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New possibilities in wound healing
Nováková, Laura ; Szotkowski, Martin (referee) ; Skoumalová, Petra (advisor)
The diploma thesis is focused on the study of fibrous wound dressings prepared by electrospinning method from natural biopolymers. Three active ingredients were added to the dressings: ampicillin, ibuprofen and collagenase, which are responsible for relieving pain, reducing the risk of infection and selectively removing necrotic tissue in the wound. The theoretical part describes the therapeutic dressings currently available on the market and the most common methods of nanofiber production. The experimental part evaluates the optimization of the preparation of gelatin, alginate and chitosan fibrous wound dressings, which were subsequently enriched with active substances and their gradual release into the model environment was determined spectrophotometrically. Antimicrobial effects against E.coli and S. epidermidis strains andantifungal activity against C. glabrata yeast were monitored. Finally, two cytotoxicity tests on the human keratinocyte cell line HaCaT confirmed the safety of the prepared products, which can serve as bioactive skin dressings in the future.
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Application of Zymomonas mobilis for bioethanol production from cellulose materials
Čalová, Iveta ; Zichová, Miroslava (referee) ; Babák, Libor (advisor)
The thesis deals with the use of Zymomonas mobilis to produce bioethanol from waste paper. There were used three types of paper made by various manufacturers to optimize the production of bioethanol. Individual papers were subjected to the same adjustments namely mechanical grinding and subsequent enzymatic hydrolysis. During cultivation of Zymomonas mobilis in different hydrolysates were taken samples for determination of bioethanol production. The analysis was carried out on high-performance liquid chromatography. Production of bioethanol and glucose consumption was evaluated as well as production of lactic acid since it was also noticeable on the chromatograms.
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Database System for Biological Data Management
Drlík, Radovan ; Burgetová, Ivana (referee) ; Jaša, Petr (advisor)
This thesis describes the problems of storage and management of biological data, particularly of Haloalkane Dehalogenase enzymes. Furthermore, the thesis aims at project HADES (HAloalkane DEhalogenase databaSe) initiated by protein engineering group of Loschmidt Laboratories, Masaryk University in Brno. This is a project whose main goal is simply to store, preserve and display a wide variety of proteins data. The result of this work is a flexible database system allowing easy extensibility and maintainability, which is built on technologies Apache, PostgreSQL and PHP using the Zend Framework.
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Preparation and characterization of wound dressings
Dzurická, Lucia ; Márová, Ivana (referee) ; Skoumalová, Petra (advisor)
The diploma thesis if focused on the study of bioactive hydrogél and nanofiber wound dressings composed of natural biopolymers, which were functionalized by active compounds in the form of analgesic, antibiotics and enzymes. Hydrogél wound dressings were constituted from alginate and chitosan and nanofibers were created from polyhydroxybutyrate. The following 7 active compounds were selected to be added to the wound dressings: ampicillin, streptomycin, ibuprofen, papain, bromelain, collagenase and trypsin. In the theoretical part the structure of the skin and types of wound injuries were described. This part also talks about types of wound dressing and their applications, as well as treatment of skin wounds using enzymes and compounds with analgesic and antimicrobial properties. In addition, this section describes safety assays, in particular cytotoxicity assays on human cells. At the beginning of the experimental part, the process of preparation of hydrogél wound dressing was optimised. Subsequently, the dressings were enriched with active compounds and the rate of gradual releasing of the substances into model environment was monitored. In the case of enzymes, their proteolytic activity was also tested after their incorporation to the wound dressings. Furthermore, the prepared bioactive wound dressings were analyzed for possible cytotoxic effect on human keratinocytes. Finally, the wound dressing with combined content of active substances was created and also characterized for the rate of substance release, proteolytic activity and cytotoxicity. Antimicrobial activity of this wound dressings, against two selected strains of microorganisms: Escherichia coli and Staphylococcus epidermidis, was also evaluated.
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Development of antimicrobial wound healing nanostructure biomaterials
Hanák, Jan ; Wikarská, Monika (referee) ; Márová, Ivana (advisor)
The bachelor thesis is focused on optimizing the production of nanoparticles with a bioactive component - especially on enzymes and antibiotics encapsulated into lipozomes. Nanoparticles with active substances could be used in the field of pharmacy or cosmetology. In the theoretical part the composition of the skin, skin injuries, and wound covers are described. The materials used for the preparation of nanoparticles and nanofibers are characterized as well as processes to produce such nanomaterials. Enzymes, antimicrobial drugs, and methods for determining antimicrobial activity are mentioned in this section, too. The practical part deals with the characterization and optimization of the formation of lipozome particles with encapsulated enzymes. The formed particles were divided into two groups. The first type of particles were pure lipozomes, while the second ones enriched by polyhydroxybutyrate to increase the encapsulation efficiency and improve the stability of the particles. The enzymes bromelain and lysozyme were selected for the highest encapsulation efficiency. The encapsulation of these two enzymes took place in two ways - by thin film hydration and by sonication method. Both methods showed high encapsulation efficiency, but the sonication method proved to be unsuitable due to enzyme denaturation. The proteolytic activity of the released enzymes from the particles was measured. The stability of the particles was monitored in a physiological and aquatic environment by measuring the zeta potential for up to 28 days. Moreover, the enzymes, ampicillin, and particles created by sonication were subsequently subjected to antimicrobial tests by using two test bacterial strains. Finally, alginate particles with encapsulated enzymes were also formed, then the encapsulation efficiency and proteolytic activity of the enzyme were measured. In this study also other methods of encapsulation of active substances are outlined, which could be possibly used to produce nanomaterials suitable for wound healing.
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The application of electrochemical impedance spectroscopy for investigation of enzyme kinetics
Pospíšilová, Michaela ; Sedláček, Jiří (referee) ; Fohlerová, Zdenka (advisor)
The application of electrochemical impedance spectroscopy (EIS) in theory and experiment for investigation of redox enzyme kinetics has been already described. The impedance studies are quite sensitive to the changes at the interface caused by adsorption of charged species and therefore can be used to study the kinetics of adsorption of macromolecules and also enables us to study enzyme kinetics since the action of the enzymes on their substrates involved modification and generation of charged species. In this work, we would explore the potential of electrochemical impedance technique to follow the kinetics of glucose oxidasesubstrate reactions on the immobilized surface. The enzyme will be allowed to interact with different concentrations of its substrate and the resulting reaction will be recorded in real time. Changes in the imaginary component of the impedance at various substrate concentrations will be expected to follow MichelisMenten kinetics. The experimental procedure should be simple and can be carried out directly in buffer solutions most suited for the required interaction to take place, without the requirement of any additional redox probes.
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Encapsulation of some enzymes into organic particles
Hazuchová, Eva ; Obruča, Stanislav (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on encapsulation of some enzymes into organic particles. The theoretical part is aimed to enzymes, their general characteristics, structure, activity and use, and to encapsulation techniques. Some methods used for analysis of particles, namely the determination of the distribution of particle size using dynamic light scattering, and determining the stability of the particles by the zeta potential, are described too. The experimental part describes methods of preparing particles, methods of determining the encapsulation efficiency and methods for determining particle size and stability. During the experiment part four types of enzymes were encapsulated, partly by manual preparation and partly by encapsulator. Encapsulation efficiency, size and stability of prepared particles were determined too. The particles were exposed to artificial intestinal, gastric and bile juices, as well as the effect of model foods. Subsequently, their long-term stability was observed.
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Enzymy pro aktivaci protinádorových proléčiv
Kubináková, Nikol
The current issue of oncological therapy lies in its effectiveness and severe side effects. Despite the most modern devices available for treatment methods, cancer treatment is very challenging. Chemotherapy uses highly toxic substances, which results in side effects that can only aggravate the patient. These side effects very often limit the dose of the drug, so that it is not possible to achieve such a concentration that would completely eradicate the tumor in the body. The answer to the question of how to overcome or reduce the occurrence of side effects in cancer treatment is prodrug therapy, which includes less toxic substances/drugs. This therapy aims to target cytotoxic drugs to a non-toxic derivative or prodrug. Prodrugs are inactive and bioreversible derivatives of active drugs that function as a concept for improving active drugs. After administration of the prodrug, this prodrug is selectively activated by enzymes (Cytochromes P450), resulting in the regeneration of the toxic parent drug at the site of the tumor.
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Teplotní adaptace trávicích enzymů želvušek žijících v trvale chladném prostředí
JOZIĆ, Nikola
This thesis presents a literature review of the phylum Tardigrada, exploring the impact of cold temperatures on living organisms in general and focusing on adaptations of organisms to cold and freezing, particularly in terms of enzymatic adaptations. The experimental part of this work involves comparing the enzymatic activity of the -glucosidase enzyme in tardigrades from Arctic glacier (Svalbard) and soil tardigrades from a temperate climate (Czech Republic) at different temperatures. The results showed higher enzymatic activity in tardigrades from a temperate climate at all measured temperatures, suggesting that the enzymes of glacier dwelling tardigrades are not cold adapted.
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Alkaloids of Dicranostigma franchetianum (Prain) Fedde and their selected biological activity
Wijaya, Viriyanata ; Opletal, Lubomír (advisor) ; Šmejkal, Karel (referee) ; Klouček, Pavel (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy and Pharmaceutical Botany Candidate: Viriyanata Wijaya, M.Sc. Supervisor: Prof. RNDr. Lubomír Opletal, CSc. Title of Doctoral Thesis: Alkaloids of Dicranostigma franchetianum (Prain) Fedde and their selected biological activity Dicranostigma franchetianum (Prain) Fedde (Papaveraceae) is one of the representatives of the small genus Dicranostigma Hook. f. & Thomson. D. franchetianum (Prain) Fedde has been selected for the phytochemical investigation according to the screening study. In the primary screening of the alkaloid extract for cholinesterases inhibition, the inhibitory value was high (hAChE/hBChE, IC50 g/ml; 1.67 0.11 and 3.85 0.31) and together at least 15 alkaloids were found in the extract. The primary ethanol extract was prepared from 11.8 kg of dry herb (Garden of Medicinal Plants, Faculty of Pharmacy in Hradec Kralove). Using common chromatographic methods, 21 isoquinoline alkaloids of various structural types were isolated. All compounds have been identified using various spectrometric techniques (GC-MS, HPLC- MS, and 1D- and 2D-NMR, optical rotatory. The alkaloids obtained in sufficient amounts were determined for human acetylcholinesterase (hAChE), human butyrylcholinesterase (hBChE), and...
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