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Electrophoretic and immunofluorescence methods for study of plant cell cultures
Klimešová, Marie ; Babula, Petr (referee) ; Provazník, Ivo (advisor)
In all organisms are rising a reactive oxygen and nitrogen species by the effects of various stress factors and these species have a negative impact on the organism. Due to this species plants have built up an efficient antioxidant system, that helps them to resist negative effects of reactive oxygen and nitrogen species. In this work was researched the effect of hydrogen peroxide and sodium benzoate on the production of hydrogen peroxide, superoxide, reactive nitrogen species and malondialdehyde, contained in the root and above-ground part of maize (Zea mays L.). By use of the fluorescence microscopy there were obtained images of cross-cut of root from which was determined the intensity of fluorescence of individual parts of the root and was examined the effect of the intensity of fluorescence markers of oxidative stress in dependence on the type of the fluorescence filter used.
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Association of genetic polymorphism of oxidative stress with diabetes mellitus type 1 and 2
Kloboučková, Lucie ; Kotrbová - Kozak, Anna Katarzyna (advisor) ; Daňková, Pavlína (referee)
Diabetes mellitus is a chronic autoimmune disease in which the immune system attacks the insulin-secreting ß-cells in the pancreas. It leads to an absolute deficiency of insulin. Chronic hyperglycemia induces increased production of reactive oxygen species, which leads to a decrease of natural antioxidant level in blood, and it contributes to genesis of diabetes complications (e.g. vascular or pulmonic). Moreover, the oxidative stress results in onset of pancreas inflammations and the damage of its ß-cells. Aims: Our aim was to assess whether or not certain genotypes or their combinations occur with higher frequency among groups of patients of type 1 diabetes (T1D) and type 2 diabetes and in a control group of healthy individuals. Methods: The study included groups of 40 T1D patients, 40 T2D patients and 45 healthy individuals. The polymorphisms of genes involved in the oxidative stress response were analyzedby using RFLP, PCR with TaqMan probes and allele specific PCR. The target genes involved superoxide dismutase SOD1 and SOD3 genes; glutathione-S-transferase GSTM1, GSTT1, GSTP1 genes; glutathioneperoxidase gene GPX1 and catalase gene CAT. The levels of plasma malondialdehyde were measured by using liquid chromatography. Results: Statistically significant differences were found in the...
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Association of genetic polymorphism of oxidative stress with diabetes mellitus type 1 and 2
Kloboučková, Lucie ; Kotrbová - Kozak, Anna Katarzyna (advisor) ; Daňková, Pavlína (referee)
Diabetes mellitus is a chronic autoimmune disease in which the immune system attacks the insulin-secreting ß-cells in the pancreas. It leads to an absolute deficiency of insulin. Chronic hyperglycemia induces increased production of reactive oxygen species, which leads to a decrease of natural antioxidant level in blood, and it contributes to genesis of diabetes complications (e.g. vascular or pulmonic). Moreover, the oxidative stress results in onset of pancreas inflammations and the damage of its ß-cells. Aims: Our aim was to assess whether or not certain genotypes or their combinations occur with higher frequency among groups of patients of type 1 diabetes (T1D) and type 2 diabetes and in a control group of healthy individuals. Methods: The study included groups of 40 T1D patients, 40 T2D patients and 45 healthy individuals. The polymorphisms of genes involved in the oxidative stress response were analyzedby using RFLP, PCR with TaqMan probes and allele specific PCR. The target genes involved superoxide dismutase SOD1 and SOD3 genes; glutathione-S-transferase GSTM1, GSTT1, GSTP1 genes; glutathioneperoxidase gene GPX1 and catalase gene CAT. The levels of plasma malondialdehyde were measured by using liquid chromatography. Results: Statistically significant differences were found in the...
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Optimization and application of assays for determination ecotoxicity of nanomaterials
Semerád, Jaroslav ; Cajthaml, Tomáš (advisor) ; Kočí, Vladimír (referee)
This thesis deals with optimization and application of assays for determination of ecotoxicity of nanomaterials based on nanoscale zero-valent iron (nZVI), which are used in remedial technologies. After in situ application of nZVI, a significant decrease in toxicity of polluted environment was detected; however, a potential negative effect of nanoparticles has not been sufficiently investigated yet. Standard used tests were found to be incompatible with nZVI for toxicity determination. Specific characteristics of nZVI, such as high reactivity and sorption, complicate determining the toxicity by routinely used ecotoxicity tests. Concentration ranging from 0,1 to 10 g/l that are used in practise for decontamination were tested. These concentrations resulted in formation of turbidity, which prevented the use of standard tests. In this work, a new method has been optimized for in vitro toxicity testing of nZVI and derived nanomaterials using bacteria. The principle of this assay is determination of oxidative stress (OS). The disbalance between formation and degradation of reactive oxygen species (i.e. OS) leads to irreversible changes in biomolecules of organisms and formation of undesirable products. A toxic and mutagenic product - malondialdehyde (MDA) is formed during lipid peroxidation and it is a...
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Electrophoretic and immunofluorescence methods for study of plant cell cultures
Klimešová, Marie ; Babula, Petr (referee) ; Provazník, Ivo (advisor)
In all organisms are rising a reactive oxygen and nitrogen species by the effects of various stress factors and these species have a negative impact on the organism. Due to this species plants have built up an efficient antioxidant system, that helps them to resist negative effects of reactive oxygen and nitrogen species. In this work was researched the effect of hydrogen peroxide and sodium benzoate on the production of hydrogen peroxide, superoxide, reactive nitrogen species and malondialdehyde, contained in the root and above-ground part of maize (Zea mays L.). By use of the fluorescence microscopy there were obtained images of cross-cut of root from which was determined the intensity of fluorescence of individual parts of the root and was examined the effect of the intensity of fluorescence markers of oxidative stress in dependence on the type of the fluorescence filter used.
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Adipokinetic hormone counteracts oxidative stress elicited in insects by hydrogen peroxide: in vivo and in vitro study
BEDNÁŘOVÁ, Andrea
The involvement of members of Adipokinetic hormone (AKH) family in regulation of response to oxidative stress (OS) was investigated in the present study. OS was elicited in the insect body by exogenous application of hydrogen peroxide. In vivo experiments reveal that injection of hydrogen peroxide into the haemocoel of the firebug, Pyrrhocoris apterus, increases the level of AKH in the central nervous system (CNS) and in the haemolymph. The injection of hydrogen peroxide also increases the mortality of experimental bugs, whereas co-injection of hydrogen peroxide with Pyrap-AKH reduces mortality to control levels. Importantly, an increase in haemolymph protein carbonyl (an OS biomarker) levels is decreased to control levels when hydrogen peroxide is co-injected with Pyrap-AKH. Similar results were obtained using in vitro experiments. OS biomarkers such as malondialdehyde and protein carbonyls were significantly enhanced upon exposure of isolated CNS to hydrogen peroxide in vitro, whereas co-treatment of the CNS with hydrogen peroxide and Pyrap-AKH reduces their level significantly. Catalase activity was measured as well and the same pattern was observed. The results of the present study provide strong support on the feedback regulation between oxidative stressors and AKH action, and implicate AKH in counteracting OS.
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