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Generation of synthetic modified mRNAs encoding cell cycle regulators
Petroušek, Filip ; Koblas, Tomáš (advisor) ; Vašková, Michaela (referee)
One of the modern therapeutic approaches is the application of in vitro transcribed mRNA into the target cells to produce the required target proteins. This method has prooved to be more effective than therapeutic application of proteins prepared in advance, because their insertion into the organism lead to denaturation. The main task of this work was to prepare mRNA in vitro and mRNA transfection into the cells and monitoring the expression of target proteins with the method of Western blotting. The target proteins were from families of cyclins and cyclin-dependent kinases, which play an important role in cell cycle regulation. The mRNAs for 8 types of molecules have been prepared, including non-mutated forms of cyclins D1, D2, and D3, and cyclin-dependent kinase 6, as well were synthesized mutated versions of cyclins D1 T286AT288A, D2 T280A, D3 T283A and cyclin-dependent kinase 4 R24C. As for cyclins, there was a substitution of aminoacid threonine for alanine. This substitution could cause slowdown degradation of proteins in comparision with those existing in natural form. Regarding the cyclin- dependent kinase 4 the exchange of aminoacid arginine for cysteine causes its bigger stability in vivo. The prepared mRNA which encodes mentioned regulators of cell cycle was transfected into isolated...
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Development of antibacterial antibodies for cystic fibrosis patients
Vašková, Michaela ; Hodek, Petr (advisor) ; Nosková, Libuše (referee)
Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the CFTR gene (CF transmembrane conductance regulator). These mutations result in absent or defective CFTR chloride channel function. The susceptibility to bacterial respiratory infections due to the accumulation of thickened mucus and altered glycosylation in lungs is typical for this disease. Bacteria Pseudomonas aeruginosa (PA) is a major cause of these infections. Among other virulent factors, the pathogenicity of these bacteria is caused by fucose-specific PA-IIL lectin which plays a role as an adhesin. The effect of anti-PA-IIL egg yolk antibodies and multivalent fucose-based PA-IIL inhibitors on PA adherence to lung epithelial cells was studied in this work. Chicken antibodies were isolated from egg yolks before and after immunization with antigen PA-IIL. Specific anti-PA-IIL antibodies were obtained by affinity chromatography using a column with an immobilized PA-IIL. Reactivity of IgY was verified by ELISA. The presence of PA-IIL in the bacterial culture of Pseudomonas aeruginosa (PAK, ST 1763) and the ability of antibodies to recognize this bacterial lectin were verified by Western blotting followed by immunodetection. Appropriate culture conditions have also been found for the expression of this lectin. The...
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Antibody against lectin PA-IIL as a tool preventing bacterial infections of cystic fibrosis patients
Vašková, Michaela ; Hodek, Petr (advisor) ; Nosková, Libuše (referee)
Cystic fibrosis is an autosomal recessive disease caused by mutation of the CFTR gene. This mutation results in the damage of protein with ion channel function. As a consequence of this damage, electrolyte equilibrium is impaired. The respiratory tract is affected the most because of the mucus thickening and changes in the glycosylation of lung epithelium cell surface structures. These changes lead to an insufficient defense function of lungs and greater susceptibility to bacterial infections. The most common pathogen of respiratory tract of CF patients is Pseudomonas aeruginosa. This bacterium contains numerous virulence factors and has the ability to form a biofilm that protects it against the host defense mechanisms and antibiotic effect. Antibiotic treatment is also complicated by the development of bacterium resistance. Chicken antibodies have a considerable potential as a tool for preventing bacterial lung infections of CF patients. The influence of specific and nonspecific antibodies on the adherence of Pseudomonas aeruginosa (PAK, ST 1763) ex vivo was monitored on a model system of lung epithelia cells from a CF patient (CuFi-1) and a healthy individual (NuLi-1). Bacterial and pulmonary cells were labeled with PKH fluorescent dyes to allow their spectrofluorimetric determination. It has...
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Support Material for Therapy Narration of a Story for Gained Communicative Ability
Vašková, Michaela ; Housarová, Blanka (advisor) ; Sýsová, Jana (referee)
Magdalény Rettigové 4, 116 39 Praha 1 tel.: +420 221 900 111 www.pedf.cuni.cz IČ 00216208 DIČ CZ00216208 Dle čl. 4 Opatření rektora č. 6/2010 o Zpřístupnění elektronické databáze závěrečných prací http://www.cuni.cz/UK-3470.html a čl. 1 Opatření děkana 17/2010 se z časového hlediska závěrečné práce dělí do tří skupin: a. "nové práce", tj. práce odevzdávané k obhajobě počínaje 29. 9. 2010, b. "starší práce", tj. práce odevzdané k obhajobě od 1. 1. 2006 do 28. 9. 2010, c. "práce před rokem 2006", tj. práce odevzdané k obhajobě před 1. 1. 2006. V tomto případě jde o "starší práci" odevzdanou k obhajobě od 1. 1. 2006 do 28. 9. 2010. Omlouváme se, ale dokument není k dispozici.
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The analysis of the retail market segment of a healthy food in Hradec Králové
Vašková, Michaela ; Olšanová, Květa (advisor) ; Němečková, Iveta (referee)
The bachelor thesis is focused on the analysis of the health food market in Hradec Králové. The work is divided into three parts. First section aims to characterize the organic food market in the Czech Republic in terms of producers, distributors and consumers with an estimate of future development. Another part is dedicated to the analysis of the market in Hradec Králové in order to define the typical customer of specialized health food stores and his shopping behavior using quantitative research.The last part deals with the analysis of the competition of selected health food shop for the purpose of choosing an appropriate competitive strategy. Competition is analyzed on the theoretical basis of Kotler's competitive analysis and Porter's five forces model.
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