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Circulating tumor DNA in advanced colorectal carcinoma patients
Kopalová, Dominika ; Benešová, Lucie (advisor) ; Saláková, Martina (referee)
Colorectal carcinoma (CRC) is worldwide known as one of the most often tumor diseases. Approximately 50 % of patients are diagnosed with advanced stage of CRC and moreover even after removing all of the tumor bearings, there is high risk of its recurrence. As most of others solid tumors the CRC also releases fragments of DNA also known as the circulating tumor DNA (ctDNA). The ctDNA analysis is a new tool for monitoring continuous tumor burden and for observing treatment response. Due to its minimal invasivity and high specificity is suitable for instance for long period postoperative follow-up of patients with CRC. Within the long period follow-up process an evaluation radicality of operation is conducted as well as an early detection of new tumor foci. This thesis focus on an origin and releasing mechanism of ctDNA into bloodstream, its features and clinical utility options for patients with an advanced colorectal carcinoma. Key words: circulating tumor DNA, ctDNA, advanced colorectal carcinoma, mCRC, marker
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Using modern molecular-genetic methods in prevention, early diagnosis, prediction of therapy response and prognosis
Benešová, Lucie ; Minárik, Marek (advisor) ; Brdička, Radim (referee) ; Klepárník, Karel (referee)
55 5 Závěr V prezentovaných pracích jsme se zabývali vývojem a praktickou aplikací metod pro rychlou, citlivou, vysokokapacitní detekci molekulárních markerů u různých typů nádorových onemocnění. Vyvinuli jsme v této souvislosti dvě nové metodiky. První je založená na cyklujícím teplotním gradientu, který poskytuje efektivnější separaci DNA molekul a možnost opakovaného dávkování v jedné analýze a tedy výrazné zvýšení celkového počtu analyzovaných vzorků 1 . Druhá metodika založená na dynamickém značení DNA molekul interkalačním činidlem během elektroforetické analýzy odstraňuje potřebu fluorescenčně značených primerů a dělá tak analýzy finančně i prakticky dostupnějšími 3 . Obě tyto metodiky jsme následně použili při analýzách klinických vzorků pacientů s nádorovým onemocněním. Výsledky uvedených publikací zaměřených na klinické aplikace ukazují, že vyšetření molekulárních markerů zahrnující mutace, alelické ztráty a jednonukleotidové polymorfismy má široké uplatnění. Charakterizace vzorků kolorekta na molekulární úrovni, jehož optimalizaci jsme provedli, může mít význam při popisu stádia transformace adenomu v karcinom nebo při odhadu prognózy onemocnění na základě detekovaných typů mutací 2 . U pacientů s podezřením na karcinom pankreatu může molekulárně-biologické vyšetření v kombinaci s cytologickým...
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Optimization of proces for detection of free tumor DNA in plasma and its clinical utility for colorectal cancer, lung cancer and pancreatic cancer patients
Belšánová, Barbora ; Benešová, Lucie (advisor) ; Tachezy, Ruth (referee)
In current days, examination of circulating tumor DNA (ctDNA) finds new use across different cancers. It is directed at tumor-derived short fragments of DNA present in peripheral blood of patiens (mainly in advanced stages). Due to its minimal invasivity, almost 100 % specificity and relatively high sensitivity in stage IV patients, this approch found its main potential clinical utility especially in early detection of disease relapse or progression after tumor resection (i.e. post-operative follow-up), prediction and monitoring of therapy response and estimation of prognosis. As a result of minute levels of ctDNA on a high background of other non-tumor DNA fragments present in plasma, a suitable method exhibiting highest sensitivity is the key for proper detection of this marker. The approach is predominantly based on initial identification of a mutation in tumor tissue and its subsequent detection in plasma. The present work is aimed at optimization of ctDNA isolation and method of its detection based on PCR amplification followed by heteroduplex analysis by denaturing capillary electrophoresis (DCE) to achieve highest sensitivity for detection of mutated fraction in plasma sample. I have applied the optimized protocol to examine ctDNA in three types of cancers, namely colorectal cancer (122...
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Protection of Employees upon Termination of the Employment
Benešová, Lucie ; Štangová, Věra (advisor) ; Vysokajová, Margerita (referee)
The theme of this thesis is the protection of the eployees upon termination of the employment. The aim is to comprehensively assess the current legislation of termination of empoyment, with regard to the degree of protection that is provided to employees. It deals with the protection of employees, their position but also the claims related to the termination of the employment relationship. This thesis is dividend into five chapters, the first of which relates to the functions of labor law with an emphasis on the function of protection as a core function of labor law. With regard to weaker position of the employee to the employer, it is clear that labor law generally tries to balance this inequality or in some way eliminate it in order to provide the highest level of security. The second charter presents the core of this thesis and deals with the varoius types of termination of employment. It contains their assessement in relation to ensure adequate protection of employees but also the positives and negatives of legislation including possible aplication problems that are occured in practice. The emphasis is placed primarily on the termination of the employment relationship in view of the fact that the need to ensure a sufficient level of protection of employees in this unilateral termination method...
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Optimization of miRNA analysis in fine-needle biopsy samples of pancreatic cancer tissue.
Čuperková, Romana ; Benešová, Lucie (advisor) ; Kuthan, Martin (referee)
Pancreatic cancer (PC) is extremely severe malignant disease with a five-year survival of less than 5%. Currently there is no reliable tool for the diagnosis of PC in its early stages. At the time of clinical symptoms most patients are in an advanced stage of the disease and the treatment does not usually have a significant effect. For these reasons emphasis is gradually shifting to the search for the suitable molecular markers for improvement of the diagnosis and assessment of the survival prognosis with respect to a possibility of surgical treatment. MiRNA represent one of the most promising markers, although, their examination in pancreatic tissue is a complicated process. One of the reasons is the very small amount of the source material coming from a fine needle biopsy. A second cause of problems is the subtle character of the pancreatic tissue resulting in significantly lower yields of molecular genetic analysis when compared to other epithelial tissues. An additional negative factor is heterogeneity of the tissue resulting in disproportionate representation of tumor cells within the sample. A suitable choice of procedures for isolation of nucleic acids (NA) and subsequent analysis including quantification of tumor cells is critical for accurate evaluation of the miRNA levels. This work is...
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Representations of femininity in Quentin Tarantino's movies
Benešová, Lucie ; Vochocová, Lenka (advisor) ; Dvořák, Tomáš (referee)
The submitted thesis is titled Representations of Femininity in Quentin Tarantino's Movies and it focuses on the process of femininity construction in six selected movies both written and directed by Quentin Tarantino. The aim of this study is to identify how Tarantino operates the notions of conventional femininity and traditional female gender roles. Furthermore, it investigates to what extent his films either support or subvert stereotypical gender identities. In relation to the research issues the key concepts are defined by the theoretical part of this work. In particular it describes the theory of social and media construction of reality, gender as construct, film as a part of patriarchal structure or feminist perspectives related to the film production. This same part also offers basic information about Quentin Tarantino's personality and his film works in general. In order to answer the research questions the thesis employs a qualitative method of critical discourse analysis which functions as a helpful tool while reconstructing the image of femininity set by media. Moreover, the selected method serves to reveal latent meanings applied by film as a form of media to affect both behavioural and cognitive individual characteristics. Keywords Femininity, gender, construct, stereotype,...
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The User's Habits of Media Usage by Children from the Children's Houses
Benešová, Lucie ; Wolák, Radim (advisor) ; Šťastná, Lucie (referee)
The work concerns with media enviroment and life style of children from the children's homes. The empirical part is based on theoretical part and it explores their media habits, media access, media owning and parent mediation. This work compares a sample of 48 children from children's homes and a sample of 62 children who live in a family. Quantitative research is supplemented with qualitative research in a form of semistructured interviews with tutors in six children's homes and with mass discusion with children in one of them. The main aim of the work is to approach which role do media play in every day life of the children from the children home's. Comparative research answers the main question: Are media habits of the children from the children's homes different from the habits of the children from families and which strategy of the parental mediation is supreme and how the mediation differs from the mediation in the families.
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