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The use of natural antibiotics and herbal medicine in the treatment of SIBO
Svitavská, Jana ; Vaníčková, Zdislava (advisor) ; Šťovíček, Jan (referee)
This bachelor thesis deals with the syndrome of small intestinal bacterial overgrowth (SIBO), a gastrointestinal disorder caused by increased total bacterial counts and abnormal microbiome composition in the small intestine. SIBO often manifests through non-specific dyspeptic symptoms, including gas bloating, abdominal pain, diarrhea, malabsorption and malnutrition. A non-invasive breath test is most commonly used to diagnose SIBO. Treatment of SIBO should include therapy for the underlying disease, bacterial eradication and nutritional support. The main aim of this bachelor thesis is to investigate the efficacy of herbal preparations and natural antibiotics in the treatment of SIBO and to highlight the potential usage of these antibacterial agents as a stand-alone or adjunctive therapy to conventional treatment. Another aim is to confirm the hypothesis that it is impossible to predict the diagnosis of SIBO based on gastrointestinal symptoms. Therefore, treating this disease empirically without confirmation by laboratory examination is not appropriate. The data was collected in the Gastroenterology Laboratory of the Institute of Medical Biochemistry and Laboratory Diagnostics, 1st Faculty of Medicine, Charles University in Prague and in VFN. The prevalence of gastrointestinal symptoms and other...
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Characterization of transgenic forms of dipeptidylpeptidase IV expressed in astrocytoma cell line U373MG
Vomelová, Ivana ; Bezouška, Karel (advisor) ; Vaníčková, Zdislava (referee)
Dipeptidyl peptidase IV (DPP-IV) is a serine protease, which executes its proteolytic activity by cleaving X-Pro dipeptides from the N-termini of its substrates. Furthermore, DPP-IV exhibits many biological functions independent of its enzymatic aktivity. Previous studies in our laboratory proved increased expression of DPP-IV in high-grade astrocytic tumours. To evaluate the enzymatic and non-enzymatic functions of DPP-IV in a glioma model, clones of asctrocytic cell line U373MG transfected by enzymatically inactive, mutated DPP-IV (mutDPP-IV) and enzymatically active, wild type DPP-IV (wtDPP-IV), were prepared. Enzymatically inactive mutDPP-IV was prepared using point mutation the active site serine residue. Cells U373MG were transfected using a doxycycline inducible Tet-On® system. For further analysis of the transgenic forms of DPP-IV, methods were used for verification of protein expression, enzymatic activity and subcellular localization. Doxycycline induced U373MG mutDPP-IV and U373MG wtDPP-IV cells, expressing mutated and wild type DPP-IV, respectivelly, exhibited increased expression of transgenic DPP-IV in a concentration and time dependent manner. Doxycycline induced U373MG wtDPP-IV cells exhibited both increased expression and enzymatic activity of DPP-IV. In contrast, DPP-IV enzymatic...
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Biochip DNA analysis in the detection of colorectal cancer related mutations
Adámek, Petr ; Vaníčková, Zdislava (advisor) ; Kovářová, Jaroslava (referee)
The colorectal cancer is one of the most common tumour diseases of all. If diagnosed at early stage, there is a reasonable chance for at least five-year survival. Common methods for screening and diagnosis are relatively simple and also quite successive, however (still) not perfect - for proper diagnosis an invasive procedure is necessary. In general there is a significant effort to make the procedures of screening and diagnosis more comfortable for patients as well (to make them) more precise - which means that there is great chance for employing methods of molecular genetics and biology including biochip methods. The purpose of this final-year thesis was to test the possibility of the optimization of this biochip method, originally developed by the manufacturer for ~ 6 ~ testing DNA extracted from faeces, for samples of genomic DNA extracted from white blood cells with the objective to detect possible mutations in four genes of the interest (K-ras, BRAF, TP53 and APC) with a strong relation to this cancer. The evaluation of samples was made on a biochip analyzer Evidence Investigator TM. In the final -year thesis is a summary of theoretical evidence in biological, clinical and also technical side of this affair. The biochip method is described together with an example of the actual procedure, presentation...
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Characterization of transgenic forms of dipeptidylpeptidase IV expressed in astrocytoma cell line U373MG
Vomelová, Ivana ; Vaníčková, Zdislava (referee) ; Bezouška, Karel (advisor)
Dipeptidyl peptidase IV (DPP-IV) is a serine protease, which executes its proteolytic activity by cleaving X-Pro dipeptides from the N-termini of its substrates. Furthermore, DPP-IV exhibits many biological functions independent of its enzymatic aktivity. Previous studies in our laboratory proved increased expression of DPP-IV in high-grade astrocytic tumours. To evaluate the enzymatic and non-enzymatic functions of DPP-IV in a glioma model, clones of asctrocytic cell line U373MG transfected by enzymatically inactive, mutated DPP-IV (mutDPP-IV) and enzymatically active, wild type DPP-IV (wtDPP-IV), were prepared. Enzymatically inactive mutDPP-IV was prepared using point mutation the active site serine residue. Cells U373MG were transfected using a doxycycline inducible Tet-On® system. For further analysis of the transgenic forms of DPP-IV, methods were used for verification of protein expression, enzymatic activity and subcellular localization. Doxycycline induced U373MG mutDPP-IV and U373MG wtDPP-IV cells, expressing mutated and wild type DPP-IV, respectivelly, exhibited increased expression of transgenic DPP-IV in a concentration and time dependent manner. Doxycycline induced U373MG wtDPP-IV cells exhibited both increased expression and enzymatic activity of DPP-IV. In contrast, DPP-IV enzymatic...
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Model of experimentally regulated expression of dipeptidyl peptidase IV in glioma cell lines
Němečková, Jana ; Šedo, Aleksi (advisor) ; Vaníčková, Zdislava (referee)
"Dipeptidyl peptidase IV Activity and/or Structure Homologues" (DASH) represent a newly defined group of multifunctional molecules, typically bearing dipeptidyl peptidase IV-like hydrolytic activity. Dipeptidyl peptidase IV (DPPIV, EC 3.4.14.5, identical to CD26) cleaves out X-Pro dipeptides from the N-terminus of peptides. Recent knowledge shows substantial role of DASH in cancer pathogenesis. Here we present (i) an overview of the issue of DASH molecules and their functional substrates in the neuroectodermal tumor and (ii) a preparation of stable transfected human glioblastoma cell lines with inducible gene expression of DPPIV. Vectors containing human DPPIV gene and its mutated form in the catalytic active site have been prepared to assess the importance of the enzymatic activity of the final product. This will enable us to study the biological role of DPPIV in genesis and progression of neuroectodermal tumors - cells growth, invasion and migration of transformed glial cells in vitro. Moreover, complex role of DPPIV will be studied using a model of homotopic application of transfected cells into the brain of immunodeficient mice. Prepared cell lines provide more consistent information about DPPIV from the point of view of its "autocrine" importance for the expressing cells, as well as its potential...
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