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Barrier properties of hydrogels
Královič, Michal ; Kalina, Michal (referee) ; Klučáková, Martina (advisor)
The main objective of this thesis is to judge the barrier properties of hydrogels. It was examined in total six kinds of agarose hydrogels, of which five kinds of gels was enriched with humic acids isolated from various sources. Humic acids were selected from IHSS standards, specifically Leonardite, Elliott Soil, Suwannee River, Pahokee Peat and Nordic Lake. Using UV-VIS spectrometry, copper ion concentrations were used to serve as a diffusion solution. Results of diffusion coefficients were evaluated by several diffusion methods. The method of diffusion in the plane of the board and three possible diffusion solutions of the constant source were used.
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Light Scattering Techniques in the Study on Formation of Polyelectrolyte Complexes in the System Biopolymer-Surfactant
Valečková, Vendula ; Smilek, Jiří (referee) ; Kalina, Michal (advisor)
This master thesis is dealing with the use of light scattering techniques in the study on formation of polyelectrolyte complexes in the system biopolymer-surfactant. Sodium hyaluronate was chosen as biopolymer and cetyltrimethylammonium bromide and karbethopendecinium bromide were selected as surfactants. The first precipitation experiments were performed to determine the optimal concentration range of surfactants for subsequent titration measurements performed on Zetasizer Nano ZS and for SEC-MALS analysis. The key parameters obtained from these measurements were the values of critical aggregation concentrations in hyaluronate-surfactant systems. It was found out that the increasing molecular weight of hyaluronan in these systems is causing reduction of obtained values of critical aggregation concentrations. Changes of Z-average particle size, zeta potential, polydispersity index, conformation plot, radius of gyration and molecular weight in the hyaluronan-surfactant system were monitored during experiments.
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Preparation and Characterization of Mixed Hydrogels Based on the System Hyaluronan-Albumin
Hájovská, Pavla ; Kráčalík, Milan (referee) ; Kalina, Michal (advisor)
This diploma thesis deals with preparation and subsequent characterization of albumin (BSA) and mixed hyaluronan-albumin (HA+BSA) hydrogels. The experimental characterization was based on the rheological measurements of the influence of ionic strength, pH and molar mass of HA on the gelation process and the properties of formed gels, as well as the time stability of these properties. Following object of the study was swelling of hydrogels in water and buffers with different pH value. Characterization was extended by thermal analysis, morphology description using scanning electron microscopy (SEM) and study of transport properties of hydrogels for selected model substance. Due to increasing ionic strength, the gel point was shifted towards higher temperature values and the resulting gels exhibited higher values of complex moduli. In case when pH was lower than isoelectric point of BSA, attractive electrostatic interactions between albumin and hyaluronan took place and the faster gelation occurred, compared to the case when pH was higher than isoelectric point of BSA. On the contrary, values of complex moduli increased with increasing pH. The gel samples prepared in the presence of low molecular weight HA reached higher values of complex moduli, compared to samples with the high molecular weight HA. Rheological properties of BSA gels were stable over time, whereas the mixed HA+BSA were characterized by significant decrease of complex moduli during the first three days of storage. Due to heating up to 100 °C, more than 80 % of the weight of studied samples was lost, as a result of water evaporation. Further heating of hydrogels, up to 250 °C, did not cause other changes associated with weight loss, in contrast with crystalline powder BSA, in which another weight loss at 220 °C was observed, as a result of the beginning degradation. Swelling studies, as well as SEM images, indicated higher crosslink density of BSA gels, compared to HA+BSA samples, which exhibited higher values of swelling ratio. The lowest values of equilibrium swelling ratio were observed at pH 4, which is close to the isoelectric point of albumin. Calculated diffusion coefficients relative to diffusivity of methylene blue in water ranged between the order of 10E-2 for diffusion into physiological solution and the order of 10E-3 for the diffusion into water. Ongoing transport of model dye from gel to solution was significantly slower, compared to its diffusion in water, as a result of gel porosity and electrostatic immobilization of positively charged molecules of methylene blue on the negatively charged BSA and HA chains.
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Optimisation of Isolation Procedure and Characterization of Amorphous PHB granules
Kratochvíl, Zdeněk ; Obruča, Stanislav (referee) ; Kalina, Michal (advisor)
First artificial PHB granules were prepared under the terms of this thesis. The effect of used PHB solvent, ultrasonic bath temperature and time, solvent evaporation temperature and stabilizing agent nature was investigated using dynamic and electrophoretic light scattering. The most proper parameters were demonstrated at samples which were prepared by dissolving of PHB in chloroform, stabilizing with CTAB or lecithin, ultrasonifying at 35 °C followed by chloroform evaporating at 60 °C. Based on ATR-FTIR and Raman spectroscopy results, it was found out that PHB within the artificial granules were in crystalline form. The native PHB granules were isolated from Cupriavidus necator using either lysosyme, deoxyribonuclease and cell disruption by ultrasonification or digestion with alcalase, SDS and EDTA. Granules obtained by both isolation procedures were characterized by ATR-FTIR, Raman spectroscopy, light scattering techniques and DSC. According to the analyses results, the second mentioned procedure turned out to be more effective for obtaining the polymer in amorphous state. Furthermore, the polymer within granules recovered by using this procedure was thermally more stable. Last but not least, the native PHB granules samples were exposed to effect of acetone, lipase and sodium hypochlorite, assuming that polymer crystallinity should be increased by these chemicals in varying degrees. The highest degree of crystallinity was achieved after their treatment with lipase.
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Transport processes in hydrogels
Sárová, Michaela ; Kalina, Michal (referee) ; Klučáková, Martina (advisor)
This master's thesis is focused on study of transport processes in hydrogels based on humic acids. For this purpose is used methods unsteady diffusion in cuvettes, which was studied the transport of organic dyes, specifically methylene blue and rhodamine 6G, in agarose hydrogel without the addition and with the addition of individual standards humic acids (Leonardite, Elliott Soil, Suwannee River II and Pahokee Peat). This method is based on spectrophotometric monitoring of concentration changes of dyes depending on space of the cuvette and on time. The aim of this thesis was to investigate the effects of interactions between diffusing dye and the particular type of gel to the resultant effective diffusion coefficient of dye. The experiments indicate that the presence of humic acid in the hydrogel greatly affects the transport of selected dyes.
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Mobile robot control system
Kalina, Michal ; Veselý, Libor (referee) ; Hynčica, Ondřej (advisor)
This thesis deals with building of two-wheeled mobile robot and consecutive creation of drivers and control for sensors, engines and controller design. The control is based on Arduino platform using ATmega328 processor. Firstly there is in this thesis recherche. Part of this thesis is modelling one where simulation model of our system is created, its linearization and then controller design. Further it contains description of the building robots body, description and functionallity of used engines, description of all sensors and detailed description of available printed circuit board. In the end there is software part where the engines control is described, operation of the sensors and functionallity of the controller. Result of this thesis is function application of controlling and operating of two-wheeled unstable mobile robot.
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Characterization of pectin by ultrasonic spectroscopy
Trtek, Jan ; Kalina, Michal (referee) ; Hurčíková, Andrea (advisor)
Pectin solutions of various concentration in water has been investigated using high resolution ultrasonic spectroscopy. Furhermore, the influence of solvent on the properties of solutions by dissolving pectin in 0.15 M aqueous solution of sodium chloride and buffer PBS at pH 7.4. For comparison the properties of the pectin solutions were compared with solutions of pectate. It was found that pectine changes characteristics in differenet solutions. These changes have been observed due to changes in the parameters of ultrasonic waves.
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Influence of conditions on hydrogel preparation
Heger, Richard ; Kalina, Michal (referee) ; Mravec, Filip (advisor)
This work is focused on testing the influence of conditions, more precisely pH, during the creation of hydrogels that were prepared through the hydration of powdery mixtures, by another name solid-mixture way. The hydrogel system was made from a polyelectrolyte, more accurately sodium hyaluronate and a cationic surfactant, CTAB. The influence of pH was studied through observation and rheology. For measuring purposes, the hyaluronan with the molecular weight of 600-800 kDa was used. The system was examined via a series of buffers with the pH of 3,5-11. The gels were dyed by Sudan III for better observability and description. It was found that the best way of adding an active substance was by adding a dye directly into a powdery mixture. The observed properties of the gels didn’t change even after the gels were modified to the ionic strength of a physiological saline solution (0,15 M).
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Study of interactions of pigments with biopolymer chitosan
Kolesa, Pavel ; Kalina, Michal (referee) ; Klučáková, Martina (advisor)
This master's thesis was focused on the study of interaction of some organic azo dyes (model diffusion probes) with cationic biopolymer chitosan. This interactions were realized via diffusion processes in hydrogel media based on thermoreversible agarose. The main aim was study of influence of pH of solution on the diffusion process. Interactions of used dyes are based on electrostatic character. The aminogroup of chitosan interacts with the functional group of chosen dyes (anionic sulfonic group) and thus affects the process of diffusion. The model diffusion compounds were chosen organic anionic dyes Chicago sky blue 6B (C.I. 24 410), Sirius red (C.I. 35 780) and Reactive blue 49 (C.I. 621 526). The important content of this diffusion method is a monitoring of the time progression of diffusion profile by UV-VIS spectrophotometry. The presented work follows the bachelor thesis and shows comprehensive view of the reactivity of chitosan and its behavior in different systems. Unsteady diffusion in cuvettes appears to be a universal method for the study of reactivity of biopolymers and for the study of transport processes in hydrogel media. The diffusion method is universal, easy and cheap.
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Study on Interconnection between Structure and Rheological Properties of Hydrogels on Macro and Microscopic Level
Lepíková, Jana ; Klučáková, Martina (referee) ; Kalina, Michal (advisor)
Diploma thesis main goal is to obtain new pieces of knowledge about relationship between hydrogel structures and its flow and transport properties. Thesis is mainly focusing on combining pertinent biopolymers into model hydrogels based on agarose. Then perform correlation of results obtained by diffusion methods, and by rheologic measurements on macroscopic and microscopic level. Properties of hydrogels were measured by selected rheologic measurements, dynamic light scattering method, and correlative fluorescence spectroscopy. From these methods various parameters (MSD modules, values of complex viscosity) were obtained. Afterwards transport properties of prepared hydrogels were studied by observing Rhodamine 6G diffusion. Here two different approaches were used. From macroscopic perspective, simple principles of mass diffusion from dye solution to cuvettes filled with hydrogels containing individual biopolymers were used. From microscopic perspective, dye was added during the sample preparation and then the mass diffusion was investigated using FCS. Based on evaluated results it was discovered that added biopolymers don’t influence properties of carrier medium, in this case agarose hydrogels. During the study of prepared hydrogels’ reactivity and barrier properties some differences were observed. Charge of biopolymer and its charge density were discovered as main factors influencing transport of charged solutes into prepared hydrogels.
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