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Clinical and pathologic correlation of complications of intensively treated hemato-oncological patients with a targeted focus to the involvement of the oral cavity and rare case of secondary neoplasia.
Vokurka, Samuel ; Boudová, Ludmila (advisor) ; Chlumská, Alena (referee) ; Faber, Edgar (referee)
Summary: In this thesis section "Oral Mucositis (OM) and oral Graft versus Host Disease (GVHD) in patients after allogeneic stem cells transplantation and with focus to FLU/MEL conditioning regimen" the author deals with following issues: 1. Literature overview: The overall negative impact of OM is mentioned and the complex pathogenesis, clinical characteristics, incidence and factors influencing both OM and GVHD are described. The histological features of GVHD and classification of it are mentioned, too. Conventional and reducedintensity conditioning transplant regimens, including FLU/MEL are presented. 2. Methodological section: Prospective observational study of 117 patients after allogeneic stem cells transplantation with BU/CY2/±ATG (26%), FLU/MEL (60%), FLU/CY (9%), Bu/FLU/ATG (2%) and HD-CY/ATG (2%) conditioning regimens. OM assed according to WHO criteria, GVHD assessed according to NIH criteria accepted by the EBMT. The statistical univariate analysis performed by means of GraphPad In Stat - Statistica Software (Man- Whitney, Fisher, t-test), p<0,05 considered significant. Multivariate analysis used in OM risk factors assessment. 3. Results: The OM incidence was significantly dependent on pre-transplant conditioning regimen (Bu/CY2/±ATG: 100%, FLU/MEL: 78%, FLU/CY: 9%, p=0,0001) in multivariable...
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Diagnostic morphological features of PDGFRA-mutated gastrointestinal
Daum, Ondřej ; Hes, Ondřej (advisor) ; Mandys, Václav (referee) ; Ehrmann, Jiří (referee) ; Zámečník, Josef (referee)
Daum O., Grossmann P., Vanecek T., Sima R., Mukensnabl P., Michal M. (2006): Diagnostic morphological features of PDGFRA-mutated gastrointestinal stromal tumors: Molecular genetic and histological analysis of 60 cases of gastric GISTs. Ann. Diagn. Pathol. In Press Summary In this study, 60 gastrointestinal stromal tumors (GISTs) of the stomach were analyzed to elucidate the possible relation of their morphology to mutation status of KIT and PDGFRA genes. The patients included 27 men and 33 women with a mean age of 63,8 years (range 12 to 92). Only one tumor occurred before the age of 21 years. KIT mutations were detected in 31 cases (51,7%), PDGFRA mutations in 22 cases (36,7%), and seven cases (11,7%) were KIT and PDGFRA wild type. When the mutation status was correlated with histological features of the tumors, epithelioid or mixed epithelioid/spindle cell pattern and mast cell infiltration were found as the most reliable signs of PDGFRA mutation. Neoplastic rhabdoid cells and multinucleated giant cells, also previously reported as features of PDGFRA mutated GISTs, seemed to be less specific but still helpful markers in our study. Finally, tumor infiltrating lymphocytes and myxoid stroma do not seem to be valuable histological signs. Daum O., Klecka J., Ferda J., Treska V., Vanecek T., Sima R., Mukensnabl...
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Graphemics in the German documents of Václav IV 1379-1399
Bartůňková, Alena ; Ebelová, Ivana (referee) ; Vodrážková, Lenka (advisor)
The goal of this thesis is twofold. On the one hand I try to determine a system in the use of the graphemes in the documents of Vac1av IV. from the years 1379-1399 and I confront the findings with the use of the graphemes in the Middle Upper German (Mittelhochdeutsch). On the other hand, I compare these results with the use ofthe graphemes in the early Upper German (Frühneuhochdeutsch) as described in the literature, in particular in 1. T. Piirainen, V. Moser, H.Bokova, L.E. Schmitt and V.E. Mourek. The first chapter describes the period of reign of Vac1av IV as weIl as the personality ofthe ruler. The second chapter deals with the court ofVac1av IV. with a particular emphasis on the language situation at the court. Further, Vac1av IV.'s chancellery is characterized in the chapter three as apart of the court and I also shortly describe its history, structure as weIl as the competence of its members. Chapter five is devoted to documents as objects of a study; thus, first I define and c1assify different types of docurnents and second, I indicate the relevant features both in the form of the document and its content. The next chapter describes the status of the early Upper German (Fruhneuhochdeutsch) as it emerges from the literature. The seventh chapter lists the documents of Vac1av IV. between the years 13...
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Cloning, detection and analysis of the gene cluster coding synthesis of the antibiotic Anthramycin
Poubová, Martina ; Černá, Milena (advisor)
he aim was to find , isolate and sequence a cluster of genes coding for the synthesis of anthramycin , antibiotics group pyrrolobenzodiazepines whose producer Streptomyces refuineus var . thermotolerans . Pyrrolo [2,1- c ] [1,4] benzodiazepines are a class of naturally occurring antibiotics anticancer effect. These are molecules that bind covalently to DNA molecules and this leads to blockage of transcription. Although anthramycin least affects the bone marrow, was not put into clinical practice as antitumor antibiotic because of the high cardiotoxicity. Better effects and good clinical features expected from a new synthetic analogues both in terms of antitumor antibiotics , and the production of antimicrobial compounds and antimalarial drugs . So far, successful antibiotic related families of antibiotics , lincosamide , was clindamycin antibacterial and antiparasitic effect. Knowledge of the genetic background of these substances allows modification simultaneously substances used , followed by a less costly production more efficient and safer substances. Using molecular biological methods were isolated part of the chromosomal DNA Producer Streptomyces refuineus var . thermotolerans containing the complete gene cluster anthramycin biosynthesis .
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Study of the role of glutathione in the anthracycline-induced cardiomyocyte injury
Vávrová, Anna ; Šimůnek, Tomáš (advisor) ; Kvasničková, Eva (referee)
The clinical usefulness of anthracycline antineoplastic drugs has been limited by the risk of serious cardiotoxicity. Its precise mechanisms have not been fully elucidated, although the oxidative stress is widely believed to play a principal role. The anthracycline toxicity to heart may result from its high accumulation in myocardial tissue, richness on mitochondria producing reactive oxygen species and relatively poor antioxidant systems including superoxide dismutase, catalase and glutathione peroxidase. This study is focused on the role glutathione in daunorubicin (DAU)-induced cardiotoxicity both in vitro and in vivo. Total, reduced and oxidized glutathione (GSH/GSSG) have been determined by a spectrophotometric method using dithiobis(2-nitrobenzoic acid) in DAU (0.1 - 10 µM) - exposed H9c2 cardiomyoblast cell line and heart tissue from daunorubicin-treated rabbits (3mg/kg weekly, 10 weeks). In H9c2 cells, DAU induced significant toxicity (IC50=0.48 µM), levels of total glutathione were slightly but insignificantly decreased, the levels of oxidized glutathione were unafected. Neither buthionine sulfoximine (BSO, an inhibitor of glutathione synthesis) nor 2-oxo-4-thiazolidine-carboxylic acid (OTC, a glutathione synthetic precursor) had any significant effects of DAU cytotoxicity. This contrasted...
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The effect of heat shock stress on enzyme activities in plants.
Doričová, Vlasta ; Hýsková, Veronika (advisor) ; Liberda, Jiří (referee)
Heat stress is one of abiotic stress factors, which fundamentally influences the growth and development of plants. Plants response to heat stress by series of cell and metabolic changes, the specificity of heat stress is synthesis of molecular chaperons, called heat- shock proteins (HSP). The influence of heat shock (in the form of 1 hour application of 40řC from 20řC) on the activity of NADP-dependent enzymes, enzymes of Hatch-Slack cycle, glycosidases and the activity of peroxidase in tobacco plants (Nicotiana tabacum L. cv. Petit Havana SR1) was studied in this work. Since HSP are involved in proper folding of immature, misfolded or partly denaturated proteins, in degradation of denaturated proteins and in induction of thermotolerance of plants, the aim of this work was to find out, if the activity of studied enzymes will be maintained or modulated within the plant defense response. The highest amount of HSP70 detected immunochemically together with higher activities of NADP-malic enzyme, phosphoenolpyruvate carboxylase, β-hexosaminidase and α- mannosidase compared to control plants was found 1 hour after application of heat shock. Peroxidase activity was most increased 1 day after HS compared to controls. Using native electrophoresis the differences in isoform content between control and...
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SPE of Organic Sulphates on Ionexchangers
Spáčilová, Jana ; Jezberová, Michaela (advisor) ; Kubíček, Vladimír (referee)
Diploma work deal with possibility of synthesis and determination 6- sulphatoxymelatonin on weak ion exchanger. First part is focused on synthesis, isolation and determination of 6-sulphatoxymelatonin from obtained publications, description SPE techniques and reports brief summary extraction of organic sulphates using SPE from literature. Second part of work presents results of synthesis 6-hydroxymelatonin and 6-sulphatoxymelatonin based on performed experiment.
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The content of components of ATP synthasome in different rat tissues and in patients with defects in ATP synthase
Mikulová, Tereza ; Houštěk, Josef (advisor) ; Kalous, Martin (referee)
The complexes of oxidative phosphorylation (OXPHOS) are situated in the inner mitochondrial membrane in higher structural and functional complexes, so-called supercomplexes, which facilitates substrate channeling. ATP synthase is also able to organize in higher structures. In mammalian mitochondria, ATP synthase is usually present in a dimeric form. There is evidence of its trimerization and even tetramerization. Furthermore, it seems that ATP synthase catalyzing the phosphorylation of ADP to ATP, adenine nucleotide translocator (ANT) ensuring the exchange of ADP for newly synthesized ATP across the inner mitochondrial membrane and phosphate carrier (PiC) allowing the import of inorganic phosphate (Pi) into the matrix of mitochondria are assembled in a supercomplex called ATP synthasome. This association among the components of phosphorylative apparatus seems to increase the efficiency of processes leading to the ATP synthesis. First, we studied amounts of the components of phosphorylative apparatus in connection with various ATP synthase contents among mitochondria isolated from nine rat tissues. Mitochondrial proteins were separated by denaturing electrophoresis (SDS-PAGE) and their content was analyzed using specific antibodies. In agreement with our expectations, the highest content of...
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Laboratory diagnostics of micrometastases in breast cancer patients
Mikulová, Veronika ; Zima, Tomáš (advisor) ; Svoboda, Marek (referee) ; Průša, Richard (referee)
Introduction: The presence of circulating tumor cells (CTC) in the peripheral blood has been associated with worse prognosis and early relapse in breast cancer patients. CTC determination in the peripheral blood has been considered as a liquid biopsy. The aim of this project was to analyze the presence of CTC followed by their molecular characterization with the potential use not only as a new biomarker for real-time monitoring of therapy efficacy but also as a suitable tool for patient's stratification and individualization of treatment for breast cancer. Methods: A total of 54 patients with diagnosed early breast cancer were enrolled into a prospective study. Ten millilitres of peripheral blood were sequentially collected to test for the presence and characterization of CTC during the follow-up of patients. CTC isolation and detection was performed by AdnaTest BreastCancer™ (AdnaGen AG, Germany), which is based on the detection of EpCAM, HER2 and MUC1 specific transcripts in enriched CTC- lysates. cDNA from isolated CTC has been further used for newly optimized qPCR assays for breast tumor and therapy resistance associated genes: TOP1, TOP2A, CSTD, ST6GAL, KRT19 and reference gene actin. qPCR results have been analyzed by Genex software (MultiD Analysis). Results: 195 blood samples have been...
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Biochemical methods as tool for study of reproductive proteins
Postlerová, Pavla ; Zigo, Michal ; Pohlová, Alžběta ; Jonáková, Věra
Study of molecular mechanisms in reproduction is essential for the understanding of this outstanding process. Our lab studies proteins secreted by reproductive organs and sperm using various biochemical methods for a long time. We have expertise in protein extraction from spermatic cells using different approaches, and by kits for proteins from the sperm surface and distinct subcellular compartments. The proteins of reproductive organ fluids are separated by chromatographic methods, such as size exclusion chromatography, high-performance liquid chromatography with reverse phase (RP-HPLC) and affinity chromatography on matrices with various ligands. Proteins are subjected to SDS- or 2D-electrophoresis for their characterization and comparison of various extraction methods, different mammalian species, and sperm in different functional development. Electrophoretically separated proteins may be transferred onto nitrocellulose membrane (Western blot) for antibody detection or binding studies with lectin-labelled ligands (lectins, polysaccharides, zona pellucida glycoproteins). We use immunoprecipitation method with specific antibody for protein determination followed by the MALDI identification. Proteins are localized by immunofluorescent techniques on/in spermatic cells and tissue sections of reproductive organs. Isolation of proteins from reproductive tissues and fluids, and the antibody detection is crucial for the studying of reproductive protein origin.
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