Ultraviolet radiation (UVR) is a complete carcinogen, inducing skin cancer via DNA photodamage that can lead to mutagenesis, and promoting its growth via photoimmunosuppression (PI). The omega-3 polyunsaturated fatty acid (n-3 PUFA) eicosapentaenoic acid was shown in murine studies to protect against PI and UV-induced skin cancer although the mechanism is uncertain. The principal objectives of this thesis were to (i) examine whether n-3 PUFA can protect against a clinical model of PI in healthy humans and (ii) explore whether the underlying mechanism could be abrogation of UV-induced depletion of antigen-presenting Langerhans cells (LC) from the epidermis, and/or impact on immunomodulatory cytokines. Nickel (Ni) allergic females (n=79) were randomized to 3 months of daily supplementation with 5g n-3 PUFA (70% eicosapentaenoic acid, EPA; 10% docosahexaenoic acid, DHA) or the placebo medium chain triglyceride, GTCC. Local PI was clinically assessed post supplementation using the nickel contact hypersensitivity (Ni CHS) model. In each volunteer, Ni patches were applied to 3 skin sites that were irradiated for 3 consecutive days with UV-doses of 1.89, 3.82 & 7.59J/cm2 respectively. CHS responses were measured and compared to responses of control patches applied on unirradiated skin using a reflectance erythema meter. In the same subjects, assessments of cellular and biochemical mediators of PI were made pre and post supplementation. At 24hr post irradiation with an erythemal UV-dose (4 minimal erythemal doses) to upper buttock skin, half the subjects (n=39) had skin punch biopsies taken and the other half (n=40) had suction blisters raised on this irradiated skin and on unirradiated skin of the contralateral buttock. Epidermal sheets were prepared from the punch biopsies and immunohistochemically stained to assess UV-induced LC numbers. Levels of immunomodulatory cytokines were analysed in the suction blister fluid using Luminex multiplex assay kits. To evaluate compliance and bioavailability, blood samples were taken from all volunteers, pre and post supplementation and EPA% weight in red blood cell membranes was examined using gas chromatography. Post supplementation, EPA %wt was significantly higher in the active group compared to control: mean 3.61% ± 0.22% (SEM) vs. 0.93% ± 0.06% (p<0.001). 3 volunteers showed evidence of non-compliance and were excluded from further analysis. Compared to placebo, evidence for protection against local PI of Ni CHS was apparent post n-3 PUFA at all UV doses, reaching statistical significance at the UV-dose of 3.8J/cm2 (p<0.05). No significant difference in post-UV epidermal LC numbers after supplementation was seen between active and placebo groups, with a % fall following UV of 76.61 ± 3.39% (SEM) in the active group and 73.52 ± 5.24% (SEM) in the control group. When intragroup comparisons were made pre vs. post supplementation, a similar increase in UV-induced LC depletion from the epidermis was seen in both groups, reaching statistical significance following n-3 PUFA (p=0.018). Levels of interleukins IL-10 and IL-8, and of TNF-α, increased post-UVR in both active and control groups pre-supplementation, with no changes occurring following supplementation. In conclusion, supplemental EPA was bioavailable and evidence of protection against clinical PI of Ni CHS was seen in the actively treated group. However, no evidence was found that this abrogation of PI was mediated through a reduced effect of UV on migration of epidermal LC or the immunomodulatory cytokines examined. This original study gives the first evidence that dietary n-3 PUFA may protect against clinical PI, and potentially skin cancer promotion, in humans. Further research is needed to confirm this finding, and to examine the underlying mechanisms, which could involve other immunoregulatory cells of the skin, such as dermal dendritic cells and T regulatory cells and other mediators of UV-immunosuppression including the prostanoids, which may be modified by n-3 PUFA.

An experiment was designed to determine whether the response of fish to graded dietary levels of essential fatty acids (n3) was affected by the total dietary lipid concentration. Juvenile coho salmon were fed practical diets varying in lipid source and total lipid content. The diets were fed in duplicate during a 27-week and a 12-week period ran in succession. The effects of the different dietary fatty acid concentrations on body fatty acids composition were determined after each period. The dietary fatty acid classes were expressed either as percent of the dry diet or percent of the dietary lipid. Analysis of the body lipid fatty acid composition was performed for neutral and polar lipid fractions. The effect of dietary fatty acid concentration on growth and mortality was determined over a 12-week growth study (period 2). A 24-hour saltwater challenge was performed at the end of period 2. It was used to examine the effect of dietary fatty acid concentration on saltwater tolerance. Dietary n6 and n3 fatty acids appeared to be selectively incorporated into the body polar lipid pool. Linoleate and linolenate underwent elongation and desaturation which resulted in the inhibition of the elongation and desaturation of 18:ln9. The neutral lipid pool served as a source of n3 fatty acids for the polar lipid when dietary intake was limited by low temperatures during period 1. The body neutral monounsaturated and 18:1 monounsaturated fatty acids consistently reflected the composition of the diet. The n3 fatty acid concentration in the neutral lipid was also directly related to the dietary fatty acid composition during period 2. The effect of dietary fatty acids on the body neutral or polar fatty acid composition did not depend on the manner in which the dietary fatty acids were expressed. There was also no significant effect of dietary total lipid concentration on the relationship between dietary fatty acids and their incorporation into the body lipids. The growth response was difficult to interpret because of the high mortality. There was a significant difference in mortality among treatments. A positive relationship between dietary concentrations of total n3 fatty acids or n3 highly unsaturated fatty acids and mortality became evident following analysis of the regression of mortality as a function of dietary fatty acid composition. The dietary fatty acid composition did not appear to alter the saltwater tolerance of the 1+ coho salmon. / Land and Food Systems, Faculty of / Graduate

The industrial uses of cottonseed oil are limited by its fatty acid composition. Genetic modification of cotton lipid profiles using seed-specific promoters could allow cotton growers to produce valuable new oils in the seed without adverse effects on fiber quality and yield, therefore making this crop more commercially profitable. Transgenic cotton callus harboring a diverged fatty acid desaturase gene (FADX) from Momordica charantia was characterized for production of alpha-eleostearic acid (conjugated double bonds: 18:3 D9 cis, 11 trans, 13 trans), not normally found in cotton. Gas chromatography (GC) in conjunction with mass spectrometry (MS) confirmed production of alpha-eleostearic acid in the transgenic cotton tissues. A second series of transformation experiments introduced the cotton fatty acid thioesterase B (FATB) cDNA, fused to the seed-specific oleosin promoter into cotton to promote the over-expression of FATB, to generate cotton with increased palmitate in the cottonseed. PCR amplification, as well as fatty acid analysis by gas chromatography, confirmed introduction of the FATB cDNA in transgenic tissues. Collectively, these results demonstrate the feasibility of manipulating the fatty acid composition in cotton via transgenic approaches and form the basis for continued efforts to create novel oils in cottonseed.

Orientador: Helena Coutinho Franco de Oliveira / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-27T08:22:09Z (GMT). No. of bitstreams: 1 Paiva_AdrieneAlexandra_D.pdf: 2285275 bytes, checksum: e9d86e12d4ac3d041953799c130a314a (MD5) Previous issue date: 2015 / Resumo: A doença hepática gordurosa não alcoólica (Nonalcoholic fatty liver disease - NAFLD) é a principal manifestação hepática na obesidade e síndrome metabólica. A história natural da doença envolve a esteatose, estresse oxidativo, inflamação, fibrose e morte celular. Níveis plasmáticos elevados de lipoproteínas ricas em triglicérides são fatores de risco independentes para doenças cardiovasculares. Estudos clínicos e experimentais mostram forte correlação entre triglicérides plasmáticos (TG) e os níveis de apolipoproteína CIII. A apolipoproteína CIII também é aumentada no plasma de pacientes diabéticos. Ao comparar camundongos apoCIII transgênicos com controles não transgênicos irmãos (NTg), mostramos aqui que a super-expressão de apoCIII, independentemente da dieta rica em gordura (High fat diet - HFD), resulta em características NAFLD, ou seja, aumento do conteúdo de lípidos no fígado, diminuição poder antioxidante, aumento da expressão de TNF'alfa', receptor de TNF'alfa', caspase-1 clivada e interleucina-1'beta', diminuição do receptor de adiponectina-2 e aumento da morte celular. Além disso, os indicadores de inflamação sistêmica de TNF'alfa' e a proteína C-reactiva estão também elevados em camundongos apoCIII. Esse quadro é agravado e características adicionais da NAFLD são diferencialmente induzidas por HFD em camundongos apoCIII. A HFD induziu intolerância à glicose, juntamente com aumento da gliconeogênese, evidenciando a resistência hepática à insulina, induziu um aumento significativo da TNF'alfa' (8 vezes) e IL-6 (60%) no plasma em camundongos apoCIII comparados aos NTg. Os outros indicadores de inflamação (proteína C reativa plasmática e TNF?, TNFr, caspase-1 e IL-1'beta' hepática) mantiveram-se superiores em camundongos apoCIII em HFD. A ativação da via intrínseca (Bax / Bcl-2) e a proteina efetora (caspase-3) de apoptose foram aumentados em ambos os camundongos apoCIII com low fat diet (LFD) e HFD. Como esperado, o tratamento com fenofibrato reverteu vários dos efeitos da dieta e da apoCIII. No entanto, o fibrato não normalizou o padrão inflamatório nos animais apoCIII, tais como o aumento do TNF'alfa', TNFr, IL1'beta' e redução do adiponectina-R2, mesmo com total correção do teor de lipídios no fígado. Estes resultados indicam que a superexpressão apoCIII desempenha um papel importante na inflamação hepática e na morte celular e aumenta a suscetibilidade e a gravidade da esteatose hepática induzida pela dieta / Abstract: Nonalcoholic fatty liver disease (NAFLD) is the principal liver manifestation in obesity and metabolic syndrome. The natural history of the disease involves steatosis, oxidative stress, inflammation, fibrosis and cell death. Elevated plasma levels of triglyceride-rich remnant lipoproteins are independent cardiovascular disease risk factors. Clinical and experimental studies show strong correlation and causal links between plasma triglycerides (TG) and apolipoprotein CIII levels. Apolipoprotein CIII is also increased in the plasma of diabetic patients. By comparing apolipoprotein (apo) CIII transgenic mice with control non-transgenic (NTg) littermates, we show here that the overexpression of apoCIII, independently of high fat diet (HFD), results in NAFLD features, namely, increased liver lipid content, decreased antioxidant power, increased expression of TNF'alfa', TNF'alfa' receptor, cleaved caspase-1 and interleukin-1'beta', decreased adiponectin receptor-2 and increased cell death. In addition, systemic indicators of inflammation TNF? and C-reactive protein are also elevated in apoCIII mice. This picture is aggravated and additional NAFLD features are differentially induced by HFD in apoCIII mice. HFD induced glucose intolerance together with increased gluconeogenesis, evidencing hepatic insulin resistance. Marked increases in plasma TNF'alfa' (8-fold) and IL-6 (60%) were induced by HFD in apoCIII mice compared to NTg mice. The other inflammatory indicators (plasma C reactive protein, liver TNF'alfa', TNFr, caspase-1 and IL1 'beta') remained higher in HFD apoCIII mice. Cell death signaling (Bax/Bcl2), effector (caspase-3) and apoptosis were augmented in both low and HFD apoCIII mice. As expected, fenofibrate treatment reversed several of the diet and apoCIII effects. However, fibrate did not normalize apoCIII inflammatory traits, such as increased TNF'alfa1, TNFr, IL1'beta' and reduced adiponectin-R2, even under fully corrected liver lipid content. These results indicate that apoCIII overexpression play a major role in liver inflammation and cell death increasing susceptibility to and the severity of diet induced NAFLD / Doutorado / Fisiologia / Doutora em Biologia Funcional e Molecular

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21802号 / 農博第2315号 / 新制||農||1065(附属図書館) / 学位論文||H31||N5174(農学部図書室) / 京都大学大学院農学研究科応用生物科学専攻 / (主査)教授 久米 新一, 教授 松井 徹, 教授 廣岡 博之 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22304号 / 医博第4545号 / 新制||医||1040(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 川口 義弥, 教授 坂井 義治, 教授 羽賀 博典 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第22472号 / 工博第4733号 / 新制||工||1739(附属図書館) / 京都大学大学院工学研究科合成・生物化学専攻 / (主査)教授 梅田 眞郷, 教授 浜地 格, 教授 跡見 晴幸 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

The ability to adapt fuel usage to nutrient availability is termed metabolic flexibility, and is influenced by activity of the pyruvate dehydrogenase complex (PDC). The Virginia lines of chickens are a unique model of anorexia and obesity that have resulted from 56 generations of artificial selection for high (HWS) or low (LWS) juvenile body weight. We hypothesized that hyperphagia and obesity in juvenile HWS chickens are associated with altered fatty acid oxidation efficiency and metabolic flexibility in tissues associated with energy sensing and storage, and relative cellular hypertrophy in white adipose tissue. Hypothalamus, liver, Pectoralis major, gastrocnemius, abdominal fat, clavicular fat and subcutaneous fat were collected from juvenile (56-65 day-old) HWS and LWS chickens for metabolic, gene expression and histological assays. The HWS chickens had reduced fatty acid oxidation efficiency in abdominal fat (P < 0.0001) and reduced rates of oxidation in abdominal fat and gastrocnemius (P < 0.0001) as compared to LWS. There was reduced citrate synthase activity in white adipose tissue (P < 0.0001) and greater metabolic inflexibility in skeletal muscle (P = 0.006) of HWS compared to LWS. Greater pyruvate dehydrogenase kinase 4 (PDK4) and forkhead box O1 (FoxO1) mRNA were found in skeletal muscle and white adipose tissue of 56-day-old HWS than LWS. Expression of peroxisome proliferator-activated receptor γ (PPARγ) in all adipose tissue depots was greater (P < 0.05) in LWS than in HWS chickens. The HWS chickens had larger (P < 0.0001) and fewer (P < 0.0001) adipocytes per unit area than LWS. These results suggest that the HWS chickens have impaired metabolic flexibility and fatty acid oxidation efficiency due to an up-regulation of pyruvate dehydrogenase activity to accommodate the influx of acetyl CoA from fatty acid oxidation in skeletal muscle and white adipose tissue. These metabolic adaptations can be linked to differences in gene expression regulation and body composition between the lines. Adipocyte cellularity data are consistent with greater oxidative efficiency in the adipose tissue of LWS, because of the greater number of unfilled cells in all depots that were sampled. Results can be extrapolated to agricultural production in the understanding of factors regulating the amount of lipid deposition in chicken carcass fat. Results may also provide insight into eating disorders and the development of obesity. / Master of Science

No description available.

This master thesis deals with the determination of bound and free fatty acids in selected types of cheeses with/without „Protected designation of the Europen Union“ and their mutual comparison. The theoretical part focuses on a milk fat, a brief overview of cheeses, protected designation of the Europen Union, lipids, fatty acids and a possibility of their determination. In the experimental part, free and bound fatty acids profile in 12 selected samples of white mold cheeses was determined. The declared amount of total fat was verified. The cheeses were compared by a content of bound and free fatty acids, the profile of fatty acids was compared based on a type of milk used. Finally, the results were statistically treated by PCA method. The method according to ČSN EN ISO 1735 was chosen for extraction of lipids from the cheese samples. A methanolic solution of boron trifluoride was used for fatty acids esterification. The fatty acids methyl esters were determined by gas chromatography with flame ionization detection.