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Repercussão do uso parenteral prévio de emulsão lipídica de óleo de peixe sob a resposta inflamatória sistêmica e sobrevida em modelo experimental de pancreatite aguda / Impact of prior use of parenteral fish oil lipid emulsion in thesystemic inflammatory response and survival in an experimental model of acutepancreatitisPriscila Casarin Garla 19 August 2015 (has links)
Introdução: A infusão parenteral de emulsão lipídica (EL) de óleo de peixe (OP), rica em ácidos graxos ômega-3 (AG n-3), está associada à diminuição do perfil de mediadores pró-inflamatórios em estudos experimentais e clínicos. AG n-3 se incorporam em poucas horas em membranas celulares e geralmente são administrados após a agressão inflamatória. A pancreatite aguda (PA) experimental é modelo de inflamação, local e sistêmico, bem estabelecido. Objetivo: O presente estudo avaliou o efeito da infusão parenteral de EL de OP por curto período, antes da indução de PA, sobre a modulação da resposta inflamatória sistêmica e sobrevida. Métodos: Após cateterização do sistema venoso central, ratos isogênicos Lewis receberam infusão parenteral de EL de óleo de peixe ou solução salina durante 48 horas, quando então foram submetidos à pancreatite aguda, pela injeção retrógrada de 0,5 mL de solução de taurocolato de sódio a 3% no duto pancreático. Após indução de PA, nos períodos de 2, 12 e 24 horas, os animais foram sacrificados para coleta de amostras de sangue e de tecidos para dosagem de marcadores inflamatórios e histopatológicos. Paralelamente, 20 animais de cada grupo foram observados até sete dias após indução de PA, para avaliação de sobrevida. Resultados: O tratamento com EL de óleo de peixe foi associado com diminuição de citocinas pró-inflamatórias IL-1beta (p=0,0006) e IL-6 (p=0,05), diminuição de IL-4 (p= 0,0019) e tendência no aumento de anti-inflamatória IL-10 (p= 0,06), após 24 horas de PA; e com aumento da expressão pulmonar e hepática de proteínas de choque térmico HSP 90, 2 e 12 horas após PA, respectivamente. Após infusão de EL de óleo de peixe, não foram encontrados efeitos sobre os níveis de malonaldeído no fígado e na histopatologia do pâncreas, no período de 2 e 12 horas pós pancreatite aguda; e na taxa de sobrevida, em relação aos demais grupos (p > 0,05). Conclusão: Nossos resultados sugerem que a infusão parenteral de EL de OP 48 horas antes da indução de pancreatite aguda experimental parece influenciar favoravelmente a produção de citocinas inflamatórias e HSP90 hepática e pulmonar, sem impactar sobre a histopatologia da lesão pancreática e a taxa de sobrevida / The parenteral infusion of fish oil (FO) lipid emulsion (LE), rich in omega-3 fatty acids (n-3 FA), is associated with the decrease of pro-inflammatory mediators profile in experimental and clinical studies. N-3 FA are incorporated in a few hours in cell membranes and are generally administered after the inflammatory injury. The experimental acute pancreatitis (AP) is an inflammation, local and systemic well-established model. This study evaluated the effect of parenteral infusion of fish oil LE for a short period before AP induction, on the modulation of systemic inflammatory response and survival. For this, after the central venous catheterization Lewis rats received parenteral infusion of fish oil LE or saline solution for 48 hours, when they were induced to acute pancreatitis by retrograde injection of 0.5 mL of sodium taurocholate at 3% in pancreatic duct. After AP induction, in periods of two, 12 and 24 hours, the animals were sacrificed to collect blood samples and tissues for measurement of inflammatory markers and histopathological. In parallel, 20 animals in each group were observed up to 7 days after induction of AP, for survival analysis. The treatment with fish oil LE was associated with decreased of pro-inflammatory cytokines IL-1beta (p = 0.0006) and IL-6 (p = 0.05), reduction of IL-4 (p = 0.0019) and upward trend of anti-inflammatory IL-10 (p = 0.06) after 24 hours of AP; and increased pulmonary and hepatic expression of heat shock proteins HSP 90 two and 12 hours after AP, respectively. After infusion of fish oil LE, there were no effects on malondialdehyde levels in the liver and the pancreas histopathology in the periods of 2 and 12 hours after acute pancreatitis; and survival rate, compared to the other groups (p > 0.05). Our results suggest that parenteral infusion of FOLE 48 hours before the induction of experimental acute pancreatitis appears to favorably influence the production of inflammatory cytokines; hepatic and pulmonary HSP90, without impacting on the histopathology of pancreatic injury and the survival rate
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Efeito de emulsão lipídica parenteral composta por mistura de triglicérides de cadeia média e óleos de soja, oliva e peixe sobre a migração e fagocitose de leucócitos de ratos / Effect of parenteral lipid emulsion containing mixture of medium-chain triglycerides and soybean, olive and fish oils on leukocytes migration and phagocytosis in ratsLetícia de Nardi Campos 04 September 2007 (has links)
INTRODUÇÃO: Emulsões lipídicas parenterais (EL) contendo óleo de peixe podem modular favoravelmente a resposta inflamatória e manter ou promover a resposta imunológica, mas há dados insuficientes sobre seu impacto em funções de células da imunidade inata. OBJETIVO: Verificar o efeito da administração endovenosa de EL composta por mistura de triglicérides de cadeia média e óleos de soja, oliva e peixe sobre a migração e fagocitose de leucócitos de ratos, em comparação à EL composta por mistura física de triglicérides de cadeia média e cadeia longa - TCM/TCL, suplementada ou não com óleo de peixe (OP). MÉTODOS: Ratos (Lewis) isogênicos (n=40) foram submetidos à cateterização da veia jugular externa para acesso parenteral. Os animais foram randomizados em quatro grupos, de acordo com sua infusão endovenosa: grupo SMOF: EL contendo 30% de óleo de soja (TCL), 30% de TCM, 25% de óleo de oliva e 15% de OP; grupo TCM/TCL: EL contendo TCM e TCL (1:1 v/v); grupo TCM/TCL/OP: EL composta por TCM/TCL com adição de OP (8:2 v/v); grupo SF: solução fisiológica. Um grupo de animais sem cateterismo venoso também foi desenvolvido (CO-NC). No quinto dia de experimento e após injeção de carvão coloidal pela veia caudal, amostras de sangue e tecido (fígado, pulmão e baço) foram coletadas para análise quimiotática de neutrófilos (câmara de Boyden adaptada) e quantificação do número de macrófagos fagocitantes do carvão coloidal (imunohistoquímica). Os dados foram analisados por ANOVA e pós-teste de Tukey. RESULTADOS: SMOF não alterou a quimiotaxia e fagocitose nos leucócitos estudados. TCM/TCL e TCM/TCL/OP aumentaram o número de macrófagos fagocitantes do fígado e pulmão e somente TCM/TCL/OP apresentou aumento no número de macrófagos fagocitantes no baço (p<0,05). CONCLUSÕES: 1) Emulsões lipídicas, independente de sua composição, não influenciaram a quimiotaxia de neutrófilos; 2) Emulsão lipídica composta por mistura de triglicérides de cadeia média e óleos de soja, oliva e peixe apresentou efeito neutro sobre a quimiotaxia, migração espontânea de neutrófilos e recrutamento de monócitos no fígado, pulmão e baço; 3) Emulsão lipídica de mistura física de triglicérides de cadeia média e cadeia longa estimulou o recrutamento de monócitos, com aumento do número de macrófagos fagocitantes no fígado e pulmão; 4) Emulsão lipídica de mistura física de triglicérides de cadeia média e cadeia longa enriquecida com emulsão lipídica de óleo de peixe, estimulou o recrutamento de monócitos, com aumento do número de macrófagos fagocitantes no fígado, pulmão e baço. / RATIONALE: Parenteral lipid emulsions (LE) with fish oil could modulate inflammatory response and promote or maintain immunologic response, but there are insufficient data about the impact on innate immunity cells functions. AIM: To evaluate the effects of endovenous infusion of LE containing mixture of medium-chain triglycerides and soybean, olive and fish oils on leukocytes migration and phagocytosis in rats, compared to a physical mixture of medium and long-chain triglycerides - MCT/LCT LE supplemented or not with fish oil (FO). METHOD: Isogenic Lewis rats (n=40) were submitted to jugular vein catheterization for parenteral access. The animals were randomized in four groups, according to their infusion: group SMOF: LE containing 30% of soybean oil (LCT), 30% MCT, 25% olive oil and 15% fish oil; group MCT/LCT: LE containing MCT and LCT (1:1 v/v); group MCT/LCT/FO: MCT/LCT LE enriched with fish oil based LE (8:2 v/v); group SS: saline. A non-surgical control (CO-NS) was also performed. In the 5th experimental day and after colloidal carbon injection in tail vein, blood and tissue (liver, lung and spleen) samples were collected for chemotaxis assay (adapted Boyden chamber) and colloidal carbon phagocyting-macrophages quantification (immunohistochemistry). ANOVA and Tukey post test were performed. RESULTS: SMOF LE didn?t influence leukocytes chemotaxis and phagocytosis. MCT/LCT and MCT/LCT/FO LE increased liver and lung resident phagocyting-macrophages number (p<0.05) and only in MCT/LCT/FO group, spleen resident phagocyting-macrophages number was increased (p<0.05). CONCLUSION: 1) Lipid emulsion, independently of composition, has no influence on neutrophils chemotaxis; 2) Lipid emulsion with a mixture of medium-chain triglycerides, soybean, olive and fish oils has neutral effect on neutrophil chemotaxis, random migration and monocyte recruitment to the liver, lung and spleen; 3) Lipid emulsion with a physical mixture of medium and long-chain triglycerides has stimulatory effect on monocyte recruitment, with increase of phagocyting-macrophages number in liver and lung; 4) Lipid emulsion with a physical mixture of medium and longchain triglycerides supplemented with fish oil, has stimulatory effect on monocyte recruitment, with increase of phagocyting-macrophages number in liver, lung and spleen.
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The effect of dietary fish oil replacement with soybean oil on growth and health of dusky kob, Argyrosomus japonicus (Pisces: Sciaenidae)Rossetti, Nani Adami January 2012 (has links)
Lipids are essential components for fish because they contain fatty acids that are vital for regular growth and health. Fish oil is rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are essential fatty acids for carnivorous fish, and therefore this product has traditionally been used as the main source of lipids in fish feeds. However, with declining fisheries resources worldwide and the rapid expansion of the aquaculture industry pressuring this finite resource, such ingredients are becoming less available and more expensive. It is therefore necessary to explore the utilization of ingredients that are sustainable and competitive alternatives to fish oil in marine finfish feeds. This work investigated the effects of the substitution of fish oil with soybean oil on the growth performance, feed efficiency, fatty acid composition of the liver tissue and some health parameters in juvenile dusky kob, Argyrosomus japonicus; an increasingly popular sciaenid marine aquaculture species in South Africa. Six diets (18 % total lipid and 46 % protein) with increasing percentage substitution of fish oil with soybean oil (1, 14, 28, 42, 56 and 70 %) were fed to juvenile kob. After 84 days of feeding these diets to the fish, no significant differences in fish length and weight between treatments were observed. However, there was a significant trend of a decrease in specific growth rate, ranging from (± standard error) 0.87 ± 0.06 to 0.72 ± 0.04 % body weight day⁻¹, and condition factor, ranging from 1.59 ± 0.03 to 1.54 ± 0.02, with increasing vegetable oil replacement in the diets between days 56 and 84. There were no differences in red blood cell count, haematocrit and haemoglobin concentration after 206 days of feeding. However, visceral fat index (VFI) increased significantly from 1.08 ± 0.17 % for fish fed diets with 28 % soybean oil, to 2.24 ± 0.15 % for fish fed diets with 70 % soybean oil. Similarly, hepatosomatic index (HSI) increased significantly from 0.84 ± 0.08 % to 1.80 ± 0.12 % in the control diet and the 56 % soybean oil diet, respectively. After 206 days of feeding, fish fed diets with 42 to 70 % soybean oil showed greater number of lipid vacuoles in the liver, which were also larger in size, and hepatocytes nuclei were displaced to the cell periphery. The fatty acid composition of the liver tissue strongly corresponded to the fatty acid composition of the diets. Linoleic acid accumulated in the liver of the fish fed increasing soybean oil in the diets. In contrast, EPA and DHA decreased from 13.63 to 1.97 %, and 14.34 to 3.28 %, respectively, in the liver tissue of fish fed diets with increasing soybean oil content; consequently the n-3/n-6 ratio was also significantly reduced with inclusion of vegetable oil in the diets. The trend of decreasing growth rate with increasing oil replacement towards the end of the trial corresponds with increases in VFI, HSI, as well as the fatty acid accumulation and lipid vacuoles in the liver. This suggests that dusky kob is less able to metabolise soybean oil at increased substitution levels which would account for the poorer growth at higher levels. The dependence of fish on dietary marine oil decreased significantly with each inclusion of soybean oil in the diets. Nonetheless, the calculations based on the nutrient ratio presented positive outcomes for all treatments, that is, values of marine oil dependency ratio were below one for all treatments. It is concluded that soybean oil can replace fish oil in formulated diets for dusky kob up to a level of 28 % of total dietary lipids, as evidenced by the good growth and feed efficiency, and no apparent negative health effects observed up to this level.
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Fizičko-hemijska i reološka karakterizacija mikrokapsula ribljeg ulja inkorporiranih u čokoladni matriks / Physico-chemical and rheological characterization of fish oil microcapsules incorporated in a chocolate matrixKalić Marina 31 July 2020 (has links)
<p>Omega-3 masne kiseline su uslovne za zdravlje ljudi i imaju značajne fiziološke uloge. Dijetetski proizvodi na bazi omega-3 masnih kiselina predstavljaju značajan izvor omega-3 masnih kiselina. Riblje ulje je dobar izvor polinezasićenih masnih kiselina (PUFA). Dnevni unos omega-3 polinezasićenih masnih kiselina je u većini delova sveta ispod preporučenog, uglavnom usled nedovoljne zastupljenosti ribe u ishrani. Zbog toga se danas riblje ulje nalazi u obliku različitih dijetetskih proizvoda i ponuda ovih preparata na tržištu je veoma široka. Problem sa unosom ribljeg ulja kao dodatka ishrani je njegov intenzivan i neprijatan ukus i miris, što može da dovede do neadekvatne suplementacije. Sušenje raspršivanjem (engl. spray drying) predstavlja tehniku koja omogućava trenutno sušenje rastvora, suspenzija ili emulzija. U pitanju je metoda koja ima široku primenu u farmaceutskoj industriji, a između ostalog se primenjuje i u cilju maskiranja neprijatnog ukusa lekova. Kao omotač mikrokapsula dobijenih sušenjem raspršivanjem je moguće koristiti proteine, ali je neophodno dobro ispitati i poznavati njihove fizičko-hemijske osobine i funkcionalnost. Inkorporiranjem mikrokapsula ribljeg ulja u čokoladu bi se dobila funkcionalna ili obogaćena hrana, što predstavlja i finalnu formulaciju u ovom radu. Obogaćivanjem čokolade sa visokim sadržajem kakao delova mikrokapsulama ribljeg ulja kreirao bi se višestruko funkcionalan proizvod. Odabir čokolade kao matriksa za obogaćivanje uslovljen je činjenicom da je ona široko konzumiran proizvod. Ciljevi ovog rada bili su da se ispita uticaj metode sušenja raspršivanjem na stabilnost preformulacije ribljeg ulja, da se utvrde karakteristike mikrokapsula dobijenih sušenjem raspršivanjem (prinos i efikasnost mikrokapsulacije, oksidativnu stabilnost ulja, morfološke osobine i veličinu mikrokapsula), zatim da se utvrdi uticaj veličine čestica na kristalizaciju u masnoj fazi suspenzije koja se koristi za izradu konditorskih proizvoda i da se utvrde fizičko-hemijske karakteristike (teksturu, boju, reološke osobine) čokolade koja sadrži inkorporirane mikrokapsule ribljeg ulja u odnosu na čokoladu bez dodatka mikrokapsula. Metode su obuhvatale karakterizaciju proteina dobijenih iz soje, graška, krompira, pirinča i surutke, njihovih rastvora, kao i emulzija ribljeg ulja u vodenim rastvorima tih proteina, određivanje prinosa i efikasnosti mikrokapsulacije i karakterizaciju dobijenih mikrokapsula. Prilikom ispitivanja uticaja veličine čestica na kristalizaciju u masnoj fazi suspenzije koja se koristi za izradu konditorskih proizvoda i fizičko-hemijskih osobina čokolade koja sadrži inkorporirane mikrokapsule ribljeg ulja primenjivane su metode za određivanje teksture, reoloških karakteristika, sadržaja čvrstih masti i boje dobijenih formulacija. Dobijeni rezultati su pokazali da se proteini ponašaju kao dobri emulgatori i da sušenje raspršivanjem predstavlja efikasan način za dobijanje mikrokapsula ribljeg ulja sa proteinima kao omotačima mikrokapsula. Kristalizacija masne faze u suspenziji koja predstavlja model čokolade zavisi od veličine čvrstih čestica. Kada je u pitanju proizvodnja čokolade sa inkorporiranim mikrokapsulama ribljeg ulja kod kojih su kao omotači korišćeni proteini soje, surutke i krompira, dodatak tih mikrokapsula ne utiče na karakteristike čokolade u meri dovoljnoj da bi se narušio proizvodni proces izrade. Sve navedeno upućuje na zaključak da bi proizvodnja čokolade sa inkorporiranim mikrokapsulama ribljeg ulja, tehnološki bila moguća.</p> / <p>Omega-3 fatty acids are essential for human health and have significant physiological roles. Dietary products based on omega-3 fatty acids are a significant source of omega-3 fatty acids. Fish oil is a good source of polyunsaturated fatty acids (PUFA). The daily intake of omega-3 polyunsaturated fatty acids is below the recommended level in most parts of the world, mainly due to the lack of fish in the diet. This is why fish oil is now found as various dietary products which are widely present in the world’s market. The problem with fish oil intake as a dietary supplement is its intense and unpleasant taste and odor, which can lead to inadequate supplementation. Spray drying is a technique that allows instantaneous drying of solutions, suspensions or emulsions. It is a widely used method in the pharmaceutical industry and is used, among other things, to mask the unpleasant taste of medicines. It is possible to use proteins as a coating of spray-dried microcapsules, but it is necessary to test and know their physicochemical properties and functionality. Incorporating fish oil microcapsules into chocolate would make functional or enriched foods, which is consider as a final formulation in this work. Enriching the high cocoa content chocolate with fish oil microcapsules would create a multi-functional product. The choice of chocolate as a base is conditioned by the fact that it is a widely consumed product. The aim of this study was to investigate the effect of spray drying method on the stability of fish oil pre-formulation, to determine the characteristics of microcapsules obtained by spray drying (yield and efficiency of microencapsulation, oxidative stability of oil, morphological properties and size of microcapsules), to determine the effect of particle size crystallization in the oil phase of the suspension used for confectionery products and to determine the physicochemical characteristics (texture, color, rheological properties) of chocolate containing fish oil microcapsules in comparison with chocolate without the addition of microcapsules. Methods included characterization of proteins obtained from soybeans, peas, potatoes, rice and whey, their solutions, as well as fish oil emulsions in aqueous solutions of these proteins, determination of the yield and efficiency of microencapsulation, and characterization of the microcapsules obtained. In the examination of the effect of particle size on crystallization in the oil phase of the suspension used for the manufacture of confectionery and physicochemical properties of chocolate containing fish oil microcapsules, methods were used to determine the texture, rheological characteristics, solid fat content and color of the formulations obtained. The results show that proteins act as good emulsifiers and that spray drying is an effective way to obtain fish oil microcapsules with proteins as microcapsule shells. The crystallization of the oil phase in the suspension representing the chocolate model depends on the size of the solid particles. In the case of production of chocolate with incorporated fish oil microcapsules using soybean, whey and potato proteins as coating material, the addition of these microcapsules does not affect the chocolate characteristics to a degree sufficient to impair the manufacturing process. All of the above points to the conclusion that the production of chocolate with incorporated fish oil microcapsules would be technologically possible.</p>
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The effects of dietary polyunsaturated fatty acids on prostate cancer-proteomic and phosphoproteomic studiesZhao, Heng 15 January 2016 (has links)
Indiana University-Purdue University Indianapolis (IUPUI) / This dissertation studies the effects of fatty acids on prostate cancer. Prostate cancer is one of the most common malignant diseases in males in the U.S. Because of the slow progression of this disease, early intervention methods, especially, dietary fatty acid interventions are considered very important to control the disease in early stages. This study describes how the depletion of the enzyme for endogenous fatty acid synthesis, fatty acid synthase, influences the expression of enzymes that metabolize dietary fatty acids and show how dietary fatty acids affect prostate cancer protein expression and function. Fatty acid synthase is an oncoprotein overexpressed in prostate cancer and its expression is suppressed with omega-3 fatty acid treatment. This study finds that the depletion of fatty acid synthase by siRNA knockdown induces suppression of cyclooxygenase-2 and fatty acid desaturase-1. Our results also show that fish oil (omega-3 fatty acid), but not oleic acid (omega-9 fatty acid), suppresses prostate cancer cell viability. Assessment of fatty acid synthesis activity indicates that oleic acid is a more potent inhibitor than fish oil of de novo fatty acid biosynthesis. In addition, the inhibition of its activity occurs over several days while its effects on cell viability occur within 24 hours. To better understand this relationship, label free LC-MS/MS based mass spectrometry was carried out to determine global proteomic and phosphoproteomic profiles of the prostate cell line PC3, with longitudinal treatment with fish oil or oleic acid. With short-term fish oil treatment, sequestosome-1was elevated. Prolonged treatment induced downregulation of microseminoprotein, a proinflammation factor, as well as proteins in the glycolysis pathway. In the phosphoproteomics study, we confidently identified 828 phosphopeptides from 361 phosphoproteins. Quantitative comparison between fish oil or oleic acid treated groups and the untreated group suggests that the fish oil induces changes in phosphorylation of proteins involved in the pathways associated with cell viability and metabolic processes, with fish oil inducing significant decreases in the levels of phospho-PDHA1Ser232 and phospho-PDHA1Ser300 and they were accompanied by an increase in PDH activity, suggesting a role for n-3 polyunsaturated fatty acids in controlling the balance between lipid and glucose oxidation.
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Lipase-catalyzed purification and functionalization of Omega-3 polyunsaturated fatty acids and production of structured lipids / Purification et fonctionnalisation d’acides gras polyinsaturés Oméga-3 par des lipases et production de lipides structurésCasas Godoy, Leticia 14 December 2012 (has links)
Les lipases sont des enzymes présentant un grand intérêt industriel. L’intérêt de ces enzymes a conduit à caractériser ces enzymes, à mieux comprendre leur mécanisme réactionnel et leur cinétique, et à établir des méthodes efficaces de production en système d’expression homologue et hétérologue. Plus récemment, l’ingénierie enzymatique permet d’améliorer les caractéristiques des enzymes. Ce thèse s’est fixé deux objectifs principaux: premièrement, la purification et la fonctionnalisation d’acides gras poly-insaturés de type Omega-3 (PUFAs), et spécialement l’acide cis-4, 7, 10, 13, 16, 19-docosahexaénoique (DHA) et deuxièmement la production de lipides structurés (SL). Un premier objectif fut de produire une molécule pharmaceutique, le nicotinyl DHA ester. Le co-substrat du DHA est le nicotinol, un alcool qui après absorption, il est rapidement converti en acide nicotinique (Vitamine B3). La trans-esterification enzymatique entre l’ester éthylique du DHA et le nicotinol a été optimisée dans le but de synthétiser un ester présentant les propriétés cumulatives des deux réactants. Après la sélection de l’enzyme optimale (lipase immobilisée de Candida antarctica; Novozyme 435) et le choix du milieu réactionnel (milieu sans solvant), le procédé a été optimisé. Une conversion supérieure à 97 % a été obtenu en 4 heures avec 45 g.L-1 d’enzyme. Dans ces conditions, une productivité de 4.2 g de produit .h-1.g d’enzyme-1 a été obtenue. Ce projet nécessite une haute pureté en DHA. Un procédé de purification enzymatique a été choisi. Les lipases sont capables de discriminer entre les acides gras en fonction de la longueur de chaine et du degré d’insaturation. Les lipases agissent par résolution cinétique, en réagissant plus efficacement avec les acides gras saturés et mono-insaturés qu’avec les PUFAs résistants. La lipase YLL2 de Yarrowia lipolytica apparait comme un bon candidat car elle est homologue à une des lipases les plus efficaces, la lipase de Thermomyces lanuginosus. YLL2 a permis d’obtenir une discrimination très efficace. Les raisons de la sélectivité de l’enzyme ont été identifiées : il s’agit du positionnement de la double liaison la plus proche de la fonction carboxylique. La concentration en DHA la plus élevée a été obtenue avec YLL2 (73%) avec un pourcentage de récupération du DHA-EE de 89%. YLL2 est par conséquent l’enzyme décrite la plus efficace pour la purification du DHA.La mutagénèse ciblée dans le site actif de YLL2 a été utilisée pour améliorer la sélectivité de cette enzyme. L’analyse de la structure 3D et les alignements avec des lipases homologues a permis de choisir les cibles de mutagénèse dirigée. Les acides aminés cibles ont été changés de manière à restreindre ou élargir le site actif. De ce premier screening de variantes deux positions ont permis d’améliorer la spécificité de l’enzyme, les positions I100 et V235. Finalement la saturation de ces 2 positions a été réalisée. Le dernier objectif de la thèse était la production de SL par acidolysis enzymatique entre l'huile d'olive vierge et les acides caprylic ou capric utilisant la lipase YLL2 immobilisé. Le SL obtenu devrait être riche en acide oléique à la position sn-2 tandis que les C8:0 et C10:0 devraient être principalement estérifiés aux positions sn-1,3. YLL2 immobilisé sur Accurel 1000 a été testé dans un système sans solvant. La réaction d’acidolysis d'huile d'olive avec C8:0 ou C10:0 a été optimisée avec la méthodologie de surface de réponse (RSM). / Lipases are enzymes with applications extended to a wide variety of industries. The variety of lipases applications led to increased research to characterize them and better understand their kinetics and reaction mechanisms and to establish methods for lipase production in homologous and heterologous expression systems. Lately enzymatic engineering allowed the improvement of lipase characteristics. This thesis project studies the use of lipases for two main objectives: lipase-catalyzed purification and functionalization of Omega-3 polyunsaturated fatty acids (PUFAs), especially cis-4, 7, 10, 13, 16, 19-docosahexaenoic acid (DHA) and production of structured lipids (SL). DHA was used for the synthesis of a pharmaceutical molecule, the nicotinyl DHA ester. The co-substrate of the reaction was nicotinol, an alcohol from the group B pro-vitamin, which after absorption is rapidly converted into nicotinic acid (Vitamin B3). The enzymatic trans-esterification of DHA ethyl esters with nicotinol was optimised to synthesise an ester presenting the cumulative properties of the two reactants. After enzyme (immobilized lipase from Candida antarctica; Novozym 435) and reaction medium (solvent-free system) selection, the process was optimised. A conversion to nicotinyl-DHA superior to 97 % was obtained in 4 hours using 45 g.L-1 of enzyme. With a productivity of 4.2 g of product .h-1.g of enzyme-1.This project requires DHA of high purity. Enzymatic purification was chosen for the production of DHA concentrates. Lipases can discriminate between fatty acids in function of their chain length and saturation degree. Lipases react more efficiently with the bulk of saturated and mono-unsaturated fatty acids than with the PUFAs. The objective was the discovery of more specific enzymes for DHA purification. The lipase Lip2 from Yarrowia lipolytica (YLL2) appears as a good candidate since it is homologous to one of the most efficient lipase, the lipase from Thermomyces lanuginosus. YLL2 enables a high discrimination to be obtained, enzyme selectivity being principally due to the positioning of the double-bond the closest from the carboxylic group. The highest concentration of DHA was obtained with YLL2 (73%) with a recovery percentage of DHA-EE of 89%. YLL2 is the most efficient described lipase for DHA purification.Site directed mutagenesis was used to improve YLL2 from Y. lipolytica. Using its three dimensional structure and alignment with homologous lipases, targets for site directed mutagenesis were chosen. Chosen amino acids were substituted by two amino acids of different sizes. From the screening of variants two positions with promising specificities where chosen, positions I100 and V235. Finally saturation of both positions and the analysis of their performances in the selected reactions were carried out. The last objective was the production of SL by enzymatic acidolysis between virgin olive oil and caprylic or capric acids using immobilized Lip2 from Y. lipolytica. The SL obtained should be rich in oleic acid at the sn-2 position while C8:0 and C10:0 should be mainly esterified at the sn-1,3 positions. Lip2 from Y. lipolytica immobilized on Accurel MP 1000 was tested in a solvent-free system. The acidolysis reaction of olive oil with C8:0 or C10:0 was optimized by response surface methodology (RSM)
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Effect of n-3 vs n-6 fatty acids and methyl ethyl ketone peroxide on adipose tissue cellularity, muscle weight, and lipoprotein lipase activity in ratsVenkateswaran, Lakshmi, 1965- 22 March 1993 (has links)
Graduation date: 1993
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Olive oil or lard? Distinguishing plant oils from animal fats in the archaeological record of the eastern Mediterranean using gas chromatography/combustion/isotope ratio mass spectrometrySteele, Valerie J., Stern, Ben, Stott, A.W. 15 December 2010 (has links)
Yes / Distinguishing animal fats from plant oils in archaeological residues is not straightforward. Characteristic plant sterols, such as ¿-sitosterol, are often missing in archaeological samples and specific biomarkers do not exist for most plant fats. Identification is usually based on a range of characteristics such as fatty acid ratios, all of which indicate that a plant oil may be present, none of which uniquely distinguish plant oils from other fats. Degradation and dissolution during burial alter fatty acid ratios and remove short chain fatty acids, resulting in degraded plant oils with similar fatty acid profiles to other degraded fats.
Compound specific stable isotope analysis of ¿13C18:0 and ¿13C16:0, carried out by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS), has provided a means of distinguishing fish oils, dairy fats, ruminant and non-ruminant adipose fats but plant oils are rarely included in these analyses. For modern plant oils where C18:1 is abundant, ¿13C18:1 and ¿13C16:0 are usually measured. These results cannot be compared with archaeological data or other modern reference fats where ¿13C18:0 and ¿13C16:0 are measured, as C18:0 and C18:1 are formed by different processes resulting in different isotopic values.
Eight samples of six modern plant oils were saponified releasing sufficient C18:0 to measure the isotopic values, which were plotted against ¿13C16:0. The isotopic values for these oils, with one exception, formed a tight cluster between ruminant and non-ruminant animal fats. This result complicates the interpretation of mixed fatty residues in geographical areas where both animal fats and plant oils were in use. / AHRC
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