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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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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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Electrophoretic determination of oxidized and reduced form of glutathion in plant material
Durychová, Eva ; Křížek, Tomáš (advisor) ; Kubíčková, Anna (referee)
This work is based on a previously published method for the determination of twenty proteinogenic amino acids by capillary electrophoresis with contactless conductivity detection and deals with possibility of its use for the determination of oxidized (GSSG) and reduced (GSH) forms of glutathione with amino acids. The theoretical part firstly deals with brief introduction of the properties and importance of glutathione and its protection against autooxidation, especially by alkylation with N-ethylmaleimide (NEM), which binds to GSH to form its alkylated form (GSH-NEM). Then, a brief summary of the available methods for determination of glutathione is given, with a focus on the already developed capillary zone electrophoresis methods. Finally, it briefly presents the electrophoretic method for the determination of twenty proteinogenic amino acids on which this work is based. In the experimental part, the integration of the determination of both forms of glutathione into the method for the determination of proteinogenic amino acids is presented. First, a simplified, less time consuming method was tested in shorter 50.0 cm capillary without hydroxyethylcellulose, where both forms of glutathione were separated at baseline within 10 minutes. Relative standard deviations of peak areas (GSH 2.1%; GSSG...
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Biotransformation aspects on novel carbocyclic nucleoside analogs.
Rozumová, Nela ; Mertlíková Kaiserová, Helena (advisor) ; Rumlová, Michaela (referee)
Carbocyclic nucleoside analogs with norbornane moiety that have been synthesized at IOCB AS CR, represent new potential chemotherapeutic agents with significant activity against Coxsackieviruses. The main objective of this work was to study the metabolism and mechanism of action of the original analog carbocyclic nucleoside MS 254, which is characterized by its antiviral and cytostatic effects. The attention was partially paid also to the two structurally related substances (MS 255, MS 320). In this work, we determined cytotoxicity of these compounds in cell culture and the effect of MS 254 on the amount of total and oxidized glutathione, activity of glutathione-S-transferase (GST), glutathione reductase (GR) and the effect on cellular oxidative stress. The kinetics of the conjugation of MS 254 by human GST was also studied. It was found that of the three substances tested MS 255 was the most cytotoxic and MS 254 was the least cytotoxic compound. It was further found that MS 254 does not cause significant oxidative stress and that it increases the activity of GST and GR in a dose-dependent manner. Michaelis-Menten constant of the conjugation of MS 254 with the glutathione (main metabolic pathway) was determined in the milimolar range, indicating a relatively low affinity of MS 254 for GST.
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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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