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The influence of humic acids methylation on the interaction of surface active agents
Bělušová, Anna ; Weidlich, Tomáš (referee) ; Smilek, Jiří (advisor)
The presented diploma thesis deals with the study of interactions between humic acids as the main component of soil organic matter with a model organic pollutant presented by cationic surfactant Septonex (which was expected to interact positively with humic acids due to its charge) through unconventional combination of dialysis and diffusion techniques realized in diffusion cells together with isothermal titration calorimetry capable of elucidating the nature of ongoing reactions from a thermodynamic point of view. The experiments were performed with a humic acid isolated by standard isolation procedure from Leonardite (1S104H) purchased from the International Society for Humic Substances, with humic acids isolated from lignite as well as modal chernozem. The aim of the submited diploma thesis is to assess the influence of carboxyl functional groups in the structure of humic acids to interactions with Septonex. For this reason, said samples of humic acids have been modified by selective methylation of their carboxyl functional groups, which can no longer participate in interactions with ionic substances. Results from diffusion cells and isothermal titration calorimetry confirmed the assumption that Septonex interacted with all types of unmodified humic acids more than with their methylated derivatives. Despite the available literature, it was surprising that the differences in the interaction of humic acids with Septonex were not diametrically different when comparing unmodified humic acids with their methylated derivatives. The extent of these interactions was assessed through the apparent equilibrium constant Kapp determined in the diffusion (dialysis) experiments and from the course of enthalpy curves from isothermal titration calorimetry. After the experiments and their evaluation, it can be concluded that humic acids and Septonex in their interactions, due to their structures, probably apply not only electrostatic interactions, especially through available carboxyl functional groups of humic acids and positively charged hydrophilic parts of Septonex molecules, but also a significant role have a hydrophobic effect, hydrogen bonds and Van der Waals forces.
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Preparation and characterization of hydrogels based on cationic biopolymer
Tesařová, Barbora ; Mravec, Filip (referee) ; Smilek, Jiří (advisor)
Presented thesis deals with the preparation and characterization of hydrogels based on positively-charged biopolymer and negatively-charged tenside. As an appropriate representative of positively-charged polymer was chosen chitosan, because of his biocompatibility and biodegrability, and N,N,N-trimethylchitosan, which was synthetised from chitosan in this diploma thesis. The final product of synthesis was characterised by SEC-MALS, elemental analysis, infrared spectroscopy and nuclear magnetic rezonation. Interactions between these polymers and representatives of negatively-charged tensides, which are sodium dodecylsuphate and sodium tetradecylsulphate, were investigated. The main method used in this thesis was rheology, through it were investigated prepared hydrogels based on chitosan and tensides and also prepared microgels based on N,N,N-trimethylchitosan and tensides. The interactions between polymers and tensides have been proved.
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Characterization of methylated humic acids
Solná, Irena ; Kislinger, Jiří (referee) ; Klučáková, Martina (advisor)
The main concern of this bachelor thesis was a characterization of the properties of methylated humic acids. Retrieval on modifications of acidic functional groups of humic substances was done. In addition appropriate experimental methods for assessing the impact of methylation of carboxylic groups of humic acids were selected. Methylation of formentioned carboxylic groups is used to improve stability of humic acids while working with them. The sample of methylated humic acids, produced by the University of Poitiers in France, was investigated by the following methods: UV-Vis spectroscopy, FT-IR spectroscopy, fluorescence spectroscopy and thermal analysis - TGA, DSC. The degree of methylation of carboxylic functional groups of methylated humic acids was considered using an indirect titration. Results were trade-off with the sample without methylation. The impact of gelation on properties of methylated and non-methylated humic acids was also studied.
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Utilization of Diffusive Techniques in Study on Reactivity of Biocolloids
Kalina, Michal ; Maršálek, Roman (referee) ; Mráček, Aleš (referee) ; Klučáková, Martina (advisor)
The main aim of this thesis is the utilization of simple diffusion techniques for the study on transport properties of copper ions in the systems containing humic acids with respect to the other parameters, which can affect the process (the structure of diffusion environment, the interactions between transported specie and diffusion matrices, selective blocking of binding sites of humic acids). The first part of experimental works was focused on characterization of studied materials (humic acids, humic sol and humic hydrogel). The main part of the thesis was dealing with the optimization of simple diffusion techniques, which were suitable for the study on transport of copper ions in matrices containing humic acids, taking into account the mutual interactions between studied components in the system. The obtained diffusion characteristics were compared to the data determined using sorption experiments. Consequently, the minor goal of the experimental works of this thesis was also the assessment of the influence of basic physico-chemical parameters of studied materials on transport phenomenon.
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Transport and sorption properties of modified humic acids studied in hydrogels matrixes
Polášková, Nikola ; Pekař, Miloslav (referee) ; Smilek, Jiří (advisor)
This diploma thesis is focused on transport and sorption properties of humic acids studied in hydrogel matrixes, which are similar to their natural environment (soil). These properties are studied by diffusion techniques. The humic acid standard isolated from Leonardite (from International Humic Substances Society, identification number 1S104H-5) was chosen as a representative of humic acids. Their transport and sorption properties were studied in agarose-based biopolymer hydrogel matrixes (agarose is a nonreactive, thermoreversible polysaccharide) with use of model pollutant Rhodamine 6G (positively charged organic dye), that simulates common pollutants. Humic acids were modified to block their specific functional groups in their structure. Then we are able to study the influence of each functional groups on the interactions with other compounds individually, that was the main goal and motivation of this work. Carboxyl functional groups were modified by methylation with a methylating agent trimethysilyldiazomethane (TMS-DM). For blocking carboxyl and phenolic functional groups together we repeated methylation of carboxyl groups with TMS-DM and then we acetylated phenolic functional groups with acetic anhydride. The successful modification was confirmed by thermogravimetry (TGA), Fourier transform infrared spectroscopy (FT-IR) and acid-base titrations (direct, indirect and carboxyl titration). The techniques of diffusion cell and non-stationary diffusion in cuvettes were chosen to study how organic dye interacts with unmodified and modified humic acids in inert hydrogel matrix. Humic acids were characterised by their structure-based reactivity, respectively by the acidic functional groups content. This thesis verifies the use of the diffusion techniques as a low-cost and universal techniques suitable for studying properties of humic acids mentioned above. Due to the finding that acidic functional groups have a minor influence on these properties, the possibilities of taking other factors into account (such as hydrophobic interactions, hydrogen bonds, van der Waals forces or specific surface area) are opened up.
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Biological age prediction using DNA analysis
Stokičová, Linda ; Vaněk, Daniel (advisor) ; Tomášek, Petr (referee)
Eye colour, hair colour and biogeographical origin can be predicted based on DNA analysis. This is used in forensic genetics when an unknown perpetrator leaves a biological material on a crime scene. Prediction of biological age is the subject of further research. Nowadays, there is no standardised model, which can be used for accurate prediction. The best option for biological age prediction is DNA methylation - stable epigenetic modification. Many markers have been evaluated. Methylation of these markers correlates with biological age positively or negatively. Methylation has tissue specificity, so one model cannot be applied on all types of tissues. Model for biological age prediction can be applied during investiganting of a criminal activity, when the identity of a person commiting a crime is unknown. Key words: methylation, chronological age, biological age, DNA analysis, forensic genetics
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The relationship between splicing and posttranslational modifications of chromatin in Saccharomyces cerevisiae
Kovaľová, Libuša ; Folk, Petr (advisor) ; Čáp, Michal (referee)
Protein Prp45, the yeast ortholog of the human transcription coregulator SNW1/SKIP, has been previously associated only with the regulation of pre-mRNA splicing. However, our laboratory found that protein Prp45 genetically interacts not only with the proteins involved in pre-mRNA splicing, but also with factors important for transcription elongation and with chromatin modifying enzymes. Our data and the information about the human ortholog SNW1/SKIP suggest that Prp45 could serve as a regulator coupling splicing, transcription and chromatin state in S. cerevisiae. The main aim of this diploma thesis was to find out whether the protein Prp45, which is essential for cotranscriptional assembly of the spliceosome, affects posttranslational modifications of chromatin on transcribed genes. Using chromatin immunoprecipitation, the influence of prp45(1-169) mutation on trimethylation of histone H3 at lysine 4 and acetylation of histone H3 at lysines 9, 14 and 18 on transcriptionally active genes was not confirmed. The other aim was to analyse the behavior of cells synchronized by α-factor by using flow cytometry. According to our results, prp45(1-169) mutation leads to the prolongation of the cell cycle. For the purpose of monitoring the dynamics of nucleosomes in S. cerevisiae strains, the system of...
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Impact of the rRNA modifications on protein synthesis
Kročová, Eliška ; Pospíšek, Martin (advisor) ; Holá, Dana (referee)
A ribosome is a supramolecular structure, which mediates synthesis of all cellular proteins, and therefore is essential for cell life. The fact, that some nucleotides of ribosomal RNA are modified, is known for forty years. However only recently, successful deeper studies on how the individual modifications are synthesized and what is their effect on ribosome synthesis and function appear. Some particular nucleotide modifications are important for the ribosome formation (like m1 acp3 Ψ1191 SSU), some others influence proper function of the ribosome (e.g. Um2921, Gm2922, Ψ2923 LSU, m1 acp3 Ψ1191 SSU). Majority of modified nucleotides in eukaryotic rRNA is being recognized by small nucleolar RNA (snoRNA). Few nucleotides is, however, recognized and subsequently modified by specific proteins. These proteins also play crucial role in ribosome maturation. In thesis presented, current knowledge on the role of ribosomal RNA nucleotide modifications during their formation and maturation, and on their function is summarized and overviewed.
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The influence of humic acids methylation on the interaction of surface active agents
Bělušová, Anna ; Weidlich, Tomáš (referee) ; Smilek, Jiří (advisor)
The presented diploma thesis deals with the study of interactions between humic acids as the main component of soil organic matter with a model organic pollutant presented by cationic surfactant Septonex (which was expected to interact positively with humic acids due to its charge) through unconventional combination of dialysis and diffusion techniques realized in diffusion cells together with isothermal titration calorimetry capable of elucidating the nature of ongoing reactions from a thermodynamic point of view. The experiments were performed with a humic acid isolated by standard isolation procedure from Leonardite (1S104H) purchased from the International Society for Humic Substances, with humic acids isolated from lignite as well as modal chernozem. The aim of the submited diploma thesis is to assess the influence of carboxyl functional groups in the structure of humic acids to interactions with Septonex. For this reason, said samples of humic acids have been modified by selective methylation of their carboxyl functional groups, which can no longer participate in interactions with ionic substances. Results from diffusion cells and isothermal titration calorimetry confirmed the assumption that Septonex interacted with all types of unmodified humic acids more than with their methylated derivatives. Despite the available literature, it was surprising that the differences in the interaction of humic acids with Septonex were not diametrically different when comparing unmodified humic acids with their methylated derivatives. The extent of these interactions was assessed through the apparent equilibrium constant Kapp determined in the diffusion (dialysis) experiments and from the course of enthalpy curves from isothermal titration calorimetry. After the experiments and their evaluation, it can be concluded that humic acids and Septonex in their interactions, due to their structures, probably apply not only electrostatic interactions, especially through available carboxyl functional groups of humic acids and positively charged hydrophilic parts of Septonex molecules, but also a significant role have a hydrophobic effect, hydrogen bonds and Van der Waals forces.
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Microfluidic reactors for chemical synthesis
Jaklová, Natalie ; Klusoň, Petr (advisor) ; Cvak, Ladislav (referee)
The main topic of the Diploma Thesis is the conversion of the batch N-alkylation reaction into a continuous regime. It is the reaction of 10α-methoxydihydrolysergic acid methyl ester to 1-methyl-10α-methoxydihydrolysergic acid methyl ester. It represents one step in the industrial synthesis of Nicergoline, a pharmaceutical used for senile dementia treatment. This methylation reaction is performed in two immiscible liquid phases. Phase-transfer catalysis is used to increase the efficiency of the reaction, which allows the reaction to proceed in the entire volume of the organic solvent, not only at the phase interface. The reaction is rapid, with an exothermic character. The advantages of continuous arrangement are better control of the reaction, increased operational safety due to handling smaller volumes of reaction components, and more intensive removal of heat of the reaction. The Thesis deals with the influence of reaction conditions on the course of the reaction in a microfluidic reactor. The glass microchip reactor was chosen to perform the reaction in a flow arrangement. This reactor could produce up to grams of the product per hour. Several series of experiments were performed on this apparatus. The conversion of the starting material, and selectivities to the product and the side products...
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