Ever since the emergence of the hypothesis that linked the aetiology of schizophrenia with abnormal membrane phospholipids composition, an increasing number of evidences have suggested reduced membrane polyunsaturated fatty acids in patients with schizophrenia. This has led to a conduct of several studies to evaluate the efficacy of omega-3 fatty acid supplement in the modification of membrane phospholipids and treatment of schizophrenia. The two main omega-3 fatty acid classes, EPA and DHA, play a vital role in membranes. This project work reviews omega-3 fatty acid studies and summarizes their outcomes. Eight original articles (nine studies) were reviewed. Six out of nine studies measured RBC membrane fatty acids levels and all six studies reported a significant increase in EPA after EPA supplement. Two studies reported increased DHA post omega-3 fatty acid and DHA supplement, respectively. One study observed a dose-dependent increment in DHA after EPA supplement. Improved symptoms were observed in seven studies, while one study found a worsening of symptoms in patients with low baseline PUFA. Moreover, out of the six studies that evaluated the correlation between symptom change and membrane fatty acids change, three studies observed a correlation between increased EPA and symptom improvement. One study reported an increased AA associated with improved symptoms, in contrast to another study, which found a correlation between increased AA and worsened symptoms. The conclusion from this project work is that EPA supplement can increase the EPA levels in membranes; however, its therapeutic effect in schizophrenia requires further investigation using larger studies. / Ända sedan tillkomsten av hypotesen som länkade etiologin av schizofreni med onormala sammansättningar av membranfosfolipider, har bevis för nedsatt membranfettsyror hos patienter med schizofreni ökat. Detta har lett till genomförandet av flera studier för att utvärdera effekten av omega-3 supplement i modifieringen av membranfosfolipider och i behandling av schizofreni. De två viktigaste omega-3 klasserna, EPA och DHA, spelar en viktig roll i membran. Detta projektarbete granskar de omega-3 studierna och sammanfattar deras resultat. Åtta originalartiklar (nio studier) granskades. Sex av nio studier mätte nivåer av RBC membranfettsyror och alla sex studierna rapporterade en signifikant ökning av EPA efter EPA behandling. Två studier rapporterade ökad DHA efter omega-3 och DHA behandling, respektive. En studie observerade en dosberoende ökning i DHA efter EPA behandling. Förbättrade symtom observerades i sju studier, medan en studie fann en försämring av symtom hos patienter med låg baseline PUFA. Av de sex studier som utvärderade sambandet mellan symtomförändring och förändring i membranfettsyror, hittade två studier samband mellan ökad EPA och symtomförbättring. En studie rapporterade en ökad AA i samband med förbättrade symtom, i motsats till en annan studie, som fann ett samband mellan ökad AA och försämrade symtom. Slutsatsen från detta projektarbete är att EPA tillägg ökar nivåer av EPA i membranfosfolipider; men dess terapeutiska effekt vid schizofreni kräver ytterligare utredning med hjälp av större studier.

O experimento foi realizado no Biotério Experimental da USP em Pirassununga - SP. Na primeira fase foram utilizados 42 coelhos machos em crescimento da raça Nova Zelândia mantidos individualmente durante todo o experimento. O ensaio biológico totalizou 150 dias. Na Fase 1 os animais receberam dietas com indução de hipercolesterolemia (0,5% de colesterol), com desequilíbrio na proporção de Omega-6/Omega-3 (n-6:n-3) de 15:1 e vice versa e balanço na proporção de n-6:n-3 de 4:1 (controle), totalizando 6 ensaios, os quais foram divididos aleatoriamente em 7 grupos de 6 coelhos cada. Óleos de girassol e de peixe foram utilizados como fontes de ácidos graxos poli-insaturados n-6 e n-3, respectivamente. Na Fase 2, os animais remanescentes continuaram recebendo as mesmas dietas, exceto no grupo B (hipercolesterolemia) que passou a receber o tratamento controle para verificar o efeito. No final da Fase 1, três animais de cada grupo foram eutanasiados com retirada da artéria aorta para determinação de placas lipídicas, histopatológica, colesterol total e imunohistoquímica para verificação da expressão da enzima LDL-receptor e determinação histopatológica do tecido hepático. Em soro foram realizadas determinações de perfil de ácidos graxos, colesterol total, LDL e HDL-colesterol e triacilgliceróis. Células endoteliais aórticas de coelhos foram isoladas para a realização do teste dose resposta com óleo de peixe para identificar a melhor relação do consumo destes ácidos graxos comparando com o experimento in vivo. O objetivo deste trabalho foi monitorar os efeitos do fornecimento de cada dieta sobre o teor de colesterol total, LDLcolesterol, HDL-colesterol e triacilgliceróis em sangue de coelhos. No geral, o perfil de ácidos graxos no soro correspondeu diretamente com a dieta consumida. Os principais efeitos dos ácidos graxos poli-insaturados Omega 3 foi a redução dos lipídios séricos quando os coelhos remanescentes da dieta hipercolesterolêmica (0,5% de colesterol e n-6:n-3 de 2:1) passaram a receber o equilíbrio da relação de ácidos graxos 4:1 de n-6:n-3. O teor de colesterol total no soro, artérias e o crescimento dos ateromas foram influenciados pelo elevado consumo de ácidos graxos n-6 e n-3 associados à adição de colesterol nas dietas. Por outro lado verificamos que o excesso de Omega 3 associado ou não ao colesterol contribuiu para o agravamento das placas ateroscleróticas inclusive deposição de cálcio nas mesmas e paredes endoteliais. Provavelmente, o excesso de n-3 tenha ocasionado efeito contrário às suas funções anti-inflamatória, antiagregatória e anti-trombótica. Dietas hipercolesterolêmicas provocaram esteatose hepática e o elevado consumo de Omega-6 em detrimento do baixo consumo de Omega -3 induziram hepatite crônica. Já o consumo elevado de Omega-3, sem colesterol, regrediu a esteatose hepática nos animais. O consumo equilibrado de n-6:n-3 reduziu a expressão da enzima LDL-receptor no grupo que anteriormente recebeu dieta hipercolesterolêmica. Ressaltando-se a importância do consumo equilibrado destes ácidos graxos já que esta enzima é controlada pelo colesterol livre circulante. No teste dose-resposta em células endoteliais da aorta de coelhos foi evidenciado que a melhor proporção de n-6:n-3 seria de 9 vezes mais do que a concentração de Omega 3 recomendada de 5:1. / The experiment was conducted at the Experimental Biotery by USP in Pirassununga - SP. In the first phase were used 42 male rabbits growing New Zealand breed individually maintained throughout the experiment. The biological assay amounted to 150 days. In phase 1, the animals received diets induced hypercholesterolemia (0.5% cholesterol) with imbalance in the ratio of Omega-6 / Omega-3 (n-6 / n-3) 15:1 and vice versa, and balance the ratio of n-6 / n-3 of 4:1 (control), totaling six tests, which were randomly divided into 7 groups of six rabbits each. Sunflower and fish oils were used as sources of fatty acids polyunsaturated n-6 and n-3, respectively. In Phase 2, the remaining animals continued to receive the same diets except in group B (hypercholesterolemia) who went on to receive the control treatment to check the effect. At the end of Phase 1, three animals from each group were sacrificed with the aorta artery removed for determination of lipid plaques, histopathology, immunohistochemistry and total cholesterol to verify the expression of LDL-receptor enzyme and determination of histopathological liver tissue. In serum profile were made determinations of fatty acids, total cholesterol, LDL and HDL cholesterol and triglycerides. Aortic endothelial cells of rabbits were isolated to perform the dose response test with fish oil to identify the best value for the consumption of these fatty acids compared to the in vivo experiment. The objective of this work was to monitor the effects of provision for each diet on total cholesterol, LDL-cholesterol, HDLcholesterol and triglycerides in the blood of rabbits. In general, the profile of fatty acids in serum corresponded directly with the diet consumed. The main effects of polyunsaturated Omega 3 fatty acids was the reduction of serum lipids when the remaining rabbits with hypercholesterolemic diet (0.5% cholesterol and n-6: n-3 2:1) have received the balance of the relationship fatty acid 4:1 n-6: n-3. The total cholesterol content in serum, the growth of arteries and atheroma were influenced by a high intake of n-6 fatty acids and n-3 associated with the addition of cholesterol in diets. On the other hand we verified that the excess of Omega 3 with or without cholesterol contributed to the aggravation of the atherosclerotic plaques including deposition of calcium in them and endothelial walls. Probably the excess of n-3 has caused the opposite effect to their anti-inflammatory, anti-thrombotic and anti-aggregatory functions. Hypercholesterolemic diets caused hepatic steatosis and high intake of Omega-6 to the detriment of low consumption of Omega -3 induced chronic hepatitis. High consumption of Omega-3, no cholesterol, decreased hepatic steatosis in animals. Highlighting the importance of the balanced intake of this fatty acids, since this enzyme is controlled by circulating free cholesterol. In dose-response test in endothelial cells of the rabbits aorta was evident that the best ratio of n-6: n-3, nine times more than the recommended concentration of Omega 3 to 5:1.

Em equinos é descrita uma variabilidade na qualidade do sêmen congelado, relacionada principalmente a variações consideráveis na composição da membrana plasmática do espermatozoide. Neste contexto, estudos investigam alternativas para aumentar a fertilidade ao se usar sêmen congelado, e a adição de ácidos graxos polinsaturados ao diluente de congelamento é sugerida. O objetivo deste estudo foi avaliar o efeito da adição de diferentes níveis de ácidos graxos poli-insaturados: ácido eicosanóico e ácido docosahexaenóico, em meio diluente específico para a espécie sobre as características do espermatozoide pós-descongelamento. Foi utilizado sêmen de quatro garanhões (quatro ejaculados por garanhão) da raça Crioula. O sêmen foi diluído em diluente de congelamento comercial a base de gema de ovo, glicerol e dimetilformamida (Botu-Crio©) ajustando a concentração para 200 x 10⁶ espermatozoides viáveis/mL (grupo controle) e, na sequência, os demais tratamentos: adição de ácido docosahexaenóico nas doses 25μm e 50μm /mL e ácido eicosanóico nas doses 25μm e 50μm /mL. Após o descongelamento foram realizadas análises de cinética espermática no sistema de Análise Computadorizada para Avaliação Espermática, das seguintes variáveis: motilidade total, motilidade progressiva, motilidade rápida, motilidade lenta, motilidade local, velocidade de trajeto, velocidade progressiva, velocidade curvilinear, amplitude do deslocamento lateral da cabeça, frequência de batimentos, retilinearidade e linearidade; e avaliação da integridade física através do uso de sondas fluorescentes, e funcionalidade de membrana pelo teste hiposmótico. Não ouve diferença nas variáveis avaliadas. Este é o primeiro estudo que descreve a adição de ácido eicosanóico ao sêmen equino. Concluí-se a adição de ácido docosahexaenóico e eicosanóico nas concentrações testadas não alterou as variáveis avaliadas no sêmen de garanhões da raça Crioula. / In equines, a variability in the quality of frozen semen is described, mainly related to the considerable variations in the composition of the sperm plasma membrane. In this context, studies investigate alternatives to increase fertility when using frozen semen, and the addition of polyunsaturated fatty acids to the freezing diluent is suggested. The objective of this study was to evaluate the effect of the addition of different levels of polyunsaturated fatty acids: eicosanoic acid and docosahexaenoic acid, in specific diluent medium for the species on the characteristics of the post-thawing spermatozoa. Semen was used of four stallions (four ejaculates per stallion) of the Crioula breed. The semen was diluted in commercial freezing diluent based on egg yolk, glycerol and dimethylformamide (Botu-Crio ©) by adjusting the concentration to 200 x 10 vi viable spermatozoa / mL (control group) and, in sequence, the other treatments: addition of docosahexaenoic acid at 25μm and 50μm / mL and eicosanoic acid at 25μm and 50μm / mL. After thawing, sperm kinetics analyzes were performed in the Computerized Analysis System for Sperm Evaluation, of the following variables: total motility, progressive motility, rapid motility, slow motility, local motility, path velocity, progressive velocity, curvilinear velocity, lateral head displacement amplitude, beating frequency, linearity and linearity; and evaluation of physical integrity through the use of fluorescent probes, and membrane functionality by the hyposmotic test. No difference in the variables evaluated. This is the first study to describe the addition of eicosanoic acid to equine semen. We concluded that with the addition of docosahexaenoic and eicosanoic acid at the concentrations tested did not alter the variables evaluated in the semen of Crioula stallions.

Cette thèse a consisté à développer une nouvelle stratégie analytique en utilisant le couplage chromatographie liquide - spectrométrie de masse (LC-MSn) afin d’identifier dans les huiles et corps gras, les triglycérides (TAGs) et leur régio-spécificité. Cette méthodologie analytique générale a été appliquée à 8 huiles de graine de résineux en particulier, à l’huile de pignon de Pinus Koraiensis car ses TAGs sont constitués d’acides gras, possédant une double liaison en position Δ5, non-méthylène alternée, susceptibles de jouer un rôle anti-cholestérol chez l’homme.La thèse se décompose en quatre chapitres: les deux premiers, bibliographiques, traitent respectivement de la taxonomie des 8 résineux étudiés, de la composition en acide gras (AG) de leur graine et des techniques analytiques déjà décrites pour caractériser les TAGs. Le troisième chapitre, expérimental, est dédié au développement de nouvelles stratégies analytiques NARP-LC-MSn mises en place pour identifier les TAGs présents dans les huiles et plus particulièrement à l’huile de pignon de Pinus Koraiensis. Grâce à l’ajout post colonne de sel d’argent, il a été possible de déterminer de manière non ambigüe la structure de tous les TAGs d’un lipide, même ceux présents en faible quantité, par Ag+-NARP-LC-MS mais aussi de déterminer la structure de chaque AG constitutif d’un TAG par fragmentation de l’adduit moléculaire par Ag+-NARP-LC-MS2. Le problème de la distinction des TAGs ayant la même masse moléculaire, les mêmes longueurs de chaine mais des positions de double liaisons différentes, a été résolu en développant une méthode d’identification des TAGs à partir des lois de rétention chromatographiques qui relient linéairement le logarithme du facteur de rétention de chaque TAG soit au nombre total de carbone soit au nombre total de double liaisons. Cette étude a permis d’identifier 22 nouveaux TAGs parmi un nombre total de 58 TAGs caractérisés. Elle a amené la preuve que le résidu AG saturé à 17 atomes de carbone est ramifié et non linéaire. Elle a mis en évidence la présence de trois AGs constitutifs qui n’ont jamais été décrit: 19:1, 19:2 et 24:0 dans l’huile de pignon de Pinus Koraiensis. Le quatrième chapitre porte sur le développement de trois méthodes de détermination de la régiospécificité des TAGs, grâce à la seule SM: (1) une méthode Ag+-NARP-LC-ESI-MS2 utilisant les rapports des ions diglycériques des TAGs. Il en ressort que cette méthode n’est pas assez fiable pour déterminer la structure des TAGs. (2) la seconde méthode fait appel à des expériences MS4/ MS5 sur les adduits argent des TAGs. Elle s’avère pertinente à condition d’avoir à disposition les couples de TAGs standards. (3) enfin une troisième méthode, utilisant la MS2, s’appuie sur le principe de la méthode de dissociation compétitive d’une paire [TAGref – Li -TAG]+. En utilisant la méthode des ajouts dosés il est montré qu’elle ne nécessite que de la disponibilité d’un seul des deux TAGs stéréoisomères comme standard.Ces méthodes originales ont permis, malgré leurs limites respectives de caractériser la régiospécificité d’un certain nombre de TAGs présents dans l’huile de graine de Pinus Koraiensis. / This thesis consisted of developing a new analytical strategy using liquid chromatography coupled with mass spectrometry (LC-MSn) to identify in oils and fats, triglycerides (TAGs) and their regio-specificity. The general analytical methodology was applied to 8 conifers seed oils, in particular, the seed oil of Pinus koraiensis. These conifer seed oils differ from common edible vegetable oils by having a series of unusual polyunsaturated fatty acids (UPIFA) with a polymethylene-interrupted (PMI) double bond system and a double bond at the 5 position which may have anti-cholesterol properties. This thesis is composed of four chapters: the two first chapters, literature review, are devoted respectively to TAGs of the 8 resinous seed oils studied and analytical techniques already used. The third chapter is dedicated to the development of a new analytical strategy combining HPLC with mass spectrometry method to identify TAGs in Pinus Koraiensis seed oil. Thanks to post column addition of silver salt, it was possible to determine the TAGs present by Ag+-NARP-LC-MS and also the fatty acids composition of the TAGs by Ag+-NARP-LC-MS2. Concerning the distinction between TAGs with the same mass, same chain length but differing positions of double bond on fatty acid chain, it was determined by chromatographic retention rules which link linearly the logarithm of retention factor of each TAG to the total carbon number or the total number of double bonds. This study has identified 22 new TAGs from a total of 58 TAGs characterized. It confirmed the knowledge that the saturated fatty acid with 17 carbon atoms is branched, not linear. It highlighted the presence of three constituent fatty acids that have never been described: 19:1, 19:2 and 24:0 in the seed oil of Pinus koraiensis. The last experimental part is devoted to the regiospecificity determination of TAGs. Three methodologies were developed. The first one used the ratios of diacylglycerol ions of TAGs but was not reliable enough. The second method used the LC-MS4 experiments (It is relevant if they have available couples of TAG standards). Finally, a third method, using MS2, based on the principle of the method of competitive dissociation of a pair [TAGref - Li-TAG] +. Using the method of standard additions it has been shown that it requires the availability of one of the two stereoisomers TAGs as a standard.These methods provide a significantly different approach to regioisomer characterization of TAGs and overcome most of the shortcomings of existing methodologies.

Submitted by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-28T15:57:27Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao Katia Alvarez Rodriguez.pdf: 22830774 bytes, checksum: 7da7c913719a287d7f9ec9ce099f951e (MD5) / Approved for entry into archive by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-09-28T15:57:43Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao Katia Alvarez Rodriguez.pdf: 22830774 bytes, checksum: 7da7c913719a287d7f9ec9ce099f951e (MD5) / Made available in DSpace on 2017-09-28T15:57:43Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao Katia Alvarez Rodriguez.pdf: 22830774 bytes, checksum: 7da7c913719a287d7f9ec9ce099f951e (MD5) Previous issue date: 2011 / Hospital Central do Exército / Nos últimos anos a obesidade mórbida tem despertado grande interesse em pesquisadores, particularmente no que diz respeito às alterações metabólicas e nutricionais e está associada a várias co-morbidades entre elas a hipertensão arterial, diabetes mellitus, dislipidemias. Dentro dessa abordagem, diversas pesquisas têm sido conduzidas visando o desenvolvimento de modalidades terapêuticas que tenham impacto positivo na obesidade. Este estudo tem como principal objetivo avaliar a ação terapêutica da farinha de linhaça na redução do peso e na diminuição do risco de doença cardiovascular em indivíduos obesos mórbidos candidatos à cirurgia bariátrica. Este ensaio clínico de caráter prospectivo, randomizado e duplo-cego cruzado comparado ao placebo, foi realizado com 21 pacientes com diagnóstico de obesidade mórbida que são acompanhados no Hospital Central do Exército através da equipe multidisciplinar do grupo de obesidade. Os pacientes foram randomizados para a suplementação da farinha de linhaça (contendo 20 g em forma de barra de cereal) e placebo e avaliados por um período de 30 dias consecutivos, com intervalo de 30 dias após a administração das barras para a posterior inversão da administração dos mesmos por mais um período de 30 dias, foram submetidos à uma avaliação clínica, dietética, antropométrica e laboratorial. No presente estudo, o estado nutricional e metabólico foi avaliado pelo Índice de Massa Corpórea (IMC), circunferências cintura (CC), quadril (CQ), relação cintura /quadril (RC/Q) e dosagens bioquímicas de glicemia em jejum, perfil lipídico e Proteína C Reativa (PCR). Os achados entre os grupos foram comparados pela análise de variância. O valor de p foi significante se p< 0,05. Observou-se redução significativa para PCR com p valor= 0,01 e nos valores antropométricos como peso, IMC, CC e R C/Q com p valor < 0,05. Na presente experiência, houve adesão e tolerância satisfatória ao consumo às barras de cereal, mostrando que a ingestão de 20g/dia de farinha de linhaça pode apresentar efeitos positivos na diminuição do peso e PCR, diminuindo assim, complicações metabólicas e cardiovasculares futuras, com consequente diminuição da mortalidade / In recent years, morbid obesity has attracted great interest by researchers, particularly regarding metabolic and nutritional changes and it is associated with multiple co-morbidities including hypertension, diabetes mellitus, dyslipidemias. Within this approach, many studies have been conducted for the development of therapeutic modalities that have positive impact on obesity. This study aims to evaluate the therapeutic action of flaxseed flour on weight reduction and decreased risk of cardiovascular disease in morbidly obese candidates for bariatric surgery. This prospective clinical trial of character, randomized, double-blind crossover comparison with placebo, was conducted on 21 patients diagnosed with morbid obesity that are followed in the Central Army Hospital by a multidisciplinary team of the group of obesity. Patients were randomized to supplementation of flaxseed meal (containing 20 g bar-shaped cereal) and placebo it is evaluated for a period of 30 consecutive days with an interval of 30 days after administration of the bars for the subsequent reversal of the administration the same for over a period of 30 days and after, underwent a clinical evaluation, dietary, anthropometric and laboratory. In this study, nutritional and metabolic status were evaluated by body mass index (BMI), waist circumference (WC), hip circumference (HC), waist / hip ratio (RC / Q) and biochemical measurements of fasting glucose, lipid profile and C-reactive protein (CPR). The findings were compared between groups by analysis of variance. The p value was significant at p <0.05.There was a significant result for PCR with p value = 0.01. Significant reduction in the assessment of weight, BMI, WC and WHR, with p value <0.05. In this experiment, there was satisfactory compliance and tolerance to the consumption of cereal bars, showing that the intake of 20g/day of flaxseed flour may have positive effects in reducing the weight and CRP, thereby reducing future cardiovascular and metabolic complications, with consequent decreased mortality

Made available in DSpace on 2015-04-22T22:01:58Z (GMT). No. of bitstreams: 1 Banny Silva Barbosa.pdf: 3257736 bytes, checksum: aa77c1eca753a92a01d30a721f2da598 (MD5) Previous issue date: 2013-01-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Seeing the importance of fish for amazonian people, it potential in the market, it nutritive value, and information shortages relative to the lipid composition of amazonian fish, it was aimed to determinate the lipid composition attendant in dorsal muscle of six amazonia fish species, jaraqui, curimatã, pacu, sardinha, pescada and surubim, through fatty acids analyses constituent of their diferent classes of lipids and attendant steroids in their unsaponifiable lipids. This study involved the development and methodology application for extraction of total lipids, separation of lipid class, extraction of unsaponifiable lipids, derivatization of fatty acids and steroids, analyses of gas chromotagraphy with detector of flames ionization and with mass spectrometry for quantitative and qualitative evaluation of steroids and fatty acids of lipids in class. And also approached the determination of lipids nutritional quality through atherogenicity, thrombogenicity index and quantity of hypercholesterolemic fatty acids. The results indicated the fish in study have interesting lipids, having bigger quantity of total lipids the fishes curimatã and pacu, with bigger participation of neutral lipids, and cholesterol as majority steroid for all fishes. The unsaturated fatty acids, essentials for human health, it was found in bigger quantity in fishes sardinha and pacu in phospholipids class and in pescada (whitefish), curimatã, jaraqui and surubim in neutral lipids class. Besides pacu fish has shown bigger quantity of omega 3 and 6 fatty acids, the fish pescada (whitefish) highlighted for nutritional quality. / Considerando a importância do pescado para o povo amazônico, o seu potencial no mercado, a sua valorização nutritiva, e à escassez de informações referentes à composição lipídica dos peixes amazônicos objetivou-se determinar a composição lipídica presente no músculo dorsal de seis espécies de peixes amazônicos, jaraqui, curimatã, pacu, sardinha, pescada e surubim, através das análises dos ácidos graxos constituintes de suas diferentes classes de lipídeos e dos esteróides presentes em seus lipídeos insaponificáveis. O estudo envolveu o desenvolvimento e a aplicação de metodologias para extração dos lipídeos totais, separação de classes de lipídeos, extração de lipídeos insaponificáveis, derivatização de ácidos graxos e esteróides, análises por cromatografia gasosa com detector de ionização de chamas e com espectrometria de massas para avaliação quanti e qualitativa de esteróides e de ácidos graxos dos lipídeos em classes. E ainda abordou a determinação da qualidade nutricional dos lipídeos através dos índices de aterogenecidade, de trombogenecidade e pela quantidade de ácidos graxos hipocolesterolêmicos. Os resultados indicaram que os peixes em estudo contêm lipídeos interessantes, possuindo maior quantidade de lipídeos totais os peixes curimatã e pacu, com maior participação dos lipídeos neutros, e colesterol como esteróide marjoritário para todos os peixes. Os ácidos graxos insaturados, essenciais para a saúde humana, foi encontrado em maior quantidade nos peixes sardinha e pacu na classe dos fosfolipídeos e nos peixes peixes curimatã, pescada, jaraqui e surubim na classe dos lipídeos neutros. Apesar de o peixe pacu ter mostrado maior quantidade de ácidos graxos de ômega 3 e 6 o peixe pescada se destacou pela qualidade nutricional.

Vitamin E exhibits anti-tumor activity by regulating pathways in cancer cells, potentially the lipoxygenase (LOX) pathway. We studied the effects of alpha tocopherol (AT), gamma tocopherol (GT), gamma tocotrienol (GT3), and an alpha-gamma tocopherol mixture (ATGT) on the production of the LOX metabolites 13-hydroxyoctadecaenoic acid (HODE), 15-hydroxyeicosatetraenoic acid (HETE), 12-HETE, and 5-HETE in colorectal cancer. These metabolites were examined in the HCT-116 cell line after 24 h treatment with select vitamin E isoforms and quantified by LC/MS/MS. Under physiological conditions, we find that treatment with varying vitamin E isoforms have different effects on the production of 13-HODE, 15-HETE, 12-HETE, and 5-HETE. GT increases 13-HODE and decreases 12-HETE. AT reverses the effects of GT regulation on the LOX pathway, while GT3 has no significant effect on the metabolites tested. GT shows superiority in regulating the LOX pathway as it increases 13-HODE and decreases 12-HETE for possible prevention of colorectal cancer.

The epidermal growth factor receptor (EGFR) is a transmembrane receptor tyrosine kinase integral in regulating cell growth, survival, and migration. EGFR signaling, which is dependent on localization of the receptor within lipid rafts, is often hijacked during colon tumorigenesis. Previous work has found that docosahexaenoic acid (DHA) is protective against colon cancer. This fatty acid is proposed to function in part by perturbing lipid rafts and thereby altering cell signaling. The overall objective of this work was to determine whether DHA alters EGFR function and signaling. We assessed EGFR localization and ligand-induced phosphorylation in YAMC cells treated with fatty acids. We found that DHA reduced the localization of EGFR to lipid rafts. Concomitant with altering receptor localization, DHA was found to increase EGFR phosphorylation. However, DHA paradoxically suppressed EGFR signal transduction. We found that DHA uniquely altered EGFR activity, and other long chain polyunsaturated fatty acid did not exert the same effect. We additionally observed similar effects on EGFR activation and signaling by feeding mice a diet enriched in fish oil (high in DHA), and this was attendant with reduced colon tumorigenesis. We next probed the mechanism by which DHA enhances EGFR phosphorylation. We found that DHA facilitates receptor dimerization to increase phosphorylation. We additionally identified Ras activation as the site of perturbation of signal transduction. DHA suppressed signal transduction by both changing the localization of EGFR within the plasma membrane and increasing receptor endocytosis and degradation. Lastly, we extended our observations into a wounding model. Although DHA uniquely altered ligand-stimulated EGFR activity, both DHA and EPA altered EGFR transactivation and signaling upon injury. This culminated in reduced wound healing in DHA and EPA treated cells. In an animal model, we found that diets enriched in either DHA or EPA altered EGFR signaling in the colonocytes of wounded animals. Overall, we found that DHA modifies EGFR signaling, which can be beneficial or detrimental for health depending on the disease state of an individual. These data help elucidate a mechanism by which DHA protects against colon cancer, as well as indicating a potential downside of n-3 PUFA therapy.

Introduction: Fast and convenience foods are abundant, relatively inexpensive, and accommodating to the fast-paced lifestyle of many Americans. One popular method of cooking used by many fast food establishments is deep-fat frying. Soybean oil is commonly used for frying and is rich in polyunsaturated fatty acids (PUFA) such as linoleic acid (LA). When soybean oil is used for deep-fat frying, LA becomes oxidized (Ox-LA). Endogenous Ox-LA has the capacity to be a ligand to peroxisome proliferator-activated receptor gamma (PPAR¥ã), a nuclear transcription factor that regulates adipocyte maturation. It is not yet known whether or not dietary Ox-LA has the same capacity with respect to PPAR¥ã. Considering the fact that dietary oxidized lipids are abundant in the typical American diet, it is important to know if they regulate weight gain and especially adipose tissue mass. In this study, we investigate the effects of fresh and heated soybean oil on weight gain and adiposity in mice fed isocaloric low fat diets. Methods: Soybean oil was heated on a hot plate, under a hood, at 190¨¬C for three hours. Fresh soybean oil served as the source of unoxidized oil (Unox-oil) and the heated oil served as the source of oxidized oil (Ox-oil). Both the Ox-oil and Unox-oil were incorporated into a low-fat (10% of calories) mouse chow by Research Diets, Inc. (New Brunswick, NJ). Sixteen C57BL/6J mice were divided into two groups and fed low fat diets with Ox-oil (low fat oxidized, LFO) or with Unox-oil (low fat unoxidized, LFU). Another group of 8 mice were pair fed to the LFO group with the Unox-oil containing chow (PLU). Mice in the LFO and LFU groups were fed ad libitum and known amounts of fresh food was added to the cages every three days. Leftover food was weighed. Body weights were measured once a week. After 16 weeks mice were euthanized and epididymal white adipose tissue (EWAT), retroperitoneal white adipose tissue (RWAT), inguinal white adipose tissue (IWAT), and intrascapular brown adipose tissue (IBAT) samples were collected, weighed and stored at -80 degrees Celsius until further analysis. Fat pads were homogenized and cytosolic and nuclear proteins were extracted by standard methods. These extracts were subjected to Western blotting to determine the amount of PPAR¥ã in the cytosol and nuclear compartments of the fat pads. Differences in group means were analyzed by Mann Whitney U test. Comparisons were considered statistically significant at a p-value of < 0.05. Results: Final mean body weights were significantly different when comparing the mice in the LFU group to the pair fed mice (PLU) (mean ¡¾ SD; 29.52 ¡¾ 1.09 grams (g) and 26.85 ¡¾ 1.44 g, respectively; p < 0.05). Mice fed a low fat diet consisting of Ox-oil (LFO) had a final mean body weight of 27.88 ¡¾ 2.03 g. Mice in the LFU group gained significantly more weight on average than did mice in the LFO or PLU groups (mean ¡¾ SD; 8.86 ¡¾ 1.37g, 7.10 ¡¾ 1.47 g, and 5.71 ¡¾ 1.13 g, respectively). Although mean food intakes were not significantly different between any of the three groups, the average food intake was greatest for the LFU mice in comparison to the LFO and the PLU mice (mean ¡¾ SD; 20.65 ¡¾ 0.09 g/week, 18.40 ¡¾ 0.05 g/week, and 18.38 ¡¾ 0.19 g/week, respectively). Feeding efficiency (g of weight gain/g of food consumed) was the highest in the LFU mice compared to the PLU mice (mean ¡¾ SD; 0.031 ¡¾ 0.005 g/g and0.022 ¡¾ 0.004 g/g) and this difference was statistically significant. The LFO mice gained less weight per gram of food consumed than did the LFU mice (mean ¡¾ SD; 0.028 ¡¾ 0.006 g/g). Mean weights of all fat pads in the LFO group were significantly greater than those of the LFU and PLU mice (mean ¡¾ SD; 0.329 ¡¾ 0.109g, 0.199 ¡¾ 0.055g, and 0.219 ¡¾ 0.041 for EWAT, 0.091 ¡¾ 0.039g, 0.050 ¡¾ 0.026g, and 0.051 ¡¾ 0.017 for RWAT, 0.221 ¡¾ 0.065g, 0.135 ¡¾ 0.053g, and 0.144 ¡¾ 0.038 for IWAT, and 0.079 ¡¾ 0.012g, 0.055 ¡¾ 0.013g, and 0.062 ¡¾ 0.011 for IBAT, respectively). PPAR¥ã protein in the cytosol of EWAT fat pads was analyzed and quantified in comparison to the amount of Glyceraldehyde-3-phosphate dehydrogenase (GAPDH; loading control) present. Mean PPAR¥ã /GAPDH ratios for LFU mice was 0.226 ¡¾ 0.082, for LFO mice was 0.264 ¡¾ 0.122, and for PLU mice was 0.234 ¡¾ 0.108. Mean PPAR¥ã:GAPDH ratios were not significantly different between any of the groups. Conclusion: It appears that the consumption of oxidized oil caused a significant decrease in weight gain and food intake (although not significant) and a significant increase in fat pad mass in mice compared to those consuming a diet with unoxidized oil. The lack of difference in the amount of PPAR¥ã among the three groups of mice suggests that the changes in weight gain and fat pad mass among the oxidized oil consuming animals is not mediated through regulation of PPAR¥ã protein. To our knowledge, ours is the first study to report that mice consuming a low fat diet inclusive of dietary oxidized lipids exhibit greater adiposity than do mice consuming a low fat diet consisting of unoxidized lipids.

Kyoto University (京都大学) / 0048 / 新制・課程博士 / 博士(農学) / 甲第19047号 / 農博第2125号 / 新制||農||1032 / 31998 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 小川 順, 教授 喜多 恵子, 教授 栗原 達夫 / 学位規則第4条第1項該当