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Application of flow cytometry for multiplex analyses in clinical biochemistry
Babušíková, Lucie ; Tremlová,, Bohuslava (referee) ; Beňovská,, Miroslava (advisor)
This thesis discusses the technique of flow cytometry for multiplex analysis and its use in conjunction with imunochemical methods. As part of this work was carried out clinical studies dealing with secondary prevention of myocardial infarction and atherosclerosis in 186 pacientů. In this time represents myocardial infarction worldwide civilizational problem. A number of possible parameters for monitoring atherosclerosis in the world is still an unresolved issue. In the practical part of this work we performed an analysis using Luminex xMAP technology for new parameters (adiponectin, resistin, osteopontin) to predict atherosclerotic disease associated with myocardial infarcion. Also we wanted to see how these parameters are changed in patients after increasing the dose of therapeutic drugs.
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Determination of biological activity of adiponectin, a recombinant protein using cell culture
Pernicová, Iva ; Rittich, Bohuslav (referee) ; Ševčík, Mojmír (advisor)
This thesis deals with the determination of biological activity of adiponectin, a recombinant protein using cell culture. First it was important to acquire the working skills for the cell culture of cell line 3T3-L1. An optimal concentration of inactivated fetal bovine serum in cell culture media was determined. A stimulation of the cell proliferation by HB-EGF, PDGF-BB and bFGF growth factors was observed at various concentration levels. Afterwards the biological activity of adiponectin was determined as an inhibition of growth stimulation with 5 ng/ml PDGF-BB. This biological activity assay for adiponectin was also conducted with lyophilized adiponectin and a growth factor bFGF (0.1 ng/ml). The lyophilization did not affect the biological activity of adiponectin.
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Circadian rhythms in adipocytes and impact of their disruption
Zavřelová, Michaela ; Sumová, Alena (advisor) ; Spišská, Veronika (referee)
This bachelor thesis deals with circadian rhythms in adipocytes and the consequences of their disruption. The thesis summarizes current scientific knowledge and explores the mechanism of rhythm control at the molecular level, which is formed by transcriptional translational feedback loops. Adipocytes control lipid metabolism through the production of adipokines, adipogenesis, lipogenesis, and lipolysis. The research focuses on how clock genes are involved in these processes. All of these events, which are characteristic of adipocytes, are highly regulated and the genes that are involved in them are transcriptionally activated by clock gene proteins. Circadian rhythms are an integral part of our lives, and desynchronization of rhythms between the main oscillator, which is the suprachiasmatic nuclei, and peripheral adipocytes can lead to metabolic disorders, which in turn can lead to obesity, insulin resistance, or type 2 diabetes. Obesity is one of the biggest problems of modern society, and disrupted circadian rhythms due to today's lifestyle are one of the causes of its development. Key words: circadian rhythms, clock genes, adipocytes, obesity, PPARγ, adipogenesis, lipogenesis, lipolysis, leptin, adiponectin
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Vplyv účinných zložiek rybieho tuku na expresiu vybraných génov v modelovom organizme
Frólo, Michal
The aim of this work was to verify the hypothesis that eicosapentaenoic acid and docosahexaenoic acid favorably influence the inflammatory response in the organism through their influence on the expression of genes associated with the inflammatory process. The experiment was carried out on pigs fed with fish oil supplement and palm oil supplement. Certain experimental groups were additionally stimulated by lipopolysaccharide to induce a rapid onset of inflammatory response, with particular concern for the ability of polyunsaturated fatty acids n-3 to inhibit this process. I have been observing changes in gene expression for AdipoR1 and AdipoR2 adiponectin receptors, as well as expression in other genes related to the inflammatory process of LBP; ICAM1; IL-6; TNFalfa; IL-12. The determination of the expression of selected genes using qRT-PCR was used to process the results from the collected liver and adipose tissue samples. I statistically evaluated the expression results of the genes and displayed them as graphs. The favorable demonstrable effect of fish oil was only observed in the ADIPOR2 gene in the liver, in the non-stimulated lipopolysaccharide group (P <0.05). There was no detectable effect of fish oil on modulation of inflamation in pigs. However, we can deduce the causes of the failure from the experimental results, and in other studies we might be able to confirm the hypothesis. However, we can deduce the causes of the failure from the experimental results and we might be able to confirm the hypothesis in other studies.
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Impact of weight loss in obese subjects on the sensitivity of adipose tissue cells in relation to stress of endoplasmatic reticulum.
Karlická, Michaela ; Rossmeislová, Lenka (advisor) ; Němcová, Vlasta (referee)
Adipocytokines released by the adipose tissue play an important role in the regulation of immune and inflammatory responses. In obesity their production is dysregulated, which is one of the major factors contributing to the onset of a chronic low-grade systemic inflammation representing a risk factor for the progression of other diseases, such as atherosclerosis or type-2 diabetes. The main goal of this thesis was to analyze the secretion of selected adipocytokines (adiponectin, IL6 and MCP1) by in-vitro differentiated adipocytes, isolated from the adipose tissue prior to and after a dietary intervention, and this under basal conditions and during stimulated lipolysis. In case of adiponectin, the secretion of its isoforms was analyzed too. The concentration of adiponectin, IL6 and MCP1 was determined by the ELISA method, the Western Blot method was used to determine the distribution of the adiponectin isoforms. The thesis also concentrates on the gene expression of ATF3, ATF4 and HSPA5, factors engaged in the ER stress in the course of the differentiation of adipocytes. The changes in the gene expression were measured by the quantitative Real Time PCR method. At the same time the development of the endoplasmic reticulum (ER) in the course of adipogenesis was monitored by indirect...
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Vliv polynenasycených mastných kyselin n-3 na markery zánětu u modelového organizmu
Pešková, Petra
The aim of this thesis was to verify the hypothesis implying that n-3 polyunsaturated fatty acids inhibit the development of mild chronic inflammation. The experiment was conducted on rats with different diets (control, with the addition of safflower oils, fish oils and oils from algae Schizochytrium). For processing the results of the collected tissues was used determination of expression of selected genes by qRT-PCR, detection of proteins and in the cytosol and nuclear fractions by Western blot and the quantification of cytokines by ELISA. Feeding oil from algae Schizochytrium has led to lowering the final weight and blood glucose, further to enhance expression of PPAR-gama and increasing the production of anti-inflammatory cytokines IL-10 and TGF-beta1. In kontrast, no difference was observed in the expression of GPR120 and adiponectin receptors or proteins in an amount of NF-kappaB and PPAR-gama between diets. Elevated plasma levels of adiponectin were found. The results of the experiment shows that it is possible to recommend oil from algae Schizochytrium as a useful supplement of human diet as prevention of chronic degenerative diseases.
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Determination of biological activity of adiponectin, a recombinant protein using cell culture
Pernicová, Iva ; Rittich, Bohuslav (referee) ; Ševčík, Mojmír (advisor)
This thesis deals with the determination of biological activity of adiponectin, a recombinant protein using cell culture. First it was important to acquire the working skills for the cell culture of cell line 3T3-L1. An optimal concentration of inactivated fetal bovine serum in cell culture media was determined. A stimulation of the cell proliferation by HB-EGF, PDGF-BB and bFGF growth factors was observed at various concentration levels. Afterwards the biological activity of adiponectin was determined as an inhibition of growth stimulation with 5 ng/ml PDGF-BB. This biological activity assay for adiponectin was also conducted with lyophilized adiponectin and a growth factor bFGF (0.1 ng/ml). The lyophilization did not affect the biological activity of adiponectin.
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Application of flow cytometry for multiplex analyses in clinical biochemistry
Babušíková, Lucie ; Tremlová,, Bohuslava (referee) ; Beňovská,, Miroslava (advisor)
This thesis discusses the technique of flow cytometry for multiplex analysis and its use in conjunction with imunochemical methods. As part of this work was carried out clinical studies dealing with secondary prevention of myocardial infarction and atherosclerosis in 186 pacientů. In this time represents myocardial infarction worldwide civilizational problem. A number of possible parameters for monitoring atherosclerosis in the world is still an unresolved issue. In the practical part of this work we performed an analysis using Luminex xMAP technology for new parameters (adiponectin, resistin, osteopontin) to predict atherosclerotic disease associated with myocardial infarcion. Also we wanted to see how these parameters are changed in patients after increasing the dose of therapeutic drugs.
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