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Lipid emulsions in lonf-term parenteral nutition
Ševela, Stanislav ; Novák, František (advisor) ; Cahová, Monika (referee) ; Lahoda Brodská, Helena (referee)
The combination of intestinal failure and parenteral nutrition is a condition associated with mild chronic inflammation and organ injury, the most significant of which is hepatic injury. This crossover-controlled trial investigated the effects of different intravenous lipid emulsions (IVLE) enriched with increased doses of omega-3 polyunsaturated fatty acids (n-3 PUFA) from fish oil (FO) in patients on home parenteral nutrition (HPNP) for chronic intestinal failure. Twelve patients were consecutively given three different IVLEs, with an FO emulsion added after each IVLE. The study also included twelve paired healthy controls for comparison. The aim of this study was to investigate how long-term administration of these IVLEs affected primarily the erythrocyte and plasma fatty acid (FA) spectrum, as well as the inflammatory response, antioxidant status, markers of liver function and bile acid spectrum in HPNP, and how these changes differed from healthy subjects. We demonstrated successful incorporation of n-3 PUFA into erythrocyte phospholipids and corresponding changes in the plasma FA spectrum, characterized in particular by significant increases in n-3 versus n-6 PUFA, with a decrease in the n-6/n-3 PUFA ratio. We also found a more prolonged effect on these changes with high dose FO...
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Influence of Storage Conditions on Metabolic Profile of Apples
Duroňová, Kateřina ; Vávrová, Milada (referee) ; Kráčmar, Stanislav (referee) ; Márová, Ivana (advisor)
The goal of presented dissertation has been complex analysis of changes in content of fatty acids, enzymatic and low-molecular antioxidants in apples and related evaluation of perception of originators of storage diseases for apples stored in various conditions. Main part of the work has been dedicated to study of the impact of storage of apples in modified atmosphere with reduced amount of oxygen and in reference “normal” atmosphere for six months. Next part of the work has been dedicated to study how apple storage in common, consumer affordable, conditions (storage in a cellar, in a refrigerator, and in room temperature) affects content of low-molecular antioxidants. For testing has been selected apple kinds Jonagored, Golden Delicious, Idared, Šampion, and Granny Smith. Within the scope of this work has been optimized the method for determination of fatty acids in plant material with higher content of wax. The measured values imply the apples are valuable source of many important nutrition substances like vitamins, provitamins and antioxidants. During the storage process these substances exhibit considerable protective function. Long-term storage, mainly in the atmosphere with reduced amount of oxygen (FAN), enables preservation of majority of these important nutrition substances depending on the kind of apple and conditions of storage. Freezing process is conservative to apples (mainly in the presence of protective substances), while in the process of drying the values of all monitored antioxidants decrease depending on temperature and conditions of drying. Upon the choice of the storage method one must consider nutrition, sensoric characteristic and consumer demands.
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Proteome and Metabolome Changes in Selected Organisms under Stress
Halienová, Andrea ; Kráčmar, Stanislav (referee) ; Čertík, Milan (referee) ; Šucman, Emanuel (referee) ; Márová, Ivana (advisor)
Living conditions of every organism are influenced by various factors at this time. Some of them have positive effect on organism, some negative. Basic condition for surviving is the ability to resist and adapt to changing metabolic and living conditions. Every single stress effect can lead to changes in metabolism but organisms have ability to develope sufficient mechanisms for stress response. Some of them are similar for all living organisms (enzyme production, endogenous primary stress metabolites) some of them are specific for certain organism or stress type. Cell stress response can be observed on different levels (proteomic, genomic, metabolomic). In proper conditions it can be used indrustrially. In this work, influences of various stress factors were studied. These factors were applied on selected organisms – carotenogenic yeast and plant materials. Yeast stress response was induced by osmotic and oxidation stress factors. Changes on proteomic level and in production of selected secondary metabolites were observed. Proteome was analyzed by 1D and 2D electrophoresis with subsequent analysis of proteins by mass spectrometry. Yeast strain Rhodotorula glutinis CCY 20-2-26 showed the best adaptation to stress factors, which was moreover accompanied by overproduction of carotenoids. This finding can be premise for next industrial production of carotenoids. In plant samples predominantly enzymes and metabolites involved in antioxidant response were studied.
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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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Changes of enzyme activities in fruits during long-term storage
Ferdová, Jitka ; Melounová,, Jitka (referee) ; Márová, Ivana (advisor)
This study is focused on study of changes of enzyme and low-molecular weight antioxidants in different fruits during long-term storage. In theoretical part individual low-molecular weight antioxidants and enzymes are described. The main causes of fruit decay and some possibilities of fruit preservation and storage are summarized. As biological material some common fruits were chosen - green and red apples, peaches, plums and white grapes. The fruits were stored in laboratory, cellar, in refrigerator and in freezer. In freezing experiments some ways of fruit preparation and processing were tested and their influence on fruit antioxidant status was compared. Shortened storage experiment was applied on blueberries, cranberries, raspberries and strawberries too. In fruits some group parameters – total antioxidant status, dry mass content, ascorbate level, total flavonoids and total phenolics were analyzed spectrophotometrically. Individual flavonoids and phenolics were determined by RP-HPLC/UV-VIS and on-line LC/PDA/ESI-MS. Antioxidant enzyme activities (superoxide dismutase SOD, catalase CAT, polyphenol oxidase PPO and lipoxygenase LOX) were measured by spectrophotometry. The surface microscopy and cultivation of moulds from fruit surface were performed too. Influence of storage conditions on biological activities is dependent on fruit sort. Freezing is the most suitable procedure for long-term storage without significant changes of active substance content. Long-term storage in controlled temperature conditions and/or atmosphere is usable for fruits with longer storage period. In these fruits stabile levels of antioxidant enzymes are stored for relatively long time. Some of enzymes act synergistically. Enzyme activities differed according to storage phase; at the beginning mainly high SOD and LOX activities were observed. CAT and PPO are probably activated as defence systems in rippened and/or damaged fruits. Levels of total as well as individual low molecular weight antioxidants varied during storage in all sorts, generally, increased course with longer storage period can be observed.
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Influence of storage conditions on content of biologically active substances in apple fruits.
Ferdová, Jitka ; Čarnecká, Martina (referee) ; Márová, Ivana (advisor)
This study deals with antioxidants in diet and their effects on human organism. Further, it summarizes the agents affect the quality of apples in the course of long-term storage and it outlines the possibility of defence against them. In the experimental part methods of determination of antioxidant enzymes superoxid dismutase (SOD), catalase and polyphenol oxidace (PPO) in apples were introduced. The enzymes were measured in apples tissues in liquid nitrogen after 158 days in normal or modified atmosphere. In Apple the quantitative and qualitative analysis of proteins was realized. Further, some low molecular antioxidants (total phenolics, total flavonoids and vitamin C) as well as total antioxidant status were measured in frozen raw juice. This values were compared with values from apples analyzed immediately after the harvesting. Artificial inoculation with fungi was made in the last part and the fruitfulness was observed.
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Sublethal effects of pesticide mixtures on crayfish
HOVORKOVÁ, Anna
Prometryn, a triazine herbicide, was banned in Europe in 2004. Metazachlor, a chloracetamide used to treat mainly oilseed crops, is one of the herbicides still in use. Individual pesticides or their metabolites can cause adverse effects in the aquatic environment, and the resulting mixtures of pesticides that can form in the environment are another issue in terms of adverse effects in the environment. The aim of this study was to assess the effect of mixtures of pesticides mixed from selected concentrations (0.01 mg.l-1, 0.1 mg.l-1, 1 mg.l-1). Pesticides were also tested individually at selected concentrations. The biochemical profile of hemolymph and antioxidant biomarkers and oxidative stress biomarker in hepatopancreatic, muscle and gill tissues of marbled crayfish were determined. Toxicity was assessed using a subchronic toxicity test performed over a period of 8 days. Metazachlor concentrations (0.1 and 1 mg.l-1) caused decreasing ALT, Cl-, Mg2+ and Ca2+ levels and increasing NH3 levels in hemolymph. The effect of all tested mixtures was observed on the increase of GLO, ALT and NH3 levels, here a synergistic effect of the mixtures was observed. The activity of antioxidant biomarkers was affected only in CAT for prometryn (0.01 mg.l-1) and metazachlor (0.01 and 1 mg.l-1). LPO levels were affected by metazachlor (0.01 mg.l-1). There was no effect of pesticide mixtures on LPO levels and antioxidant enzyme activities.
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Vliv metazachloru a jeho metabolitu metazachloru OA na raná vývojová stádia raka mramorovaného
ZÁVORKA, Milan
Metazachlor is a selective herbicide used for controlling monocotyledonous and dicotyledonous weeds in oilseed rape and other cruciferous plants. It is mainly used as a pre-emergence herbicide. Metazachlor OA is one of the major degradation products of metazachlor. The aim of this study was to evaluate the effect of metazachlor and its major metabolite metazachlor OA on the early live stages of marbled crayfish (Procambarus virginalis). The effect of metazachlor and metazachlor OA was evaluated on the basis of behaviour, growth, biomarker of oxidative stress, antioxidant biomarkers, ontogenetic development and occurrence of morphological anomalies. The toxicity test was performed at five concentrations, two concentrations of metazachlor (3.2 micrograms/l and 22 micrograms/l) and two concentrations of metazachlor OA (3.2 micrograms/l and 22 micrograms/l) and one control (C). The test exposure was 40 days under defined laboratory conditions. The exposure of metazachlor and metazachlor OA at all tested concentrations caused statistically significantly higher mortality, delayed ontogenetic development and slower growth of crayfish. Metazachlor exposure in concentration 22 micrograms/l and metazachlor OA in concentrations 3.2 micrograms/l and 22 micrograms/l caused statistically significant reduction of antioxidant enzymes (SOD, CAT, GR, GST and GSH) levels in whole-body crayfish homogenate. Metazachlor OA showed a more pronounced adverse effect on the early live stages of marbled crayfish than its parent compound, metazachlor. Based on the results of this study we can point out the suitability of crayfish, especially their early live stages for toxicity tests and the possibilities of their use for biomonitoring of aquatic environment herbicides and their metabolites.
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Effect of fungicides and biological control agens on glutathione metabolism.
Vašková, Marie ; Hýsková, Veronika (advisor) ; Jaklová Dytrtová, Jana (referee)
Triazole fungicides are widely used in agriculture to treat a large number of crops. When they accumulate in soil, plants or water sources, they can also affect non-target organisms, in which they can have a negative effect on the endocrine system or re- production. Much less is known about the effect of triazoles on plants, specifically their antioxidant and detoxification systems. In this work, the effect of penconazole (P), tebuconazole (T) or their combination (PT) on tomato plants Solanum lycopersicum, cv. Cherrola was studied. In contrast to the conventional method of fungicide application by foliar spraying, the effect of soil drench containing P, T or PT was also studied. Soil drenching of fungicides had a worse impact on glutathione metabolism: on the thirty-fifth day after weekly fun- gicide application, the content of reduced thiols and glutathione peroxidase activity were reduced in roots, and the key conjugation enzyme of phase II biotransformation, glutathione-S-transferase (GST), was reduced in leaves by 43 to 20 % (depending on the fungicide) compared with untreated plants. In contrast, the content of reduced thiols and glutathione peroxidase activity were increased in leaves. In the case of spraying with both fungicides (PT), although at the same concentration as P and T alone, a...
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Antioxidant system of tomato plants exposed to triazole fungicides
Žufić, Antoniana ; Hýsková, Veronika (advisor) ; Jaklová Dytrtová, Jana (referee)
Fungal pathogens cause severe crop losses worldwide. Recently, however, chemical fungicides have become dangerous environmental contaminants affecting non-target organisms. The aim of this study was to investigate the effect of triazole fungicides in the form of penconazole (P), tebuconazole (T) or their combination (PT) on tomato plants Solanum lycopersicum L. cv. Cherrola. No significant change in the activity of a key enzyme of the ascorbate-glutathione cycle, ascorbate peroxidase and activity of cytosolic peroxides, was detected in leaves, roots or fruits at the intervals studied, i.e. after two and five weekly treatments of P, T or PT, either as a foliar or a soil application. While slightly increased guaiacol peroxidase activity was found in roots due to T spraying and PT combination, the activity of this enzyme was reduced in leaves similar to catalase. The greatest changes in leaf antioxidant capacity correlated with increased phenolic and flavonoid content were observed after five treatments of penconazole (P) as a soil application both P alone and in combination as PT. The studied triazoles significantly affected the abundance of isoforms of superoxide dismutase and partially total peroxides in leaves, roots and fruits and of acorbate peroxidase in leaves. The highest content of HSP70 was found in...
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