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Study on the induction of intestinal conjugation enzymes by food supplements
Hrdinová, Johana ; Hodek, Petr (advisor) ; Bárta, František (referee)
Cytochrome P450s (CYP) are enzymes, that play an important role in the metabolism of foreign compounds (e.g. drugs, carcinogens, pollutants) and they also participate in the metabolism of hydrophobic endogenic substrates (e.g. sterols, fatty acids, prostaglandins). Cytochromes of subfamily CYP1A are involved in activation of precarcinogens. Whereas the consumption of dietary supplements containing chemopreventive substances (e.g. flavonoids) that can prevent the process of carcinogenesis is significantly increasing in recent years, although these substances may also have a negative effect on the organism. That because these substances can induce the cytochrome P450s and thus increase the risk of activation precarcinogens. In this study had been investigated the inductive effect of some flavonoids compounds - β-naphthoflavone, myricetin, dihydromyricetin and a drug containing an extract of red vine leafs - Antistax®.
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Detection of circulating antibodies to determination of autoimmuno diseases by means of immunofluorescence.
Sieglová, Kateřina ; Nachtigal, Petr (advisor) ; Laco, Jan (referee)
Kateřina Sieglová Detection of circulating antibodies to determination of autoimmuno diseases by means of imunofluorescence. Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Medical Laboratory Technician Background: The aim of this thesis is to explore of coherence between the presence of antibody against anti brush border antibody (ABBA) and idiopatic bowel disease (Crohn's disease and ulcerative colitis). Methods: Evidence of connection idiopatic bowel disease with positive ABBA test is examine by two perspectives, which are used for selecting and comparising of data. The first way is comparison of the number ABBA positive patients with their diagnosis of endoscopic biopsy. The second way check the positivity or negativity of ABBA examination of all patients with diagnose of idiopatic bowel disease. Experimental part summarize by this way the results, which was done at the Fingerland institute of pathology since 2009 to 2011. Results: The survey showed that among all the ABBA-positive patients is a large group of those who don't have the morphological signs of Inflammatory Bowel Disease - 72,47%. 66.67% of patients with a clinical diagnosis of Crohn's disease, and 39.37% of patients with ulcerative colitis are ABBA positive. Conclusion: The results shows, that...
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The role of human papillomavirus in the etiology and prognosis of squamous cell carcinoma of the oropharynx
Rotnáglová, Eliška ; Klozar, Jan (advisor) ; Šlapák, Ivo (referee) ; Zámečník, Josef (referee)
Tonsillar cancer (TC) presents an important part of head and neck cancers and its incidence is rising in the Czech Republic. Cigarette smoking and alcohol abuse are risk factors responsible for approximately two thirds of all head and neck cancers. Furthermore, a role of human papillomavirus (HPV) as an independent risk factor has been demonstrated recently. The aims of the present study were to find demographical differences between the group of HPV-positive and negative patients, to determine the prevalence of HPV in TC of Czech patients and also to find out whether the virus in HPV DNA positive tumors is transcriptionally active. Another aim was to test different markers of HPV infection for a precise selection of patients with HPV associated disease and to determine the prognostic role of these markers. A set of 109 patients with primary TC was examined for HPV DNA presence in tumor tissue and for the presence of HPV specific antibodies in serum. Data regarding risk factors were obtained by questionnaires. Forty-five samples were available for E6 and E2 mRNA analysis and for immunohistochemical analysis of p16 and p53 expression. The overall prevalence of HPV DNA in TC was 65%. In 93% of HPV16 DNA positive samples the virus was transcriptionally active. The E2 expression status has shown that in...
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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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Development of tool for passive immunization of cystic fibrosis patients
Pacholíková, Lenka ; Hodek, Petr (advisor) ; Švédová, Martina (referee)
Cystic Fibrosis (CF) is one of the most common heriditary diseases. This congenital condition, caused by CFTR gene mutation, affects gastrointestinal and respiratory system especially. The affection of respiratory system is considered the most serious life-threatening symptom. Pacients suffer from reccurent infections proceeding to the development of chronic inflammation and progressive pulmonary tissue destruction. A typical specific microoganism colonizating pulmonary tissue of thouse suffering from CF is recognised as Pseudomonas aeruginosa. Pulmonary infections caused by this microorganism are the most often cause of death in patients suffering from CF. Antiobiotics are the first-line therapy of this condition currently. Nevertheless, the need to find alternatives occurs due to antibiotics resistance development. Passive immunization by specific hen-egg-yolk antibodies against P. aeruginosa is a possible alternative. An observation of IgY influence on a bacterial adhesion to pulmonary epithelial cells required an appropriate model. In this context an appropriate adhesion testing method based on P. aeruginosa and pulmonary epithelial cells visualisation was searched. At first bacterial cells labelling was tested by CellTracker, resazurin, FITC and consequently PKH 26. P. aeruginosa fluorescent...
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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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Immunity in hosts repeatedly exposed to sand fies and the effect on pathogen transmission
Pohanková, Lucia ; Kolářová, Iva (advisor) ; Fialová, Anna (referee)
6 Abstrakt During the feeding of infected sand flies are inoculate into the host also sand fly saliva, which can strongly modulate the response of the immune system. If the host is naive, the course of infection is usually worse. In cutaneous leishmaniasis, the lesions developed early, are more destructive and persist longer. The hosts living in endemic areas of leishamniasis and their vector hosts are often exposed to feeding uninfected sand flies. For host are the saliva antigenic and induces specific cellular and antibody responses. This responses induce the protection against leishmania infection and differ for different hosts, attempts were made most frequently in murine and canine models. In humans, as hosts is difficult to monitor developments leishmania infection after previous exposure, because in humans mainly monitors the levels of antibodies, by which we can determine the degree of sand fly bites and the risk of transmission of leishmaniasis. Keywords: Lutzomyia, Phlebotomus, DTH, antibody, IgG,
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Toxoplasma gondii manipulation in the host
Dittrichová, Anna ; Langrová, Iva (advisor) ; Nechybová, Stanislava (referee)
The aim of this bachelor thesis is to collect so far discovered findings about manipulation of behaviour caused by Toxoplasma gondii. The first part of this thesis introduces basic characteristic of the parasite and toxoplasmosis, which is a disease caused exactly by this parasite. In the second part we summarize outcomes of many different scientific essays that deal with manipulation of intermediate host behaviour. Toxoplasma gondii is a widespread parasite that can modify the behaviour of the intermediate host, which can lead to serious changes in psychological profile of infected people. Toxoplasmosis is one of the most common zoonosis in the Czech Republic, the prevalence reaches 25-30 %. Toxoplasmosis is usually an asymptomatic infection, however it modifies the behaviour and acting of the intermediate host in some special situasions, it also changes excretion of important substances and affects the secondary sex ratio too. Toxoplasmosis is a very important risk especially for imunodeficit people and pregnant women. The principle of prevention is very strict personal and food hygiene.
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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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