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Synthesis of quantum dots for in-vivo imaging
Ferdusová, Helena ; Majzlíková, Petra (referee) ; Pekárková, Jana (advisor)
The aim of this work was to synthesise water-soluble QDs using different precursors and stabilizers and to determine the toxicity of the synthesized QDs by in vivo imaging. Experiments were performed on water-soluble QDs (MPA-CdTe, MPA-CdTe/ZnS, MSA-CdTe, MSA-CdTe/ZnS, GSH-CdTe, GSH-CdTe/ZnS, TGA-CdTe, TGA-CdTe/ZnS, GSH-ZnSe and GSH-ZnSe/ZnS ) and toxicity was measured. Synthesized QDs were characterized by high intensity (fluorescence spectroscopy), FWHM and zeta potential (ZS Zetasizer) were selected due to their suitability for this task. The toxicity of QDs was determined by the MTT assay on the cell line HEK 293. The experiments show that a core/shell structure is less toxic than a core structure. The results indicate that the toxicity of our synthesized QDs is the lowest for MPA-CdTe (core structure) and MPA-CdTe/ZnS (core/shell structure).
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Synthesis of core/shell quantum dots for diagnostics
Mihajlović, Ana ; Vaculovičová, Markéta (referee) ; Pekárková, Jana (advisor)
This thesis deals with biosensors based on modified semiconductor core/shell quantum dots (QDs) for diagnosis. The work is divided into four main parts. The first one discusses the theory required for the use of QDs in bioaplications, there are described methods of synthesis, modification, application and bioconjugation of QDs. In the experimental part, CdTe/ZnS QDs with core/shell structure were prepared, in which the core was modified by MPA, GSH and TGA. In the next step, these QDs were further modified using CDI, EDC and NHS as mediators in order to increase affinity to BSA (bovine serum albumine) and IgG (imunoglobuline G). Prepared conjugates were characterized by fluorescence spectroscopy (Infinite M200Pro, Tecan) and capillary electrophoresis (Agilent 7100).
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Biomolecules influence on optical properties of deposited quantum dots
Roček, Vojtěch ; Mravec, Filip (referee) ; Drbohlavová, Jana (advisor)
This thesis is focused on creation of quantum dots of titanium dioxide with help of nanoporous alumina mask and on funcionalization of their surface in purpose to increase the quantum yield of its. This thesis is devided into 6 main chapters, where the glossary topic of quantum dots creation process and ways of practical usage of these quantum dots are made. The theoretical knowledge is used for creation of titanium dioxide quantum dots, which was deposited by gold and funcionalized with glutation. The results are shown and discussed in the end of this work.
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Synthesis of quantum dots for proteins detection
Šibíková, Anna ; Fohlerová, Zdenka (referee) ; Pekárková, Jana (advisor)
This thesis is focused on synthesis of quantum dots (QDs) for protein detection. It comprises three parts. The first part summaries the theory of QDs, their synthesis, functionalization, interactions and applications in medicine. In the second part synthesis of CdTe/ZnS core/shell QDs modified by glutathione (GSH) is described, followed by the conjugation with biomolecules BSA and IgG. Several coupling agents such as EDC with NHS and CDI were used. In the last part, the final products were characterized by fluorescence spectroscopy and capillary electrophoresis. The results show the dependence of the fluorescence intensity of the QDs on pH range, concentration of BSA and IgG concentrations using different crosslinkers.
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Evaluation of glutathione content in plants as a marker of heavy metals environmental contamination
Borková, Marie ; Stoupalová, Michaela (referee) ; Opatřilová,, Radka (advisor)
Dependence of glutathione concentration on the amount of thallium in the plant was studied. Observed plant was maize (Zea mays) which was divided to two parts – root and overground. Two culture procedures were elaborated where seeds and young seedlings were cultivated in a solution of thallium of concentration 0, 1, 3, 5, 8, a 10 µmol/l. Extraction agents used during extraction were phosphate buffer and solution of ascorbic acid. Determination of glutathione was realized by capillary electrophoresis (CE) and high performance liquid chromatography (HPLC). Diode array detector (DAD) was used in both methods. Quantification of the thallium amount in the plant was done by method of inductively coupled plasma-atomic emission spectrometry (ICP-AES).
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Determination of glutathion in plant samples using capillary electrophoresis
Skála, Michael ; Křížek, Tomáš (advisor) ; Kubíčková, Anna (referee)
This work is based on a previously published method for determination of both glutathione forms by capillary zone electrophoresis equipped with UV spectrometric detector. Theoretical part described biological and biochemical functions of glutathione as a molecule. It mentioned historically used methods for glutathione determination, their accuracy, precision and reliability and focused on electrophoretic methods, especially capillary zone electrophoresis. Basic principles of capillary zone electrophoresis were described, followed by an overview of approaches to treatment of different types of samples, with special focus on plant samples. At the end, method this work was based on was described. The experimental part describes the development and optimization of this method. Firstly, optimum chemical composition and ionic strength of background electrolyte were determined - two electrolytes were compared, with NaCl and without NaCl - no significant differences were observed, so NaCl was excluded from BGE. Next, optimum concentration of hexadimethrine bromide was determined. The effect of HDB concentration on electroosmotic flow was investigated. Calibration curve for different concentrations of analytes in water was constructed and repeatability, LOD and LOQ of the method were determined....
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Role cytokininů v regulaci odpovědi na nedostatek síry u Arabidopsis thaliana
Štuříková, Helena
Sulfur deficiency stress induces a number of adaptive responses which must be coordinated. Phytohormone cytokinin has been previously shown to regulate sulfur aquisition and utilizati-on, however, the cytokinin-regulation of sulfur nutrition remains elusive. In this diploma the-sis, it was reported that cytokinin induces a sulfur-deficiency-like gene expression which is accompanied by a decrease in pool of the key sulfur-containing compound glutathione. Further, it was found that the sulfur deficiency responses involve altered cytokinin status to coordinate developmental and/or metabolic adaptations. Arabidopsis plants deficient in cytokinin signaling showed disrupted root elongation in response to sulfur starvation and, unexpectedly, the root elongation in response to sulfur starvation was further stimulated by a low cytokinin concentration. Further, the role of cytokinin in sulfur nutrition was indicated in cytokinin signaling-deficient plants which accumulated more glutathione under normal nu-tritive conditions in comparison to wild-type, and vice versa, the decrease of glutathione pool was exaggerated during sulfur starvation. Altogether, results of this diploma thesis suggest that cytokinins participate in nutritional sulfur homeostasis control.
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Vliv taurinu na biochemické ukazatele v orgánech laboratorních potkanů
Mrázková, Martina
The aim of this dissertation was to valorize the effect of various levels of taurine on the antioxidant status in rats body, monitor serum taurine and individual aminoacids in selectedorgans. This dissertation valorize the influence of taurine in the rats liver and brain after its application into feed mixtures. The effect of taurine was monitored on the ratio of GSH / GSSG,concentration of metallothionein and taurine. To this experiment were used 48 Wistar male rats, which were divided into eight groups (n = 6). During four weeks feed mixtures were given to these rats enriched with taurin eat a dose of 0, 1000, 1500, 2000 2500, 3000, 3500 and 4000 mg / kg / diet. Animals accepted diet and drinking water ad libitum. Average consumption of feed mixtures were 35 grams per day. Application taurine in creased concentration of taurine in the organism, which has an positive effect on antioxidant mechanisms of the organism. In deed we can not confirm that the addition of taurine in to the diet increases the concentrations of metallothionein and glutathione in the organism. In the brain was found increasing ratio of GSH / GSSG and reduced concentration of metallothionein. The other way around in the liver was reduction ratio of GSH / GSSG and increasing concentration of metallothionein.
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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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