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Proposal of systems for targeted transoprt of compounds of sweet taste
Demová, Radoslava ; Hlaváček, Viliam (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on possibility of immobilization sweeteners on polymeric system. In the theoretical part information about sweeteners, carrier systems and the possibility of immobilization technique were reviewed. In the experimental part preparation of nanoparticles of polyhydroxybutyrate (PHB) was optimized. The PHB particles were characterized by dynamic light scattering (DLS). PHB particles were activated by plasma treatment. The surface morphology was studied using infrared and Raman spectroscopy. Immobilization of following substances with a sweet taste was tested: glucose, fructose, sucrose, mannitol, sorbitol and erythritol. From these compounds only sucrose was immobilized on PHB particles successfully. Prepared immobilized particles were exposed to the artificial stomach juice, intestinal juices and bile acids and amount of released sucrose was monitored. Finally, long-term stability prepared particles was measured and also the amount of sucrose released was determined.
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Preparation of nanoparticles and nanofibers with antimicrobial components
Kubišová, Veronika ; Slaninová, Eva (referee) ; Skoumalová, Petra (advisor)
This thesis addresses the problem of inadequate current wound therapy and presents a solution in the form of nanomaterial-based wound dressings (coverings). Specifically, it focuses on the development and characterization of various nanofibrous materials with integrated liposome particles that could serve as a source of therapeutic agents and be useful specifically in the field of a wound therapy. The review in the introductory part of the thesis first focused on the mentioned problem, which describes the shortcomings of existing conventional dressing materials. A description of human skin from an anatomical and functional point of view was not omitted, nor was the wound healing process itself. Different types of covering materials were also presented. However, a large part was focused on nanomaterials and their use in the field of the skin wound therapy. The nanomaterials mentioned were mainly liposome particles and nanofibres, as the experimental part of the work was focused on these structures. The description dealt with the characterization of these structures as well as the biopolymers used for their preparation. However, the aim was to prepare nanofibers with liposome content, so the method of forming such systems was described. Various therapeutic agents are also an integral part of the covering materials, especially those that suppress the development of infection and reduce wound pain; therefore, the search focused on the antibiotic ampicillin and the analgesic ibuprofen. The experimental part was devoted to the actual production of nanofibers with liposomes and also to the qualitative demonstration of the presence of liposomes in electrostatically prepared nanofibers. The selected polymeric components of these systems were polyhydroxybutyrate (PHB) and gelatine. However, the nanofibers and liposome particles (as well as combined liposomes with PHB) were first prepared in their own form and characterized mainly in terms of the gradual release of the drug substances. The results obtained were then compared with those of therapeutic drug delivery via combined nanofibrous structures with liposomes. For these combined structures, the aim was to achieve synergy in drug delivery between these systems. The aim of this work was to create a biomaterial covering with the controlled drug release. The drugs contained in these materials were the aforementioned ampicillin and ibuprofen. An important part of the work was then to determine the safety of the prepared materials which were tested for cytotoxicity, where the tests performed were MTT test and LDH test. And the actual wound healing ability of the nanofibers was then monitored in a scratch test or a "wound healing" test. At the end of the paper, recommendations for future work on this topic are given.
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Influence of biodegradable polymers microplastics on properties of soil organic matter
Denková, Pavla ; Řezáčová, Veronika (referee) ; Kučerík, Jiří (advisor)
The microplastics contamination is an issue observed in all environmental compartments. Currently, the trend is to replace the synthetic polymers by biodegradable ones, which are expected to degrade faster due to omnipresent microorganisms. However, this assumption may not correspond with the reality. In nature, biodegradation can last long or not whatsoever and the microparticles of biodegradable polymers can remain in soil the same way as particles of synthetic polymers. The effect of microparticles of biodegradable polymers on soil´s biota has already been studied, but the effect on the properties of soil and water in the soil has not been elucidated yet. This thesis deals with the influence of microparticles of biodegradable plastics on the physico-chemical properties of soil organic matter, especially on the behavior of water in the soil system under arid and semiarid conditions. Microparticles of poly(R-3-hydroxybutyrate), abbreviated as P3HB, were used as a model biodegradable polymer and were introduced into the soil in various concentrations. To determine the evaporation enthalpy of water and stability of water bridges that affect (stabilize) the physical and chemical structure of soil organic matter Differential Scanning Calorimetry (DSC) was used. Simultaneously was also measured their effect on water holding capacity of soil organic matter. Under experimental conditions, P3HB microplastics were found to reduce evaporation enthalpy, which facilitates drying of soil organic matter. Nevertheless, their effect on final water holding capacity was relatively low. Besides, we observed a negative effect on the stability of water molecule bridge that connect segments of soil´s organic components. Thus, P3HB microparticles in soil could pose a risk due to their influence on the water retention mechanism in soil under arid and semiarid conditions.
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Preparation and characterization of nanofiber wound dressings
Jiroušková, Pavla ; Uhlířová, Renata (referee) ; Skoumalová, Petra (advisor)
The presented bachelor thesis deals with the preparation and characterization of nanofiber wound dressings. The materials used for the preparation of nanofibers were poly(3-hydroxybutyrate) and gelatin. Active substances, such as ampicillin and ibuprofen, were incorporated into the wound dressings. In the theoretical part of the work skin, wound healing, and nanofibers and their properties were described. This part also deals with the use of nanofibers as wound dressings. Additionally, it depicts materials suitable for the preparation of nanofiber wound dressings. Furthermore, this section contains a review focused on various methods of preparation of nanofibers, drugs that can be incorporated into the fibers and the subsequent determination of the safety of the prepared dressings. In the experimental part, nanofiber wound dressings containing ampicillin and ibuprofen were produced. The maximum possible amount of active substance feasible to be incorporated into the nanofibers has been determined. Subsequently, combined wound dressings were prepared to study the release rate of active substances into the model environments. The release in the time intervals was determined spectrophotometrically. Finally, antimicrobial activity assays were performed to test the effect of dressings containing ampicillin on selected strains of microbiirganism: S. Epidermidis, E. Coli and M. Luteus
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Utilization of physicochemical and spectroscopic techniques in study of stress-response of cyanobacteria
Skoryk, Maksym ; Šedrlová, Zuzana (referee) ; Sedláček, Petr (advisor)
Tato bakalářská práce je soustředěná na zkoumaní cyanobakterií, vystavených hyper a hyposmotickým stresům. Na základě literární rešerše byly navžené vhodné analytické metody určené ke studiu dvou modelových organismu - Synechocystis sp. PCC 6803 a Synechocystis sp. salina Wislouch CCALA 192. Celkem čtyři metody byly použité k charakterizaci osmoticky zatížených bakterií. Průtoková cytometrie byla použita k vyhodnocení viability. Fluorescenční sonda SYTOX Blue poskytla důvěryhodnou informaci ohledně viability bakterií. Propidium jodid naopak poskytl nezřetelnou informaci. Optické vlastnosti cyanobakterií byly prozkoumané pomoci UV-VIS absorpčních a turbidimetrických měření. Termogravimetrická analýza byla použita pro mapování změn obsahu vody v osmoticky stresovaných buňkách. Tato metoda ukazala. že PHB-positivní Synechocystis sp. PCC 6803 jsou pravděpodobně vice odolné vůči hyperosmotickým stresům než PHB-negativní. Plynová chromatografie byla použita ke kvantifikaci vnitrobuněčného PHB, který činil přibližně 1-2 % suché hmoty PHB-positivních buněk.
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Preparation and application of nanoparticles and nanofibres with natural UV filters
Plachá, Monika ; Skoumalová, Petra (referee) ; Márová, Ivana (advisor)
The presented diploma thesis is focused on preparation of nanoparticles and nanofibres with natural UV filters. Liposomes with encapsulated aqueous, ethanol and lipid extracts were prepared. Nanofibers from PHB containing lipid extract were prepared too. As a part of this work, an overview of natural sources with potential effects as UV filters were introduced. Moreover, nanoparticles and nanofibers and methods of their characterization were described. Size, polydisperse index and colloid stability of prepared nanoparticles were characterized via DLS. In experimental part aqueous, ethanol and lipid extracts were prepared from roasted coffee, green coffee and cascara. These extracts were spectrophotometrically characterized for the content of polyphenols, flavonoids, antioxidant activity, tannins and their SPF. Liposomes and liposomes containing PHB with these extracts were prepared and the encapsulation effectivity, short–term and long–term stability as well as SPF of nanoparticles were determined. Nanofibers from PHB containing lipid extracts were prepared via electrospinning and forcespinning methods. Prepared nanofibers were examined via FTIR–ATR. Antioxidant activity, short–term and long–term stability were determined spectrophotometrically. From selected nanoparticles, emulsions and gels were prepared and their SPF was also determined. Three types of emulsions with the best SPF were selected and tested on volunteers. Sedimentation stability of emulsions was tested by analytical centrifuge. Finally, cytotoxicity of selected nanoparticles and nanofibers was tested via MTT assay using human keratinocytes.
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Analytical methods for determination of polyhydroxyalcanoates content in bacterial cells
Černayová, Diana ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
This bachelor thesis deals with methods that are used for determination of polyhydroxyalkanoates (PHA) in cells of bacteria Cupriavidus necator H16. To the applied methods belong Fourier transform infrared spectroscopy, Raman spectoscopy, turbidimetry and thermal analysis (thermogravimetric analysis and differential scanning calorimetry). The results of each method were compared to amount of PHA determinated by gas chromatography. Fourier transform infrared spectroscopy and Raman spectroscopy revealed a linear relation between ratio of characteristic peaks and amount of PHA. FTIR enabled to determine not only the quantitative amount of PHA, but the quality of the sample (cristallinity, amorphous state) as well. Another method was turbidimetry, which has shown correlation between dispersion of UV-VIS light to amount of PHA in bacterial cells. Thermogravimetric analysis was the most effective technique for determination of PHA. The dependence of weight loss of PHA on its content (found by gas chromatography) was determined with the highest accuracy to gas chromatography. Tha last method- DSC was not alternative replacement for quantitative PHA determination. However, DSC indicated crystalline and amorphous state of the polymer.
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Study of selected antibiotics sorption on polyhydroxybutyrate
Amrichová, Anna ; Doležalová Weissmannová, Helena (referee) ; Řezáčová, Veronika (advisor)
The bachelor thesis deals with the issue of sorption of antibiotics on microplastics. Antibiotics and microplastics are now a highly studied topic, due to the positive and negative properties. Antibiotics with groups of sulfonamides and quinolones were selected for the experiment. From sulfonamides, sulfamethoxazole and trimethoprim were selected, and from the quinolones, enrofloxacin and ciprofloxacin, which have already been the impetus for some microplastic studies. Biodegradable polyhydroxybutyrate was used as the microplastic. Two experiments were performed in the laboratory. The first aimed to monitor the time dependence of antibiotic sorption on polyhydroxybutyrate, the second the concentration. The concentration dependence was subsequently evaluated using the Freundlich and Langmuir isotherms. The method of liquid chromatography with mass spectrometry was used for analyzes.
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Crystallization of microbial polyesters - methods and biological consequences
Liczka, Jan ; Kalina, Michal (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis was to test suitable methods for preparation of amorphous form of microbial polyesters and to perform model experiment to study a kinetics of crystallization of prepared polyesters. The theoretical part deals with PHA characteristics and various methods of isolating PHB granules from bacterial cells. In the experimental part, measuring techniques such as analytical centrifugation, dynamic light scattering (DLS) and Fourier transform infrared spectroscopy (FTIR) were tested to characterize the properties of PHB granules. Furthermore, an experiment was performed to isolate native PHB granules from Cupriavidus necator using lysozyme, deoxyribonuclease and cell disruption by ultrasonification. The results of the analyzes showed that the granules were either isolated amorphous only in the wet and crystallized upon drying, or that the granules were completely amorphous but not isolated from the bacterial cell.
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Biodegradation of polyesters and other polymers in soil and compost
Papala, František ; Smilek, Jiří (referee) ; Obruča, Stanislav (advisor)
This diploma thesis studies the biodegradation of polyhydroxybutyrate (PHB) and polylactic acid (PLA) within soil and compost environments. The experimental part of this work is concerned with the differences in degradation across environments and the effects of degraded polymers on the environment itself. Experiments revealed that higher temperature and potentially also higher humidity substantially affect the rate of biodegradation. A significant degradation of all the polymers was observed in the compost medium, while polymers within the soil medium did not demonstrate any visible change nor change in mass after the 8th week of experiment. Even though SEM images revealed slow microbial degradation of soil PHB, this was not seen in any of the soil-incubated PLA samples. This might indicate primarily abiotic mechanism of PLA degradation. Germination and grow tests, which used the prepared polymer-enriched compost as a substrate and fertilizer, respectively, did not in most cases reveal any significant differences between individual plants across substrates. This seems to imply that neither the examined polymers nor their degradation products generated during composting are phytotoxic.
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