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Use of copolymer polyhydroxyalkanoates in development of intelligent packaging
Kreuzerová, Monika ; Kalina, Michal (referee) ; Sedláček, Petr (advisor)
This bachelor thesis deals with the preparation and characterization of copolymers P(3HB-co-4HB). The theoretical part discusses polyhydroxyalcanolates (PHA), where their structure, properties and synthesis are described and subsequently the selected copolymers, which contain monomeric units 3HB, are listed. Furthermore, the review part is processed for the current state of the issue, which states the use of copolymers PHA and current research work on the incorporation of active substances into copolymers. The aim of this work was to characterize and compare the created P(3HB) and P(3HB-co-4HB) foils with a 4HB monomeric unit of 65,25 % and 43,61 % and subsequently determine the potential of foil usability as intelligent packaging materials. First, PHA were subjected to gas chromatography, SEC-MALS, FTIR and DSC analysis. In the second part of the thesis, the permeability of foils to water vapor and the release of model active substances from PHA foils was studied. For this purpose, two active substances (curcumin and salicylic acid) were selected, which were incorporated into a film of homopolymer P(3HB) and two copolymers with a proportion of 4HB of 65,25 % and 43,61 %. In foils prepared from copolymers, the active substances diffusion was faster than in P(3HB). Within the copolymers, P(3HB-co-43,61%4HB) released substances at a higher speed, due to the greater crystallinity of a copolymer containing 65,25 % 4HB.
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Program-controlled freeze drying for the preparation of the delivery system of native hyaluronan and hydrophobic molecules
Waczulíková, Kristína ; Sedláček, Petr (referee) ; Enev, Vojtěch (advisor)
The amphiphilic nature of the hyaluronan biopolymer facilitates interactions between its hydrophobic portions of the chain and the hydrophobic ligands, making it one of the promising compounds that can be used as a suitable carrier for drug delivery. Therefore, the aim of this diploma work was by means of a program-controlled freeze-drying to prepare a system based on native hyaluronan, which would be capable of transporting hydrophobic drugs. Perylene, pyrene, prodan and 1,6-diphenyl-1,3,5-hexatriene fluorescence probes were used as model compounds to simulate hydrophobic drug-hyaluronan interactions. Freeze-drying efficiency was studied using infrared spectrometry and scanning electron microscopy. ATR-FTIR spectrometry confirmed for all probes but pyrene the hydrophobic interactions between the hyaluronan and probe molecules based on the presence of the C=C bond valence vibration absorption band in the aromatic probe rings. From the IR spectra, it was not possible to confirm with certainty the presence of residual tert-butyl alcohol in the samples. SEM photographs revealed that increasing the pressure during drying process had a positive effect on the quality of the lyophilized sample in the second series of samples. Interactions of individual fluorescent probes with hyaluronan were studied using steady-state and time-resolved fluorescence spectrometry. Positive results have been obtained, indicating the presence of fluorescent probes interacting with the hydrophobic regions of the hyaluronan chains. The results of this work can contribute to the development of carrier systems in the field of targeted drug distribution.
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Physico-chemical characterization of 3-hydroxybutyrate with respect to its bioprotective effects
Kahanovská, Kristína ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
The main aim of the work was to verify bio-protective properties of 3-hydroxybutyrate (3HB) in terms of its effect on protein denaturation (especially lipase) and determine the physicochemical characteristics of 3HB that could lead to the explanation of the mechanism of its protective effect. The literature search focused primarily on the mechanism of the protective effect of the compatible solutes. The 3HB belongs to the group of compatible solutes. The protective effect of the 3HB was monitored by Dynamic Light Scattering (DLS). As the other parameters of the aqueous solution 3HB were determined: the dependence of the surface tension on the concentration of the 3HB, the effect of different 3HB concentrations on the zeta potential of the lipase. Hydrophobic and hydrophilic character of the 3HB was characterized with the log Kow parameter. A simple method for quantity determination of the 3HB aqueous solution was optimized. The method is based on conversion of the 3HB into crotonic acid by reaction with sulphuric acid at higher temperatures and on subsequent psectrophotometric determination of the amount of crotonic acid.
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Methodology for experimental study on the release of humic acids from hydrogel application forms.
Palanová, Veronika ; Klučáková, Martina (referee) ; Sedláček, Petr (advisor)
Hlavnou náplňou bakalárskej práce bolo navrhnúť a optimalizovať metódu experimentálneho štúdia uvoľňovania huminovej látky z gelových matríc, ktoré boli vytvorené pomocou bežných kozmetických a farmaceutických zahusťujúcich látok. Pre štúdium uvoľňovania tejto aktívnej látky bola zvolená difúzna aparatúra vo vertikálnom usporiadaní, tzv. Francova cela. Výluhy gelových vzoriek boli podrobené spektrofotometrickým meraniam. Vzniklé interakcie medzi roztokmi lignohumátu a rôznych typov zahusťujúcich látok boli experimentálne stanovené. Celý proces optimalizácie metódy pre uvoľňovanie huminovej látky z gelových foriem bol diskutovaný.
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Preparation and Characterization of Mechanical Properties of Artificial Synovial Liquids
Hrochová, Eliška ; Sedláček, Petr (referee) ; Kalina, Michal (advisor)
This bachelor thesis deals with the optimization of preparation of artificial synovial liquids. The main subjects of study are the mechanical properties of real and artificial synovial fluid samples. DLS microrheology, thermogravimetric analysis (TGA) and infrared spectroscopy (FTIR) were used for characterization. The theoretical part of this thesis is represented by a literature research of a methods of preparation of artificial synovia and summary of definitions of rheological terms. The experimental part focuses on the preparation and characterization of the artificial synovial liquids originating from the published patent US 8716204. In the framework of the bachelor thesis, this preparation was optimized in several partial steps (method of dispersion of components, choice of molecular weight of hyaluronic acid, nature and ionic strength of used dispersion medium). The prepared optimized sample of synovial fluid was in following experimental characterization steps compared in terms of material characteristics and mechanical properties with the real horse synovial fluid sample. By modifying the process based on the aforementioned patent, an artificial synovial fluid could be formed. That meets the viscoelastic nature of the real matrix and that is stable over time.
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Development and characterization of hydrogel carriers for modern agricultural applications
Mai, Thuy Ha ; Kalina, Michal (referee) ; Sedláček, Petr (advisor)
The bachelor thesis is focused on characterization of hydrogel carriers and their application in agriculture. Entrapment of active phase within the hydrogel matrix would allow release of its content in controlled rate. The theoretical part describes encapsulation techniques and suitable biocompatible materials involved in encapsulation. For the experimental part, the release of soil bacteria Cupriavidus necator H16 from alginate matrix was examined. The focus was on the release rate of bacteria via diffusion and later, on the release rate of bacteria encouraged by chemical or enzymatic reagents. Further modification of carrier was suggested by adding hydrophobic barrier composed of polyhydroxybutyrate (PHB). The modified alginate beads seemed to be more resistant toward enzymatic or chemical agents, thus the process of the release was to some extent suppressed. This fact might suggest PHB as a possible vessel for optimization of controlled release system of hydrogel carriers.
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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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Preparation and characterization of hydrogel delivery systems from poly-gamma-glutamic acid
Mai, Thuy Ha ; Smilek, Jiří (referee) ; Sedláček, Petr (advisor)
The presented diploma thesis focuses on the development and preparation of model poly--glutamic acid based hydrogel carrier systems. For the hydrogel carrier preparation, methods based on physical interactions between polyelectrolyte and oppositely charged multivalent ion or oppositely charged macromolecule were proposed. Based on the pilot experiments, trivalent iron cation and chitosan were selected as suitable crosslinking agents. By crosslinking -PGA via ferric cations a successful bead formation has been accomplished and attempts have been made for a preparation of beads smaller than 1 000 m by employing microencapsulation techniques. Rapid disintegration of -PGA beads occurred upon immersion of the beads in water making these systems unsuitable for drug carrier applications. Interactions between -PGA and chitosan lead to polyelectrolyte complexes formation and by optimizing concentrations and pH of polyelectrolyte solutions, a formation of homogenous looking hydrogel with high water content (>0,9) has been achieved.
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Self-encapsulation of Plant Growth Promoting Rhizobacteria as a way towards new generation of bioinoculants
Gašparová, Dominika ; Obruča, Stanislav (referee) ; Sedláček, Petr (advisor)
This bachelor thesis focuses on Plant Growth-Promoting Rhizobacteria that can be used as a suitable ecological alternative to conventional fertilizers. However, most of today's approaches to incorporating PGPR into the process of soil fertilization are convoluted and costly. Crosslinking of alginate self-produced by PGPR offers a new possibility of plantgrowth- promoting bacteria encapsulation. The aim of this thesis consists of preparing gel matrices with incorporated bacteria, followed by testing the bacteria's viability and quantity. Firstly, experiments were carried out to determine various properties of gels produced by the chosen representative bacteria Azotobacter vinelandii. These properties include the molecular weight of alginate (with the highest molecular weight of strain CCM 289 (329,78± 6,8) kDa), the alginate's amount in the gel, the content of PHB (34%-44% CDW), and the gel's overall rheological properties. The complex modulus (as a representation of rheological properties) of the firmest gel reached the value of 13,34 kPa. The bacteria content was examined by flow cytometry, the CFU method, and spectrophotometry. The viability of bacteria was determined by analyzing the fluorescence intensity via fluorescent dyes (propidium iodide, fluorescein diacetate, rhodamine 123, calcein AM), then by using the CFU method, and also via the plate method. Furthermore, the effects of various factors such as different pH levels (pH 3, 7, and 10) or adding the specific alginatedegrading enzyme alginase were also examined. Releasing of the bacteria was observed over the course of seven days using flow cytometry and the CFU method. The plate method proved the bacteria's survival after gelation and without any source of carbon. The largest disintegration of gels occurred in acidic pH. Moreover, the effect of alginase on crosslinked bacterial gels and synthetic alginate were very similar. The survival of bacterial cultures in gel and their continuous release are significant findings in the development of novel bioinoculants based on this new concept.
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The stress and modal analysis in parametric adaptive CAD programs
Sedláček, Petr ; Ondrůšek, Čestmír (referee) ; Kuchyňková, Hana (advisor)
This thesis is focusing on the use of the finite element method (FEM) in program environment of Autodesk Inventor Professional (AIP) 2010. For the analysis of the components or assemblies with FEM serves the Strength analysis module of the AIP 2010, whose environment and functions are detailed in this thesis. At the conclusion practical examples of modal and static analysis on the synchronous generator rotor model are demonstrated including a brief evaluation of the results.
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