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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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Heat Shock Proteins - Focus on Therapeutic Approaches in Oncology
Rutkowski, Radoslaw ; Hromadníková, Ilona (advisor)
Since the discovery of heat shock proteins in 1960s and their immunogenic properties 20 years later in 1980s they have been under rigorous investigation with great hopes for advent of immunotherapy in oncology as well as other pathologies. So are the HSPs a breakthrough in therapy of cancers or will they be just another form of palliative treatment of these notorious and deadly diseases? HSPs is a family of proteins expressed virtually by all living organisms. These proteins have a number of functions that are crucial to organism's intracellular and extracellular homeostasis. Due to the significance of HSPs, over the eons of the evolution, they have been highly conserved in our genetic code. The human HSPs shares almost 50 percent homology with simple unicellular prokaryotes to well over 95 percent homology with higher species of living organisms. Despite of the fact that HSPs have been knows for approximately 50 years and despite of the intensive resears in the field, currently Russia is the only place where HSP immunotherapy has been approved as of 2008 for treatment in renal cell carcinoma. The HSPs immunotherapy has proven benefitial as an adjuvant treatment in some stages of a selected cancers. The advantage of HPSs immunotherapy is that it has less side effects compared to standart chemotherapy...
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Plasmid DNA vaccines
Machan, Radoslav ; Chroboková, Maria (referee) ; Rittich, Bohuslav (advisor)
Plasmid DNA vaccines are the new generation of vaccines with a great potential in prevention of many diseases. Recent studies and clinical test are aimed at prevention against cancer, hepatitis, malaria, HIV, influenza and other diseases. Recent main challenges covering plasmid DNA vaccines are associated with optimalization of each step of production and mainly purification steps allowing production of pDNA at kilogram levels. Main purification techniques used are based upon chromatographic methods, but research and development also shows other potential methods, like two-phase aqueous systems or magnetic microparticles as carriers. In experimental part of this thesis isolation of pUC 19 plasmid from Escherichia coli JM 109 (pUC 19) cell culture was performed via method of alkaline lysis. Isolation was verified by agarose gel electrophoresis. Isolated samples were purified in four repetitions with lithium chloride and magnetic microparticle carriers and the extent of purification was verified spectrophotometrically. Purified samples were visualised via agarose gel electrophoresis and results were compared.
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Use of selected fluorescence techniques to characterization of microbial cells
Vaněk, Martin ; Pekař, Miloslav (referee) ; Márová, Ivana (advisor)
Carotenoids are one of the most abundant natural pigment. Furthermore, they have great potential for wide industrial utilization. Nowadays, prefered (and cheaper) synthetic method counters progressively improved biotechnological production. Far more cheaper production should be reached via implementing methods of on-line optimization, such as flow cytometry. In this thesis the dependence of yeasts autotofluorescence intensity on carotenoids concentration in dry mass was studied. The dependence was established by means of linear regression. Moreover, the dependence between light scatter and the amount of dry mass was established by means of model utilizing forward scatter, side scatter and number of cells in unit volume. Yeasts' cells was also studied under the lenses of laser scanning confocal microscope. The goal was to localize carotenoids inside cells. Experiments undertaken suggest that they are localized inside the lipidic granules, but more experiments are required for a proof.
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"Features of the tumor microenvironment and their utilisation for drug delivery"
Větvička, David ; Hovorka, Ondřej (advisor) ; Reiniš, Milan (referee) ; Drbal, Karel (referee)
- David Větvička, M.Sc . Many researchers have, in the past, focused on pathophysiological features of tumor tissue, various tumor-nonmalignant cell interactions, and secretion of active molecules within the tumor mass. All these aspects of tumor structure are known as tumor microenvironment. The composition of particular tumor ecosystem is highly variable, with differences between various tumor types, even between patients with the same diagnosis, and in separate areas of the same tumor. Moreover, further changes in tumor microenvironment often occur during the progression of the disease. Studies of tumor microenvironment have revealed both novel targets for therapy and new prognostic markers. New therapy modalities are being developed to target these discovered features, including drugs functioning to boost anti-malignancy immunity, to block pro-metastatic potential, or to utilize the unique features of this pathological environment established by the tumor. These are obviously of great interest and harbor high potential for better management of malignant diseases. The focus of this thesis is to study the interactions of polymeric drug delivery systems within the tumor microenvironment and to utilize various features of this specific niche for drug delivery research. We have followed three...
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Biological effects and metabolism of cis-zeatin-type cytokinins in plants
Gajdošová, Silvia
9 Summary Cytokinins (CKs) are plant hormones that affect a wide range of developmental processes. The most important group of isoprenoid CKs represent zeatin and its derivatives occurring in two, cis and trans, positional isomers. Whereas trans-zeatin (transZ) was found to be a highly bioactive substance, cis-zeatin (cisZ) has been viewed for years as inactive or weakly active adjunct to its corresponding trans counterpart playing only an insignificant physiological role in plants. The occurrence of cisZ-type CKs was found in a great number of plant species with especially high levels identified in species of family Poaceae. All tested derivatives of cisZ, surprisingly including also zeatin-N9-glucoside, delayed dark-induced chlorophyll degradation in oat and wheat leaf segments. Additionally, cisZs effectively induced cell division in CK-dependent tobacco callus. The most pronounced activity was exhibited by cisZ riboside (cisZR) in the two types of CK bioassays. Metabolism of both zeatin isomers differed in short-term as well as in long-term experiments, which was supported also by various affinity of CK degrading enzyme, CK oxidase/dehydrogenase (CKX), to individual cis and trans isomers. Primary root enlargement of Arabidopsis seedlings was inhibited by cisZR in the same or similar way as by transZR,...
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Tailoring the Morphology of Microcellular Polymer Foams
Nistor, A. ; Rygl, A. ; Sajfrtová, Marie ; Toulec, M. ; Vonka, M. ; Kosek, J.
In this work, we focus on polymer foams with improved heat and/or sound insulation properties, in particular on microcellular foams with the cell sizes below 10 μm. By decreasing the cell size below 10 μm or even lower we can highly improve the foam heat and sound insulation properties. Depending on the foam structure (open-, closed-cell or dentritic) the foam application can vary. The structure-property relation is thus of great interest as well as the preparation of foams with various morphologies and scale-up of the manufacturing of microcellular foams.
Fulltext: content.csg -  PDF
Plný tet: 115_nistor - PDF
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Artery tracking in CT data
Doubek, Miloš ; Pelikán, Josef (advisor) ; Felkel, Petr (referee)
This thesis focuses on the processing of CT data received during the CT examination of the vessels - CT angiography. The area of interest is the extracranial course of the carotic arteries. Regarding the major role of these vessels in the blood supply of the brain cells this examination is frequently performed and influences the diagnostic decision and following therapy in case of pathological changes of the vessels. The contemporary clinical software included in the tomographs does not provide sufficient comfort for easy 3-dimensional orientation which is necessary for the diagnostics. The automatic segmentation and classification of the vessels might be a possible solution to this problem and also a great challenge. For the segmentation of the extracranial part of the carotic arteries the parallel and robust model fitting algorithm was used. The parameters of the model make it possible to classify the vessel and to draw attention to its stenosis, for example. The algorithm was tested on both synthetic and real data. The core of the system is implemented as a set of separate ITK filters. It exploits all the advantages of the software architecture of the ITK framework (object-oriented design, parallel and stream data processing, multiplatform, reusable components). The user friendliness is ensured by the...
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Metabolic Effects of L-Carnitine Supplementation in Hemodialyzed Patients
Cibulka, Roman ; Racek, Jaroslav (advisor) ; Zima, Tomáš (referee) ; Eiselt, Jaromír (referee)
Chronic kidney failure is associated with many kinds of metabolic disorders caused by the kidney disease and also attributable to dialysis treatment. Phenomena such as accumulation or deficit of various substances and dysregulation of metabolic pathways participate in the pathogenesis of these changes. One of these disorders, which we studied in more detail, is a deficit of carnitine. Carnitine is a substance which plays an essential role in beta-oxidation of fatty acids by catalyzing their transport into the mitochondrial matrix. It enables obtaining of energy, namely in muscle cells including myocardium. Patients with chronic kidney failure treated by hemodialysis are known to have decreased carnitine concentration in plasma and tissues due to its impaired synthesis in kidneys and the great loss across the hemodialysis membrane during dialysis sessions. A single hemodialysis session reduces plasma free-carnitine concentrations to about one-third of their predialysis values because of small molecule of free-carnitine. On the other hand, renal elimination of acylcarnitine, which is physiologically ineffective, may be impaired in chronic kidney failure, leading to increased blood concentrations of acylcarnitine. Considerable evidence suggests that carnitine deficiency and abnormalities of carnitine...
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The Emersonian Pynchon
Naser, Safwan ; Robbins, David Lee (advisor) ; Roraback, Erik Sherman (referee)
Ralph Waldo Emerson and his postmodemist colleague Thomas Pynchon ... a seemingly incongruous connection. The aim of this thesis is to explore the unusual relationship between these prominent authors and advert to the great influence which Ralph Waldo Emerson unquestionably had not only on authors who were not separated from him by such a noticeable temporal abysm, but also a most recent author who, according to the vast majority of the definitions of the postmodern, should be entirely free of any Emersonian influence. This intricate relationship will be assessed mainly through Mason and Dixon, the most recent novel by Thomas Pynchon which reflects many aspects of what Emerson found absolutely central. The summation of what seems to be propounded throughout the entire novel is represented by the idea of determining boundaries, in both the literal and the abstract sense. Emerson himself devoted much attention to this subject matter and it is clear that Pynchon and Emerson have much in common from this perspective, which holds true to such an extent that the boundary between the postmodem and romantic is itself facing the pressure of redefinition, which is in turn a fundamental concept which both authors share.
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