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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
141

Efeito do tratamento antioxidante sistêmico em touros Bos taurus taurus submetidos ao estresse térmico e suplementados com dieta rica em ácidos graxos poliinsaturados sobre a capacidade de ligação de espermatozóides à membrana vitelínica de ovos de galinha / Effect of systemic antioxidant treatment in Bos taurus taurus bulls under heat stress and supplemented with polyunsaturated fatty acids on sperm binding capacity to the chicken egg perivitelline membrane

Losano, João Diego de Agostini 02 July 2013 (has links)
Diversos estudos indicam que touros europeus (Bos taurus) apresentam uma maior susceptibilidade ao estresse térmico com um consequentemente aumento do estresse oxidativo. Este, por sua vez, leva a uma diminuição na qualidade espermática. O espermatozoide possui, fisiologicamente, uma grande quantidade de ácidos graxos poli-insaturados (PUFAs) em sua membrana. As ligações duplas entre carbonos desses PUFAs confere a membrana plasmática uma maior fluidez, necessária aos processos fisiológicos do espermatozoide como motilidade e fertilização. No entanto essas ligações são mais facilmente oxidadas e, portanto, mais susceptíveis a peroxidação lipídica. Sendo assim, a suplementação com PUFAs promoveria uma melhoria na qualidade espermática, porém, tornaria o espermatozoide ainda mais susceptível à peroxidação lipídica. A vitamina E é um antioxidante chave na prevenção do estresse oxidativo através da interceptação do radical hidroxila, protegendo os PUFAs da membrana espermática contra a peroxidação lipídica. Portanto, uma alternativa para melhorar a qualidade espermática de touros europeus submetidos ao estresse térmico, seria a suplementação com PUFAs associada a um tratamento com vitamina E para prevenir um possível efeito deletério oxidativo desses ácidos graxos. Estudos sugerem uma importante relação entre os testes convencionais de avaliação de sêmen e fertilidade. No entanto, a limitação destes testes também tem sido reconhecida, principalmente em casos de infertilidade idiopática. Estudos demonstram que os testes funcionais de avaliação do sêmen, podem ser uma alternativa importante para a avaliação da capacidade fecundante de uma determinada amostra seminal. A interação entre espermatozóide e zona pelúcida ocorre através da ligação de receptores acrossomais às proteínas ZP3 e ZP2 do oócito. A membrana perivitelínica de ovos de galinha possui uma homologia com essas proteínas, permitindo sua ligação com espermatozóides de diversas espécies. Sendo assim, o teste de ligação espermática a essa membrana parece ser bastante eficiente para avaliar a capacidade fecundante do sêmen. No presente estudo, 16 touros Bos taurus foram submetidos à insulação testicular durante 4 dias e submetidos a um arranjo fatorial 2x2, sendo os fatores: dieta rica em PUFAs (Megalac®, 4 Kg ao dia) e tratamento com Vitamina E (Monovin E®, 3000 UI a cada 13 dias) durante 60 dias. As amostras seminais criopreservadas/descongeladas foram submetidas à avaliação da análise computadorizada da motilidade (CASA; Hamilton Thorne, Ivos 12.3, USA), integridade de membrana (Eosina Nigrosina), integridade acrossomal (Fast Green Rosa Bengala), atividade mitocondrial (33 diaminobenzidina), susceptibilidade ao estresse oxidativo (Substâncias reativas ao ácido tiobarbitúrico - TBARS), integridade de DNA (SCSA) e o teste de ligação de espermatozóides à membrana vitelínica de ovos de galinha, que foi previamente padronizado. O tratamento com vitamina E apresentou efeitos benéficos para algumas características espermáticas, sendo o contrário observado pelo tratamento com PUFAS, quando ambos foram analisados isoladamente. No entanto, a associação entre estes tratamentos não foi eficiente para melhorar a qualidade espermática assim como a capacidade fecundante avaliada através do teste de ligação espermática em membrana perivitelínica de ovos de galinha. / Several studies suggest a high susceptibility of European bulls (Bos taurus) to the heat stress, which will lead to an increase on oxidative stress and consequent impairment on sperm viability. Sperm membranes are rich in poly-unsaturated fatty acids (PUFAs). The carbon-carbon double bounds (insaturations) that characterize these PUFAs confer the necessary fluidity to the sperm membrane, essential to motility and fecundation capacity. However, these insaturations are more unstable and, therefore, more susceptible to the lipid peroxidation (i.e., attack of reactive oxygen species to the lipids). Hence, PUFA supplementation would improve certain sperm characteristics but increase the already high susceptibility to the oxidative stress. Vitamin E is a key antioxidante on oxidative stress prevention by intercepting hydroxyl radical and protecting sperm membrane PUFAS against lipid peroxidation. Therefore, an alternative to improve sperm quality in heat stressed european bulls would be a PUFA supplementation associated to a Vitamin E therapy in order to avoid a possible deleterious oxidative effects of these fatty acids. Estudies suggest an important relationship between conventional tests for sperm evaluation and fertility. However, limitations have been also verified, especially in cases of idiopathic infertility. Thus, tests to evaluate sperm functionality may be an interesting alternative to assess fecundation capacity of semen samples. The binding of sperm to the zona pellucida occurs initially by the interaction between acrosomal receptors and oocyte proteins ZP3 and ZP2. The perivitelline membrane of chicken eggs present homologies to zona pellucida proteins, which allows sperm from several species to bind to this membrane. Thus, the sperm binding test to the perivitelline membrane may be an efficient method to asses the fecundation capacity in semen of bulls. In the present study, 16 Bos taurus bulls were submitted to testicular insulation for 4 day. Animals were then submitted to a 2x2 factorial design, with the factor represented by a PUFA oral supplementation (Megalac®, 4 Kg for day) and the treatment with Vitamin E (Monovin E®, 3000 UI, each 13 days) for 60 days. Samples were cryopreserved/thawed and analyzed for motility variables by the computer assisted sperm analysis (CASA; Hamilton Thorne, Ivos 12.3, USA), membrane integrity (Eosin / Nigrosin), acrosome integrity (Fast Green / Bengal Rose), mitochondrial activity (33 diaminobenzidine), susceptibility to the oxidative stress (tiobarbituric acid reactive substances - TBARS), DNA integrity (SCSA) and the sperm binding test to the chicken egg perivitelline membrane, which was previously validated. Results demonstrated a beneficial effect of Vitamin E on some sperm characteristics, with the inverse effect observed for PUFA when both treatments were considered individually. However, the association between both treatments were not efficient to improve sperm quality as well as the fecundation capacity as evaluated by the sperm binding test to the chicken perivitelline membrane.
142

Efeito do ômega-3 sobre biomarcadores cardiometabólicos clássicos e emergentes em indivíduos com diferentes níveis de risco cardiovascular / Effect of omega-3 on classic and emergent cardiometabolic biomarkers in individuals with different levels of cardiovascular risk

Marlene Nuñez Aldin 31 March 2014 (has links)
Introdução: As doenças cardiovasculares são a principal causa de morbimortalidade mundial, tendo o Brasil um perfil semelhante. Embora diversos fatores de risco sejam associados com os eventos cardiovasculares, esses não conseguem justificar a manifestação de todos os eventos clínicos, indicando que outros componentes estão envolvidos. Diante desse cenário, a dieta ocupa papel de destaque na modulação dos fatores de risco modificáveis e por isso tem sido foco de inúmeros programas de políticas públicas. Objetivo: Avaliar o efeito da suplementação de ômega-3 sobre os fatores de risco cardiometabólicos clássicos e emergentes em indivíduos com diferentes níveis de risco cardiovascular. Métodos: Este estudo foi do tipo clínico baseado em intervenção nutricional, prospectivo, randomizado, duplo cego e placebo controlado. Foram selecionados indivíduos adultos e idosos (n=146), de ambos os sexos, dentre os quais 77 faziam parte do grupo intervenção w-3 e 69 eram do grupo placebo. O risco cardiovascular foi estimado por meio do escore de risco de Framingham (ERF), os grupos foram subdivididos em escore de risco baixo, intermediário e alto. Nos tempos basal e após 4 e 8 semanas de intervenção [3,0 g/dia w-3, contendo 60% de ácido eicosapentaenoico-docosahexaenoico (EPA-DHA) ou placebo] foram coletadas informações socioeconômicas e clínicas, antropométricas, consumo alimentar, nível de atividade física e coleta de sangue após jejum de 12-14h. A partir do plasma foram analisadas as concentrações dos marcadores lipídicos [Colesterol Total, lipoproteínas de baixa densidade (LDL) e alta densidade (HDL), Triacilglicerol (TG), Apolipoproteínas AI e B, ácidos graxos não esterificados (NEFAs) e LDL eletronegativa (-)], ácidos graxos plasmático e aspectos fisicoquímicos de lipopartículas (tamanho e concentração). Os resultados foram analisados por meio do programa SPSS 16.0, sendo adotado o nível de significância de p < 0,05. Resultados: Houve incorporação de 4% do DHA plasmático e consequente redução de 34% do TG em indivíduos com ERF intermediário e de 0,2% da APO AI, 20% de NEFAs e de 0,6% de HDLINTERMEDIÁRIA em indivíduos com alto ERF. Ao longo da intervenção, houve diminuição do CT (p = 0,002), da LDL (p = 0,003), do TG (p < 0,001), do colesterol não HDL (p < 0,001), da HDLPEQUENA (p = 0,011) e da LDL (-) (p < 0,001). Houve aumento da HDL-c (p = 0,003), das partículas de HDLGRANDE (p < 0,001) e HDLINTERMEDIÁRIA (p < 0,001) Conclusão: A suplementação com w-3 promoveu redução dos biomarcadores de risco cardiovascular clássicos e emergentes. Esse conjunto de resultados amplia o papel cardioprotetor do w-3. / Introduction: Cardiovascular diseases are the leading cause of morbidity and mortality worldwide, with Brazil a similar profile. Although many risk factors are associated with cardiovascular events, these can not justify the manifestation of all clinical events, indicating that other components are involved. Given this scenario, the diet takes prominent role in the modulation of modifiable risk factors and so has been the focus of numerous programs of public policies Objective: To evaluate the effect of supplementation of omega3 (n-3) on cardiometabolic risk factors and emerging classics in individuals with different levels cardiovascular risk. Methods: This study was based on clinical type of nutrition intervention, prospective, randomized, double-blind and placebo controlled. We selected adult and elderly subjects (n=146), both sexes, among which 77 were in the intervention group n - 3 and 69 were in the placebo group. Cardiovascular risk was estimed using the Framingham risk score (FRS), the groups were divided into low-risk score, intermediate and high. At baseline and after 4 and 8 weeks of intervention [3.0 g/ day n-3 containing 60% eicosapentaenoic + docosahexaenoic acid (EPA + DHA) or placebo] socioeconomic and clinical, anthropometric data, dietary intake, physical activity level and blood samples were collected after fasting for 12 -14h. Were analyzed from the plasma concentrations [total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides (TG), apolipoprotein (Apo) AI and B, non-esterified fatty acids (NEFAs) and electronegative LDL (-)], plasma fatty acids and physicochemical aspects of lipoparticles (size and concentration). The results were analyzed using the SPSS 16.0 program, adopting the level significance p < 0.05. Results: There was incresed of 4 % of plasma DHA and consequent 34% reduction of TG in individuals with intermediate FRS and 0.2% of the APO AI, 20 % of NEFAs and 0.6 % HDLINTERMEDIATE in subjects with high FRS. During the intervention, CT decreased (p = 0.002), LDL (p = 0.003), TG (p < 0.001), non-HDL cholesterol (p < 0.001) of the HDLSMALL (p = 0.011) and LDL (-) (p < 0.001). There was an increase in HDL-c (p = 0.003), HDLLARGE particles (p < 0.001) and HDLINTERMEDIATE (p < 0.001). Conclusion: Supplementation with n-3 promoted reduction of c1assic and emerging biomarkers on cardiovascular risk. This set of results extends the cardioprotective role of n-3.
143

Caractérisation de métabolites oxygénés dérivés des acides arachidonique et docosahexaénoïque dans le cerveau de rat / Characterization of oxygenated metabolites derived from arachidonic and docosahexaenoic acids in rat brain

Jouvène, Charlotte 15 September 2016 (has links)
Les acides docosahexaénoïque (DHA) et arachidonique (ArA), qui appartiennent respectivement aux familles n-3 et n-6, sont présents en grande quantité dans les tissus cérébraux, en particulier dans les phospholipides membranaires. Ces deux acides gras polyinsaturés (AGPI) jouent des rôles essentiels dans le fonctionnement cérébral, notamment dans le neurodéveloppement et la neuroinflammation. De plus, ces AGPI sont des précurseurs de plusieurs médiateurs lipidiques oxygénés impliqués dans divers processus physiologiques et pathologiques.Dans ce contexte, l'objectif de cette thèse était de caractériser les métabolites oxygénés dérivés du DHA et de l'ArA dans le cerveau, étape cruciale pour une meilleure compréhension de leurs rôles biologiques dans cet organe. Ainsi, une technique d'analyse à haute performance a été utilisée, la chromatographie liquide couplée à de la spectrométrie de masse en tandem (UHPLC-MS/MS), pour identifier les différents métabolites oxygénés dérivés du DHA et de l'ArA présents dans les cerveaux de rats exsanguinés et non exsanguinés, que ce soit en conditions basales ou inflammatoires. Ainsi, plusieurs produits oxygénés dérivés du DHA et de l'ArA ont été identifiés et quantifiés dans les cerveaux exsanguinés et non-exsanguinés, à la fois à l'état libre dans le cerveau mais également estérifiés dans les phospholipides. Les métabolites mono-hydroxylés sont les principaux dérivés oxygénés du DHA et de l'ArA, cependant, des quantités mesurables de produits di-hydroxylés, tels que le 8,15-diHETE et la protectine DX, ont également été détectées. L'exsanguination permet de différencier les métabolites oxygénés du tissu cérébral de ceux présents dans le sang. De plus, le métabolisme oxygéné du DHA et de l'ArA est impacté lors d'une inflammation cérébrale. En effet, dans ces conditions, la synthèse de certains métabolites est augmentée, notamment celle de la protectine D1, molécule connue pour ces fortes propriétés anti-inflammatoires. Ces résultats méritent de plus amples recherches dans des conditions pathophysiologiques, notamment lors de maladies neurodégénératives, afin d'observer leur impact sur le métabolisme oxygéné de ces AGPI / Docosahexaenoic (DHA) and arachidonic (ArA) acids, two PUFA which belong to n-3 and n-6 families respectively, are both present at high amount in brain tissues, and especially in membrane phospholipids. These two polyunsaturated fatty acids (PUFA) play important roles in brain functioning, notably in neurodevelopment and neuroinflammation. Moreover, these PUFA are precursors of various oxygenated lipid mediators involved in diverse physiological and pathological processes. In this context, the aim of this work was to characterize oxygenated metabolites derived from DHA and ArA in brain, crucial step for better understanding their biological roles in this organ. For this purpose, a high performance analytical approach was usd, the liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS), in order to identify the different oxygenated metabolites derived from DHA and ArA in exsanguinated and non-exsanguinated brains, both in basal and inflammatory conditions. Several oxygenated products from DHA and ArA were identified and measured in exsanguinated and non-exsanguinated brain, both in the free form and esterified in phospholipids. Mono-hydroxylated metabolites were the main oxygenated derivatives from DHA and ArA, however, measurable amounts of di-hydroxylated products such as 8,15-diHETE and protectin DX, were also detected. Exsanguination allowed discriminating oxygenated metabolites from brain tissue against those in blood brain. Moreover, DHA and ArA oxygenated metabolism was impacted during brain inflammation. Indeed, in these conditions, the synthesis of some metabolites was increased, including that of protectin D1, molecule known for its strong anti-inflammatory properties. These results deserve further research in pathophysiological conditions, especially in neurodegenerative diseases, in order to observe their impact on the oxygenated metabolism of these PUFA
144

Study of the effects of docosahexaenoic acid (DHA) and a structured phospholipid containing DHA on physiological and pathological conditions of neurogenesis in vitro / Etude des effets de l'acide docosahexaénoïque (DHA) ou d'un phospholipide structuré contenant le DHA sur des conditions physiologiques et pathologiques de neurogenèse in vitro

Lo Van, Amanda 12 January 2017 (has links)
L'acide docosahexaénoïque (DHA, 22:6n-3) est un acide gras polyinsaturé (AGPI) oméga-3. Il est particulièrement abondant dans le cerveau et la rétine et est nécessaire pour le bon développement et fonctionnement du cerveau. Tandis qu'une déficience en DHA a été montrée être liée à l'émergence de maladies cérébrales (i.e. maladie d'Alzheimer ou maladie de Parkinson), des études ont également montré qu'un apport alimentaire en AGPI oméga-3 pouvait empêcher ou atténuer les perturbations neurologiques liées au vieillissement ou aux maladies neurodégénératives. Il est alors primordial de transporter efficacement le DHA au cerveau. Le laboratoire français a synthétisé auparavant une forme stabilisée de la lysophosphatidylcholine-DHA, qui est le vecteur principal d'apport de DHA au cerveau, de structure 1-acétyl,2-docosahexaénoyl-glycérophosphocholine, brevetée et nommée AceDoPC®. L'injection d'AceDoPC ou de DHA après un accident vasculaire cérébral ischémique provoqué expérimentalement a montré que ces deux molécules étaient neuroprotectrices. Ces effets sont supposés être dus en partie à la conversion du DHA en métabolites oxygénés. Notre étude vise à examiner les effets du DHA et de ses métabolites dérivés, estérifiés ou non dans des phospholipides structurés sur un modèle de neurogenèse in vitro en conditions physiologiques ou pathologiques. Le premier objectif de ce travail a été de synthétiser le phospholipide structuré contenant du DHA, l’AceDoPC®, la protectine DX (métabolite oxygéné du DHA), et un nouveau phospholipide structuré contenant la protectine: 1-acétyl,2-protectine DX-glycérophosphocholine (AceDoxyPC). Le second objectif était d’étudier les effets du DHA, de l'AceDoPC et de la PDX sur la neurogenèse en utilisant un modèle in vitro de neurogenèse, constitué de cultures de cellules souches progénitrices neurales (NSPCs) dérivées de cerveaux de souris adultes, dans des conditions physiologiques ou pathologiques (ischémiques ici). Enfin, le troisième objectif de cette thèse a été d'identifier les mécanismes impliqués dans la réponse des cellules aux conditions ischémiques. La synthèse du phospholipide structuré AceDoxyPC a été réalisée avec succès par une double lipoxygénation enzymatique de l'AceDoPC, et l'identification du produit a été possible grâce à l'utilisation de techniques avancées de chromatographie liquide couplée à la spectrométrie de masse (LC/ESI/MS). De futures études sur ce transporteur de molécule neuroprotectrice potentielle doivent être réalisées prochainement. Les cellules incubées en présence d’AceDoPC présentent une augmentation de neurogenèse comparativement à celles cultivées avec addition de DHA non estérifié ou du véhicule contrôle, notamment sous conditions pathologiques. Les études préliminaires des mécanismes potentiellement impliqués dans la neuroprotection indiquent que les effets neuroprotecteurs et régénératifs de l'AceDoPC pourraient être en partie dus à des effets anti-oxidants. / Docosahexaenoic acid (DHA, 22:6n-3) is an essential omega-3 polyunsaturated fatty acid (PUFA). It is specifically enriched in the brain and the retina and it is required for visual acuity, proper brain development and cerebral functions. While DHA deficiency in the brain was shown to be linked to the emergence of cerebral diseases (i.e. Alzheimer’s disease or Parkinson’s disease), studies showed that a dietary intake of omega-3 PUFA could prevent or attenuate neurologic disturbances linked with ageing or neurodegenerative diseases. In this context, it is primary to deliver DHA efficiently to the brain. Targeting the brain with DHA might offer great promise in developing new therapeutics for neurodegenerative diseases. The French host laboratory previously synthesized a stabilized form of lysophosphatidylcholine-DHA, which is main vector of DHA transportation to the brain, of structure 1-acetyl,2-docoshexaenoyl-glycerophosphocholine, patented and named AceDoPC®. Injection of AceDoPC or DHA after experimental ischemic stroke showed that both molecules also had neuroprotective effects. These potential neuroprotective effects are expected to be due, in part, to DHA conversion into oxygenated metabolites. This study aims to investigate the beneficial effects of DHA and its derived metabolites either unesterified or esterified within structured phospholipids on a model of neurogenesis in vitro under physiological or pathological conditions. The first objective of this work was then to synthesize the DHA-containing structured phospholipid AceDoPC®, DHA oxygenated derivative protectin DX (PDX) and a novel structured phospholipid of protectin: 1-acetyl,2-protectinDX-glycerophosphocholine (AceDoxyPC). The second objective was to investigate the effects of DHA, AceDoPC and PDX on neurogenesis using an in vitro model of neurogenesis, namely cultures of neural stem progenitor cells (NSPCs) derived from the adult mouse brain under physiological or pathological conditions (ischemic conditions). Following this, the third objective of this work was to identify the mechanisms involved in such response to stress induced under pathological conditions. Synthesis of the novel structured phospholipid AceDoxyPC was successfully performed by double enzymatic lipoxygenation of AceDoPC and identification of the product was possible using advanced techniques of liquid chromatography (LC)/electrospray ionization (/ESI)/mass spectrometry (/MS). Future studies on this potential neuroprotective molecule transporter are to be investigated in the near future. Neurogenesis study of cell cultures with AceDoPC showed enhanced neurogenesis compared to addition of unesterified DHA or vehicle control, especially under pathological conditions. Preliminary studies of the potential mechanisms involved in neuroprotection hinted that AceDoPC neuroprotective and regenerative effects might be due in part to its anti-oxidative effects.
145

Rôle des acides gras polyinsaturés n-3 sur la régulation de l’inflammation et le processus de tumorigenèse déclenché par Helicobacter pylori / The role of n-3 polyunsaturated fatty acids in Helicobacter pylorimediated gastric inflammation and tumorigenesis

Correia, Maria Marta de Ascensao Teixeira 24 September 2012 (has links)
La bactérie Helicobacter pylori est responsable de l’infection la plus répandue dans la population mondiale. Cette bactérie est considérée comme le principal agent étiologique de la gastrite chronique, de l’ulcère duodénal et du cancer gastrique non-héréditaire. La thérapeutique prescrite pour l’éradication de cette infection est inefficace pour un nombre de plus en plus élevé de patients, dû à l’induction constante de souches résistantes aux antibiotiques habituellement prescrits.H. pylori a aussi d’autres façons d’assurer sa survie dans le milieu gastrique que l’induction de résistances aux antibiotiques. En particulier en intéragissant avec le cholestérol des cellules épithéliales gastriques. De ce fait, l’utilisation de molécules inhibitrices de la croissance de H. pylori, autres que les antibiotiques classiques, est une stratégie importante pour combattre cette infection.L’objectif majeur de ce travail a été de mettre en évidence de nouvelles molécules inhibitrices de la croissance et de la viabilité de H. pylori, permettant ainsi le développement de solutions alternatives à la thérapeutique classiquement utilisée. Les acides gras polyinsaturés et l’acide docosahexaenoic (DHA) se mettre en lumière pour réunir incontestable propriété anti-inflammatoires et anti-tumoral. Nous avons émis l’hypothèse que le DHA influence la survie de H. pylori, et peut moduler la disponibilité des acides gras et du cholestérol cellulaires.Nos résultats montrent que le DHA inhibe la croissance de H. pylori in vitro et affecte sa capacité à coloniser la muqueuse gastrique dans le modèle souris. Dans ces conditions, le DHA diminue la réponse inflammatoire gastrique induite par l’infection. Au niveau des cellules épithéliales gastriques, des modifications du profil des acides gras et du cholestérol avec des conséquences sur le métabolisme et la signalisation cellulaire sont observées. De plus, un traitement antibiotique classique combiné à une administration de DHA aux souris infectées diminue de façon drastique la récidive de l’infection. En conclusion, cette étude démontre un effet inhibiteur du DHA sur l’infection par H. pylori et sa récidive. Ces résultats justifient la proposition du DHA comme coadjuvant thérapeutique, constituant ainsi une stratégie prophylactique alternative de l’éradication de l’infection par H. pylori. / H. pylori infection is extremely common worldwide and is recognized as a major etiological factor in chronic active gastritis, gastric duodenal ulcers and gastric cancer development. H. pylori eradication treatment has not changed to a large extent in the last decades and can raise some concern mainly due to recurrence of infection, and most importantly, acquired resistance to classically used antibiotics. In this context, the use of compounds other than antibiotics that could decrease H. pylori infection in a safe way could provide an alternative to tackle this problem. It is known that H. pylori extracts cholesterol from host cell-membrane rafts, modifies it into an α-glycosylated form, and uses this mechanism to increase its survival. The main aim of this thesis work was to explore the role of different non-antibiotic molecules in inhibiting H. pylori growth. Among molecules known to affect in vitro H. pylori growth and viability are certain polyunsaturated fatty acids (PUFAs). Within the many molecules available, we concentrated our efforts on the study of docosahexaenoic acid (DHA). We also pursued the hypothesis that DHA affects survival of H. pylori by modulating the host epithelial cell levels of fatty acids and cholesterol availability.Our results show that DHA inhibits H. pylori growth both in vitro and in vivo, and attenuates the host inflammatory response. Additionally, we demonstrate that DHA induces morphological and cell wall protein composition changes that altogether decrease bacteria-gastric epithelial cell adherence, inflammation and survival. Also, we demonstrated that DHA alters cholesterol levels in epithelial cells, thereby influencing H. pylori ability to uptake and use epithelial cholesterol. This will ultimately impair H. pylori survival. Importantly, the combination of DHA and antibiotic standard treatment decreased the recurrence of H. pylori infection in a mouse model. Our results have gathered important evidence to pave the way for DHA use in the clinical setting and in prophylactic/preventive strategies against H. pylori infection.
146

Etude de la production et de l’extraction couplée de fucoxanthine et d’acides gras polyinsaturés par des microalgues / Study of the coupled production and extraction of carotenoids and polyunsaturated fatty acids from microalgae.

Delbrut, Antoine 01 June 2018 (has links)
Cette thèse s’inscrit dans le cadre du projet SMILE de Microphyt qui vise à proposer une gamme d’ingrédients naturels issus de microalgues afin de répondre à deux enjeux sociétaux majeurs : le surpoids/obésité et le déclin des fonctions cognitives liées à l’âge (DCLA). Les acides gras polyinsaturés oméga-3 (AGPI-w3) et la fucoxanthine (Fx) ont des activités prouvées contre les voies métaboliques liées à la prise de poids et aux DCLA. C’est dans ce contexte que les efforts se sont tout d’abord focalisés vers la sélection de deux espèces de microalgues brunes dont le critère principal a été la productivité en ces molécules d’intérêts. Deux souches des espèces Tisochrysis lutea et Phaeodactylum tricornutum ont alors été sélectionnées du fait de leurs fortes teneurs et productivités en AGPI-w3 et Fx, pour être par la suite cultivées à l’échelle de 5000 L dans des photobioréacteurs (PBR) industriels Camargue©. Dans un second temps, les travaux ont porté sur l’optimisation des procédés post-culture pour les deux espèces, c’est-à-dire, de la récolte de la biomasse à l’obtention d’un extrait, afin de garantir la sécurité, la stabilité et l’efficacité des produits finis. L’étude des procédés de récolte/concentration de la culture a montré leur influence sur les classes lipidiques et l’apparition d’acides gras sous forme libre (AGL), engendrés par les stress mécaniques et thermiques subis par la biomasse lors de cette étape. La formation de ces AGL a pu être limitée par l’utilisation de la filtration tangentielle et de la centrifugation dans certaines conditions. De plus, le lien entre procédé et activité a été souligné via une étude sur modèle cellulaire CaCo2 : les AGL pourraient être à l’origine d’une cytotoxicité accrue des extraits. Enfin, l’optimisation des conditions de co-extraction des AGPI-w3 et de la Fx a abouti à la sélection d’un solvant éco-compatible (éthanol) et a mis en lumière le fort impact de la granulométrie et du ratio solvant/biomasse sur les rendements d’extractions. / This thesis is part of Microphyt's SMILE project, which aims to propose a range of natural ingredients derived from microalgae. The main objective of this project is to respond to two major societal issues: overweight/obesity and the decline of age-related cognitive functions (DCLA). Omega- 3 polyunsaturated fatty acids (PUFAs) and fucoxanthin (Fx) have proven activities against the metabolic pathways associated with weight gain and DCLA. In this context, the first efforts were initially directed towards the selection of two species of brown microalgae with their productivity in these molecules of interest as the main criterion. Two strains of the species Tisochrysis lutea and Phaeodactylum tricornutum were then selected as raw materials rich in PUFAs and Fx and were grown at a 5,000 L scale in Camargue© photobioreactors. The work then focused on optimizing the downstream processing steps to the cultivation of microalgae in order to guarantee the safety, stability and efficiency of the final products. Harvesting processes had, in particular, through the mechanical stress generated, an impact on lipid classes and the appearance of free fatty acids (FFA) in the studied species. A CaCo2 cell model study has suggested that FFA may be responsible for increased cytotoxicity of extracts. The formation of FFA during the concentration stage could be limited by the use of tangential filtration and centrifugation under certain conditions. Finally, the optimization of the co-extraction conditions of PUFAs and Fx resulted in the selection of an eco-friendly solvent (ethanol) and highlighted the strong impact of particle size and solvent/biomass ratio on extraction yields.
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Avaliações ultrassonográficas, morfométricas e histológicas testiculares de touros Bos taurus taurus submetidos a insulação escrotal sob o tratamento sistêmico com antioxidante e suplementado com ácidos graxos poli-insaturados / Testicular ultrasound, morphometry and hystology of Bos taurus taurus bulls submitted to testicular warming, systemic antioxidant treatment and polyunsaturated fatty acids supplementation

Rocha, Carolina Camargo 12 April 2013 (has links)
Vários estudos indicam uma maior susceptibilidade ao estresse térmico de touros europeus quando criados em regiões tropicais, levando a efeitos deletérios à reprodução (e.g., diminuição da qualidade espermática, degeneração testicular). Tal fato pode causar expressivo prejuízo econômico visto que, nestas regiões, a estação de monta é realizada no verão. Um dos principais mecanismos propostos para explicar este efeito seria um aumento do estresse oxidativo. Assim, uma alternativa promissora seria o tratamento com antioxidantes e ácidos graxos poli-insaturados. O presente estudo tem como objetivo: Avaliar os possíveis efeitos benéficos da suplementação oral com ácidos graxos poli-insaturados (com vs. sem) e a ação protetora do tratamento sistêmico com vitamina E (com vs. sem) contra os danos oxidativos causados pelo estresse térmico testicular em touros taurinos insulados através das avaliações ultrassonográfica, morfométrica testicular (i.e, volume, temperatura, consistência, circunferência) e histológica dos testículos. Para isso, foram utilizados 16 touros Bos taurus taurus jovens (em torno de 2 anos de idade) divididos em 4 grupos (fatorial 2x2) com perfil espermático normal. Todos os animais foram submetidos à insulação testicular por um período de 96 horas. Foi realizada a ultrassonografia testicular a cada 13 dias por todo o período e mais dois meses após o período da insulação. Como controles foram avaliados os testículos dos touros antes da insulação. Concomitante à insulação, os animais foram divididos aleatoriamente em 4 lotes que foram submetidos a um arranjo fatorial 2x2, sendo um dos fatores a dieta rica em PUFA (Ácidos graxos poli-insaturados Megalac E&reg;) todo o dia durante dois meses pós insulação. Já o tratamento sistêmico com Vitamina E (Monovin E&reg;) foi realizado através da injeção subcutânea de 5 ml de &alpha;-tocoferol a cada 13 dias pós insulação com duração de dois meses pós insulação. Aos animais que não receberam a vitamina E, foi administrado o mesmo volume em placebo (óleo inerte). No final deste período, os animais foram castrados e os testículos utilizados para a histologia. Os resultados foram analisados através do SAS system for Windows e indicaram que tanto a suplementação oral de ácidos graxo poli-insaturados como o tratamento sistêmico com vitamina E, não alteraram as características ultrassonográficas, morfométricas e histológicas testiculares de touros de origem europeia submetidos a insulação testicular. / Several studies indicate an increased susceptibility to the heat stress in European bulls raised under tropical conditions, which leads to deleterious effects on reproduction (e.g., testicular degeneration, impaired sperm quality). This may cause significant economic losses since, in these regions, breeding season usually occurs during summer. The main mechanism proposed to explain such event is the oxidative stress. Therefore, an alternative to overcome this effect would be the treatment with antioxidant combined with a supplementation with poly-unsaturated fatty acids (PUFA). The present study aimed to evaluate the possible beneficial effect of the oral PUFA supplementation combined with a protective antioxidant treatment to overcome the deleterious effects of the heat stress on testicular echogenicity, morphometry (i.e., volume, circumference, consistency) and histology. Towards this aim, 16 young but sexually mature Bos taurus taurus (around 2 years old). All animals were submitted to testicular warming for 96 hours. Simultaneously to the insulation, animals were randomly allocated into 4 groups in a 2x2 fatorial design. One of the factors was a PUFA supplementation (supplemented vs. nonsupplemented; Megalac E&reg;) everyday for two months. The other factor was a systemic treatment with a subcutaneous administration of 5 mL of &alpha;-tocopherol (treated vs. non treated) every 13 days for two months. Animal that did not receive the treatment were injected with 5 mL of inert oil. Testicular ultrasound evaluation was performed each 13 days from the beginning until two months after testicular warming. At the end of the period experimental period, animals were castrated and the testicles were submitted to histological evaluation. Data were statistically analyzed using the SAS system for Windows. Results indicated that oral PUFA supplementation combined with the systemic -tocopherol treatment had no influence on testicular echogenicity, morphometry and hystology in European bulls submitted to testicular warming.
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Fontes marinhas e vegetais de PUFAs na dieta de galinhas poedeiras: efeito na composição lipídica da gema do ovo e tempo de incorporação dos ácidos graxos / Marine and vegetal sources of PUFAs for laying hens: effect on egg yolk composition and fatty acid incorporation time

Pita, Maria Carolina Gonçalves 06 July 2007 (has links)
Foram utilizadas 144 galinhas poedeiras da linhagem Shaver White por um período de quatro semanas, com o intuito de verificar o efeito da adição de 3% dos óleos de soja, milho, canola, linhaça, salmão ou da mistura de sardinha e atum na dieta das aves, sobre a composição dos ácidos graxos na gema dos ovos e no plasma sanguíneo das aves, bem como o tempo de incorporação total de cada ácido graxo na gema dos ovos durante o período experimental. Paralelamente foram avaliados, o desempenho das aves e a qualidade externa dos ovos. Para análise estatística dos resultados empregou- se modelo fatorial 6X6 em blocos casualizados sendo as aves distribuídas em 72 gaiolas. A adição de 3% de óleo de salmão ou de milho à ração determinou redução significativa no consumo alimentar. O tratamento que recebeu 3% de óleo de salmão promoveu menor peso da casca dos ovos (g) bem como menor espessura da casca (mm). A inclusão dos óleos de linhaça, soja ou milho à dieta, determinou aumento dos ácidos graxos poliinsaturados na gema e no plasma das galinhas. A adição de óleo de sardinha e atum proporcionou maiores concentrações de ácidos graxos saturados na gema. Os teores de PUFAs n-3 foram maiores nas gemas do tratamento a base de óleo de sardinha, enquanto que no plasma a maior concentração foi observada no tratamento com óleo de linhaça. O óleo de linhaça proporcionou o maior teor de ácido &alpha;-linolênico incorporado à gema e no plasma sanguíneo. As quantidades de EPA na gema e no plasma mostraram-se maiores no grupo que recebeu 3% de óleo de sardinha e atum que, por sua vez, também foram responsáveis pelos mais elevados teores de DHA na gema dos ovos. Já com relação ao plasma, as maiores concentrações de DHA foram observadas nos grupos alimentados com óleos de salmão e sardinha/atum. Os PUFAs diminuíram na gema até o oitavo dia experimental, enquanto que os PUFAs n-3 aumentaram até este dia. Os teores de ácido &alpha;-linolênico foram crescentes até o décimo dia de experimento, enquanto que as concentrações de EPA e DHA aumentaram até o oitavo dia de experimento. / Hundred forty-four Shaver White laying hens were used over a 4 week experimental period to investigate the effect of 3% of soybean oil, corn oil, canola oil, flaxseed oil, salmon oil or tuna and sardine oil added to the diets, upon the fatty acid egg yolk composition, blood plasma levels and incorporation time of each fatty acid into the egg yolk. Reproductive performance of hens and external egg quality were evaluated. Hens were allocated into 72 cages and the experimental design was a 6X6 randomized factorial model. Hens fed 3% salmon or corn oil diet showed a significant reduction of food consumption, eggshell weight (g) and eggshell thickness (mm). The addition of flaxseed, soybean or corn oil into the diet increased the polyunsaturated fatty acids levels into the egg yolk and in the blood plasma. Adding tuna and sardine oil into the diet increased the concentration of yolk saturated fatty acids. The levels of PUFAs n-3 were increased in the tuna and sardine oil treatment, while the flaxseed oil increased the plasma fatty acids. The deposition of &alpha;-linolenic fatty acids was higher in the group fed flaxseed oil The percentage of EPA into the yolk and plasma was higher for the hens fed 3% tuna and sardine oil diet, as well as the levels of yolk DHA. The concentration of DHA into the plasma was higher for the salmon and tuna/sardine oil treatments. The PUFAs yolk decreased during the first eight days of experiment, while the PUFAs n-3 increased during the same period. The concentration of &alpha;-linolenic acid increased until ten days of experiment, while the percentage of EPA and DHA increased up to the eighth experimental day.
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Efeito da dieta contendo alta inclusão de soja extrusada e fonte orgânica de selênio sobre a composição, teor de CLA, perfil de ácidos graxos e estabilidade oxidativa do leite de vacas holandesas / Effect of extruded soybean diet and organic selenium on milk composition, CLA content, fatty acid profiles and oxidative stability of milk

Paschoal, Juliana Jorge 20 April 2007 (has links)
A formulação de dietas para ruminantes pode alterar o perfil de ácidos do leite, potencializando o valor nutritivo e nutracêutico do mesmo. Estas alterações podem, no entanto, aumentar a susceptibilidade do leite à oxidação, perdendo qualidade em sabor e tempo de prateleira. O objetivo deste trabalho consistiu em avaliar o efeito do fornecimento de dieta com alto teor de soja integral extrusada, associada ou não à suplementação com selênio orgânico, sobre a produção, a composição, a concentração de CLA, o perfil de ácidos graxos e a estabilidade oxidativa do leite. Vinte e quatro vacas holandesas foram agrupadas em três blocos, de acordo com o número de lactações e alimentadas com dieta total contendo 56% de volumoso e 44% de concentrado. Os animais foram distribuídos aleatoriamente nos seguintes tratamentos: A) dieta controle, B) dieta contendo 21% de soja extrusada e C) dieta contendo 21% de soja extrusada + 5 mg de selênio orgânico. As dietas foram balanceadas para obtenção dos mesmos níveis de energia e proteína. O fornecimento de soja extrusada não afetou o consumo de matéria seca, o peso vivo dos animais e a produção de leite. As vacas alimentadas com soja extrusada produziram leite com menor (P<0,01) porcentagem de gordura, menor (P<0,01) concentração de ácidos graxos de cadeias curta e média e maior (P<0,01) concentração de ácidos graxos de cadeia longa, quando comparadas às vacas alimentadas com a dieta controle. Com relação ao teor de saturação, produziram leite com menor (P<0,01) concentração de ácidos graxos saturados e maior (P<0,01) concentração de ácidos graxos poliinsaturados e de ácido linoléico conjugado (CLA). A concentração de colesterol total foi maior (P<0,01) no soro sanguíneo das vacas alimentadas com soja extrusada, quando comparado ao controle. A suplementação com selênio aumentou (P<0,05) significativamente a concentração do mineral no leite e retardou a oxidação do mesmo, avaliada pelo índice de peróxido (P<0,01). O fornecimento da dieta contendo 21% de soja alterou o perfil de ácidos graxos do leite, diminuindo a concentração de ácidos graxos saturados e aumentando a concentração de ácidos graxos poliinsaturados e ácido linoléico conjugado, no entanto, produziu um leite mais susceptível à oxidação. O enriquecimento do leite com selênio conseguiu minimizar este efeito influenciando positivamente a estabilidade do leite. / The manipulation of ruminant diet can modify the fatty acid profiles increasing its nutritive and therapeutic value. However, this change in fatty acid composition can increase milk oxidation and can affect its flavor and shelf life. The objective of this study was to evaluate the effect of extruded soybean diet and organic selenium supplementation on milk composition, CLA content, fatty acid profiles and oxidative stability of milk. Twenty four Holstein cows were grouped into three blocks according to number of parturition and were fed diets containing 56% forage and 44% concentrate. The animals were allocated to three treatments: A) control diet, B) 21% extruded soybean diet and C) 21% extruded soybean diet + 5 mg of organic selenium. The diets were formulated to provide the same energy and protein levels. The extruded soybean did not affect the dry matter intake, body weight and milk production. The cows fed extruded soybean diet produced milk with lower (P<0,01) concentration of fat, lower (P<0,01) concentration of short and medium chain fatty acids and higher (P<0,01) concentration of long chain fatty acids when compared to control group. Regarding to saturation level, those animals produced milk with lower (P<0,01) concentration of saturated fatty acids and higher (P<0,01) concentration of polyunsaturated fatty acids and CLA content. The serum concentration of cholesterol was higher (P<0,01) for treatments containing extruded soybean when compared to control. The organic selenium supplementation increased (P<0,05) milk selenium concentration and delayed milk oxidation measured by peroxide value (P<0,01). The inclusion of 21% of extruded soybean in dairy cattle diet resulted in production of milk fat with modified fatty acid profile with lower concentration of saturated fatty acids and higher concentration of polyunsaturated fatty acids and CLA content. However the milk fat was more susceptible to oxidation. The organic selenium supplementation was efficient to overcome this problem and therefore could maintain the stability of milk.
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Effets des acides gras polyinsaturés n-3 sur les processus cibles des rétinoïdes impliqués dans la plasticité synaptique et la mémoire au cours du vieillissement / Effects of n-3 LC-PUFAs on retinoid target processes involved in synaptic plasticity and memory during aging

Létondor, Anne 16 December 2013 (has links)
Les acides gras polyinsaturés à longue chaîne (AGPI-LC) de la série n-3 jouent un rôle essentiel dans le fonctionnement cérébral, notamment dans le maintien des processus de plasticité synaptique et de mémoire au cours du vieillissement. Il est maintenant bien admis que ces acides gras peuvent moduler la transcription de gènes impliqués dans les processus de plasticité synaptique sous-tendant les performances mnésiques via leur liaison à des récepteurs nucléaires tels que les PPAR (peroxisome proliferator-activated receptor) et les RXR (retinoid X receptor). Les RXR sont les partenaires communs d’hétérodimérisation de nombreux autres récepteurs, dont le récepteur nucléaire de l’acide rétinoïque (AR), RAR (retinoic acid receptor), métabolite actif de la vitamine A. Ainsi, les RXR jouent un rôle majeur dans la régulation des voies de signalisation des AGPI n-3 et des rétinoïdes.Dans ce contexte, l’objectif de notre travail était de mieux comprendre les mécanismes mis en jeu dans l’action des AGPI-LC n-3 sur les processus neurobiologiques qui sous-tendent les performances mnésiques au cours du vieillissement, notamment en abordant de manière spécifique les mécanismes mis en jeu dans les interactions entre les voies de signalisation des AGPI-LC n-3 et des rétinoïdes. Les approches expérimentales mises en place ont consisté notamment à évaluer chez le rat âgé les effets de supplémentations nutritionnelles en AGPI-LC n-3 et/ou vitamine A sur les performances de mémoire, ainsi que l’action du DHA administré seul sur différents types de mémoire dépendants de l’hippocampe.Nos principaux résultats montrent une altération du métabolisme des acides gras et de la vitamine A au cours du vieillissement. Ces changements métaboliques sont associés à une hypoexpression des voies de signalisation des AGPI n-3 et des rétinoïdes, accompagnée de déficits mnésiques. Nous montrons par ailleurs un effet synergique d’une supplémentation nutritionnelle en AGPI-LC n-3 et en vitamine A sur le maintien des performances de mémoire chez l’animal âgé. De plus, cette supplémentation permet de prévenir, dans l’hippocampe, les changements de composition en AGPI n-3 ainsi que l’hypoexpression des ARNm de RXRγ et de kinases régulées par l’AR et les AGPI n-3.Ces résultats plaident en faveur d’une action synergique des AGPI-LC n-3 et de la vitamine A sur le maintien des performances mnésiques au cours du vieillissement, via une action combinée sur leurs voies de signalisation, lesquelles participeraient ainsi au maintien de certains processus de plasticité synaptique sous-tendant la mémoire et qui se trouvent être altérés avec l’âge. / N-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFAs) play a critical role in brain functioning, notably in the maintenance of synaptic plasticity and memory processes during aging. It is now well accepted that n-3 PUFAs can modulate transcription of genes involved in synaptic plasticity processes underlying the memory performances through binding and activating nuclear receptors such as PPARs (peroxisome proliferator-activated receptors), and RXRs (retinoid X receptors). RXRs are the common heterodimerization partner of numerous nuclear receptors, among them the retinoic acid receptor (RAR), which binds retinoic acid (RA), the active metabolite of vitamin A. Thus, RXRs play a key role in the regulation of n-3 PUFA and retinoid signaling pathways.In this context, the aim of this work was to study the mechanisms involved in the action of n-3 PUFAs on neurobiological processes underlying the memory performances during aging, and more particularly by assessing specifically the molecular mechanisms involved in interactions between n-3 PUFA and retinoid signaling pathways. For this purpose, we studied the effects of dietary supplementations in n-3 PUFAs and/or vitamin A on memory performances in aged rats. We also studied the specific effect of unesterified DHA pharmacological treatments on different hippocampal-dependent memory tasks.Our main results showed impairments in fatty acid and vitamin A metabolism during aging. These modifications were associated with an hypoexpression of n-3 PUFA and retinoid signaling pathways, and memory deficits. Furthermore, we demonstrated a synergetic effect of the joint n-3 LC-PUFA and vitamin A dietary supplementation on the maintenance of memory performances in aged rats. Moreover, in the hippocampus, this supplementation prevented the n-3 PUFA compositional changes, and also the mRNA hypoexpression of RXRγ and of several kinases regulated by RA and n-3 PUFAs which were observed during aging.These results suggest a beneficial synergetic effect of n-3 LC-PUFAs and vitamin A on the maintenance of memory performances during aging, through a combined action on their signaling pathways, which could be involved in the maintenance of synaptic plasticity processes underlying memory performances impaired during aging.

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