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Introduction of Western blotting method for detection of AMPK cascade activation in LS174T cell line
Dubecká, Michaela ; Pávek, Petr (advisor) ; Nachtigal, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Michaela Dubecká Supervisor: Doc. PharmDr. Petr Pávek, Ph.D. Title of diploma thesis: Introduction of Western blotting method for detection of AMPK cascade activation in LS174T cell line LKB1/AMPK is the main cellular energetic pathway, which acts as the sensor of various extra- and intracellular signals. In response to the signals, it regulates energy metabolism and the maintenance of cellular homeostasis, so its major role lies in the survival, growth and development of the whole organism. This pathway is of significant importance because it has the potential to suppress tumoral progress in the cell. Metformin, the widely used drug in the treatment of type 2 diabetes, exerts a significant antitumour activity. However, the direct mechanism of metformin's action is still unknown. Metformin induces cellular stress similar to the metabolic stress via inhibiting mitochondrial respiratory chain complex I, which results in an increase of AMP levels in plasma. AMP then binds to the AMPKγ subunit, so metformin mediates the activation of AMPK. AMPK is suddenly phosphorylated on Thr172 of the AMPKα kinase domain via LKB1, which mediates the downregulation of many downstream kinases. The...
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Regulatory mechanisms of WNT signalling
Pospíchalová, Vendula ; Kořínek, Vladimír (advisor) ; Trka, Jan (referee) ; Bryja, Josef (referee)
AB T mu hom dise β sign mu liga stab tran sign T the focu disc cate of t cell targ the inte con sec I sign BSTRACT The Wnt si lticellular o meostasis. A eases, most β-catenin is nalling). In ltiprotein c ands when t bilized and nscription f nalling is tig This thesis knowledge uses on seq cusses the enin signall the Wnt pa ls of intest geted mous thesis des eraction wit nditional Hi retory cell t In conclusi nalling path T ignalling pa organisms Accordingly notably can s a central m n unstimula complex and they engage d transloca factors and ghtly regula is based on e of the reg quential po positive ro ling outcom athway whic tinal epithe e strains th scribes unp th members ic1 deletion types and en ion, our fin hway in dev athway is o ensuring s y, mutations ncer. mediator of ated cells d degraded e their recep tes to the to drive th ated at vario n four origin gulation of sttranslation ole of nucle me. The third ch reduces lium. Final at enable st ublished da s of the Wn n in the inte nhanced tum ndings contr velopment an one of the m successful s in the pat f canonical W β-catenin d in the pro ptors, degrad nucleus t he transcrip ous levels b nal articles f the Wnt s nal process ear protein d study repo the levels o lly, the las tudying the ata on the nt pathway, estinal epith mourigenesi ributed to t...
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Issues of direct taxes
Čerňanský, Michal ; Bakeš, Milan (advisor) ; Kohajda, Michael (referee)
The thesis is focused on issues of direct taxes mainly on the most discussed problems in the direct taxation area. The introductory section is devoted to basic financial law categories, such as the very definition of tax, the notion of direct taxes, tax functions, tax systems and tax schemes. The history of taxes from the very beginning to the present legislation is outlined in the thesis. The main central theme of the diploma work is mentioned in the chapters that deal with the issues of direct taxes. These chapters are devoted to the characteristics of direct taxes in a broader sense. According to a significant extent of the chosen theme, the thesis deals with income taxes within "de ferenda" legislation. The intention of the thesis was not the complex processing of all relevant aspects of income taxes, but especially expresses the most discussed problems such as direct tax issues, tax progression and relation to social aspects. In this thesis, tax issues are identified at EU level and compared to the level of taxation in other EU countries. In conclusion the tax system of our nearest EU neighbour Germany is also briefly mentioned.
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Proteomic Analysis of Gamma-Irradiated Human Leukemic Cells
Šalovská, Barbora ; Řezáčová, Martina (advisor) ; Šebela, Marek (referee) ; Ebhardt, H. Alexander (referee)
(ENG) Title of the dissertation: Proteomic analysis of gamma-irradiated human leukemic cells In the presented doctoral thesis, we aimed to elucidate molecular mechanisms underlying radiosensitization of MOLT-4 cell line (T-ALL) by specific inhibition of kinases from the phosphatidylinositol-3 kinase-related kinases (PIKKs) family. We tested two highly potent inhibitors of ATR and ATM, VE-821 and KU55933, respectively, for their effects on proliferation, viability, and cell cycle of sham-irradiated and irradiated MOLT-4 cells. Both inhibitors proved to radiosensitize MOLT-4 cells and furthermore, 10 µM VE-821 was shown to act as a strong antiproliferative agent in sham-irradiated MOLT-4 cells. To further describe cellular mechanisms underlying the VE-821-mediated radiosensitization of MOLT-4 cells, we employed high-resolution mass spectrometry to identify and quantify changes in proteome and phosphoproteome of irradiated VE-821-treated cells. As the detection and quantification of phosphorylated peptides in complex biological samples is challenging due to their low stoichiometry, we first compiled and optimized protocol for their enrichment. The protocol was then applied to study changes in radiosensitized MOLT-4 cells. In concordance with our expectations, VE-821 did not cause any significant...
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The name of a company, the concept of its legal regulation and protection
Hegerová, Kristína ; Černá, Stanislava (advisor) ; Zahradníčková, Marie (referee)
Resumé The aim of my thesis, as anticipated in the introduction, was to introduce to the reader the topic of company-name law with all its key institutes headed by the company name as such. In the process of executing the thesis I worked with older, as well as current literature, legislation and judicature while simultaneously using internet sources. I have made effort to provide a complex view of company name and the legal regulation related thereto, including the regulation on international and foreign (Slovak) level. Therefore, having carried out a general discourse on the concept and meaning of company name, I introduced the thesis with a brief historical excursion into the development of this insitute in the area of the Czech Republic which, as I believe, has provided at least an elementary base for understanding the current progress trends, as well as the current regulation in this field. This was followed by a part focused on the current Czech regulation, in which I closely adressed myself to the principles of company-name law and consequently to the particular means of protecting the company name which where, at the same time, the focal point of my studies of this matter. As I believe, that also in this field it is impossible to default the importance of the international regulation, I made an...
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Model of experimentally regulated expression of dipeptidylpeptidase IV in glioma cell lines
Trylčová, Jana
Model of experimentally regulated expression of dipeptidyl peptidase IV in glioma cell lines Abstract "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...
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Editor matematických výrazů
Holaň, David ; Lidický, Bernard (referee) ; Jelínek, Vít (advisor)
In the present work we study the concept and implementation of "MaEd for LATEX", a portable graphical user interface program for creating and editing of LATEX formulae. The program is designed to allow a beginner to create even complex formulae without any knowledge of underlying LATEX source code. The user can also import his own source code and the program doesn't make unnecessary changes to the imported source code, like removing comments and indentation. The concept describes how was the program's GUI designed. The LATEX commands available for creation of math formulae are described along with their syntax and level of support in the program. The work also analyzes structure of LATEX source code.
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Distropine - Distributed System for Anotation of Web Pages
Kaprál, Ondrej ; Zavoral, Filip (advisor) ; Nečaský, Martin (referee)
The aim of this bachelor thesis is to design and implement peer-to-peer version of a service for commenting of internet resources. From a functional point of view it is an implementation of P2P discussion forum. We attempt to achieve the highest possible degree of decentralization while meeting these requirements: usage of distributed hash table to store data content and deployment of user identity concept while protecting this identity against theft. In the analysis we came to realize that in the environment where users do not establish any bindings and content distribution is broadcast in nature there is need for globally trusted authority. For this reason it is deployed PKI with certication authority (CA). Our pursuit of maximum decentralization is reached by multiple measures in the design of the application. We allowed simultaneous operation of multiple CA instances, we have also minimized the power of CA to monitor user activities or to censor the content of the system. User interface of the software is integrated into web browser what helps us to shield user from complexity caused by P2P design.
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English language teaching in the primary school in the context of competences for learning
Krausová, Jarmila ; Kasíková, Hana (referee) ; Krykorková, Hana (advisor)
The competency for learning is one of the key competencies which are set out within broadly educational programmes, are of supra-subject form and are attainable as a result of a comprehensive education. The development of competencies for learning while learning a foreign language is closely associated with the level of language competency of learning individuals and their feel for the language. The whole process of the development of a pupil's competencies is associated with a complex assessment of the pupil, with the development of not only his/her cognitive abilities and skills, but also the development of his/her personalityrelated and social attitudes. The dissertation "English language teaching in the primary school in the context of competences for learning" consists of three parts. The first part covers theoretical outcomes of learning and language competence, the second part describes strategies for learning a foreign language with the emphasis on metacognitive strategy and psychodidactic application of metacognition in a school environment. The third part presents the results of research conducted in a primary school during English language lessons aimed at educational activities focused on developing of competencies for learning.
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Detection of Malicous Traffic in Local Network
Šabík, Erik ; Kováčik, Michal (referee) ; Žádník, Martin (advisor)
This bachelor's thesis discusses monitoring local networks using IP flows. It describes Nemea framework which is used for building complex systems for detecting malicious traffic. Analysis of data from three different networks was performed by using this framework. Based on this analysis a design for detection of malicious traffic in local network was created. The detection method monitors network traffic for suspicious communication targeting IP or URL addresses that are listed in public blacklists. The detection method is evaluated on various traffic samples and the results show that three analysed samples belong to networks that are well managed and secured since the communication with the blacklisted entities is rare.
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