CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O diabetes mellitus tipo 2 (DM2), doenÃa multifatorial, heterogÃnica, resulta de suscetibilidade genÃtica associada a fatores ambientais, especialmente sedentarismo e dieta rica em gorduras saturadas, e a obesidade. Caracteriza-se por resistÃncia à insulina (RI) e pela diminuiÃÃo da secreÃÃo desse hormÃnio. InflamaÃÃo moderada e crÃnica no tecido adiposo branco disfuncional, denominada âmeta-inflamaÃÃoâ, parece ser o elo entre obesidade, RI e DM2. Papel de adipocinas produzidas pelo tecido adiposo nessas afecÃÃes vem sendo investigado. Objetiva-se neste estudo verificar se suplementaÃÃo oral (SO) de mistura de Ãleos (MXO) com relaÃÃo de Ãmega 9 / Ãmega 6 (ω9/ω6) elevada e de Ãmega 6 / Ãmega 3 (ω6/ω3) baixa, de diferentes fontes de ω3, interfere em adipocinas plasmÃticas de camundongos com DM2. Depois de alimentados ad libitum com dieta da AIN-93G atà ficarem adultos, camundongos Swiss (CSW) machos receberam, por onze semanas, dieta AIN-93HA, hiperlipÃdica adaptada, para induÃÃo de DM2, confirmado em 90% deles. Mantida a dieta AIN-93HA, os CSW com DM2 receberam, em grupos, por sete dias, SO com MXO: GA: H₂O (controle nulo); GB: MXO [ω9:ω3 0,4:1;ω6:ω3 8:1 (controle neutro)]; GC: MXO [ω9:ω3 3,7:1; ω6:ω3-ALA 1,4:1]; GD: MXO [ω9:ω3 3,7:1;ω6:ω3-EPA+DHA de peixe 1,4:1]; GE: MXO [ω9:ω3 3,7:1; ω6:ω3-DHA de algas 1,4:1]. Por imunoensaios, realizou-se dosagem plasmÃtica de insulina e de adipocinas, fator de necrose tumoralâalfa (TNF-α); interleucina-6 (IL-6); interleucina-1 beta (IL-1β); fator ativador de monÃcitos (MCP-1); resistina (RES); leptina (LEP); inibidor do fator ativador de plasminogÃnio 1 (PAI-1) e adiponectina (AdipoQ). Constatou-se diferenÃa estatÃstica significante de adipocinas do grupo GE (ω3-DHA de algas), em relaÃÃo aos outros grupos, com aumento de IL-6 em relaÃÃo ao GC e GD; diminuiÃÃo de LEP em relaÃÃo ao GA; aumento de TNF-α em relaÃÃo aos grupos GB, GC e GD; e diminuiÃÃo de AdipoQ em relaÃÃo ao GB; assim como de RES entre os grupos GC (ω3-ALA) e GD (ω3-EPA+DHA). NÃo houve diferenÃa estatÃstica significante em nenhuma das variÃveis entre grupos controles. Continuidade de dieta rica em gordura saturada pode ter comprometido a eficÃcia da suplementaÃÃo de MXO ricos em ω3 e ω9. O âestado da arteâ demanda outros estudos para esclarecer o papel do DHA na âmeta-inflamaÃÃoâ. / Diabetes mellitus type 2 (DM2), a multifactorial disease, heterogenic results of associated genetic susceptibility to environmental factors, especially sedentary lifestyle and a diet rich in saturated fats, and obesity. It is characterized by insulin resistance (IR) and by decreasing the secretion of this hormone. Moderate and chronic inflammation in white adipose tissue dysfunctional, called "meta-inflammation," seems to be the link between obesity, IR and DM2. Role of adipokines produced by adipose tissue in these diseases has been investigated. Objective of this study was to verify whether oral supplementation (SO) of oil blend (MXO) compared with omega 9 / omega 6 (ω9/ω6) and high omega 6 / omega 3 (ω6/ω3) low, from different sources of ω3 interferes with adipokines plasma of mice with T2DM. After fed ad libitum with AIN-93G diet until they become adult Swiss mice (CSW) males received by eleven weeks AIN-93HA, hyperlipidic adapted to induce DM2 confirmed in 90% of them. Maintained the AIN-93HA, the CSW with T2DM were in groups of seven days, with MXO SO: GA: H ₂ O (null control) GB: MXO [ω9: ω3 0.4:1; ω6: ω3 8: 1 (neutral control)]; GC: MXO [ω9: 3.7:1 ω3, ω6: ω3-ALA 1.4:1]; GD: MXO [ω9: 3.7:1 ω3, ω6: ω3-EPA + DHA from fish 1.4:1]; GE: MXO [ω9: ω3 3.7:1; ω6: ω3, DHA from algae 1.4:1]. Why immunoassays, held measurement of plasma insulin and adipokines, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), monocyte activating factor ( MCP-1), resistin (RES), leptin (LEP), an inhibitor of plasminogen activator 1 (PAI-1) and adiponectin (ADIPOQ). It found a statistically significant difference of adipokines group GE (ω3-DHA from algae), compared to the other groups, with increased IL-6 compared to GC and GD, fewer LEP compared to GA; increase of TNF- α in relation to groups GB, GC and GD, and ADIPOQ decrease compared to GB, as well as RES between GC (ω3-ALA) and GD (ω3-EPA + DHA). There was no statistically significant difference in any of the variables between control groups. Continuity diet high in saturated fat may have compromised the effectiveness of supplementation MXO rich in ω3 and ω9. The "state of the art" demand further studies to clarify the role of DHA in the "meta-inflamaÃÃoDM2, Adipokines, saturated fatty acids, omega fatty acids 3, 6 and 9 and" meta-inflammation

Os ácidos graxos trans competem com os ácidos graxos essenciais inibindo as enzimas envolvidas na síntese dos ácidos graxos polinsaturados de cadeia longa. Quantidades adequadas de ácidos graxos essenciais na dieta minimizam este efeito. No entanto, pouco se conhece da ação dos isômeros trans, sob condições de restrição ou deficiência dietética dos ácidos graxos essenciais, sobre o percentual de deposição tanto dos próprios isômeros trans, como dos ácidos graxos polinsaturados nos lipídios dos tecidos. Foram avaliadas dietas ricas em ácidos graxos trans e com diferentes concentrações de ácidos graxos essenciais em ratos. A incorporação de ambos foi proporcional à concentração na dieta, mas variou entre os tecidos estudados. Verificou-se que as quantidades de ácidos graxos trans incorporadas pelo tecido adiposo foi a maior, enquanto que o cérebro incorporou a menor quantidade. A despeito da incorporação dos ácidos graxos trans, o perfil de ácidos graxos do cérebro manteve-se estável em relação às variações dietéticas. Não se observou acúmulo dos ácidos graxos trans em função do prolongamento do consumo da dieta. Os ácidos graxos essenciais incorporados aos tecidos também foram modulados pela dieta, e em concentrações adequadas influenciaram na menor deposição dos isômeros trans no tecido adiposo, plasma e tecido cardíaco. Avaliando-se a composição dos ácidos graxos nos fosfolipídios do tecido cardíaco, observou-se que a fosfatidiletanolamina incorporou a maior porcentagem dos isômeros trans seguida da fosfatidilcolina e da cardiolipina, respectivamente. Embora tenha incorporado quantidades intermediárias dos ácidos graxos trans a fosfatidilcolina teve uma maior alteração no perfil de ácidos graxos em comparação aos demais. A reduzida concentração de ácidos graxos trans incorporados na cardiolipina das mitocôndrias e no cérebro sugere a existência de um mecanismo protetor para a manutenção da composição lipídica necessária às atividades funcionais. / Trans fatty acids compete with essential fatty acids inhibiting the enzymes of the long chain polyunsaturated fatty acids synthesis. Appropriate amounts of dietary essential fatty acids minimize this effect. However, little is known about the action of trans isomers on the deposition percentage of these own trans isomers itself or as polyunsaturated fatty acids in tissue lipids, when under dietary restriction or deficiency conditions of essential fatty acids. Diets high in trans fatty acids and with different concentrations of essential fatty acids were assessed. The incorporation of trans isomers and of essential fatty acids was proportional to their dietary concentration, but varied among the tissues studied. It was verified that the adipose tissue incorporated the largest amount of trans fatty acids while the brain incorporated the smallest. In spite of the trans fatty acids incorporation, the profile of brain fatty acids was maintained stable in relation to dietary variations. No accumulative deposition of trans fatty acids was observed in relation to extended length of time of dietary consumption Essential fatty acids incorporated in tissues were also modulated by diet and in moderate concentrations influenced the lower deposition of trans isomers in adipose tissue, blood plasma and heart. By assessing fatty acids composition in phospholipids of heart, it was verified that phosphatidylethanolamine incorporated the largest percentage of trans isomers followed by phosphatidylcholine and cardiolipin, respectively. Phosphatidylcholine presented the greatest change in fatty acids profile when compared to the others phospholipids, although incorporating intermediate amounts of trans isomers. The reduced concentration of trans fatty acids incorporated in cardiolipin and brain suggests that there is a protector mechanism for maintenance of the lipids composition required for functional activities.

The current dietary recommendations for optimal health are designed to increase our intake of two bioactive omega-3 (n-3) fatty acids, eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), abundant naturally in fatty fish such as salmon. Health authorities recommend that the general population consume two to three fatty fish meals per week (1) for optimal health and for the prevention of cardiovascular disease. However, some modern Western societies consume only modest amounts of fish and seafood (2;3). Land based vegetable oils may provide an alternative to meet these needs. Linseed and canola oils are rich in alpha-linolenic acid (ALA, 18:3n-3) (4). ALA can be converted endogenously to EPA and DHA and suggests that increasing the dietary intake of ALA may increase the conversion and accumulation of DHA in tissues and plasma. However, elevated dietary intakes of ALA in animals and humans results in an increased level of EPA in tissues yet there is little or no change in the level of DHA (5-7). The current consensus is that the synthesis of DHA from ALA in humans is limited yet the mechanisms involved in regulating the accumulation of DHA in tissues are poorly understood. The reputed rate-limiting enzyme in the conversion of fatty acids is delta 6 desaturase (D6D). ALA is a substrate for D6D and undergoes a series of desaturation and elongation reactions to yield n-3 long chain polyunsaturated fatty acids (LCPUFA). The final step in the synthesis of DHA from ALA involves translocation of its immediate fatty acid precursor, 24:6n-3 from the endoplasmic reticulum to the peroxisome to be partially beta-oxidised to yield DHA. The involvement of multiple enzymes in the desaturation-elongation pathway, and the integration of other pathways, such as phospholipid biosynthesis, suggests there are various steps that may regulate the accumulation of DHA in cell membranes. This thesis aimed to examine the possible regulatory steps in the conversion of fatty acids to LCPUFA, particularly in the synthesis of DHA from n-3 fatty acid precursors. The human hepatoma cell line, HepG2, was used as an in vitro cell system to examine the accumulation of individual fatty acids and their metabolites in isolation from other competing fatty acid substrates. The accumulation of linoleic acid (LA, 18:2n-6) and ALA in HepG2 cell phospholipids following supplementation with increasing concentrations of each respective fatty acid correlated with that described in vivo, as was the accumulation of their conversion products. The accumulation of DHA in cells supplemented with ALA reached a plateau at concentrations above 5 micro g/ml and paralleled the accumulation of 24:6n-3 in cell phospholipids, suggesting that the delta 6 desaturation of 24:6n-3 was prevented by increasing concentrations of ALA, thereby limiting the accumulation of DHA. The accumulation of DHA in cells supplemented with eicosapentaenoic acid (EPA, 20:5n-3) or docosapentaenoic acid (DPA, 22:5n-3) was significantly greater than the level of DHA that accumulated in cells supplemented with ALA. However, regardless of substrate, the level of DHA in cell membranes reached a plateau at substrate concentrations above 5 micro g/ml. This thesis further aimed to examine the effect of fatty acid supplementation on the mRNA expression of D6D in HepG2 cells. The expression and activity of D6D mRNA is subject to nutritional and hormonal regulation. The mRNA expression of D6D in HepG2 cells following supplementation with oleic acid (OA, 18:1n-9), LA, ALA, arachidonic acid (AA, 20:4n-6) or EPA was examined by real time RT PCR. The expression of D6D mRNA was reduced by up to 50% in cells supplemented with OA, LA, ALA , AA or EPA compared with control cells and suggests that fatty acids modulate the expression of the key enzyme involved in the conversion of fatty acids. The effect of fatty acid co-supplementation on the fatty acid composition of HepG2 cell phospholipids was also examined in an attempt to gain insights into the role of D6D and the enzymes involved in peroxisomal beta-oxidation on the accumulation of DHA from n-3 fatty acid precursors. The reduction in the accumulation of DHA in cells co-supplemented with DPA and docosatetraenoic acid (DTA, 22:4n-6) was greater than in cells co-supplemented with DPA and LA, suggesting that peroxisomal beta-oxidation may have a greater role in determining the accumulation of DHA from DPA than the activity of D6D. Further investigation should be directed towards understanding the role that peroxisomal beta-oxidation may play in the synthesis of DHA from precursor fatty acids. The fatty acid composition of cell membranes in vivo is a result of several physiological processes including dietary intake, phospholipids biosynthesis and fatty acid conversion as well as catabolic processes. This thesis demonstrates that a greater understanding of the regulation of the conversion of fatty acids will help to define dietary approaches that enhance the synthesis of n-3 LCPUFA from n-3 fatty acid precursors to lead to improved outcomes for health.

Periodontitis is an infective disease caused predominantly by gram negative anaerobic bacteria. However it is apparent that alveolar bone loss, which characterises periodontitis, is a result of the host inflammatory response to pathogenic bacteria, not the infectious agents directly. Omega-3 polyunsaturated fatty acids (O-3 PUFAs) are recognised, and used widely, for their anti-inflammatory effects. Evidence is emerging that their oxygenated derivatives are key chemical mediators in the resolution of inflammation. We hypothesised that dietary supplementation with fish oil rich in the O-3 PUFA docosahexaenoic acid would modify inflammatory reactions within the periodontium and thus reduce alveolar bone loss in mice infected with periodontopathic bacteria. Eighty mice were fed experimental diets containing either 10% tuna oil (40) or a sunola oil (40) which contained no traceable O-3 PUFAs for 57 days. After two weeks each dietary set was split into four groups of ten mice, with these groups being inoculated with either a) Porphyromas gingivalis b) P. gingivalis and Fusobacterium nucleatum (combined inoculum) c) Carboxymethylcellulose (control) or d) No inoculations (control). Of the twenty mice which received no inoculations, half were sacrificed after fifteen days and half at the end of the experiment to enable comparative fatty acid analysis of the oral soft tissues. Results demonstrated that eicosapentoic acid and docosahexaenoic acid were found in significantly higher proportions in the oral soft tissues of mice fed a tuna oil diet, and that animals receiving this diet exhibited an average of 54 - 72% less alveolar bone resorption in response to the different bacterial infections. Irrespective of diet, the combined inoculum resulted in slightly more alveolar resorption than P. gingivalis alone. The findings of this study suggest that fish oil dietary supplementation may have potential benefits as a host modulatory agent in the adjunctive management of periodontitis. Given its advantages in terms of safety, cost effectiveness and widespread availability, this dietary supplement warrants further research in human trials to assess its ability to modulate alveolar bone loss in individuals with periodontitis. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1313252 / Thesis (D.Clin.Dent.) - University of Adelaide, School of Dentistry (Periodontics), 2008

Thesis (Ph. D.)--West Virginia University, 2006. / Title from document title page. Document formatted into pages; contains xi, 183 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references.

Diets formulated to contain varying ratios of omega 6 to omega 3 fatty acids were fed to exercising yearlings to evaluate bone activity and inflammatory response. Nine Quarter Horse yearlings were arranged within a triplicated 3 X 3 Latin Square experimental design and fed one of three diets. Exercise protocol was designed to stimulate sub-clinical inflammation and normal bone response. Body weight and physical growth measurements were not different between groups (P > 0.05), and feed intake was similar between groups (P > 0.05). Horses consuming soybean oil (SBO) diet had lower fatty acid profiles (% by weight) of C16:0 and C16:1 (P < 0.05) when compared to horses consuming either corn oil (CO) or menhaden/corn oil (MCO) diets. Though numerically different, percentage changes in C16:0 and C16:1 were not different between diets (P < 0.05). Horses consuming MCO had significantly higher measurements of C20:4, C20:5 and C22:6 over the 28 day period when compared to horses consuming SBO or CO. Percent change in mean concentrations of C20:5 were significantly different between the MCO group and the SBO group (P < 0.05) with no observed difference between MCO and CO treatment groups. Overall mean carboxyterminal telopeptide of type I collagen (ICTP) concentrations did not differ between diets (P > 0.05) nor was there a significant change from baseline values when compared to day 28 of the period. Mean Osteocalcin (OC) concentrations did not differ between treatments (P > 0.05). Numerically, OC levels were lower after 14 days, with subsequent increases occurring from day 14 to day 28; however, there was no significant day effect (P > 0.05). Mean measurements of PGE2 and fibrinogen, the two inflammation markers evaluated, did not differ among groups (P > 0.05). However, when fibrinogen data were normalized, horses consuming SBO had a significantly lower change in baseline values of fibrinogen compared to horses fed CO or MCO diets (P< 0.05). In general, horses fed SBO exhibited reduced levels of the inflammatory marker fibrinogen (P< 0.05). No other variable evaluated was influenced by the supplementation of varying ratios of polyunsaturated fatty acids into the equine diet.

Essential fatty acids nutrition and its effects on immune responses of the juvenile grouper, Epinephelus malabaricus Feng-Cheng Wu (Advisor: Dr. Houng-Yung Chen) Institute of Marine Biology, National Sun Yat-sen University Kaohsiung 804 Taiwan. A series of three experiments was conducted to study the essential fatty acids nutrition and its effects on immune responses (IR) of the juvenile grouper, Epinephelus malabaricus. All experimental diet contained 10 g/100 g diet supplemental lipids from various sources. A reference diet was used in all experiments and contained natural oil mixture of cod liver oil, linseed oil, and safflower oil at a rate of 2:1:1 (wt/wt/wt). In experiment 1, juvenile grouper (11.8 ¡Ó 0.7 g) were fed for 12 wks on one of the seven experimental diets, control diet and the reference diet to investigate dietary requirement for docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) and effects on IR of grouper. Seven experimental diets contained 1 g/100 g diet of DHA and EPA in various combinations and 9 g/100 g diet of tristearin. The control diet contained 1 g/100 g diet of trilinolenin and trilinolein (3:1, wt/wt). The results indicate that there was a significant difference among dietary treatments in growth, phagocytosis and leucocytes proliferation, when stimulated by Con A and PHA-P but not by LPS. However there was no difference in survival rate and relative liver weight. Enhanced growth was observed when the dietary DHA/EPA was greater than 1, indicating that DHA was superior to EPA in promoting fish growth. DHA is the only member in the family of (n-3) highly unsaturated fatty acid (HUFA) that stimulates phagocytic functions of leucocytes and T cell proliferation of the juvenile grouper. In experiment 2, juvenile grouper (11.3 ¡Ó 0.6 g) were fed for 12 wks on one of the eight experimental diets or the reference diet to investigate dietary requirement for linolenic acid (LNA) and linoleic acid (LA), as well as effects on nonspecific IR of grouper. The test diets were supplemented with LNA or LA at a rate of 1 or 2 g LNA or LA/100 g diet or 2 g/100 g diet of LNA and LA in various ratios (3, 1.4, 0.7 and 0.4). Tristearin was used to fill the lipid supplemental level to 10 g/100 g diet. The results show that enhanced growth and optimal non-specific cellular IR were observed when the grouper were fed on the diet having the highest LNA/LA ratio (3:1) or on the diets supplemented with LNA. But the enhancement was not different (P>0.05) from that of the reference diet group. Thus, incorporating 2 g/100 g LNA/LA (3:1, wt/wt) in diet ensures adequacy of the grouper for essential fatty acid. In experiment 3, juvenile grouper (13.2 ¡Ó 0.9 g) were fed on one of the six experimental diets in the 2 ¡Ñ 3 factorial design or on the reference diet for 12 wks to investigate dietary requirement for (n-3)HUFAs and arachidonic acid (AA), as well as effects on IR of grouper. Two levels of (n-3)HUFAs (1 or 2 g/100 g ) in combination with 3 levels of AA (0, 1 or 2 g/100 g) were tested. The results show an enhanced growth when optimal concentrations of AA and (n-3)HUFA were incorporated to the diets. Liver (n-6)HUFAs concentration reflects IR of the juvenile grouper. Interaction of AA and (n-3)HUFAs in affecting fish growth and IR was insignificant (P>0.05), and concentrations of dietary (n-3)HUFAs or AA did not significantly affect fish survival rate. The results of the 3 experiments show that the grouper will benefit most in growth and IR when their diets contain 1 g/100 g diet of (n-3)HUFA and 1 g/100 g diet of AA, when (n-3)HUFA is a mixture of DHA and EPA at a ratio of 3:1 (wt/wt).

Thesis (M.Sc.)--Memorial University of Newfoundland, 2002. / Bibliography: leaves 144-173.

Natural variation in the relative proportions of saturated and unsaturated fatty acids in seed oils of plants is enormous when considered across a broad taxonomic range of oil seeds. It has been shown that this variation follows a latitudinal cline where the proportions of unsaturated fatty acids increases with increasing latitude as the unsaturated fatty acids in seeds provide energy at a faster rate to germinating seeds at higher latitudes. This variation which follows a latitudinal cline suggests that there may be an adaptive role for this variation. We tested this hypothesis in Arabidopsis thaliana which followed the same trend seen in Helianthus and other angiosperms. In order to understand the underlying genetics of the regulation of the relative proportions of fatty acids and their role in plant evolution, we mapped quantitative trait loci (QTLs) and candidate genes. Here we identified 67 major QTLs responsible for fatty acid synthesis in A. thaliana in Ler-0 x Sha, Ler-0 x Col-4, Ler-2 x Cvi and Ler-0 x No-0 RIL populations. Eight candidate genes were identified based on what is known about seed oil biosynthesis in A. thaliana. Six of the candidate genes collocated to most of the major QTLs. In order to demonstrate that a particular allelic variant is indeed causally related to the phenotype, we investigated DNA polymorphisms in the parental and the RIL line alleles of the collocating candidate genes. Single nucleotide polymorphisms (SNPs) were identified in the collocating candidate genes to study the correlation between the sequence variants and the particular phenotype. We identified 232 SNPs with 77 in the putative regulatory regions upstream of the 5’UTR, 61 in the introns, 18 in the 5’UTR regions, 2 in the 3’UTR regions, and 45 occurring in the exons with 10 non-synonymous substitutions affecting the amino acid residues. We also detected 44 insertions/deletions in the coding, non-coding, 5’UTR, 3’UTR and the regulatory regions. Sequence variation in the fatty acid genes due to SNPs and insertions/deletions should be valuable in tests of association to investigate how the relative proportions of saturated and unsaturated fatty acids are regulated in wild plants and what role they have played in plant evolution and also in breeding oil seed crops that are healthier or have two types of fatty acids in proportions appropriate for different uses. / text

published_or_final_version / Chemistry / Doctoral / Doctor of Philosophy