Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Ivana Turňová Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Consultant: prof. MUDr. Zdeněk Zadák, CSc. Title of Diploma Thesis: Determination of fatty acids in human tissues The fatty acids are non-negligible component of lipids as one of the basic nutrients. This thesis in its theoretical part presents above all the group of polyunsaturated fatty acids (PUFA), which are important structural units of the cell membranes, they are also the precursors to several significant biologically active substances. In the human organism PUFA participate in many physiological and pathological processes this way, where they cause the large spectrum of actions. Onwards the thesis describes the gas chromatography method that is used in chemical analysis of lipids for the fatty acid determination as the gold standard. Experimentally, the fatty acid representation was determined in the blood, muscle, heart, liver and kidney among 26 cadavers divided into two groups according to the presence/absence of inflammatory process at the time of death. The data obtained were statistically analysed. The results were descriptively evaluated and there were indicated possible explanations of differences...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Author of Diploma Thesis: Bc. Barbora Kuříková Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Consultant: prof. MUDr. Zdeněk Zadák, CSc. Title of Diploma Thesis: Determination of the fatty acids spectrum in patients undergoing treatment with taxanes The diploma thesis deals with the determination of fatty acids spectrum in erythrocytes in patients treated with taxanes. The theoretical part describes general features of fatty acids, their synthesis and degradation. Then it is focused on the topic of breast cancer, taxane treatment and negative side effects associated with taxane treatment, especially polyneuropathy. The gas chromatography, which is commonly used in practice, is also described in this part. The experimental part is divided into three parts. The first part describes working process, chromatographic analysis and evaluation of this analysis. In the second part there is comparing of spectrum of fatty acids of patients without and with polyneroupathy. And in the last part there is a comparing of spectrum of fatty acids of patients before taxane treatment, closely after treatment and about month after the treatment. Results of the experimental part are evaluated at the end...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Alena Čunátová Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Title of Diploma Thesis: The fluctuation of fatty acids levels in the tissues of suddenly deceased persons and comparison with values in pathological states Fatty acids and their metabolites are significantly involved in many physiological and pathological processes. This thesis monitors the levels of selected fatty acids in human tissues and the effect of long-term disease on their stores. In addition to the general characteristics, the theoretical part focuses on the intake of polyunsaturated fatty acids and their importance in the human body. It also deals with the effects of eicosanoids and other fatty acid metabolites. Methods used in fatty acid bioanalysis are also described. In the experimental part, the proportion of fatty acids in seven tissue types was compared between two groups of donors. One of them included 8 relatively young, healthy, suddenly deceased individuals. The second group included 12 elderly polymorbid patients. The obtained tissue samples were adjusted using extraction and derivatization techniques. Gas chromatography with flame ionization detection was used for analysis. The measured...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Kamila Kynclová Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Title of Diploma Thesis: Determination of fatty acids levels in selected human tissues Fatty acid are an essential part of many metabolic processes in the body. The theoretical part of the the thesis presents primarily omega-3 and omega-6 fatty acids, their general characteristics, metabolism and their effect on various disease. The roles of fatty acid in cancer, neuropsychiatric and inflammatory diseases are described here. The thesis also mentions the method of gas chromatography, which was used to determine the levels of fatty acids. The experimental part of the thesis includes method, chromatographic analysis and results. 13 fatty acids in 7 different tissues were analyzed: subendocardial tissue, liver parenchymal tissue, kidney and adrenal tissue, skeletal muscle tissue, adipose and brain tissue. Tissues were collected from 20 healthy, suddenly deceased, cadavers. Donors were divided into two groups according to age. The obtained data were statistically processed and divided according to individual fatty acids. In the end the results between the two groups were evaluated with a possible explanation of the...

Résumé : L’apolipoprotéine E (apoE) joue un rôle important dans le transport des acides gras (AG) via les lipoprotéines. Cependant, il existe possiblement une perturbation dans l’homéostasie des AG au niveau des lipoprotéines chez les porteurs du génotype de l’apolipoprotéine E epsilon 4 (E4+). L’objectif de cette étude est de déterminer le profil en AG dans les lipoprotéines de hautes et de faibles densités (HDL et LDL) chez les E4+ et les non-porteurs (E4-), pendant une supplémentation en AG oméga-3 (n-3) de 28 jours. Matériels et Méthodes: 80 participants (34 hommes et 46 femmes) en santé, âgés entre 20-35 ans, ont consommé 1,6 g/jour d’AG n-3 sur une période de 28 jours. Des prélèvements sanguins à jeun ont été récoltés chaque semaine. Les lipoprotéines ont été séparées par ultracentrifugation sur gradient discontinu de sucrose. Les lipides totaux des particules de HDL et de LDL ont été analysés par chromatographie en phase gazeuse. Les génotypes de l’APOE (E4+ ou E4-) ont été déterminés par la méthode de polymorphisme de longueur des fragments de restriction (RFLP) et les données ont été analysées par logiciel SAS à l’aide d’une procédure MIXED. Résultats: Les caractéristiques anthropométriques et habitudes de vies ne variaient pas significativement entre les E4+ et E4-. Le ratio d’AG n-6/n–3 était environ 17% plus élevé chez les E4+ dans les LDL (P = 0.043) pendant la supplémentation. Ceci peut être attribuable au niveau plus élevé d’AG n-6, sans changement dans le niveau d’AG n-3 chez les E4+. Une interaction génotype × temps a été trouvée pour l’acide linoléique (LA) dans les HDL ainsi qu’un effet génotype pour les AG n-6 totaux dans les HDL et LDL (P ≤ 0.05). De plus, l’acide palmitique (PA) et palmitoléïque (PAL) est plus bas chez les E4+ comparativement aux E4-. Conclusion: Le débalancement de la distribution des AG dans les HDL et LDL chez les E4+ peut être causé par une altération de la spécificité de la β-oxydation des AG chez les E4+. Plus d’investigation doit être faite à cet égard afin de confirmer ces hypothèses.

Com a utilização de 18 éguas, adultas, sem raça definida, com peso médio de 521±56 Kg, em delineamento inteiramente casualisado com três tratamentos e seis repetições por tratamento com medidas repetidas no tempo. Os objetivos deste trabalho foi verificar os efeitos da suplementação dietética com ácidos graxos ômegas 3 e ômega 6 sobre a composição do colostro e leite de éguas e transferência imunitária para os potros. Diferenças significativas encontradas foram na qualidade do leite das éguas onde foi observado efeito de tempo (semanas) sobre as variáveis de gordura, proteína, lactose e sólidos totais. Para a variável sólidos totais observou-se efeito do tratamento, semana e da inclusão de óleo na dieta. Observou-se nos potros uma atividade linfoproliferativa, no grupo óleo de soja semelhante ao encontrado no grupo controle, em ambos os períodos analisados. O grupo óleo de linhaça apresentou atividade linfoproliferativa maior do que a encontrada nos dois outros grupos no dia sete. No dia trinta, o grupo de linhaça continuou a apresentar uma maior atividade proliferativa, no entanto sem diferença estatística. A suplementação fonte de óleo aumentou a energia dietética disponível e a resposta proliferativa das éguas do dia sete. Observou-se que a suplementação com óleo de linhaça promoveu um aumento das concentrações dos isótopos IgGa e IgGb no colostro. A concentração média de IgGa no grupo óleo de linhaça foi quase 4 vezes superior aquela encontrada no colostro das éguas do grupo controle e quase 3 vezes superior aquela encontrada no grupo suplementado com óleo de soja com uma significância de. Assim concluímos que o ômega 3 melhorou a posta linfoproliferativa e aumentou a concentração de IgGb, porem não alterou os componentes do leite. / With the use of 18 mares, adult mongrel with a mean weight of 521 ± 56 kg in completely randomized design with three treatments and six replicates per treatment with repeated measures. The objectives of this study was to determine the effects of dietary supplementation with fatty acids omega 3 and omega 6 on the composition of colostrum and milk of mares and foals for immune transfer. Significant differences were found in milk quality mares where was no effect of time (weeks) on the variables of fat, protein, lactose and total solids. For variable total solids was observed effect of treatment week and oil inclusion in the diet. It was observed in foals lymphoproliferative activity in the soybean oil group similar to that found in the control group in both periods analyzed. The group linseed oil lymphoproliferative showed activity greater than that found in the other two groups on day seven. On the thirtieth day, the flaxseed group continued to show a higher proliferative activity, however no statistical difference. The source of oil supplementation increased dietary energy available and the proliferative response of mares of day seven. It was observed that supplementation with flaxseed oil promoted increases in concentrations of isotopes and IgGa IgGb in colostrum. The average concentration of IgGa group in linseed oil was almost 4 times higher than that found in colostrum of mares in the control group and almost 3 times higher than that found in the group supplemented with soybean oil with a significance. Thus we conclude that omega 3 put lymphoproliferative improved and increased the concentration of Ig, but it does not change the components of milk.

Linoleic acid, with a DRI of 12-17g/d, is the most highly consumed polyunsaturated fatty acid in the Western diet and is found in virtually all commonly consumed foods. The concern with dietary linoleic acid, being the metabolic precursor of arachidonic acid, is its consumption may enrich tissues with arachidonic acid and contribute to chronic and overproduction of bioactive eicosanoids. However, no systematic review of human trials regarding linoleic acid consumption and subsequent changes in tissue levels of arachidonic acid has been undertaken. In this study, we reviewed the human literature that reported changes in dietary linoleic acid and its subsequent impact on changing tissue arachidonic acid in erythrocytes and plasma/serum phospholipids. We identified, reviewed, and evaluated all peer-reviewed published literature presenting data outlining changes in dietary linoleic acid in adult human clinical trials that reported changes in phospholipid fatty acid composition (specifically arachidonic acid) in plasma/serum and erythrocytes within the parameters of our inclusion/exclusion criteria. Decreasing dietary linoleic acid up to 90% was not significantly correlated with changes in tissue arachidonic acid levels (p=0.39). Similarly, when dietary linoleic acid levels were increased six fold, no significant correlations with tissue arachidonic acid levels were observed (p=0.72). However, there was a positive relationship between dietary gamma-linolenic acid and arachidonic acid on changes in tissue arachidonic levels. Our results do not support the concept that modifying current intakes of dietary linoleic acid has an effect on changing tissue levels of arachidonic acid in adults consuming Western-type diets.

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.

Two different studies were conducted to reduce the overall amount of omega-6 fatty acids in broiler chickens. The first experiment was performed to determine the effects of dietary conjugated linoleic acid (CLA) and omega-3 fatty acid combination on the omega-6 fatty acid accumulation in broiler chicken breast and thigh meat. Eight broilers from each treatment were processed at 4 and 6 weeks of age, respectively. Regarding the diets containing five different fat sources, broiler chickens fed CLA and fish oil diet had a lower C20:4 (arachidonic acid, AA, n-6) deposition but showed a higher n-3/n-6 ratio in breast and thigh meat than those fed a flaxseed oil diet and CLA and flaxseed oil diet (P < 0.05). The C20:4 and n-3/n-6 ratio of breast and thigh samples from fish oil diet was similar to those of the conjugated linoleic acid and fish oil combination diet (P > 0.05). However, the addition of CLA and fish oil to the diet resulted in a increase of polyunsaturated fatty acid (PUFA) concentration in broiler chicken breast and thigh meat when compared to that of fish oil diet (P<0.05). The second experiment was conducted based on six different combination of n-3 and n-9 fatty acids. One bird per pen was processed, and each bird was weighed, and blood, liver, breast and thigh samples from the bird were collected. Although the generation of prostaglandin E2 (PGE2) was not affected due to combination of n-3 and n-9 fatty acids in our diets, the deposition of n-6 fatty acids including C18:2 and C20:4 was decreased in broiler chicken breast and/or thigh muscles as n-3 fatty acids were supplied to broiler chickens for 9 weeks. Eicosapentaenoic acid (C20:5, EPA, n-3) addition to poultry diet (FEO) did not reduce the deposition of C18:2 and/or C20:4 as much as C22:6 (FDO) did. When C20:5 and C22:6 were blended to poultry diet (FHO) and fed to broiler chickens for 9 weeks, synergistic effects were observed. Reduction of C20:4 was obtained when FHO diet was fed to broiler chickens, and it may be induced due to decreased expression of delta-6 desaturase mRNA.

Ulm, Univ. Diss., 2006.