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Hyaluronan-albumin complexes behavior at physiological conditions.
Šuleková, Nikola ; Kratochvílová, Romana (referee) ; Chytil, Martin (advisor)
This thesis discusses the interactions of low molecular weight and high molecular weight hyaluronic acid (HA) with bovine serum albumin (BSA) in physiological environment. It is based on previous work dealing with HA and amphiphilic substances which are currently widely studied for its potential applications in medicine. Solutions of HA with fixed concentration and albumin of concentration in the range of 0,25 – 20 g/l were prepared at room temperature and at 37°C and examined by rheometry and turbidimetry. It was observed that the maximum absorbance increases with increasing concentration of BSA and does not show variations depending on the temperature of preparation. On the other hand, the turbidity occurs in significant differences between the samples prepared at ambient temperature and at 37°C. Overall, the turbidity increases with the concentration of BSA. Rasing the temperature caused the increase in viscosity of the samples, which was further varied with the concentration of BSA. Mixed solutions of high molecular weight HA and BSA showed the highest turbidity, maximum absorbance and viscosity, while the solutions of BSA showed the lowest values. The results indicate formation of complexes, of which properties are dependent on BSA concentration, the molecular weight of HA, and the temperature of preparation.
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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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critical Evaluation of metrhods for determination of Sulphates in waters
Havelková, Ludmila ; Řezáčová, Veronika (referee) ; Sommer, Lumír (advisor)
The aim of this work was the evaluation and method optimalization for determination of sulfates in aqueous solutions with application on different kinds of waters. For determination of sulfates were used two different procedures, turbidimetric and volumetric. In turbidimetric procedure was use optimized conditions applicable determination of sulfates in range from 0 – 20 mg.l-1. For stabilization of the colloid was use glycerine, himself sulfates sedimentation is discharged in BaSO4 form. By volumetric methods are used titration on the dithizone like indicator while using Pb(NO3)2 like volumetric reagent. Other indicator bromsulfonazo III, whitch are used for determination of sulfates by the help of volumetric solutions Ba(ClO4)2 and BaCl2. All volumetric determinations proceeded in acidic medium. During research of interferences ions on the individual titraction was detected substantial effect Ca2+, Mg2+ and Fe3+, which was needing divide off on the exchanger resin. Ions aforementioned don't interfere turbidimetric determination of sulfates.
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Preparation and characterization of triple-IPA for the formation of vesicular systems
Novotná, Ludmila ; Pekař, Miloslav (referee) ; Mravec, Filip (advisor)
This thesis deals with the preparation and characterization of vesicular systems consisting of triple-IPA (ion pair amphiphile with three hydrophobic chains). For the preparation of tripleIPA were used double-chain cationic surfactants dimethyldimyristylammonium bromide (DMSAB), dimethyldipalmitylammonium bromide (DPAB), dimethyldioctadecylammonium bromide (DODAB) and dimethyldioctadecylammonium chloride (DODAC) and the singlechain anionic surfactant sodium dodecyl sulfate (SDS). Among the cationic surfactants, DMSAB and DPAB were selected to be studied more thoroughly. Triple-IPAs were prepared by mixing solutions of oppositely charged surfactants in equimolar ratio, and the resulting triple-IPA precipitate was filtered and dried. From the obtained powder, vesicular systems were prepared by rehydration and sonication. Cholesterol and cationic surfactants, from which the triple-IPAs were prepared, were used to stabilize the cationic vesicles. The properties of the vesicular systems were evaluated by dynamic and electrophoretic light scattering measurements and turbidimetry. The dispersion solutions with the addition of cholesterol were less turbid; therefore, it can be assumed that they supported the formation of cationic vesicles. The optimum cholesterol content is 20 mol. %. At this concentration, the effect of cholesterol on vesicle membrane reorganization becomes apparent and the cationic system remains monodisperse. A positive zeta potential in the stable region was determined for all prepared solutions, which was further increased by the addition of a positive surfactant. The most suitable cationic surfactant for the preparation of triple-IPA is DPAB, which is easy to work with, produces the most monodisperse systems and has the lowest measurement uncertainties.
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Preparation and characterization of quatsomes
Havlíčková, Anna ; Szabová, Jana (referee) ; Mravec, Filip (advisor)
This bachelor thesis deals with preparing and characterizing quatsomes and similar vesicular systems. The preparation consisted of weighing an equimolar amount of sterol and surfactant, hydration with deionized water, and subsequent sonication with an ultrasonic probe with an energy of 5, 10 and 15 kJ. In this work, two types of quatsomes were prepared, the first contained equimolar amounts of cholesterol and cetyltrimethylammonium bromide (CTAB), and the second consisted of cholesterol carbethopendecinium bromide (Septonex). The optimal dispersion energy of 10 kJ was selected for both types based on turbidity measurements, visual observation and dynamic and electrophoretic light scattering analysis. Furthermore, the observed properties were found to be constant from day 7 after sonication. In addition to quatsomes, similar vesicles were prepared, which contained a surfactant in the form of a sulfate salt. Once again, two types were prepared, the first containing sodium dodecyl sulfate (SDS) and cholesterol, the second sodium tetradecyl sulfate (STS) and cholesterol. Based on the same measurements as for quatsomes, optimal dispersion energy of 10 kJ was selected. Furthermore, mixtures with different ratios of cholesterol and a given surfactant were prepared, in which the effect of cholesterol addition was monitored. The analysis of these mixtures was performed by the same methods as the analysis of quatsomes.
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Utilization of various methods of porosimetry in investigation of internal structure of biopolymer hydrogels
Zahrádka, Jan ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
The main aim of the thesis was to select, optimize and test suitable methods for studying inner structure of hydrogel materials. The study of the size of pores with potential use in fundamental and applied research of these materials is the central focus of the thesis. Firstly, a literature search was done in order to collect basic information about usefulness of the individual methods. On the basis of this literature search, the experimental part of the work was divided into several steps. First, selected methods of conventional porosimetry (mercury intrusion porosimetry, BET analysis and DSC thermoporometry) were applied on reference porous materials and the results obtained by the individual methods were compared. The methods optimized with the use of these reference materials were then utilized in the analysis of porosity of model hydrogels based on polyvinyl alcohol. Based on the results of this analysis, the usability and benefits of the individual methods were discussed with respect to the sample preparation, time and financial demands, and also from the viewpoint of compliance of the determined pore sizes with the range of sizes declared for the physical PVA gels.
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Microbiological Aspects of Infectious Diseases Therapy
Paterová, Pavla ; Buchta, Vladimír (advisor) ; Kolář, Milan (referee) ; Jílek, Petr (referee)
Background: The method of serum bactericidal assay represents an alternative possibility of optimization of anti-infectious therapy and administration of antibiotics. It mirrors the real activity of one or more administered antibiotics in the complex system of the antibacterial effect of patient's serum. The paper aimed to confirm non-inferiority of bactericidal testing using the broth dilution method according to CLSI M21-A Guidelines (time to results 48, 72 hours) in comparison with modified methods of testing on the basis of turbidimetry (time to result 6, 8, 24 hours) and resazurin color (time to results 8, 24 hours). Methods: Four antibiotics were tested: gentamicin, amikacin, piperacillin/tazobactam and meropenem with 30 Escherichia coli strains isolated from blood cultures of 29 pacients hospitalised in different wards, University Hospital in Hradec Kralove. Human blood sera (n = 76) from ten hematological patients (4th Department of Clinical Medicine, University Hospital, Hradec Kralove) were tested to establish bactericidal titer. Patients' blood was withdrawn prior to and in the course of the first and third day of antibiotic therapy of febrile neutropenia. Testing employed the reference strain Escherichia coli ATCC 25922. Results: A comparison with the standard CSLI showed that the...
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Dry gel forms - aerogels, xerogels - preparation, properties and potential use.
Sedlář, Marian ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
The content of this bachelor thesis was to find out what influence has the drying of hydrogel on its internal structure on an appropriate model hydrogel system. In the theoretical part of this thesis selected methods of structural analysis that are commonly used for characterization on gels in their original hydrated state and in dry state were described. In the practical part, agarose based hydrogels with different concentrations and with various additions of polyelectrolytes were chosen as the model hydrogel system. Cryo-scanning electron microscopy and turbidimetry were used for the structural analysis of hydrogels. For the dry gels, scanning microscopy and mercury porosimetry were used. The obtained data and results for our model system can be use in analyse of other hydrogel systems and their dry forms.
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Complex structural characterization of hydrogels based on semi interpenetrating biopolymer networks.
Trudičová, Monika ; Krzyžánek, Vladislav (referee) ; Sedláček, Petr (advisor)
The content of this diploma thesis was testing the applicability of available structural analysis techniques on a appropriate model hydrogel system. The main aim was to evaluate the advantages and disadvantages of selected structural analysis techniques and the influence of the composition change of the hydrogel system on its internal structure. Semi-interpenetrated hydrogels based on agarose of different concentration were chosen as model system, this type of material was chosen for simple and repeatable preparation and also for its application potential. Electron microscopy (SEM, cryoSEM), mercury porosimetry and turbidimetry were chosen as structural analysis techniques. Experimental results could be used to improve knowledge about the influence of hydrogel composition on its structure and the comparison of chosen techniques will be used for the choice of appropriate structural analysis in the future, which will be applicable to other hydrogel systems as well.
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Hyaluronan-Micelle Aggregates and their Potential for Nanomedicine Applications
Pilgrová, Tereza ; Burgert, Ladislav (referee) ; Reháková, Milena (referee) ; Pekař, Miloslav (advisor)
This thesis deals with the study of preparation and characterization of hyaluronan-micelle aggregates. The theoretical part deals with drug delivery systems, characterization of used materials and methods especially fluorescence spectroscopy, dynamic light scattering and turbidimetry. Methods of determination of measured data are summarized in the experimental section. The result section is divided into two subsections dedicated to different preparation methods of hyaluronan-surfactant complexes. Induced aggregates of hyaluronan with Septonex are characterized in terms of their origin and stability, and the results are compared with previously studied surfactants CTAB. In the second part are discussed so-called decorated micelles, their formation, properties and stability.
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