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Connection between stress resistance of bacteria to temperature-mediated stress and PHA accumulating ability
Nováčková, Ivana ; Benešová, Pavla (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis deals with the involvement of polyhydroxyalkanoates (PHA) into the stress response of bacteria, specifically when they are exposed to temperature-mediated stress. The theoretical part focuses on the stress response of bacteria in general, the mechanism of response to temperature-mediated stress and PHA involvement into the stress response. The aim of experimental work was to study the correlation between the amount of PHA in the biomass of the bacteria and their viability after exposure to temperature-mediated stress. PHA producing bacterial strains Cupriavidus necator H16, Burkholderia cepacia and Burkholderia sacchari and non-producing mutant strain Cupriavidus necator PHB4 were used for experimental work. Bacteria accumulated poly(3-hydroxybutyrate) (PHB), the amount of PHB in the biomass was determined by gas chromatography with FID. Viability of live bacterial cells and cells after exposure to temperature-mediated stress was determined by using flow cytometry. It turned out that the content of PHB in biomass assists bacteria to resist the freezing shock. By comparing the viability of bacteria C. necator H16 and C. necator PHB4 after exposure to multiple stresses it also has been observed that PHB increased the resistance of cells against temperature-mediated stress and stress caused by low pH.
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Preparation and stability of core-shell type organic micro- and nanoparticles
Byrtusová, Dana ; Benešová, Pavla (referee) ; Márová, Ivana (advisor)
This bachelor thesis is focused on possibilities of encapsulation polyphenolic compounds in core-shell type organic micro- and nanoparticles. The theoretical part is aimed at characterization of polyphenolic compounds, their biological effects and their possibilities of encapsulation into various types of particles. The experimental part deals with the content of polyphenolic compounds, flavonoids and anthocyanins in guarana, ginseng ang goji. Extracts from these samples were packed into liposomes and polysaccharide particles. Encapsulation´s effectiveness was determined spectrophotometrically. The stability of particles was monitored for size and stability by dynamic light scattering. The particles were exposed to the model body fluids and model food.
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Microcalorimetric study of bacterial degradation of oil and its products
Bola, Tomáš ; Benešová, Pavla (referee) ; Krouská, Jitka (advisor)
This bachelor thesis deals with a study of the oil degradation as one of the possible products of petroleum using bacterial stems Pseudomonas Putida a Pseudomonas Chlororaphis. The work is motivated by the research of usability of a method which has not been used for those purposes so far – calorimetric analysis. Using IČ spektrometry (ATR), calorimetry and UV-VIS spectrometry was studied a degree and a character of interactions. Calorimetric analysis showed that the Pseudomonas Putida has bigger abbility of the degradability than Pseudomonas Chlororaphis. There was also detected that the optimal concentration for more effective degradation of oil fuel is 5 g/l and 20 g/l. By measurement of the absorbance deppending on time was verified that the bigger is the concentration of the sample, the higher is the resulted absorbance. Using UV-VIS analysis was determined a quantity of biomass. Using IČ spectometry in the ATR mode (reduced reflectance) was detected structure of the sample and also verified that the bacterial cultures Pseudomonas Putida and Chlororaphis degradated entire amount of petroleum.
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Characterization of active substances in several kinds of beer
Benešová, Pavla ; Obruča, Stanislav (referee) ; Márová, Ivana (advisor)
The aim of this bachelor thesis is to analyse biologically active compounds, especially of phenolic character, in 10 kinds of alcohol-free beer in comparison with alcoholic beer. In theoretical part basic steps of brewery technology are described including alcohol-free beer production. Characteristic compounds occurring especially in Czech beer e.g. proteins, phenolics, bitter substances and also methods for their analysis are discussed as well. Microfluidic electrophoresis (Experion, BioRad) was used to beer protein analysis. Phenolic compounds, flavonoids, technological characteristics and antioxidative activity were measured by spectophotometry. Individual phenolics of selected beers were identified and quantified by LC/ESI-MS analysis too. In alcohol-free beers similar concentrations of biologically active compounds were found in comparison with alcoholic beers, especially in the case of beer phenolics.
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Biotechnological production of polyhydroxyalkanoates using wastes of coffee production
Vašíčková, Monika ; Benešová, Pavla (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis is focused on study of production of polyhydroxyalkanoates (PHA) by bacteria Burkholderia cepacia and Burkholderia sacchari. Production of PHA has been studied using spent coffee grounds hydrolysates which have been prepared by different aproaches. In the thesis is compared standard method of coffee ground hydrolysis by mineral acid followed by enzymatic hydrolysis. Hydrolysates which have been produced were analyzed in terms of concentration of saccharides and efficiency of hydrolysis. The best producer of PHA (based on results) was bacterium Burkholderia cepacia, in the medium which has been prepared by hydrolysis of spent coffee grounds extracted by 60% solution of ethanol. Biomass yield of this cultivation was 3,553 g/l with 32,472% PHA content. This PHA contained 6,09% 3-hydroxyvalerate. In the other experiment, we verified alternative hydrolysis of spent coffee grounds – by commercially available enzymes. We used cellulase, hemicellulase and the enzymatic cocktail which has been produced by mould. Hemicellulase was the most effective hydrolytic enzyme and its application resulted in production of the highest amount of biomass – 5,708 g/l. In this cultivation, only homopolymer PHB has been which is probably caused by the fact, that during the enzymatic hydrolysis levulinic acid or any other potential precursor of 3HV is formed.
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Trough media education to media literacy
Benešová, Pavla ; Kasíková, Hana (advisor) ; Valenta, Josef (referee)
The theme of bachelor thesis "Through Media Education to Media Literacy" is a new subject in field of education: media education. The first part is briefly overlooking the history of media education worldwide and implementation of media education in Czech environment. The thesis introduces questions of media literacy and presents media education as one of Cross-curricular Subjects as it is understood by Framework Education Programme of Ministry of Education of the Czech Republic. From strictly didactic point of view the media education is presented in chapter Principal Elements of Methodology Construction and in chapter Method of Media Education. Second part deals with education of teachers in field of media education and projects of non-governmental agencies providing their education. This part of thesis also offers an overview of methodical handbooks published to be used while teaching media education in Czech system of schools. The last chapter compares three most important handbooks mentioned above.
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School on the move
Benešová, Pavla ; Kuhnová, Věra (advisor) ; Pokorný, Ladislav (referee)
The thesis examines physical activities in a primary school both inside and outside of the classroom. In several randomly selected schools, I watched how many hours a day the pupils spend time moving while learning and taking breaks. Research has shown that pupils keep the activity at a minimum in school, mostly only to scheduled moves. My endeavour was to propose ways to enrich the school day by physical activity. Based on this finding, in a selected school, I conducted an experiment that was supposed to bring more physical activity into classes and during school breaks. Whether the active time spent in the school environment managed to increase through the new proposition, shows the results of the experiment. Part of this work are practical ideas for implementation of physical activities into the classroom, and tips for classrooms and school corridors equipment.
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Nurses' duty of conifidentiality and the consequences of its breach
Benešová, Pavla ; Prošková, Eva (advisor) ; Konečná, Jana (referee)
The aim of my thesis was to find out what is the legal awareness of nurses in the issue of confidentiality. Furthermore, it was examined whether nurses know what are the consequences for unauthorized access to the medical records. The theoretical part of this thesis describes the mandatory confidentiality issue and its consequences. And it defines the concepts of personal and sensitive personal data. In addition the conditions under which it is required to break confidentiality and the consequences of breach of legal obligation are mentioned. Describe the legal responsibility of nurses. In the practical part of the work I use qualitative methods of data collection, interview, for me to respond to major and minor research questions. I use interviews conducted with nurses, for doing analysis of the case and to describe the measures taken by the employer at the Municipal Hospital in Litoměřice.
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The probation and mediation as a form of work with juvenile offenders
Benešová, Pavla ; Lorenzová, Jitka (advisor) ; Sedláčková, Daniela (referee)
The aim of diploma thesis "Probation and mediation as a form of work with juvenile offenders" is to elaborate the social pedagogic aspects of probation and mediation activities. It is intended to be a theoretic study and is divided into five chapters. The opening chapter is concerned for causes of delinquent beaviour and offers an enumeration of factors causing the uprise of this kind of behaviour among youth. The second chapter aims at possibilities of juvenile criminality prevention. The third chapter is employed with an institution of Probation and Mediation Services, which plays an important role on the field of terciary prevention of juvenile criminality. Fourth chapter treats with concrete ways of work of probation officers with their clients. Their work is analysed in the last chapter emphasising the social pedagogic potential of probation and mediation activities. It also mentions a possibility of social support in the work of probation officers.
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Bioproduction, Characterization and Application of Selected Biopolymers
Benešová, Pavla ; Pekař, Miloslav (referee) ; Čertík, Milan (referee) ; Márová, Ivana (advisor)
Proposed doctoral thesis deals with microbial production of biopolymers (polysaccharides and polyesters) using renewableble inexpensive resources as carbon and nitrogen source. At first, production of extracellular metabolites, mainly pullulan and poly-L-malic acid, by polymorphic microorganism Aureobasisium pullulans was studied. Further part of thesis was focused on production of bacterial biopolyesters polyhydroxyalkanoates (PHA). PHA are produced in the form of intracellular granules by various bacteria species. During the study of PHA production, sythesis of poly(3-hydroxybutyrate) and technologically significant copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) was investigated as well with regard to waste substrate utilization in biorafinery concept. Spent coffee ground seemed to be applicable waste in biorefinery proces. One of the coffee wastes used for PHA production was spent coffee ground hydrolysate which was converted to PHA by Burkholderia cepacia. Moreover, copolymer of 3HB and 3HV was accumulated when SCGH was used as a sole carbon source, no precursor had to be added. Another coffee waste, which was utillized by Cupriavidus necator H16, is represented by oil extracted from spent coffee grounds. Hydrolysis proces of waste substrates had to be optimized, to reach the highest possible saccharide concentration. Due to the hydrolysis process several toxic compounds can be formed, hence, influence of hydrolysis procesings on polyphenols content in hydrolysates was monitored. Waste substrates were also tested as potentional complex nitrogen source for PHA production. Hydrolysates of cheese whey and chicken feather were used as inexpensive complex nitrogen source for PHA production by C. necator, when waste frying oil was used as a carbon substrate. The finnal part of the thesis is dealing with possibilities how the PHA can be processed into atractive aplication forms. Liposomes with partial content of PHB had been prepared, moreover, for improving of antioxidant activity and antimicrobial effect the waste coffee oil was added. Micro-fibrous PHB materials were prepared for the purposes of water filtration. Higher filtration effectivity was observed when composite PHB fibrous material containing active charcoal or metal oxides was used. Important part of thesis is represented by study of degradation processes of prepared PHA-based aplication forms. Various microorganisms were tested on PHA depolymerases production. Biodegradability of PHA materials was tested in of composting conditions by standard composting test IS/ISO 20200. Changes in mechanical properties of polyurethan, where polyol was partialy replaced by PHB, were tested depending on the exposure of composting condition.
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