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Qualitative analysis of selected glycosphingolipids subclasses isolated from human blood plasma using HILIC-ESI-MS/MS
ŠIKLOVÁ, Michaela
This bachelor thesis deals with the qualitative analysis of selected subclasses of glycosphingolipids isolated from human blood plasma using hydrophilic interaction liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). Glycosphingolipids are ubiquitous and structurally very diverse components of cell membranes involved in many cellular functions. It has been shown that changes in the structures of glycosphingolipids and their distinct expression can pose a key factor in the development of numerous diseases and different types of cancer.The theoretical part of this thesis is dedicated to a summary of fundamental knowledge about glycosphingolipids, their biosynthesis, classification, biological functions and their association with various types of diseases and cancers. Furthermore, methods of isolation, purification and analytical possibilities for the determination of glycosphingolipids in biological samples are described here.In the experimental part, blood samples were taken and processed together with the preparation of the necessary solutions, plasma samples, and standards. Moreover, a test analysis, optimization of mobile phase gradient and comparison of the most frequently used extraction methods according to Folch, Bligh and Dyer, Matyash, and Alshehry alongside fractionation of lipid extract is described in this part. An integral part of the experimental section was the identification and structural characterization of glycosphingolipids within selected subclasses, including the generation of GSL profiles in human blood plasma. In conclusion, the results of the work and answers to the stated research questions are summarized, including the evaluation of possible improvements in this field.
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Effect of physical exercise on adipose tissue characteristics: influence of obesity and aging
Čížková, Terezie ; Šiklová, Michaela (advisor) ; Horníková, Daniela (referee)
Obesity and ageing are associated with the changes in adipose tissue characteristics, which are con- sidered as one of important contributors to development of complications, such as cardiovascular diseases or type 2 diabetes. Physical exercise is known to be effective in prevention of diseases rela- ted to obesity and ageing. However, the effect of exercise on adipose tissue characteristics is only few elucidated in humans. The goal of this bachelor thesis is to summarize the main effects of exer- cise on adipose tissue characteristics. This thesis subsequently discuss detrimental changes (dysfunction) of the adipose tissue in obesity. Recent findings about the effects of some types of physical training on health of the elderly people are also mentioned. The initial chapter deals with adipose tissue in general, concretely the morphology, metabolism and endocrine function of the adipose tissue are described. Next part is focused on obesity, the concept of a chronic low-grade inflammatory state is introduced. The key section is the fourth chapter, in which the effects of exer- cise on adipose tissue, i.e. effects on metabolism of adipocytes and activation of immune cells, are reviewed. At the end of this thesis, the pathophysiological changes in inflammatory state and adi- pose tissue distribution associated...
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Metabolipidomic profiling of white adipose tissue by UPLC-MS/MS
Rombaldová, Martina ; Kuda, Ondřej (advisor) ; Šiklová, Michaela (referee) ; Čajka, Tomáš (referee)
Obesity is a serious problem in society today [1,2]. It might seems to have been caused simply by excess consumption of food compared to energy expenditure but obesity is actually a complex metabolic disorder centred on adipose lipid metabolism and cellular signalling systems linked to it [3]. Understanding the biology of adipose tissue (AT) is very important for the identification of novel and potential therapeutic targets in order to prevent and treat obesity-related disorders [4]. We utilized analytical approach liquid chromatography coupled to mass spectrometry (LC-MS) to study adipose tissue metabolism. Also, we were especially interested in the effect of omega-3 polyunsaturated fatty acids (PUFA) on that metabolism. Rodent and cell line experiments were performed and analyses were done of white adipose tissue (WAT), serum/plasma samples or cells as well as milk samples from mothers. At first, we established several ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) methods for analysis of acylcarnitines (AC), amino acids (AmA) and other metabolites. Importantly, these methods were able to distinguish isobaric species of AC which is not usually possible. Using these approaches we uncovered several acylcarnitines, i.e. long chain AC, carnitine, acylcarnitine C4 and...
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Metabolipidomic profiling of white adipose tissue by UPLC-MS/MS
Rombaldová, Martina ; Kuda, Ondřej (advisor) ; Šiklová, Michaela (referee) ; Čajka, Tomáš (referee)
Obesity is a serious problem in society today [1,2]. It might seems to have been caused simply by excess consumption of food compared to energy expenditure but obesity is actually a complex metabolic disorder centred on adipose lipid metabolism and cellular signalling systems linked to it [3]. Understanding the biology of adipose tissue (AT) is very important for the identification of novel and potential therapeutic targets in order to prevent and treat obesity-related disorders [4]. We utilized analytical approach liquid chromatography coupled to mass spectrometry (LC-MS) to study adipose tissue metabolism. Also, we were especially interested in the effect of omega-3 polyunsaturated fatty acids (PUFA) on that metabolism. Rodent and cell line experiments were performed and analyses were done of white adipose tissue (WAT), serum/plasma samples or cells as well as milk samples from mothers. At first, we established several ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) methods for analysis of acylcarnitines (AC), amino acids (AmA) and other metabolites. Importantly, these methods were able to distinguish isobaric species of AC which is not usually possible. Using these approaches we uncovered several acylcarnitines, i.e. long chain AC, carnitine, acylcarnitine C4 and...
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Effect of regular physical exercise on metabolism of adipose tissue in elderly
Čížková, Terezie ; Šiklová, Michaela (advisor) ; Horáková, Olga (referee)
Ageing is associated with changes in body composition, which is caused by decreases in muscle mass and a progressive increase of fat mass. It is also associated with redistribution of adipose tissue with increased visceral and ectopic fat deposition. These changes are related to insulin resistance and other metabolic disturbances. Adipose tissue dysfunction is important contributor to the disturbed metabolic status of elderly and it is characterized by altered adipogenesis, metabolic flexibility and changes in secretory activity. Regular exercise is indisputably a key part of a healthy lifestyle. It was shown to improve muscle function, cardiovascular fitness and metabolic health of elderly. The aim of this thesis was to investigate whether a regular physical exercise can influence characteristics of an adipose tissue in elderly women, and if these changes in the adipose tissue can contribute to an improvement in an insulin sensitivity. In the two groups of elderly women, a long-term trained and a sedentary, the subcutaneous adipose tissue was analyzed for a gene expression of number of metabolic genes, an adipocyte size and a lipolytic rate at the basal state and after the isoproterenol stimulation. Anthropometric parameters and insulin sensitivity were also determined. Outcomes of this thesis...
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Effect of physical exercise on adipose tissue characteristics: influence of obesity and aging
Čížková, Terezie ; Šiklová, Michaela (advisor) ; Horníková, Daniela (referee)
Obesity and ageing are associated with the changes in adipose tissue characteristics, which are con- sidered as one of important contributors to development of complications, such as cardiovascular diseases or type 2 diabetes. Physical exercise is known to be effective in prevention of diseases rela- ted to obesity and ageing. However, the effect of exercise on adipose tissue characteristics is only few elucidated in humans. The goal of this bachelor thesis is to summarize the main effects of exer- cise on adipose tissue characteristics. This thesis subsequently discuss detrimental changes (dysfunction) of the adipose tissue in obesity. Recent findings about the effects of some types of physical training on health of the elderly people are also mentioned. The initial chapter deals with adipose tissue in general, concretely the morphology, metabolism and endocrine function of the adipose tissue are described. Next part is focused on obesity, the concept of a chronic low-grade inflammatory state is introduced. The key section is the fourth chapter, in which the effects of exer- cise on adipose tissue, i.e. effects on metabolism of adipocytes and activation of immune cells, are reviewed. At the end of this thesis, the pathophysiological changes in inflammatory state and adi- pose tissue distribution associated...
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