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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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Utilization of biosurfactants for incorporation of hydrophobic domains into modern controlled-release systems
Nešpor, Tomáš ; Pekař, Miloslav (referee) ; Sedláček, Petr (advisor)
This work deals with the current topic of carrier systems. Since the biggest problem is the passage of hydrophobic drugs through the bloodstream, or through universal body barriers (eg blood-brain), it is necessary to chemically modify these carriers in order to be able to administer hydrophobic substances effectively. Based on a literature search, several systems are designed and subsequently studied, in which there is a presumption of possible use for carrier systems and at the same time they have biosurfactants incorporated in them due to their ability to solubilize hydrophobic molecules. The theoretical part of this work will describe the individual biosurfactants, the process of their production, their physicochemical properties, and the possibility of their use in carrier systems. At the same time, the individual carrier systems, the procedure of their preparation, the possibilities of their use are described, and their advantages and disadvantages are also compared. In the practical part, the screening of both individual substances and their mutual interactions, as well as methods used to study the emerging structures is then performed. The study of molecular interactions is primarily performed using the technique of dynamic light scattering. The next part describes the optimization of hydrogel formation with incorporated biosurfactants in their structure and then the formed gels are subjected to rheological and solubilization tests. The study of the internal structure of these gels is performed using a scanning electron microscope.
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Plasticizing of polyhydroxyalkanoates in microbial cells - inspiration for development of artificial controlled release systems
Liczka, Jan ; Obruča, Stanislav (referee) ; Sedláček, Petr (advisor)
The diploma thesis is focused on the study of the mechanism of plasticization of polyhydroxyalkanoates (PHA) in vivo, focusing mainly on methods of isolation native PHA granules and physical and chemical methods of initiation their phase transitions. The literature search prepared in this work deals with this microbial polyester and further focuses on individual methods of isolating PHA from bacterial cells. The main task of the experimental work was to optimize the isolation process of native PHA granules, as well as to design and test processes that initiate the PHA crystallization in isolated granules, as well as analytical procedures to correctly detect this phase transition. Isolation of PHA granules from Cupriavidus necator was performed by enzymes, centrifugation several times and sonication of the cells. Fourier transform infrared spectroscopy (FTIR) and an enzymatic method in which amorphous PHA is selectively degraded by PHA-depolymerase with turbidimetric detection of its loss in dispersion was used to detect PHA crystallization. Techniques based on fluorescence staining of granules (fluorescence spectroscopy and flow cytometry) were used in a more detailed study of the mechanism of PHA plasticization in granules. The results of the analyzes confirmed, among other things, that the optimized procedure makes it possible to isolate granules in the amorphous state, which remain amorphous even after drying and are comparable to native granules in their basic physicochemical properties.
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Structural aspects of interaction between humic substances and charged organic xenobiotics
Prisažný, Adam ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
This diploma thesis was focused on studying the interaction of humic substances with ionic organic xenobiotics and its structural aspects. The method was chosen from my bachelor thesis, steady-state fluorescence spectroscopy, which is suitable for substances with weak fluorescence. The results showed that the interaction between humic acids and representatives of ionic organic xenobiotics (Septonex) was reflected in fluorescence quenching of humic acids and the shift of emission maximum to lower wavelength, hypsochromic (blue) shift. From the measurement results, we can assume that the interaction that is formed between the aromatic structures in humic acids and Septonex could be -cation interaction.
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Study of decomposition of organic matter in soil using the method of "tea bags"
Lišková, Kateřina ; Sedláček, Petr (referee) ; Enev, Vojtěch (advisor)
The main aim of this master thesis was to introduce an optimal method which could be used for study of a decompostion of an organic matter in a soil. For this purpose was chosen based on a literaly review so-called tea bag method. With regard to the completeness of the work, a physico-chemical characterization of the soil environment and HS isolated from it was also performed. The degraded organic material was then further characterized using thermogravimetric analysis, elemental analysis, FT-IR spectroscopy, and the total content of water-extractable polyphenolic substances in the samples was also determined.
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Preparation and application particles from copolymer of P(3HB-co-4HB)
Ostanina, Kseniia ; Sedláček, Petr (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with preparation of particles from poly(3-hydroxybutyráte-co-4-hydroxybutyráte). Copolymer was dissolved in acetone and then the solution had been dosed into the anti-solvents. Acetone and anti-solvent are perfectly miscible. Theoretical part focuses on the properties of the copolymer, its production and potential use in medical applications. In the experimental part the copolymer was dripped into selected anti-solvents by syringe and pipette above and below the surface of anti-solvent. The following anti-solvent were chosen: water, methanol, dimethylsulfoxid (DMSO), 50 % acetone-water, 75% acetone-water, 50 % methanol-water, 75 % methanol-water mixtures. There were three copolymers used with different compositions of 4HB units (63 %, 87 % a 33 %). Copolymer composition was analyzed by gas chromatography. Copolymers were prepared by bacterial strain Aneurinibacillus sp. H1. Due to the results particles of size approximately 100-500 nm can be prepared by methods described in this work. The produced particles can be successfully used in medical applications.
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Interaction of drugs with soil organic matter - calorimetric study
Papko, Nadezhda ; Sedláček, Petr (referee) ; Klučáková, Martina (advisor)
This bachelor thesis is focused on the study of the interaction of soil organic matter with selected drugs using isothermal titration calorimetry. Two fulvic acids and drugs were used in the experiment: ibuprofen, pyridine sulfate and diclofenac. The scope of the work was to test the method for studying the interactions between fulvic acids and drugs and to optimize it for specific humic substances and drugs. Isothermal titration calorimetry includes the possibility to determine very important thermodynamic parameters, which were determined during the measurement of the bachelor's thesis. The main goal was to measure and assess whether the interactions take place and what the thermal effects are.
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Development and optimization of ointment preparation for burn healing
Müllerová, Kristýna ; Sedláček, Petr (referee) ; Enev, Vojtěch (advisor)
This bachelor's thesis aims to prepare ointments based on an ointment base with the addition of a bioactive component of chlorophyll according to a recipe from an unknow doctor. The basic characteristic of emulsions was performer in the theoretical part, their instability and possible methods of stabilization were described. Furthermore, the reader is acquainted with the bioactive substance of chlorophyll, selected ointments for the healing of burns and wounds and finally with the use in medicine. In the Experimental part, the preparation of emulsion bases of ointment was described. Based on a performed analysis of stability using analytical centrifuge, two ointments bases were selected, to which other components were added – chlorophyll, beeswax, and chamomile extract. The stability of completely prepared ointments was studied at 25 °C and 37 °C, this analysis was performed even after a longer time interval. Using ATR-FTIR spectrometry the present functional groups were identified at prepared ointments, individual components of the ointments base and chlorophyll. Based on the stability measurement of the prepared ointments and rheological measurements (oscillation and rotational measurements), it was evaluated that the best properties were shown by the prepared ointment with an ointment base consisting of 30 wt. % white vaseline and 70 wt. % synderman.
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Hydrogel carriers of microorganisms for modern environmental aplications.
Súkeník, Martin ; Slaninová, Eva (referee) ; Sedláček, Petr (advisor)
This bachelor thesis deals with the optimization of cultivation and gelation procedures by bacterium Azotobacter vinelandii, which belongs to a PGPR group, capable of synthesis of intracellular polyhydroxyalkanoates and extracellular alginate. The alginate produced by the bacteria is crosslinked with a calcium chloride solution to form a hydrogel carrier containing the described bacterium. This unconventional concept simplifies bacterial encapsulation and production of hydrogel carriers, furthermore expands its usability in modern environmental applications. In the first part of this thesis, three strains (DSM 85, 87, 720) were selected to measure a concentration of produced alginate, its molecular weight was determined by a SEC-MALS technique and M/G ratio was analysed by infrared spectroscopy. The Azotobacter vinelandii strain DSM 87 reached the highest concentration of the alginate (4.9 ± 0.6) g/l by gravimetric determination on the 5th day of cultivation. The concentration of produced PHB ranges from 48 to 6 % of CDW, which was determined by gas chromatography. Nevertheless, the strain DSM 720 showed the best gelation properties and was used in the second part of this thesis for further optimization of the cultivation process, and for the analysis the formation of gelation procedure under different temperatures, gelation time and concentrations of crosslinking solution. The formed gels were compared by rheological measurements of amplitude and frequency tests.
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