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Study on protective effect of 3-hydroxybutyrate and its derivatives
Černá, Klára ; Smilek, Jiří (referee) ; Obruča, Stanislav (advisor)
The aim of this thesis is the study of chemical chaperones, as a preservative enzymes before denaturing processes. In the experimental part of the study we investigated the protective effects of five selected potential protective compounds – derivatives of 3-hydroxybutyrate – two model enzymes: lipase and lysozyme. Protective effects of potential chemical chaperones were investigated by dynamic light scattering (DLS), which was determined by the size distribution of the aggregates lipase produced depending on the temperature at the heat-induced denaturation. Further we have been used differential scanning calorimetry (DSC), which was determined by the temperature of the denaturation of lysozyme in the presence or absence of potentially protective substances. The last parameter was to determine the effect of potentially protective agents at different concentrations on the catalytic activity of lipase and determining the residual lipase activity after application of denaturing two factors – the high temperature and freezing. Of all tested structural analogues of 3-hydroxybutyrate was the highest protective effect observed (stabilization of enzyme molecules) with the succinate, which has two carboxyl groups. Conversely, 1,3-butanediol showed virtually no protective activity, indicating that the presence of carboxyl groups on the protective effect of essential. The low protective efficacy butyrate further indicates that it is essential that an effective protectant in addition to the carboxyl group contained as a further functional group – either carboxyl or hydroxyl. More significant protective effect was observed in 3-hydroxybutyrate than in 2-hydroxybutyrate. Interestingly, the effective protectant while at higher concentrations strongly inhibit the enzyme activity of the model enzyme, which is probably related to the solvation and conformations of the protein in the presence of protectant and the availability of the active site for the substrate.
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Study of interactions of hyaluronan-amino acids
Jugl, Adam ; Smilek, Jiří (referee) ; Hurčíková, Andrea (advisor)
The master´s thesis deals with the study of the interaction between the polysaccharide hyaluronan of diffrerent molecular weights with the amino acids arginine, lysine, arginine hydrochloride and 6-aminocaproic acid. They are expected interaction between carboxyl groups of hyaluronan and amino groups of amino acids. These interactions were investigated by using ultrasonic spectroscopy, DLS, measuring pH and conductivity. Obtained results were compared with sodium polystyrene sulfonate. With ultrasonic spectroscopy was observed a change of concentration inkrement for titration of amino acid to water or polymers solutions especially for high molecular weight hyaluronan and for NaPSS in combination with 6AKK in concentration range of added amino acid 0–30 mM. The size of this change could mean a degree of interaction between polymers and amino acids. This theory has not been confirmed by other methods. By pH and conductivity measurements interations between arginine and low molecular weight hyaluronan and NaPSS were only confirmed. There was no possibility to make unequivocal conclusions from determination of particle size and zeta potential by DLS. Overall, the issue of the interaction of amino acids with polyanions was proved above expectations complex and will be appropriate to further expand the observations made in this thesis.
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Characterization of polyelectrolyte complexes using structural and thermal analysis
Řiháčková, Barbora ; Smilek, Jiří (referee) ; Krouská, Jitka (advisor)
This master thesis deals with study of chitosan-lignohumate, chitosan-polystyrenesulfonate, chitosan-alginate and chitosan-carrageenan polyelectrolyte complexes. The work was motivated by research of finding suitable alternative substance for lignohumate. The molecular weights of substances were characterized using SEC-MALLS. A degree and a character of the interactions between polyelectrolyte were studied by isothermal titration calorimetry and dynamic light scattering method. The calorimetric experiments proved that decreasing concentration of samples causes decreasing of heat flow. The best calorimetric measurements were provided by adding chitosan into polymer solution. The interactions between chitosan and polyanions and influence of mixing order were proved also by measuring intensity of zeta potential, Z-average of particle size and turbidity. New chitosan-based materials have a big potential in agriculture and medicine.
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New Methods of Study of Reactivity and Transport Properties of Biocolloids
Smilek, Jiří ; Weidlich,, Tomáš (referee) ; Pabst, Willi (referee) ; Klučáková, Martina (advisor)
The main aim of doctoral thesis was the study on reactivity, transport and barrier properties of biocolloidal and synthetic polymeric substances by simple diffusion techniques. It was studied mainly the influence of basic physic-chemical parameters (temperature, concentration, pH and modification of material) on the reactivity and barrier ability of chosen compounds. Further substances were chosen as a model compounds: biocolloids (humic acids, alginate, chitosan, hyaluronate) and synthetic polymer (polystyrenesulfonate). Reactivity, barrier and transport properties of chosen substances were studied by interactions with oppositely charged basic organic dyes (methylene blue, rhodamine 6G, amido black 10B respectively) in hydrogels medium based on linear polysaccharide (agarose). The attention was also paid to basic physic-chemical characterisation (infrared spectroscopy, rheology, elemental analysis, thermogravimetry and scanning electron microscopy) of chosen materials and also hydrogels. Key part of the whole doctoral thesis was the optimization of selected diffusion techniques (diffusion cell technique and non-stationary diffusion in cuvettes) designated for the study on reactivity and barrier properties of wide range compounds (optimized method should be used as an universal method for simple and fast determination of reactivity of different compounds at given or changing conditions). The rate of reactivity, transport and barrier properties was determined based on fundamental diffusion parameters such as diffusion coefficients, break-through time so called lag time, interfacial concentration of chosen organic dye, apparent equilibrium constant, tortuosity factor, partition coefficient.
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Reactivity-mapping studies on biopolymers in hydrogel forms
Smilek, Jiří ; Kislinger, Jiří (referee) ; Sedláček, Petr (advisor)
The diploma thesis is focused on the study of interactions between biopolymers and model probes in a hydrogel media utilizing diffusion processes. The main aim was to test and to optimize selected combination of diffusion methods (diffusion cells and unsteady diffusion) in study of interactions between biopolymers and organic dyes. Anionic (humic acids) and cationic (chitosan) representatives of biopolymers were utilized and methylene blue and methylene orange were used as a reactivity probe in the case of humic acids and chitosan, respectively. Another appreciable part of the work is represented by in-depth characterization of applied biopolymers and hydrogel samples by a wide range of analytical and physico-chemical methods (rheometry, UV-VIS, FT-IR etc.). Besides particular experimental results on studied systems, the diploma thesis provides both theoretical and practical background for subsequent applications of the innovative and full-automated method for reactivity mapping of biopolymers.
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Transport of Biomedical Active Compounds Through Porous Membranes
Vašíčková, Kateřina ; Smilková, Marcela (referee) ; Smilek, Jiří (advisor)
This bachelor thesis deals with a study of transport of biomedical active compounds through porous membranes. The main goal is to optimize a penetration method of micellar solutions with a solubilized dye and subsequently with solubilized drug. Cetyltrimethylammonium bromide is chosen as a surfactant and a nile red dye is used as a diffusion and fluorescence probe. Selected active compound is a nonsteroidal anti-inflammatory drug ketoprofen with analgesic and antipyretic effects. The diffusion processes are realized using Franz diffusion cells with polycarbonate membrane with a 2 µm size of poruses. The samples are withdrawn in a chosen time intervals and afterwards evaluated by a fluorescence correlation spectroscopy. Using this method, it is possible to screen the diffusion in time even in nanomolar concentrations of a fluorescence probe chosen. This thesis provides an overview of nile red behaviour in the micellar solutions of surfactant during diffusion processes. The experiment can be extended using an active compound, for instance using a ketoprofen as it is mentioned above.
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Influence of properties of humic acids on diffusion of cupric ions in humic hydrogels
Král, Jan ; Smilek, Jiří (referee) ; Klučáková, Martina (advisor)
Presented bachelor's thesis focuses on the study of diffusion of cupric ions in humic acid gels. The objective of the work was to assess how the properties effect diffusion in humic gels. In the experimental part, three samples of humic acids of different purity were studied. Method of instantaneous planar source diffusion was used to compare them. This methods was based on monitoring temporal evolution of diffusion profiles of cupric ions in gels. Copper ions were elected as diffusing medium because of their high affinity and strong bonds to humic acids.
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Polysaccharide-protein gels
Hájovská, Pavla ; Smilek, Jiří (referee) ; Chytil, Martin (advisor)
This bachelor thesis deals with gelation of albumin solutions and mixed solutions of albumin and the high molecular weight hyaluronic acid due to heating. Study was carried out by measuring rheological properties of the solutions, formed gels and gelation process. For the protein samples in water, the effect of concentration, ionic strength and pH were tested. For mixed solutions, the effect of concentration and a composition of the polymeric component were tested. With increasing concentration of the polymeric component, the gel point is shifted towards lower temperature values, with the concentration difference of 2 % (w/v) up to 5°C. The resulting gels exhibit higher values of loss and storage moduli. Similar behavior was observed also in the samples influenced by the ionic strength as, a result of alternation of the electrostatic interactions between the charged groups of the polymer chains and as a result of change of this charge due to the pH reduced below the pI value of the protein.
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The behavior of superabsorbent polymers in artificial soils
Männlová, Adriana ; Smilek, Jiří (referee) ; Kratochvílová, Romana (advisor)
This bachelor thesis deals with the study of superabsorbent polymers for environmental and agricultural applications. Generally thessis preparations are to perform the funciton of irrigation ,so that thanks to its unique properties , can absorb water in tha rainy season and keep it arend the root system as long as possible. Superabsorbent polymers can also perform the fiction of fertilizer carries and thus ensure its gradual release. These gel structures were subjected to testing of their swelling in environments with different ionic strenght. Subsequently, in the long term monitored thein ability to trap water in the soil environment model.
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Hyaluronan-albumin system dynamics
Jarábková, Sabína ; Smilek, Jiří (referee) ; Chytil, Martin (advisor)
The bachelor thesis deals with the hyaluronan and albumin system dynamics. In this study was to use these methods: interfacial rheology, tensiometry, method of maximum pressure in the bubble and UV-VIS spectrophotometry. The measurements were used with various concentrations of albumin and a constant concentration of hyaluronan( high molecular and low molecular too) and then conversely. All solutions were measured and the obtained data were compared with the results of BSA in buffer without HA. Solutions were prepared with citrate-phosphate buffer of pH about 4. Were observed interaction between HA and BSA.
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