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Study of Activity of Enzyma and Low Molecular Weight Antioxidant Systems
Macuchová, Simona ; Kráčmar, Stanislav (referee) ; Čertík, Milan (referee) ; Márová, Ivana (advisor)
Oxidative processes play important role in cell physiology and pathology as well. Balance of these processes is supplied by cooperating antioxidative systems; function of antioxidant defense systems depens on high levels of antioxidants in organism. Presented work is focused on developement and optimization of methods for analysis of important enzyme and non-enzyme antioxidants as well as total antioxidant capacity of selected types of biological material. Extractions and analyses of vitamin E, carotenoids, superoxide dismutase, catalase, peroxidase and lipoxygenase in barley and malt were optimized. RP-HPLC and HPLC/ESI-MS were used for analysis of vitamin E, phenolic and carotenoid content, spectrophotometry was used for enzymes activity analysis. A new methods for catalase and lipoxygenase activities were developed and compared with direct UV methods. Superoxide dismutase activity was determined by commercial diagnostic kit. A colorimetric method was used for peroxidase activity determination. Some kinetic parameters of enzymes were provided too. Optimized methods were used in the analyses of antioxidants in plant material - in barley and malt - in sets of samples of 6 varieties cultivated in four different locations for two years. Content of individual antioxidants differed depending on the variety, but usually were not found significant differences in the levels, depending on growing location. Perhaps climatic conditions have the greatest influence on levels of low molecular weight and enzymatic antioxidants at the specific location; oxidation processes are influenced both the quantity of moisture, both by sunlight, which induces oxidative processes in cultivated plants. The activity of antioxidants in barley caryopses is rapidly increasing during the malting process; an elevated temperature and moistness first induces activation the enzyme systems including antioxidant. In caryopsis is metabolic activity increased during which we can expect an increased production of radicals; for this purpose can antioxidant systems be activated that protect cells from damage by oxidative stress. In the second part of work optimized methods were applied in two clinical trials focused on study of the influence of exogenous antioxidants intake on metabolic and antioxidant status in human organism. In the first clinical study influence of food supplement containing polyunsaturated fatty acids and vitamin E on metabolism of hyperlipidaemics was evaluated. After 3-month supplemenation a lipid profile was improved and serum antioxidant levels increased. The second experiment was focused on enzyme and non-enzyme antioxidant levels in healthy subjects after temporarily intake of specific foods rich in antioxidants. After two-month intake plasma phenolic substances were slightly increased. Total antioxidant capacity and activities of enzyme antioxidants were not affected. Results of both clinical exeriments showed that supplying of antioxidants in natural form or in the form of food supplements does not markedly affect metabolism of healthy subjects, while in patients with chronic diseases antioxidant supplementation can positively influence metabolic status. Results of this work showed that optimized methods are suitable for analyses of antioxidant status parameters and also for monitoring of exogenous antioxidant intake.
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Study of Activity of Enzyma and Low Molecular Weight Antioxidant Systems
Macuchová, Simona ; Kráčmar, Stanislav (referee) ; Čertík, Milan (referee) ; Márová, Ivana (advisor)
Oxidative processes play important role in cell physiology and pathology as well. Balance of these processes is supplied by cooperating antioxidative systems; function of antioxidant defense systems depens on high levels of antioxidants in organism. Presented work is focused on developement and optimization of methods for analysis of important enzyme and non-enzyme antioxidants as well as total antioxidant capacity of selected types of biological material. Extractions and analyses of vitamin E, carotenoids, superoxide dismutase, catalase, peroxidase and lipoxygenase in barley and malt were optimized. RP-HPLC and HPLC/ESI-MS were used for analysis of vitamin E, phenolic and carotenoid content, spectrophotometry was used for enzymes activity analysis. A new methods for catalase and lipoxygenase activities were developed and compared with direct UV methods. Superoxide dismutase activity was determined by commercial diagnostic kit. A colorimetric method was used for peroxidase activity determination. Some kinetic parameters of enzymes were provided too. Optimized methods were used in the analyses of antioxidants in plant material - in barley and malt - in sets of samples of 6 varieties cultivated in four different locations for two years. Content of individual antioxidants differed depending on the variety, but usually were not found significant differences in the levels, depending on growing location. Perhaps climatic conditions have the greatest influence on levels of low molecular weight and enzymatic antioxidants at the specific location; oxidation processes are influenced both the quantity of moisture, both by sunlight, which induces oxidative processes in cultivated plants. The activity of antioxidants in barley caryopses is rapidly increasing during the malting process; an elevated temperature and moistness first induces activation the enzyme systems including antioxidant. In caryopsis is metabolic activity increased during which we can expect an increased production of radicals; for this purpose can antioxidant systems be activated that protect cells from damage by oxidative stress. In the second part of work optimized methods were applied in two clinical trials focused on study of the influence of exogenous antioxidants intake on metabolic and antioxidant status in human organism. In the first clinical study influence of food supplement containing polyunsaturated fatty acids and vitamin E on metabolism of hyperlipidaemics was evaluated. After 3-month supplemenation a lipid profile was improved and serum antioxidant levels increased. The second experiment was focused on enzyme and non-enzyme antioxidant levels in healthy subjects after temporarily intake of specific foods rich in antioxidants. After two-month intake plasma phenolic substances were slightly increased. Total antioxidant capacity and activities of enzyme antioxidants were not affected. Results of both clinical exeriments showed that supplying of antioxidants in natural form or in the form of food supplements does not markedly affect metabolism of healthy subjects, while in patients with chronic diseases antioxidant supplementation can positively influence metabolic status. Results of this work showed that optimized methods are suitable for analyses of antioxidant status parameters and also for monitoring of exogenous antioxidant intake.
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Health and social risks in children predisposed to metabolic syndrome
SLADKÁ, Alena
Metabolic syndrome was defined for the first time in 1988 by Reaven, who defined it as a hormonal-metabolic syndrome - with symptoms of resistance to insulin , hyperinzulinismus, hypertension and with laboratory-proven higher level of lipoproteid VLDL and lower level of HDL and LDL cholesterol. Obese children have big troubles concerning social relationships. Overweight children are often an object of ridicule, teasing, verbal bullying, physical bullying, cyberbullying and sexual abuse. First goal of this thesis is to find out, whether obesity (mainly of android type) in children and adolescents (aged 10-18) having BMI higher than 28 and waist size for boys over 103 and for girls over 93, overall cholesterol level over 6,5, blood pressure higher than 130/70 are predispositions of manifestation of MS in adulthood and to find out what are the social and health impacts of the obesity in the childhood. Second goal of this thesis is to find out, whether patients with metabolic syndrome (MS) had predispositions for obesity or were obese in the childhood and what are the health and social impacts of MS in their life. The first research group consisted of ten children aged 10-18 with predispositions to manifestation of MS. Methods of accomplishing goals of this thesis: Survey of health and family anamnesis. All ten children aged 10-18 were repeatedly checked during six mounths for their body weight, waist size, blood pressure and cholesterol. Surveyed values are shown in the table. I Used a structured interview to survey relationships of obese ones with same-age children, whether they are an object of ridicule and bullying and what are other troubles caused by the disease. At the same time I gave questionnaires to parents to ascertain the eating habits and life style of the family. Health and social impacts for life of people with diagnosed MS, when they were diagnosed MS and their childhood condition were examined using structured interviews in the metabolic advice centre. The research group consisted of ten patiensts of metabolic advice centre. After analysis of data and health anamnesis it was proven that 60% of researched children suffer from higher morbidity (breathing issues, upper respiratory tract infection, and easier fatigue during physical exertion). 60% of children have serious issues to establish relationships and suffer from verbal and physical bullying. 40% of adult patients of metabolic advice centre had troubles with obesity already in their childhood. All of them suffer from type 1 diabetes. Two of them recieve disability pension due to MS. 90% of them told, that they regulary go see a doctor und regulary take cures. These facts cause worse financial situation (lower income, expenses for cures and travel to a doctor). Two respondents mentioned discriminating behaviour of medical staff, however they didn't mention the type of discrimination. According to the questionaries only one person considers the MS is caused by his lifestyle. Research group consisted all the time of ten children and ten adults with MS, because 'Snowball sampling' research methods didn't survey any new facts, which would be needed to verify.
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