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Host immune response in neuroinfections caused by parasitic helminths.
Macháček, Tomáš ; Panská, Lucie (advisor) ; Štáfková, Jitka (referee)
Among helmiths, there are some parasites which may dwell in the central nervous system of humans during their ontogenic development. Contrary to the former concept of so called immune privilege of this site, hosts develop the immune response against them. Both immune cells and resident immunocompetent cells (astrocytes, microglia) participate in these immune reactions. The host immune response consists of cellular as well as humoral immunity. This bachelor's thesis gives the general features of the immune response in the central nervous system and describes the immune response during particular helminthic neuroinfections: neuroschistosomiasis, neurocysticercosis and neurotoxocarosis.
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Enrichment techniques employed in phosphoproteomics
Fíla, Jan ; Honys, David (advisor) ; Zdráhal, Zbyněk (referee)
Reversible protein phosphorylation represents one of the most rapid and dynamic posttranslational modifications. Phosphorylation plays a key role in many cellular processes, e.g. cell division, signal transduction, cytoskeleton dynamics, translation regulation, protein targeting and metabolism. Unfortunately, the importance of phosphoproteins is often not mirrored in their abundance so enriching strategies are mostly inevitable. The enrichment can be performed either at the stage of phosphoproteins or at the stage of phosphopeptides. Alternatively, a combination of both strategies can be used. Most enriching strategies are based on chemical modifications, affinity of phosphates to positively-charged chromatography matrix or antibodies. Every method used has its advantages as well as disadvantages. Moreover, satisfactory coverage of phosphoproteome is very often reached by combination of several methods. In the past five years many protocols have been improved or alternatively new approaches have appeared. So in this bachelor thesis, the available enrichment strategies are reviewed. Key words: phosphoproteomics, phosphoprotein enrichment, phosphopeptide enrichment, IMAC, MOAC, titanium dioxide TiO2, aluminium hydroxide Al(OH)3, antibodies
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Determining the occurrence of autoantibodies in the diagnosis of celiac disease
BUDSKÁ, Veronika
The goal of my thesis was to determine an incidence of autoantibodies important for a diagnosis of celiac disease and determine their dynamics in a course of the disease. The determination of antibodies is performed by using serological laboratory tests called immunofluorescence and ELISA. For the diagnosis of celiac disease is important to detect antibodies against endomysium, deaminated gliadin and tissue transglutaminase. Results were discussed on the basis of findings from available literature. In a theoretical part I focused especially on the characteristics of celiac disease. I mentioned the cause of the disease - gluten, I focused more detail on genetics, which is strongly related to the emergence of the celiac disease. Last but not least, I outlined a laboratory diagnosis of celiac disease and the treatment. In a practical part I devoted to laboratory examination of celiac disease. Laboratory tests include tests for antibodies against endomysium IgA by immunofluorescence method. The outcome is evaluated under a fluorescence microscope. Antibodies against deaminated gliadin and tissue transglutaminase are determined in IgA and IgG antibodies by ELISA method. The resulting sample is evaluated photometrically. I conducted antibody testing a total of eighteen samples. Two patient samples were based on the positivity of antibodies against endomysium, deaminated gliadin and tissue transglutaminase proven celiac disease. Eleven patient samples were negative for all antibodies assayed. In the remaining five patient samples were detected only antibodies against deaminated gliadin. Dynamics of antibodies during the disease were determined by a patients who were diagnosed with celiac disease. The graphs shows when the patients properly adhere a gluten-free diet, ie. determined antibodies decline. If patients do not comply gluten-free diet, IgA and IgG rise again. Celiac disease belongs, because of its high prevalence, to one of the major diseases of a digestive tract. Insufficient diagnosis associated with an occurrence of atypical symptoms for celiac disease. Very important measure to early diagnose celiac disease is targeted screening, aimed at groups with a higher probability of occurrence of this disease.
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Synthesis of quantum dots for proteins detection
Šibíková, Anna ; Fohlerová, Zdenka (referee) ; Pekárková, Jana (advisor)
This thesis is focused on synthesis of quantum dots (QDs) for protein detection. It comprises three parts. The first part summaries the theory of QDs, their synthesis, functionalization, interactions and applications in medicine. In the second part synthesis of CdTe/ZnS core/shell QDs modified by glutathione (GSH) is described, followed by the conjugation with biomolecules BSA and IgG. Several coupling agents such as EDC with NHS and CDI were used. In the last part, the final products were characterized by fluorescence spectroscopy and capillary electrophoresis. The results show the dependence of the fluorescence intensity of the QDs on pH range, concentration of BSA and IgG concentrations using different crosslinkers.
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Methods of assesment and treatment of coeliac disease
Valkus, Martin ; Omelková, Jiřina (referee) ; Vránová, Dana (advisor)
This diploma thesis deals with methods of determining and treatment of coeliac disease. In the theoretical part of the work sums up informations about coeliac disease - history, prevalence, etiopathogenesis, immunopathogenesis, possibilities of diagnosis, discusses about gluten-free diet, mentions legislation of the Czech Republic and the European Union and compares expensiveness of gluten-free diet and normal diet. The experimental part of this thesis compares PCR, ELISA and indirect immunofluorescence methods for assesment of determination of genetic predisposition and antibodies in coeliac disease (greatest emphasis was placed on the comparison of antibodies against gliadin and deamidated gliadin antibodies in IgA and IgG).
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Imunomagnetic separation of lactic acid bacteria using magnetic microparticles functionalised by antibodies
Vaňásek, Jakub ; Španová, Alena (referee) ; Trachtová, Štěpánka (advisor)
Immunomagnetic separation is based on binding of antibody with antigen, where antibody is bound to magnetic particle. In this thesis there were used particles of magnetic pearl cellulose with antiLactobacillus and antiBifidobacterium antibodies. Immunomagnetic separation method was optimalized and verified for its efficiency and specifity with bacterial and yeast cells. This cells were identified by polymerase chain reaction. Efficiency of immunomagnetic separation was verified on probiotic meat product, where Lactobacillus cells were isolated. With DNA from isolated Lactobacillus cells the high resolution melting was performed. The results show presence of several bacterial strains of Lactobacillus species.
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