790 bytes, checksum: 476ac8accf8427c699effbc54192a57d (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-08-29T19:51:06Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_giana_carla_gaboardi.pdf: 846790 bytes, checksum: 476ac8accf8427c699effbc54192a57d (MD5) / Made available in DSpace on 2017-08-29T19:51:13Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_giana_carla_gaboardi.pdf: 846790 bytes, checksum: 476ac8accf8427c699effbc54192a57d (MD5) Previous issue date: 2015-01-15 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O Brasil produziu 12,2 milhões de toneladas de arroz em casca na safra 2013/2014, sendo a região sul responsável por 60% da produção. O arroz parboilizado representa 50% das exportações nacionais de arroz e aproximadamente 20% do arroz total comercializado no Brasil. No processo de parboilização, são gerados 2 L de efluente para cada quilograma de grão processado, gerando grandes volumes de efluente que contém nutrientes, como nitrogênio e fósforo, e matéria orgânica. As metodologias de tratamento utilizadas para o tratamento deste efluente são as lagoas aeróbias e anaeróbias, lodos ativados, precipitação química e reatores UASB. O cultivo de leveduras em efluentes tem sido empregado como forma de tratamento, gerando simultaneamente biomassa que pode ser usada como probiótico ou fonte de proteína unicelular. A levedura Saccharomyces boulardii é capaz de crescer em efluente de arroz parboilizado quando este é suplementado com uma fonte extra de carbono. O objetivo deste estudo foi avaliar o uso de Saccharomyces boulardii no tratamento do efluente de arroz parboilizado e a produção de biomassa. Realizaram-se ensaios para a seleção de um meio de cultivo de Saccharomyces boulardii, testando a adição de sacarose ou glicerol de biodiesel ao efluente da parboilização de arroz, coletado em uma indústria de beneficiamento de arroz. Os melhores resultados obtidos foram no cultivo em efluente suplementado com 1% de sacarose, que gerou viabilidade celular de 2,4 x 108 UFC.mL-1 e a remoção de 72% do fósforo, 50% do nitrogênio (N-NTK) e 56% da demanda química de oxigênio (DQO) em 24 h de cultivo. A partir do meio de cultivo selecionado, analisou-se em biorreator o crescimento e a produção de biomassa da levedura e sua influência na redução de parâmetros ambientais. Após 48 h de cultivo, foi produzido 3,8 g.L-1 de biomassa e 1,8 x 10 11 UFC. L-1. A eficiência de remoção foi de 74% da DQO e 78% do fósforo, alcançando as taxas de remoção de fósforo estabelecidas pelo Conselho Estadual do Meio Ambiente (CONSEMA 128/06). Além disso, a biomassa seca produzida e a viabilidade celular alcançada permitiriam obter probiótico suficiente para adicionar a 36 toneladas de ração a cada tonelada de grão processado. Estes resultados ressaltam os benefícios do cultivo de Saccharomyces boulardii em efluente de arroz parboilizado suplementado com sacarose como sistema de tratamento e produção de probiótico. / Brazil produced 12.2 million tons of paddy rice in 2013/2014, and the south region was responsible for 60% of the production. The parboiled rice represents 50% of the national rice exports and about 20% of the total rice sold in Brazil. During the parboiling process are generated 2 liters of effluent per kilo of processed grain, generating large volumes of effluent which contains nutrients such as nitrogen and phosphorus, and organic matter. The treatment methodologies used for the treatment of this effluent are aerobic and anaerobic lagoons, activated sludge, chemical precipitation and UASB reactors. Cultivation of yeasts in wastewater has been used as a treatment technique while generating biomass that can be used as probiotic or unicellular protein source. The yeast Saccharomyces boulardii is able to grow in parboiled rice effluent when it is supplemented with extra carbon source. The aim of this study was to evaluate the use of Saccharomyces boulardii on treatment of parboiled rice effluent and biomass production. Trials were carried out for the medium selection to Saccharomyces boulardii cultivation, testing the addition of sucrose or glycerol by-product of biodiesel industry into parboiled rice effluent, collected in a rice processing industry. The best results obtained were on cultivation in effluent supplemented with 1% sucrose, generating 2.4 x 108 CFU.mL-1 of cell viability and phosphorus, nitrogen and COD removal of 72%, 50% and 56%, respectively, after 24 h of culture. After medium selection, the cell growth, yeast biomass production and its influence on the reduction of environmental parameters was analyzed in bioreactor. After 48 h of culture, it was produced 3.8 g L-1 of biomass and 1.8 x 10 11 CFU. L-1. The removal efficiency was 74% for COD and 78% for phosphorus, reaching the phosphorus removal rates required by Conselho Estadual do Meio Ambiente (CONSEMA 128/06). Furthermore, the produced dry yeast biomass and cell viability reached would allow to obtain sufficient probiotics to add to 36 tons of feed for each ton of processed grain. These results show the advantages of Saccharomyces boulardii cultivation in parboiled rice effluent supplemented with sucrose as a treatment system and production of probiotic.

Dans de nombreuses pathologies digestives dont les maladies inflammatoires chroniques intestinales (MICI), l'intégrité de la barrière épithéliale est rompue. Cette perte d'intégrité est notamment due à la réduction ou la perte d'expression des jonctions adhérentes composées du complexe E-cadhérine/caténines. Il est donc important d'identifier de nouvelles molécules capables de réguler ce complexe dans les MICI. C'est dans ce contexte que nous nous sommes intéréssés à une levure non pathogène, Saccharomyces boulardii (Sb) utilisée dans la prévention et le traitement de désordres gastro-intestinaux et qui présente des bénéfices thérapeutiques chez les patients atteints de MICI, notamment en régulant l'intégrité de la barrière intestinale. L'étude que nous avons réalisée sur des explants tissulaires provenant de patients atteints de MICI nous a permis de mettre en évidence que le surnageant de Sb (Sbs) protège la morphologie tissulaire et maintient l'expression de la E-cadhérine à la membrane. In vitro nous avons également pu montrer que Sbs accélère la ré-expression de la E-cadhérine à la membrane en régulant son recyclage par les endosomes (Rab11A), entrainant ainsi la restauration et le renforcement de la barrière épithéliale intestinale. / Some intestinal pathologies including inflammatory bowel disease (IBD) are associated with an altered barrier function. The reduction or the lost of adherens junctions composed by E-cadherin/catenins complex are linked to changes in the barrier integrity. Characterization of molecules targeting the E-cadherin/catenins complex during IBD is crucial for the development of alternative therapies. From this perspective, we focus ours studies on a non pathogenic yeast, Saccharomyces boulardii, used to prevent and treat gastro-intestinal disorders and may have beneficial effects in IBD treatment, including the regulation of barrier integrity. Ours studies on colonic explants from IBD patients showed that Sb supernatant (Sbs) protects epithelial morphology and maintains E-cadherin expression at the cell surface. In vitro study pointed out that Sbs accelerated the recovery of E-cadherin at the cell membrane. This process involved the modulation of the recycling of E-cadherin by endosomes (Rab11A), leading to restoration and strengthening of intestinal barrier function.

Zugl.: Bonn, Univ., Diss., 2009

Penicillium roqueforti é um fungo deteriorante que ocasiona perdas principalmente em queijo parmesão, devido alterações no sabor e na cor do produto final. O controle biológico com leveduras do gênero Saccharomyces tem capacidade antifúngica e algumas têm efeito probiótico. Desta forma, o objetivo do presente trabalho foi utilizar leveduras como biocontrole do fungo P. roqueforti em queijo parmesão. Inicialmente, realizou análises in vitro para verificar a capacidade das leveduras S. cerevisiae UFT 5962, UFT 5992, UFT 5976, YEF 186 e S. boulardii em reduzir a quantidade de esporos produzidos por P. roqueforti e como também investigar o potencial de substâncias GRAS (bicarbonato de sódio, carbonato de sódio, carbonato de cálcio, cloreto de cálcio nas concentrações 1, 3 e 5% e cloreto de potássio nas concentrações 0,1; 0,5 e 1%) na inibição deste fungo. Foi realizada a segunda etapa do experimento através de testes em queijos parmesão utilizando o controle biológico clássico aplicando as leveduras e o controle biológico integrado onde utilizou a substância GRAS selecionada associado às leveduras antagonistas. Os resultados foram calculados pela área de contaminação e realizado o tratamento estatístico com teste Scott-Knott. Além disso, foi averiguado o tempo de permanência das leveduras na superfície dos queijos parmesão, através da inoculação e re-isolamento das leveduras no queijo. Nos resultados in vitro a S. cerevisiae UFT 5992 reduziu 92% dos esporos no 7o dia, e não foi observado modificações nas estruturas dos esporos. Das cinco substâncias GRAS testadas o bicarbonato de sódio a 1% obteve 100% de inibição do crescimento fúngico, e as leveduras antagonistas (S. boulardii e S. cerevisiae UFT 5992) sobreviveram por 60 dias nesta solução. Os resultados obtidos dos ensaios de controle biológico clássico constataram que a levedura S. cerevisiae YEF 186, reduziu em 17,5% o crescimento do P.roqueforti comparado com o controle no 13º dias. No controle integrado (S. cerevisiae UFT 5992+ BS), a redução fúngica foi de 18%. Nos dois processos, as leveduras sobreviveram por 50 dias demostrando uma boa adaptação das leveduras em queijo parmesão e podendo agregar valor probiótico ao produto final, estendendo sua vida de prateleira por tempo considerado. / Penicillium roquefortiis a filamentous fungus that can to produce loses, mainly in parmesan cheese, due the alterations on organoleptic characteristics. The biological control, using yeasts from Saccharomycesgenus can to control this pathogen and some species from this genus are considered probiotic. In this way, the objective of this work was to use yeasts to control P. roqueforti in parmesan cheese. Initially, were performed in vitro assays to verify the S. cerevisiae, strains UFT 5962, UFT 5992, UFT 5976, YEF 186 and S. boulardii capacity in to reduce the spore production by P. Roqueforti. At the same time, were verified the activities of some GRAS substances (sodium bicarbonate, calcium carbonate and calcium chloride at 1, 3, and 5 % and potassium chloride at 0.1, 0.5, and 1 %) over this pathogen. After this, were realized a classical biological control (using only the yeasts to control the pathogen) and integrated biological control (using yeast plus the selected GRAS substances) assays in the cheese. The results were obtained measuring the growing area of pathogen and the values were analyzed by Scott-Knott test. The time of endurance of the yeasts in the parmesan surface was obtained by re-isolation of the yeasts inoculated on cheese surface. S. cerevisiae UFT 5992 reduced 92 % of spore production after 7 days, but were not observed morphological alterations in the spores. Sodium bicarbonate at 1 % reduced totally the pathogen growth, and the antagonistic yeasts (S. boulardii e S. cerevisiae UFT 5992) keep alive in sodium bicarbonate (1 %) by 60 days. In the classical biological control, S. cerevisiae YEF 186, reduced the pathogen growth in 17.5 %after 13 days. The integrated control (S. cerevisiae UFT 5992+ BS) reduced the pathogen in 18 %.In both process, the yeasts keep alive by 50 days in parmesan surface, showing a good possibility of their use as probiotic and increasing the shelf life of this product.

Made available in DSpace on 2017-07-10T18:08:06Z (GMT). No. of bitstreams: 1 Barbara Lepretti de Nadai.pdf: 10475653 bytes, checksum: 7df346ebd0d9a5c7cd7b241864ca6506 (MD5) Previous issue date: 2015-02-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Whey is considered one of the most polluting waste from the food industry, and its treatment a highly expensive process. An alternative for its reuse is applied to obtain biotechnological products. The objective of this study was to optimize the cell biomass production conditions of S. boulardii yeast in a batch system, using whey permeate. An experimental design was applied DCCR (a central composite design), and in response to cellular biomass, in order to evaluate the effect of four variables on cell biomass response. Fermentations were carried out at different pH, ammonium sulfate concentrations, and permeate. Two experiments with aeration to the fermentation medium were performed, aiming to increase the production of cell biomass. The highest cell biomass obtained in DCCR was 40,06 gL-1, under the following conditions: 7,5 gL-1 of ammonium sulfate, 1,25 gL-1 of magnesium sulfate, 1,5 gL-1 of monobasic potassium phosphate and 180 gL-1 of permeate, 30°C, agitation 100 rpm and pH 5,5. The investigated pH that favored the production of cell biomass was 4,5. The permeate concentrations which favored the production of biomass are included in the range of 180 to 220 gL-1. Thus, the study demonstrated that the whey permeate is a residue having great potential in producing cellular biomass using the yeast S. boulardii, showing how to dispose of this alternative, reducing negative impacts on the environment and decreasing costs for wastewater treatment industries. / O soro do leite é considerado um dos resíduos mais poluentes da indústria alimentícia, sendo seu tratamento um processo altamente dispendioso. Uma alternativa para a sua reutilização é a aplicação para a obtenção de produtos biotecnológicos. O objetivo deste trabalho foi otimizar as condições de produção de biomassa celular da levedura S. boulardii em sistema descontínuo, utilizando permeado de soro de leite. Foi aplicado um planejamento experimental DCCR (delineamento composto central rotacional), tendo como resposta a biomassa celular, com o intuito de avaliar o efeito de quatro variáveis sobre a resposta de biomassa celular. Foram realizadas fermentações com diferentes valores de pH, concentrações de sulfato de amônio e permeado. Foram realizadas dois ensaios com aeração ao meio fermentativo, objetivando o aumento na produção de biomassa celular. O maior valor de biomassa celular obtido no DCCR foi de 40,06 gL-1, sob as seguintes condições: 7,5 gL-1 de sulfato de amônio, 1,25 gL-1 de sulfato de magnésio, 1,5 gL-1 de fosfato monobásico de potássio e 180 gL-1 de permeado, 30ºC, agitação de 100 rpm e pH 5,5. O valor de pH investigado que favoreceu a produção de biomassa celular foi 4,5. As concentrações de permeado que favoreceram a produção de biomassa estão compreendidas na faixa de 180 à 220 gL-1. Assim, o estudo demonstrou que o permeado de soro de leite é um resíduo com grande potencial na produção de biomassa celular utilizando a levedura S. boulardii, mostrando-se como alternativa ao descarte deste, reduzindo os impactos negativos ao meio ambiente e diminuindo os custos de tratamento deste efluente para as indústrias.

In zahlreichen Studien konnte an Mausmodellen gezeigt werden, dass dendritische Zellen eine wichtige Rolle im Rahmen der mukosalen Immunabwehr spielen. Eine unkontrollierte Aktivierung immunologischer Effektorzellen durch antigenpräsentierende Zellen ist die Folge, welche die Antigene der luminalen Flora folglich falsch erkennen und damit zu einer Schädigung des Gewebes führen. In der Arbeit wurden humane CD1c+CD11c+CD14-CD19- myeloide dendritische Zellen (mDCs) aus dem peripheren Blut und der intestinalen Mukosa von CED Patienten sowie von gesunden Probanden phänotypisch und funktionell näher charakterisiert. mDCs von Patienten reagieren auf LPS im Gegensatz zu DCs von Gesunden mit der Ausbildung eines aktivierten Phänotyps und der Sekretion pro-inflammatorischer Zytokine. Die Daten lassen vermuten, dass ihre tolerogene Rolle gestört ist und die Zellen so möglicherweise aktiv zum Entzündungsgeschehen durch eine Fehlreaktion auf die kommensale Flora beitragen. Es konnte gezeigt werden, dass zirkulierende mDCs von Erkrankten mehr LPS aufnehmen. Des Weiteren ist die Häufigkeit von mukosalen und aktivierten mDCs bei CED Patienten signifikant erhöht. Die vermehrte Häufigkeit von aktivierten mDCs in der entzündeten Mukosa ist ein Hinweis auf intestinales „homing“, also ein Wiedereinwandern der gereiften Lymphozyten in die Darmwand. Es ist bekannt, dass die Hefe Saccharomyces boulardii (Sb) eine Wirksamkeit bei entzündlichen sowie infektiösen Erkrankungen des Gastrointestinaltraktes hat. Kulturexperimente von mDCs mit Zellkulturüberständen von Sb (SbS) und LPS zeigten eine deutliche Reduzierung in der Expression von CD40 und CD80 sowie des Reifemarkers CD197. SbS reduzierte die Sekretion von TNF- und IL-6. Während es die Sekretion von IL-10 bei gesunden Probanden erhöhte, konnte bei CED Patienten eine leichte Abnahme verzeichnet werden. SbS vermindert die Proliferation von naïven T-Zellen in einer gemischten Lymphozytenreaktion mit gesunden mDCs signifikant. / Various animal studies have provided insights that mucosal dendritic cells play a key role in this process. However, the specific function of certain dendritic cells in IBD is still unknown. Primary CD1c+CD11c+CD14-CD19- myeloid blood (mDCs) and mucosal DCs from IBD patients and healthy controls were compared. More mDCs from IBD patients exhibited an activated phenotype shown by expression of co-stimulatory molecules. mDCs from patients secrete higher levels of pro- and anti-inflammatory cytokines. Circulating mDCs from IBD patients take up more LPS and the frequency of mucosal mDCs and the number of activated, i.e. CD40 and CD80 expressing mucosal mDCs, is significantly greater in CED. The increased frequency of activated mDCs in the inflamed mucosa suggests intestinal homing of mDCs in acute stages of IBD. Further, the data suggests an aberrant LPS response of mDCs in patients suffering from IBD which results in an inflammatory phenotype. The most widely accepted hypothesis for the cause of IBD is a disturbed interaction of the host immune system with commensal microflora and other luminal antigens. The well controlled balance of the intestinal immune system is disturbed and luminal antigens like LPS gain access to the underlying mucosal tissue via the leaky barrier. It was investigated whether the yeast preparation Saccharomyces boulardii (Sb) modulates dendritic cell function which has shown efficacy in inflammatory and infectious disorders of the gastrointestinal tract. Culture experiments of mDCs in the presence of Sb culture supernatant (SbS) significantly reduced the expression of CD40 and CD80 as well as the DC maturation marker CD197 (CCR7) induced by the prototypical microbial antigen LPS. SbS reduced secretion of TNF- and IL-6, while the secretion of anti-inflammatory IL-10 increased. IBD patients showed also a reduction in their secretion level of IL-10. SbS inhibited proliferation of naïve T cells in a mixed lymphocyte reaction with healthy mDCs.