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Remote control and visualization of bus-systems KNX and DALI for building management
Holub, Jiří ; Macháček, Jan (referee) ; Bátora, Branislav (advisor)
The thesis is focused on bus systems and their control by GUI. Basic energy management principles and the mostly known bus systems are analysed in the first – theoretical part of the paper. The theoretical part is then focused on software tool ETS 4, which is used for activation of system installation KNX. Software tool SmartServer, which is used for communication between bus system KNX and web interface, is described at the end of the first theoretical part. Two laboratory boards assembled of KNX and DALI parts were constructed in the second – practical part. Three laboratory tasks, which are prepared so they are related to theoretical description in previous part, were submitted in next step. Aim of the first task is to introduce basics of the board and PriOn interface. The second task is focused on DALI components and temperature control with PriOn regulating device. Creating of basic visualization and controlling of the board with SmartServer is described in the last task.
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Large Extracellular Vesicles in Cell Culture and Blood: Role in Prion Transmission and Detection by Flow Cytometry
Soukup, Jakub
Prions (PrP) are the main cause of neurodegenerative diseases such as Scrapie in sheep, bovine spongiform encephalopathy, chronic wasting disease in deer, and Creutzfeldt-Jakob disease in humans. Although the cellular PrP (PrPC ) is involved in many cellular processes, its precise function still needs to be discovered. The disease is caused by the accumulation of a pathological form of PrP (PrPTSE ), which is caused by direct contact of PrPTSE and PrPC . PrP is anchored in the membrane by GPI and can be transmitted by cell-to-cell contact, tunnelling nanotubes, or extracellular vesicles (EVs). EV factions are divided by different biogenesis into exosomes, microvesicles, and apoptotic bodies. PrPTSE was found in exosomes and microvesicles, but these fractions were never compared to each other. The first aim of the doctoral thesis is a comparison of PrP content, prion-converting activity and infectivity in these fractions on CAD5 and N2a-PK1 cellular models of infection. We isolated a fraction of large EVs (20,000× g) and small EVs (110,000× g) by centrifugation from a conditioned medium. We characterised EVs by cryo-electron microscopy and western blot with Alix, TSG-101, CD63, CD9, and HSP70 markers. The contamination from other cellular compartments was checked by calnexin. EV fractions differed...
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Large Extracellular Vesicles in Cell Culture and Blood: Role in Prion Transmission and Detection by Flow Cytometry
Soukup, Jakub ; Holada, Karel (advisor) ; Šebestová Janoušková, Olga (referee) ; Živný, Jan (referee)
Prions (PrP) are the main cause of neurodegenerative diseases such as Scrapie in sheep, bovine spongiform encephalopathy, chronic wasting disease in deer, and Creutzfeldt-Jakob disease in humans. Although the cellular PrP (PrPC ) is involved in many cellular processes, its precise function still needs to be discovered. The disease is caused by the accumulation of a pathological form of PrP (PrPTSE ), which is caused by direct contact of PrPTSE and PrPC . PrP is anchored in the membrane by GPI and can be transmitted by cell-to-cell contact, tunnelling nanotubes, or extracellular vesicles (EVs). EV factions are divided by different biogenesis into exosomes, microvesicles, and apoptotic bodies. PrPTSE was found in exosomes and microvesicles, but these fractions were never compared to each other. The first aim of the doctoral thesis is a comparison of PrP content, prion-converting activity and infectivity in these fractions on CAD5 and N2a-PK1 cellular models of infection. We isolated a fraction of large EVs (20,000× g) and small EVs (110,000× g) by centrifugation from a conditioned medium. We characterised EVs by cryo-electron microscopy and western blot with Alix, TSG-101, CD63, CD9, and HSP70 markers. The contamination from other cellular compartments was checked by calnexin. EV fractions differed...
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Knowledge of Pupils Secondary School in the Area of Selected Zoonoses
Svobodová, Barbora ; Pavlasová, Lenka (advisor) ; Říhová, Dagmar (referee)
The subject of the dissertation deals with the secondary school pupil's awareness of zoonoses. Zoonoses are the diseases transmited from animals to human beings. Those are one of the topics of biology study currently. The main aim of the dissertation is theoretical definition of the matter in the first place. For knowledge testing have been chosen these four illness: Toxoplasmosis, Lyme boreliossis, Rabies and Creuzfeldt-Jakobo disease. With these chosen illness the main research goal is to provide a comprehensive collection of information about the details such as disease transfer, spread of the disease, disease development, cure and avoidance. The research part of the dissertation tries to find out the extent of knowledge about the chosen kinds of zoonoses among the pupils at specialized secondary schools. As a tool of quantitative research has been used a pupil's didactic test. As based on the evaluated results we may say that the pupil's knowledge at explored schools is relatively satisfactory.
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Specific prion protein antibodies characterisation and use in diagnostic
Šafaříková, Eva ; Holada, Karel (advisor) ; Mareš, Michael (referee) ; Živný, Jan (referee)
Transmissive spongiform encephalopathies (TSEs) are neurodegenerative diseases characterized by depositions of abnormally folded prion protein (PrPTSE ) in brain. PrPTSE is at present the only specific biochemical marker of human and animal TSEs. Diagnostic tests are based on the detection of PrPres after proteinase K digestion of brain homogenate using Western blot or on the immunohistochemistry of fixed brain tissue, which are both difficult and time consuming. In this work we focused on development of a new type of tests based on PrP detection without need of proteinase K digestion. As deposits of PrPTSE remain in the body for a long time, there is a substantial chance of them being nonenzymatically modified by glycation. The detection of glycated PrPTSE may have a potential to serve as a diagnostic marker. We prepared monoclonal antibodies specific for carboxymethyl lysine/arginine modified prion protein. Bacterially expressed and purified recombinant human prion protein (rhPrP) was modified by glyoxylic acid that introduces carboxymethyl groups on lysine and arginine residues present within the molecule of the protein. Modified rhPrP (rhPrP-CML) was used for immunization of laboratory mice and hybridoma cells were prepared. Screening of cell supernatants resulted in the selection of 4...
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Study of the photodynamic inactivation of prions by phthalocyanines.
Kostelanská, Marie ; Holada, Karel (advisor) ; Zimčík, Petr (referee) ; Kolářová, Hana (referee)
Transmissive spongiform encephalopathies, also called prion disorders, are fatal neurodegenerative diseases affecting mammals. In patients, the pathological prion protein (PrPTSE ) accumulates in CNS and causes death. Prions possess high binding affinity to surfaces. Moreover, they are highly resistant to conventional sterilization procedures which rise the risk of nosocomial transmission from patients in subclinical stage of prion disease through medical tools. In the thesis, we evaluate the efficiency of photodynamic inactivation (PDI) for prion decontamination. The PDI is induced by photoactivation of phthalocyanine (Pc) derivates AlPcOH(SO3)2, SiPc(OH)2(SO3)1-3 or ZnPc(SO3)1-3. Pc exposed to light generate reactive oxygen species, mainly singlet oxygen (O2(1 ∆g)). Production of O2(1 ∆g) in aqueous solution was confirmed by iodide method, quenching by NaN3 and oxidative degradation of uric acid. The photoactivation of Pc in infectious brain homogenate led to elimination of PrPres signal (= proteinase K-resistant PrPTSE fragment) below the detection limit of western blot by using nanomolar AlPcOH(SO3)2 concentration. The complete elimination of PrPres signal was accompanied with total protein concentration decrease by a maximum of 20% in brain homogenate No signs of protein fragmentation or...
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Vliv šlechtění geneticky podmíněné odolnosti vůči scrapie na užitkové a reprodukční vlastnosti plemene suffolk
Jahodová, Marie
Submitted thesis describes breeding Suffolk breed as to scrapie problematics. The thesis summarizes the population of Suffolk breed in the Czech Republic and describes development of reproductive and productive properties. It describes occurrence of alleles influencing the resistance sheep against scrapie. The thesis describes reason, pathogenesis, symptoms, diagnostics and prevention of scrapie disease. It also mentions other prion diseases. The selection to scrapie-resistant alleles positively influences population of Suffolk breed in occurrence of resistant alleles and concurrently with improvement in productive and reproductive performance over the last fifteen years. It can be said that selection for the PRNP genotype does not have a negative impact on these indicators.
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Vliv frekvence genotypů prionového proteinu na reprodukční a produkční vlastnosti ovcí v ČR
Žváčková, Pavla
The aim of the diploma thesis was to find out how the frequency of the prion protein genotypes affects the production and reproductive characteristics of sheep in the Czech Republic. We also used graphs to evaluate changes in the frequency of allele of prion protein genotypes from 2003 to present in ten selected breeds of sheep genotyped within the breeding program for scrapie resistance in the Czech Republic. We also dealt with administrative interventions in the genotyping process by the state and recognized breeders' associations. In the next part we evaluated the changes in the frequency of the allele in relation to the development of the sheep properties in the Czech Republic. The results of the research were processed by appropriate mathematical and statistical methods. We have found that the sheep production properties are influenced by the allele frequencies of the prion protein genotypes, and that administrative interventions have contributed to changes in the frequencies of the risk groups in some breeds. Further, we have discovered that most of the highest values of utility properties are associated with the risk group R1.
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Knowledge of Pupils Secondary School in the Area of Selected Zoonoses
Svobodová, Barbora ; Pavlasová, Lenka (advisor) ; Říhová, Dagmar (referee)
The subject of the dissertation deals with the secondary school pupil's awareness of zoonoses. Zoonoses are the diseases transmited from animals to human beings. Those are one of the topics of biology study currently. The main aim of the dissertation is theoretical definition of the matter in the first place. For knowledge testing have been chosen these four illness: Toxoplasmosis, Lyme boreliossis, Rabies and Creuzfeldt-Jakobo disease. With these chosen illness the main research goal is to provide a comprehensive collection of information about the details such as disease transfer, spread of the disease, disease development, cure and avoidance. The research part of the dissertation tries to find out the extent of knowledge about the chosen kinds of zoonoses among the pupils at specialized secondary schools. As a tool of quantitative research has been used a pupil's didactic test. As based on the evaluated results we may say that the pupil's knowledge at explored schools is relatively satisfactory.
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Role of expression of cellular prion protein in the differentiation of neuronal cell lines
Kučerová, Johanka ; Holada, Karel (advisor) ; Janoušková, Olga (referee)
Cellular prion protein (PrPC ) is a membrane bound glycoprotein. The protein is expressed in all vertebrates, mainly in the nervous system, but it is present also in the cells of gastrointestinal tract, bone marrow, germ cells and heart. PrPC is necessary for pathogenesis of prion diseases, which are deadly and without the possibility of therapy. The pathogenic isoform of prion protein is formed by changing of secondary structure of PrPC and it's the main constituent of infectious prion particles. Pathological form of prion protein accumulates in brain of infected patients and this process is associated with neurodegradation. Physiological function of PrPC is poorly understood. Knock-out of the PrPC gene (PRNP) is not connected with any noticeable phenotype. Potential functions of PrPC are dispersed, protein may have antiapoptotic effect, it can be involved in ions metabolism or in protection against oxidative stress. Latest results show, that PrPC can play important role in cell differentiation. During the differentiation PrPC can influence the development of cells and their typing. It could affect cell cycle and have an influence on formation of nervous system. Aim of the present study was to elucidate, whether the down-regulation of PrPC or infection with prions has an impact on differentiation of...
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