|
|
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.
|
|
|
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.
|
|
|
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
|
|
| |
|
|
Selected Nutrients in Etiology, Prevention and Treatment of Obesity
Sedláček, Pavel ; Müllerová, Dana (advisor) ; Rajdl, Daniel (referee) ; Fiala, Zdeněk (referee)
SELECTED NUTRIENTS IN ETIOLOGY, PREVENTION AND TREATMENT OF OBESITY The 21st century pandemic of obesity is given by to the obesitogenic environment that helps to develop a positive energy balance, where the energy intake of an individual chronically exceeds his energy need, ie energy expenditure. There is an increase in body weight by increased or abnormal body fat accumulation in the body. Preventing the development of obesity is aimed at achieving energy balance, in the case of existing overweight or obesity, the treatment lies in inducing and maintaining a negative energy balance for some time, ie stimulating energy expenditure and reducing energy intake. Selected diet determines not only energy intake, but its individual nutrients can also partially affect energy expenditure and fat tissue physiology. The aim of this work is to describe and experimentally verify some nutrients that could help in weight reduction and physiological function of adipose tissue. Based on the literature data, 19 active substances or mixtures were considered. A mixture of ω-3 polyunsaturated fatty acids (ω-3 PUFA), eicosapentaenoic acid and docosahexaenoic acid, in a daily dose of 0.6 g, in the form of fish oil in a certified food supplement was chosen as the most suitable for the experiment. In a 12-week, three-arm, parallel...
|
|
|
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...
|
|
| |
|
|
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.
|
|
|
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.
|
guest ::
