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The Study of Biopolymers Stability Using Light Scattering Techniques
Kratochvíl, Zdeněk ; Sedláček, Petr (referee) ; Kalina, Michal (advisor)
The stability of chosen biopolymers (or simple carbohydrates and amino acids) was in-vestigated using light scattering methods. Samples were prepared by dissolving the sub-stance in deionised water or in diluted acetic acid. Firstly, the effect of increasing concen-tration on zeta potential was observed and the optimal concentration for the next measure-ments was determined. Afterwards, the stability and the molecular weight of used sub-stances were studied during long-time storage in different time intervals. It was found out that chitosan, CMC, hyaluronate and sodium alginate were subjected to degradation whereas increase of the molecular weight and the zeta potential was observed in case of BSA. In the end, the samples were exposed to several different external effects, namely the influence of different temperatures, UV radiation and increasing ionic strength. The results showed that rather higher temperature has a significant impact on biopolymers stability. However, used biopolymers were hardly affected by UV radiation except BSA whose molecular weight increased considerably. Decrease of the zeta potential absolute values and the molecular size usually occurred with increasing ionic strength. The change of molecular weight was not detected.
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Utilzation of physico-chemical analyses in study on microbial stress-response
Slaninová, Eva ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
This diploma thesis deals with the utilization of physico-chemical analysis in study on microbial stress–response. The main content of this work was to propose and optimize techniques and methods which are generally used in different industries. Two bacterial strains, Cupriavidus necator H16 and its mutant strain Cupriavidus necator PHB-4 producing polyhydroxyalkanoates (PHA) under certain conditions, were used as model microorganisms. Initially, microscopic teques such as transmission electron microscopy (TEM), cryo scanning electron microscopy (cryo SEM) and atomic force microscopy (AFM) were proposed and tested for the characteriozation of morphological differences of bacteria. Furthermore, viscoelastic properties of bacteria were determined and compared by the oscillatory tests of the rheology method considering another type of samples. Thermal analysis methods, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) in particular, focused on water transport and behavior of intracellular water influenced by presence of the granules of PHB in bacteria. The last technique, utilized in the study, was dynamic and electrophoretic light dispersion during pH changes for characterization of the surface properties of bacteria such as size, zeta potential and the isoelectric point.
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Rigorous Simulation of Light Interaction with Cells
Dršata, Martin ; Kalousek, Radek (referee) ; Petráček, Jiří (advisor)
This bachelor thesis focuses on rigorous simulations of light scattering by living cells. The first part is dedicated to brief introduction to the given issues and the basic description of the often used computational methods and models of cell structures. Experimental part deals with light scattering simulations using the finite difference time domain method (FDTD). Models of spherical cell and red blood cell are used in these simulations. The aim of the calculations for the first model is to assess the accuracy of the FDTD method with respect to the analytical method using Mie theory of light scattering.
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Phase-resolved Brillouin light scattering: development and applications
Wojewoda, Ondřej ; Dubroka, Adam (referee) ; Urbánek, Michal (advisor)
Spinové vlny mají potenciál být použity jako nová platforma pro přenos a zpracování dat, protože mohou dosáhnout vlnových délek v rozsahu nanometrů a frekvencí v rozsahu terahertzů. K tomu, aby bylo možné navrhnout zařízení a logické obvody založené na spinových vlnách, je zapotřebí získat informace o prostorovém rozložení intenzity spinové vlny a pokud je to možné, také o jejich fázi. To lze měřit pomocí fázově rozlišeného fokuso-vaného Brillouinova rozptylu světla (µ -BLS). Předložená práce se zabývá rozšířením stávající optické sestavy o možnost měření fáze, kde doposud bylo možné měřit pouze intenzitu. Toto rozšíření sestavy je důkladně popsáno a charakterizováno. Schopnosti optické sestavy jsou demonstrovány ve studii šíření spinových vln skrz Néelovu doménovou stěnu. Získané 2D mapy intenzity spinových vln ukazují, že propagace přes doménovou stěnu je ovlivněna topologicky vynucenou kruhovou Blochovou čarou ve středu doménové stěny a že režim propagace závisí na frekvenci spinových vln. V prvním režimu propagace se vytvoří dva svazky spinových vlny šířící se kolem kruhové Blochovy čáry, zatímco ve druhém režimu se spinové vlny šíří pouze středem. Fázově rozlišené µ-BLS měření odhaluje fázový po- sun spinových vln pro oba režimy. Mikromagnetické modelování spinových vln ukazuje rozrušení jejich fázových vlnoploch, které je třeba brát v úvahu při interpretaci měření a navrhování potenciálních zařízení. Mikromagnetické simulace ukazují, že vnější magnetické pole může být použito k pohybu kruhové Blochovy čáry ve stěně domény, a tedy k manipulaci spinových vln.
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Photoprotective properties of PHB granules
Hesko, Ondrej ; Obruča, Stanislav (referee) ; Mravec, Filip (advisor)
The objective of this work is study and design of physical-chemical methods for verification of photoprotective properties PHB inclusions using bacteria Cupriavidus necator H16 and its mutated bacteria strain Cupriavidus necator PHB-4 which differs in production capability of polyhydroxyalkanoates. Basic information about of polymer inslusions and methods used for proving photoprotectivity are discussed in theoretical part. Methods such as nephelometry and continous fluorescence microphotolysis (CMF), which is used to characterize the mechanism of interaction of light with inclusions, were optimized. In experimental part properties of light scattering in bacteria were determined and characteristic curves were obtained. Further the photostability of fluorescence probe specific for nucleic acids were compared in bacteria strains. This method should prove bigger tolerance of bacteria H16 for UV light. It should also define possible interaction of nucleic acids with PHB inclusions.
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Mucin hydrogels - artificial models of native mucus systems
Mikušová, Janka ; Obruča, Stanislav (referee) ; Sedláček, Petr (advisor)
The scope of this masters thesis is the preparation of a model mucin system and its utilization as an artificial model of the native mucus system. The creation of this model system, according to several designed methods was a part of experimental part of the thesis. The preparation of mucin system comprised of physical and chemical methods of hydrogel formation, screening and characterisation of the various physical conditions of the mucin properties on its molecular level, and the preparation of sorbent with sorption surface containing mucin. Methods of light scattering, namely dynamic light scattering (DLS), used for mucin particles size change monitoring, and electroforetic light scattering (ELS), used for Zeta potential change monitoring, were used for the screening of the impact of physical factors on the properties of mucin.For the characterisation of impact of the temperature on changes in mucin sctructure was, apart from monitoring of light scattering, used also a diferential scanning calorimetry (DSC), which registered temperature value, at which mucin thermal denaturation occurs. In the next part of the thesis we subdued the created sorption surfaces to various physical-chemical analyses, which task is the characterisation and projection of surface and confirmation of mucin presence.Substancial part in monitoring and characterisation of changes in surface sctructure of sorption surface was accomplished by Fourier transform infrared spectroscopy (FTIR). Scanning electron microscophy (SEM) was used for the final, more detailed, projection of the mucin enriched, sorbent surface structure. Suggested methods of mucin hydrogel, didnt prove sufficient results for the possibility of application of hydrogel as a artificial model of real mucus system, but the sorbent application was indicated as a suitable alternative and an instrument for the further mucin behaviour research and possibly subsequent bacterial adhesion, which represents the first step in the formation of the bacterial biofilm.
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Procedurally Generated Landscape in Fragment Shader
Leitner, Denis ; Milet, Tomáš (referee) ; Chlubna, Tomáš (advisor)
This thesis deals with rendering of procedurally generated landscape without the use of input geometry or textures. It describes techniques for generation and realistic rendering of natural outdoor scenes. These techniques include terrain generation and realistic atmosphere and cloud rendering. Thesis also describes the use of raymarching for terrain and shadow rendering and realistic lighting for terrain which includes ambient and indirect light approximation.
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Preparation and Characterisation of Polyelectrolyte Complexes in the System Dextran-Surfactant
Klímová, Eliška ; Krouská, Jitka (referee) ; Kalina, Michal (advisor)
This bachelor thesis deals with the study on preparation and characterization of the polyelectrolyte complex formation in the dextran-surfactant system. The aim of the thesis was the investigation of the influence of the ionic strenght on the formation of the interactions in the studied system. For these purposes, components having opposite charges (DEAE-dextran hydrochloride and anion-active sodium dodecyl sulphate) were selected. For individual measurements mainly the methods of light scattering, dynamic and electrophoretic light scattering in particular, have been used. In the thesis have been studied interactions and aggregation concentrations of individual components having the potential to form polyelectrolyte complexes. It was verified in all the studied samples, by measuring the intensity of scattered light and the size of Z-diameter of particles, that the increase in sizes of particles is caused by the formation of these complexes. There were also measured additional characteristicts of the system such as pH or conductivity. Their dependences provide comprehensive view on the formation of complexes in studied system and can be utilized for other measurements and possible future applications, especially for purposes in medical applications.
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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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