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Regulation of triglyceride metabolism in circulation in postprandial phase.
Zemánková, Kateřina ; Kovář, Jan (advisor) ; Kazdová, Ludmila (referee) ; Zlatohlávek, Lukáš (referee)
Increased triglyceride (TG) concentration has been generally accepted as a risk factor for ischemic heart disease and, therefore, lowering TG is therapeutic target that should reduce cardiovascular disease risk. Traditionally, concentration of TG is measured in the fasting state (8-12 hours after an overnight fasting) mainly because the rise in TG levels after meal leads to the high variation in TG values. However, human beings spend larger portion of the day in a postprandial state and postprandial hypertriglyceridemia may then play a substantial role in determination of cardiovascular disease risk. The increased and prolonged postprandial lipemia has been found in patients with coronary heart disease. Moreover, recent data from Copenhagen Heart Study point out that the non-fasting TG concentration is associated with cardiovascular disease risk more tightly than the fasting TG concentration. Importantly, concentration of non-fasting TG is substantially affected by individual behavioural habits such as diet composition and physical activity. It remains to be determined whether it would be appropriate to identify individuals at higher risk of cardiovascular disease due to increased postprandial TG using tolerance test analogous to glucose tolerance test. The protocol of standardized fat tolerance...
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Lipoprotein study by SEC/LLSD method
Šinglárová, Simona ; Tvrzická, Eva (advisor) ; Staňková, Barbora (referee)
The disorders of the lipid metabolism have to be monitored for a number of socially significant diseases. For the diagnosis of these disorders, it turned out in the last 10 years as very beneficial to determine the particle size of the individual lipoprotein classes, in particular the low-density lipoprotein. Among several known methods, we have chosen gel filtration with the light scattering detector. Lipoprotein fractions were separated by ultracentrifugation from the plasma of four donors. Three biochemical parameters - total cholesterol, triacylglycerol, and total protein - were routinely determined for plasma and for each fraction. The silica column BioBasic SEC 300x7,8 mm was calibrated by the proteins thyroglobulin, ovalbumin and angiotensin II with UV detection (280 nm). These proteins are not suitable (due to the size) for the calibration method SEC-LLSD. Therefore, latex beads of various modifications and sizes were tested. The only usable measurement was obtained for the deep-blue latex beads of the size 55 nm, the others remained retained in the column. The lipoprotein fractions of VLDL and LDL were examined by the SEC-LLSD method at different pH (7.3, 7.5, 7.7, 8.0) and ionic strength (0.1M phosphate buffer, 0.1M phosphate buffer and 0.9% NaCl, 0.05M phosphate buffer and 0.9% NaCl)....
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Serum markers of cholesterol 7α hydroxylase activity
Bohdanecká, Alena ; Leníček, Martin (advisor) ; Kovář, Jan (referee)
Cholesterol 7-hydroxylase (CYP7A1) is the rate limiting enzyme of the classical pathway of bile acid (BA) synthesis, which catabolizes approximately half of cholesterol in man. Determination of CYP7A enzymatic activity is a key subject of lipid metabolism research. Direct determination of CYP7A1 activity in hepatic biopsy is mostly not allowed for ethical reasons, so indirect methods are used with serum markers such as 7α-hydroxy-4-cholestene-3- one (C4). The first, methodical aim of the work was to convert the introduced HPLC method for the determination of C4 to LC-MS in order to increase the sensitivity. We focused on the solid phase extraction step, adjusting the composition and volumes of the washing and elution solution. By converting the method from HPLC to LC-MS, the sensitivity was increased approximately 7 times (LD = 1.39 ng/ml). In the second, clinical part of our work, we attempted to confirm the preliminary results of our laboratory on the distribution of C4 in lipoprotein fractions (LPP) in order to find parameter that would correlate with CYP7A1 activity better than C4 level itself. Preliminary results (performed in healthy individuals) showed that most of C4 is carried on HDL, and that the C4 distribution within LPP fractions is similar among examined subjects. We repeated the...
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Influence of Diet on Blood Lipid Levels in Dyslipidemic Patients
Paroha, Josef ; Zlatohlávek, Lukáš (advisor) ; Tůmová, Eva (referee)
The primary aim of this thesis is to determine whether the hypolipidemic diet has an influence on blood lipid levels in patients with dyslipidemia. The secondary aim is to find out the eating habits of patients with dyslipidemia and to determine whether their eating habits are in line with the basics of a hypolipidemic diet. To achieve these targets a questionnaire, biochemical examination of blood and education of dyslipidemic patients on the appropriate diet were used. My research contained check-in and check-out laboratory blood examinations. After check-in examination each patient filled in the questionnaire with questions targeted at eating habits and then the patient was educated about the appropriate diet. After three months' time the patients proceeded to check-out laboratory blood examination. The necessary data were acquired by comparing the results of the check-in and check-out blood examinations and by the questionnaires evaluation. This thesis in its research confirmed the positive influence of hypolipidemic diet on blood lipids levels in patients with dyslipidemia. At the end of the research the average level of total cholesterol was 10,5 % lower, the average level of LDL-cholesterol was 18,9 % lower and the average level of triacylglycerol was 10,6 % lower. On the contrary the...
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Regulation of triglyceride metabolism in circulation in postprandial phase.
Zemánková, Kateřina
Increased triglyceride (TG) concentration has been generally accepted as a risk factor for ischemic heart disease and, therefore, lowering TG is therapeutic target that should reduce cardiovascular disease risk. Traditionally, concentration of TG is measured in the fasting state (8-12 hours after an overnight fasting) mainly because the rise in TG levels after meal leads to the high variation in TG values. However, human beings spend larger portion of the day in a postprandial state and postprandial hypertriglyceridemia may then play a substantial role in determination of cardiovascular disease risk. The increased and prolonged postprandial lipemia has been found in patients with coronary heart disease. Moreover, recent data from Copenhagen Heart Study point out that the non-fasting TG concentration is associated with cardiovascular disease risk more tightly than the fasting TG concentration. Importantly, concentration of non-fasting TG is substantially affected by individual behavioural habits such as diet composition and physical activity. It remains to be determined whether it would be appropriate to identify individuals at higher risk of cardiovascular disease due to increased postprandial TG using tolerance test analogous to glucose tolerance test. The protocol of standardized fat tolerance...
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Separation and identification of lipopeptides by chromatography and mass spectrometry
Pravdová, Adéla ; Cvačka, Josef (advisor) ; Kozlík, Petr (referee)
5 Abstract Protein lipidation occupies an important place in the post-translational modification group. It plays an important role in the processes of cell differentiation or synaptic transmission. Changes in the presence of lipidated proteins may in some cases indicate the appearance of human diseases such as Huntington's disease, schizophrenia, or cancer. For this reason, their analysis has become desirable, but also problematic due to its specific characteristics. This work is focused on the development of conditions for measurement of lipopeptides produced by cleavage of proteins containing lipomodification. Two separation procedures were tested, in which the aim of work was to enrich the lipopeptides and remove as many other molecules as possible from the samples. The first test method - phase interface separation, using water and chloroform, was able to concentrate the lipopeptides on the phase interface and separate some of the added unmodified peptides from the mixture. The second method of separation was a solid phase using a C18 stationary phase. As a more suitable solid phase extraction variant, elution with solutions containing acetonitrile with trifluoroacetic acid was chosen as compared to solutions containing isopropyl alcohol. With the modified elution procedure, it was possible to separate...
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Regulation of triglyceride metabolism in circulation in postprandial phase.
Zemánková, Kateřina
Increased triglyceride (TG) concentration has been generally accepted as a risk factor for ischemic heart disease and, therefore, lowering TG is therapeutic target that should reduce cardiovascular disease risk. Traditionally, concentration of TG is measured in the fasting state (8-12 hours after an overnight fasting) mainly because the rise in TG levels after meal leads to the high variation in TG values. However, human beings spend larger portion of the day in a postprandial state and postprandial hypertriglyceridemia may then play a substantial role in determination of cardiovascular disease risk. The increased and prolonged postprandial lipemia has been found in patients with coronary heart disease. Moreover, recent data from Copenhagen Heart Study point out that the non-fasting TG concentration is associated with cardiovascular disease risk more tightly than the fasting TG concentration. Importantly, concentration of non-fasting TG is substantially affected by individual behavioural habits such as diet composition and physical activity. It remains to be determined whether it would be appropriate to identify individuals at higher risk of cardiovascular disease due to increased postprandial TG using tolerance test analogous to glucose tolerance test. The protocol of standardized fat tolerance...
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Comparing values of sedimentation of erythrocytes of lipid complex
ŠTEFKOVÁ, Michaela
The aim of this work is to compare the values of erythrocytes sedimentation rate (ESR) of the ESR obtained by the stop flow method with the individual values of lipid complex. Verify that ESR values are affected by hyperlipoproteinemia. Correlation coefficients are calculated and correlation dependence graphs between ESR values and lipid assay parameters (TAG, TC, HDL and LDL) are compiled.
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Regulation of triglyceride metabolism in circulation in postprandial phase.
Zemánková, Kateřina ; Kovář, Jan (advisor) ; Kazdová, Ludmila (referee) ; Zlatohlávek, Lukáš (referee)
Increased triglyceride (TG) concentration has been generally accepted as a risk factor for ischemic heart disease and, therefore, lowering TG is therapeutic target that should reduce cardiovascular disease risk. Traditionally, concentration of TG is measured in the fasting state (8-12 hours after an overnight fasting) mainly because the rise in TG levels after meal leads to the high variation in TG values. However, human beings spend larger portion of the day in a postprandial state and postprandial hypertriglyceridemia may then play a substantial role in determination of cardiovascular disease risk. The increased and prolonged postprandial lipemia has been found in patients with coronary heart disease. Moreover, recent data from Copenhagen Heart Study point out that the non-fasting TG concentration is associated with cardiovascular disease risk more tightly than the fasting TG concentration. Importantly, concentration of non-fasting TG is substantially affected by individual behavioural habits such as diet composition and physical activity. It remains to be determined whether it would be appropriate to identify individuals at higher risk of cardiovascular disease due to increased postprandial TG using tolerance test analogous to glucose tolerance test. The protocol of standardized fat tolerance...
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Serum markers of cholesterol 7α hydroxylase activity
Bohdanecká, Alena ; Leníček, Martin (advisor) ; Kovář, Jan (referee)
Cholesterol 7-hydroxylase (CYP7A1) is the rate limiting enzyme of the classical pathway of bile acid (BA) synthesis, which catabolizes approximately half of cholesterol in man. Determination of CYP7A enzymatic activity is a key subject of lipid metabolism research. Direct determination of CYP7A1 activity in hepatic biopsy is mostly not allowed for ethical reasons, so indirect methods are used with serum markers such as 7α-hydroxy-4-cholestene-3- one (C4). The first, methodical aim of the work was to convert the introduced HPLC method for the determination of C4 to LC-MS in order to increase the sensitivity. We focused on the solid phase extraction step, adjusting the composition and volumes of the washing and elution solution. By converting the method from HPLC to LC-MS, the sensitivity was increased approximately 7 times (LD = 1.39 ng/ml). In the second, clinical part of our work, we attempted to confirm the preliminary results of our laboratory on the distribution of C4 in lipoprotein fractions (LPP) in order to find parameter that would correlate with CYP7A1 activity better than C4 level itself. Preliminary results (performed in healthy individuals) showed that most of C4 is carried on HDL, and that the C4 distribution within LPP fractions is similar among examined subjects. We repeated the...
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