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Changes in plant metabolism due to phytoremediation of pharmaceuticals
Polívková, Linda ; Bělonožníková, Kateřina (advisor) ; Chmelík, Josef (referee)
As a result of human activity, the rate and amount at which various pollutants enter the environment are increasing. Among them, pharmaceuticals and their metabolites, whose fate in nature has not yet been fully understood, have a key role to play. Phytoremediation, as an alternative method to the traditionally established methods of remediation of pollutant- contaminated wastewater and soils, has an increasing potential to become an important and ecologically friendly tool for environmental restoration and conservation. In this bachelor thesis, the effect of the pharmaceuticals buspirone, dextromethorphan, ibuprofen, the combination of dextromethorphan and ibuprofen, paroxetine and sulpiride on the antioxidant system of maize (Zea mays L., DKC 3969) cultivated in vitro in hydroponics under sterile conditions was investigated. The activity and isoenzyme composition of peroxidases, which are among the key components of the antioxidant system, were determined in maize plants. The amount of total soluble proteins in plants was reduced in most cases due to the influence of the aforementioned pharmaceuticals. In contrast, the specific activity of total peroxidases, ascorbate peroxidase and guaiacol peroxidase was increased in most cases, most notably by buspirone, sulpiride and the combination of...
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Phytoremediation of pharmaceuticals
Forróvá, Barbora ; Bělonožníková, Kateřina (advisor) ; Kubíčková, Božena (referee)
Anxiolytics, sedatives, hypnotics and antidepressants are some of the most prescribed pharmaceuticals today. As their consumption increases, there is a growing risk of them leaking into surface water and soil, where they can be accumulated, interfere with key biological processes in animal cells or contaminate the food chain. With this is related an increasing need to find ways to ensure the removal of these substances from contaminated sites. There is a wide range of methods for decontamination, however not all of them are affordable and sufficiently environmentally friendly. Phytoremediation is one of the most environmentally and economically friendly ways used for removing pharmaceuticals and other xenobiotics from the environment. In this work, the effects of the drugs diazepam, rohypnol and trazodone on the antioxidant system of maize (Zea mays, L., cultivar DKC 3969) plants grown under sterile conditions hydroponically in vitro were studied. The effects these drugs had on the plants were monitored through the activity of important antioxidant enzymes, peroxidases and glutathione-S-transferases, as well as through non-enzymatic markers of antioxidant capacity and the content of substances indicative of oxidative stress, malondialdehyde and H2O2. A significant increase in phenolic content was...
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Steatosis of liver and mitochondrial dysfunction
Páleníčková, Eliška ; Novák, František (advisor) ; Balková, Patricie (referee)
Aim: To determine the effect of diet-induced steatosis in the development of mitochondrial dysfunction in the liver and hepatic sensitivity to the partial ischemia. Methods: Male Wistar rats (361 ± 8.8 g) were fed standard (SD) or high-fat diet (HFD). Partial ischemia was induced by short-term clamp (20 min) of vein porta two days before the end of the experiment. Results: Ten-week HFD administration lead due to increased ketogenesis the altered glucose tolerance elevated serum NEFA. We demonstrated the inhibitory effect of HFD on the respiratory capacity of mitochondria in vitro. HFD negatively affected the activity of antioxidant systems and stimulated the formation of lipoperoxides. Partial ischemia had no efect on the mitochondrial oxidative capacity but significantly elevated the oxidative stress. Conclusion: HFD administration lead to the development of fatty liver that was still not accompanied by biochemical markers of liver injury. Nevertheless, we proved the impairment of to mitochondrial respiratory capacity, signs of structural damage of mitochondria and the increased sensitivity to oxidative damage of the liver. Subject words: biochemistry, physiology Keywords: mitochondria, HFD, ischemia, respiratory chain, antioxidant systém, ROS
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Use of biochemical methods in the study of plant metabolism with emphasis on the defense system
Bělonožníková, Kateřina ; Ryšlavá, Helena (advisor) ; Luhová, Lenka (referee) ; Plchová, Helena (referee)
As sessile organisms, plants cannot avoid being exposed to various biotic and abiotic stress factors. To defend themselves plants have developed complex and sophisticated defense mechanisms. In agriculture, pesticides are used for plant protection, but they can have undesirable side effects, and therefore attention is being paid to biological control agents. One such agent is Pythium oligandrum, a soil oomycete that has a positive effect on plants at several levels. On the one hand, it secretes elicitins, which stimulate defensive responses and prepare the plant for possible infection (i.e., priming effect), and on the other hand, it produces tryptamine, a precursor of auxin, thus supporting the plant's growth. Furthermore, P. oligandrum directly acts as pathogen mycoparasite or evokes antibiosis, or competition for nutrients and space with other microorganisms. The key research topic of this thesis was the analysis of the biochemical properties of eleven Pythium strains and their impact on plant metabolism. Our results showed that even closely related P. oligandrum strains significantly differ in the content of compounds secreted into the medium, including proteins, amino acids, tryptamine, and hydrolytic enzymes capable of degrading cell walls (endo-β-1,3-glucanase, chitinase, and cellulase),...
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Use of biochemical methods in the study of plant metabolism with emphasis on the defense system
Bělonožníková, Kateřina
As sessile organisms, plants cannot avoid being exposed to various biotic and abiotic stress factors. To defend themselves plants have developed complex and sophisticated defense mechanisms. In agriculture, pesticides are used for plant protection, but they can have undesirable side effects, and therefore attention is being paid to biological control agents. One such agent is Pythium oligandrum, a soil oomycete that has a positive effect on plants at several levels. On the one hand, it secretes elicitins, which stimulate defensive responses and prepare the plant for possible infection (i.e., priming effect), and on the other hand, it produces tryptamine, a precursor of auxin, thus supporting the plant's growth. Furthermore, P. oligandrum directly acts as pathogen mycoparasite or evokes antibiosis, or competition for nutrients and space with other microorganisms. The key research topic of this thesis was the analysis of the biochemical properties of eleven Pythium strains and their impact on plant metabolism. Our results showed that even closely related P. oligandrum strains significantly differ in the content of compounds secreted into the medium, including proteins, amino acids, tryptamine, and hydrolytic enzymes capable of degrading cell walls (endo-β-1,3-glucanase, chitinase, and cellulase),...
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Glutathione and glutathione-dependent enzymes in various pathophysiological states.
Kodydková, Jana
Backround: Oxidative stress (OS) has been implicated in pathogenesis of human disorders such as depressive disorder, sepsis, cardiovascular disease, acute and chronic pancreatitis, and cancer. Increased OS is result of imbalance between increased reactive oxygen and nitrogen species (RONS) production and / or insufficient activity of antioxidant defence system. Antioxidant system, which is composed of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidases (GPX), glutathione reductase (GR) and non- enzymatic antioxidant reduced glutathione (GSH) plays an important role in the protection of cells against enhanced OS. The aim of this study was to assess the OS markers and antioxidant enzymes in different pathophysiological states. Materials and methods: Activities of erythrocyte glutathione peroxidase (GPX1), GR and concentration of GSH as well as levels of OS markers were analysed in six different pathophysiologic states. These parameters were measured in 35 women with depressive disorder (DD), 40 patients with metabolic syndrome (MetS), 30 septic patients (S) followed up in the course of sepsis; 15 non-septic critically ill patients (NC), 13 patients with acute pancreatitis (AP), 50 with chronic pancreatitis (CP) and 50 patients with pancreatic cancer (PC), compared to...
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The role of antioxidant system in cardioprotective models
Galatík, František ; Žurmanová, Jitka (advisor) ; Benák, Daniel (referee)
Ischemia/reperfusion injury is the main cause of death in patients suffering from cardiovascular disease. The efforts of tackling this problem have lead to the discovery of cardioprotective adaptations and subsequently - of several cardioprotective models. One of the mechanisms of the adaptations is the stimulation of antioxidant system as a protection against reactive oxygen species formed during reperfusion phase of ischemia/reperfusion. The aim of this thesis is to review the current knowledge regarding the role of antioxidant system in cardioprotective models and comparing them by this property. Key-words: cardioprotective models, antioxidant system, heart
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Vliv triazinových pesticidů a jejich metabolitů na necílové vodní organizmy
KOUTNIK, Dalibor
Pesticides used in agro-ecosystems and forests enter aquatic environments such as streams, rivers, and lakes if applied in adjacent areas or if an accidental spill occurs. For the studies, we chose the early life stages of common carp (Cyprinus carpio L.) and marbled crayfish (Procambarus fallax f. virginalis). In selected organisms, we observed long-term effect of triazine herbicides (prometryne, terbuthylazine, metribuzine) and their metabolites (terbuthylazine-desethyl and tebuthylazine-2-hydroxy) in concentrations commonly occurring in Czech rivers. The first part is about monitoring the effects of triazines (prometryne, terbuthylazine) on the early development stages of carp and marble crayfish. Subchronic prometryne exposure of early-life stages of common carp at concentrations of 1 200 and 4 000 ?g/L affected their survival, growth rate, early ontogeny, and histology. Terbuthylazine in concentration 1 400 and 3 000 ?g/L caused significant decrease of mass, total length, delayed in development and cause of alternation of tubular system of caudal kidney of carp. Prometryne in concentration 144, 1 444 and 4 320 ?g/L caused decrease of weight, specific growth rates and caused histopathological changes in gill of crayfish. Moreover concentration 4 320 ?g/L of prometryne caused delay in ontogenetic development of crayfish. Second part of the work has included effects of low concentrations of metabolites (terbuthylazine-desethyl and tebuthylazine-2-hydroxy) on the early development stages of carp and marble crayfish. Chronic terbuthylazine-desethyl exposure in concentrations 180, 900, and 1 800 ?g/L affected growth, ontogenetic development, and the antioxidant system and caused pathological changes in the caudal kidney of early life stages of carp. Chronic terbuthylazine-2-hydroxy exposure in concentrations 2.9, 70, 1 400 and 3 500 ?g/L caused decreased of total superoxide dismutase activity of early life stages of carp. Moreover concentration 1 400 and 3 500 ?g/L of terbuthylazine-2-hydroxy caused delay in ontogenetic development and pathological changes in the caudal kidney of carp. Chronic terbuthylazine-2-hydroxy exposure in concentrations up 75 ?g/l affected growth, ontogenetic development, antioxidant system, caused oxidative stress and pathological changes in hepatopancreas of early life stages of marbled crayfish. The last part of our study examined the effect of metribuzine on signal crayfish (Pacifastacus leniusculus Dana). Crayfish were exposed to metribuzine concentrations of 0.52 ?g/L and 3.06 mg/L for 30 days and a 30-day depuration period. In the thiobarbituric acid reactive substances, superoxide dismutase, and catalase were observed differences in all examined tissues (gill, muscle, hepatopancreas) compared to the control group. Differences from control were observed in glutathione reductase activity in hepatopancreas after 10 days for both exposure concentrations and after 30 days at 3.06 mg/L. Histological examination revealed extensive focal autolytic disintegration of tubular epithelium in hepatopancreas of crayfish exposed to metribuzin. These studies provided important results for the evaluation of long-term impact and effect of (prometryne, terbuthylazine and metribuzine) and their metabolites (terbuthylazine-desethyl and tebuthylazine-2-hydroxy) on the water non-target organisms, even in real concentrations founded in surface waters.
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Glutathion a glutathion dependentní enzymy za různých patofyziologických stavů.
Kodydková, Jana ; Tvrzická, Eva (advisor) ; Kazdová, Ludmila (referee) ; Vejražka, Martin (referee)
Backround: Oxidative stress (OS) has been implicated in pathogenesis of human disorders such as depressive disorder, sepsis, cardiovascular disease, acute and chronic pancreatitis, and cancer. Increased OS is result of imbalance between increased reactive oxygen and nitrogen species (RONS) production and / or insufficient activity of antioxidant defence system. Antioxidant system, which is composed of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidases (GPX), glutathione reductase (GR) and non- enzymatic antioxidant reduced glutathione (GSH) plays an important role in the protection of cells against enhanced OS. The aim of this study was to assess the OS markers and antioxidant enzymes in different pathophysiological states. Materials and methods: Activities of erythrocyte glutathione peroxidase (GPX1), GR and concentration of GSH as well as levels of OS markers were analysed in six different pathophysiologic states. These parameters were measured in 35 women with depressive disorder (DD), 40 patients with metabolic syndrome (MetS), 30 septic patients (S) followed up in the course of sepsis; 15 non-septic critically ill patients (NC), 13 patients with acute pancreatitis (AP), 50 with chronic pancreatitis (CP) and 50 patients with pancreatic cancer (PC), compared to...
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