National Repository of Grey Literature 6 records found  Search took 0.01 seconds. 
Lipidomic analysis of adipose tissue in patients with ischemic heart disease
Tomášová, Petra ; Vecka, Marek (advisor) ; Poledne, Rudolf (referee) ; Neckář, Jan (referee)
The ever-increasing incidence of coronary artery disease (CAD) in Czech Republic and worldwide is one of the most important health problem today. Obesity and type 2 diabetes mellitus (DM2T) are risk factors for the development of CAD. Lipid metabolism is greatly affected by these diseases and adipose tissue, previously considered a mere energy store, also plays an important role in these changes. Today adipose tissue is discussed also as an endocrine organ, which effect whole body metabolism significantly. Lipidomic analysis is an effective tool for monitoring lipid metabolism. Lipidomics combines the advantages of determining a large number of analytes and a small amount of sample and its rapid preparation. In the first part of study, I optimized the HPLC-MS method of untargeted lipidomic analysis in adipose tissue. Then, this method was used to compare subcutaneous and epicardial adipose tissue. Finally, I evaluated the effect of CAD and DM2T on the lipid profiles of subcutaneous and epicardial adipose tissues. Optimization of the pre-analytical phase enabled the analysis of 206 lipid molecules (in total) in both types of adipose tissue. Compared to subcutaneous adipose tissue, epicardial adipose tissue contained a higher proportion of phosphatidylcholines (e.g. PC(32:0), PC(32:1), PC(36:4)) and...
Mitochondria as a target of anticancer therapy.
Dvořák, Aleš ; Ježek, Petr (advisor) ; Poučková, Pavla (referee) ; Vecka, Marek (referee)
Mitochondrial isocitrate dehydrogenase 2 (IDH2) catalyzes reductive carboxylation (RC, reverse Krebs cycle pathway) and 2HG synthesis (2HG) - metabolite of which many scientists are interested. 2HG may be concurrently synthetized in cytosol by IDH1. RC is involved in anabolic reactions necessary for cell proliferation - produces citrate, fatty acid precursor - especially in hypoxia. IDH2 and IDH1 are not the only enzymes that are involved in 2HG synthesis. Recently, several enzymes, which participate in 2HG production, have been discovered. 2HG is useful in cancer diagnostics due to its overproduction by transformed cells. Moreover, 2HG may cause epigenetic changes via inhibition of 2-oxoglutarate dependent dioxygenase. In this work, the importance of RC and 2HG synthesis in cancer and healthy cells was investigated by gas chromatography with mass spectrometry detection as well as IDH2 influence. We found that IDH2 significantly participates in reverse RC and 2HG synthesis in breast cancer cell lines and uses glutaminolysis as a supplementary anaplerotic pathway. RC is increased by hypoxia, inhibition of respiration, and decreased by activation of respiration or hypocapnia. We confirmed 2HG synthesis and RC in healthy cells (fibroblasts, breast epithelial cells etc.) as well as in cancer cells....
The use of a gas chromatography for determination of fatty acids and certain xenobiotics
Jaček, Martin ; Tůma, Petr (advisor) ; Pacáková, Věra (referee) ; Vecka, Marek (referee)
The submitted thesis is concerned with the use of gas chromatography (GC) to determine fatty acids (FA) in clinical samples and study of the effect of FA on the occurrence and prevention of selected diseases. The work is focused on FA metabolism in relation to essential FA and highly beneficial polyunsaturated fatty acids (PUFA) of the omega-3 group, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are insufficiently represented in the nutrition of western populations. The experimental part is concerned with the development of GC methods for determining FA as their methyl esters and their use for analysis of samples in intervention clinical studies of OMEGA and VEGGIE-2. The study of OMEGA concentrates on the incorporation of beneficial omega-3 PUFA into the food chain through microalgae synthesizing EPA and linseed containing mainly α-linolenic acid. These sources were employed to prepare a mixture for feeding poultry and the obtained eggs and chicken meat enhanced in omega-3 FA were used as ingredients for preparing food. Following 8-week dietary intervention performed on healthy volunteers, a statistically significant increase was found in the content of DHA in erythrocytes, equal to 4.4 % before and 5.1 % after the dietary intervention in the test group. The determined...
Mitochondria as a target of anticancer therapy.
Dvořák, Aleš ; Ježek, Petr (advisor) ; Poučková, Pavla (referee) ; Vecka, Marek (referee)
Mitochondrial isocitrate dehydrogenase 2 (IDH2) catalyzes reductive carboxylation (RC, reverse Krebs cycle pathway) and 2HG synthesis (2HG) - metabolite of which many scientists are interested. 2HG may be concurrently synthetized in cytosol by IDH1. RC is involved in anabolic reactions necessary for cell proliferation - produces citrate, fatty acid precursor - especially in hypoxia. IDH2 and IDH1 are not the only enzymes that are involved in 2HG synthesis. Recently, several enzymes, which participate in 2HG production, have been discovered. 2HG is useful in cancer diagnostics due to its overproduction by transformed cells. Moreover, 2HG may cause epigenetic changes via inhibition of 2-oxoglutarate dependent dioxygenase. In this work, the importance of RC and 2HG synthesis in cancer and healthy cells was investigated by gas chromatography with mass spectrometry detection as well as IDH2 influence. We found that IDH2 significantly participates in reverse RC and 2HG synthesis in breast cancer cell lines and uses glutaminolysis as a supplementary anaplerotic pathway. RC is increased by hypoxia, inhibition of respiration, and decreased by activation of respiration or hypocapnia. We confirmed 2HG synthesis and RC in healthy cells (fibroblasts, breast epithelial cells etc.) as well as in cancer cells....
The significance of the analysis of noncholesterol sterols and profile of fatty acids in various pathophysiological states
Vecka, Marek ; Tvrzická, Eva (advisor) ; Zadák, Zdeněk (referee) ; Kazdová, Ludmila (referee)
At present time, there is growing interest in two classes of lipids - sterols and fatty acids, which posses many different roles in living organisms. The concentrations of cholesterol biosynthetic precursors (lathosterol, desmosterol) reflect level of cholesterol biosynthesis, whereas the plasma phytosterol (campesterol, β-sitosterol) concentrations can be used for estimation of fractional absorption of dietary cholesterol. These groups are sometimes called together as noncholesterol sterols. The dissertation thesis dealt with the significance of analysis of noncholesterol sterols and fatty acid profile in various pathophysiological states. The methodology part of the thesis covered some aspects of analysis of these compounds with capillary gas chromatography. We found that for the minor fatty acid analysis, both the type of the column and the software used play an important role. The gas chromatography step contributes by the least part to the total error of the procedure, whereas the effect of other steps (extraction, thin-layer chromatography and methylation process) is more pronounced. The comparison of fatty acid profiles in lipoprotein particles with low and very low density revealed differences, but these differences were highly interindividual. We also found that it is possible to replace the...

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