Graduated bachelor students

The study is focused into evaluation of in vitro and in vivo properties of de novo prepared choline derivatives in order to determine their potential therapeutical and/or diagnostic value in cancer patients.

Aberrant regulation at epigenetic level can directly and/or indirectly influence the genetic integrity and gene expression pattern of cells, resulting in the development of various types of disorders, including cancer. The local hypermethylation of tumor-suppressor genes and the global hypomethylation of genomic DNA often occur in human cancers. Multiple DNA methylation changes in the cancer methylome could be associated with the acquisition of drug resistance. Recently at our institute the cancer cell lines resistant to 2‘-deoxy 5-azacytidine (decitabine) were prepared. Decitabine, a demethylation drug, is used in clinical practice for treatment of hematological malignancies. Since epigenetic therapy is in clinical use or trials for several cancers, efficient methods for epigenetic profiling are needed. Our specific aim is to screen a genome-wide methylation profile of 2-deoxy 5-azacydine resistant cancer cell line HCT116. Our long-term aim is to elucidate the mechanism of drug resistance in cancer cells.

Attempt to find the mechanizm of action in cytotoxic active triterpenes . The main goals of this work is to prepare a set of triterpenoid derivatives that have shown high selective cytotoxicity and synthesize biotinylated probes from them that may be used in pull down essays in order to try to find molecular targets.

The aim of this thesis is to validate protein changes which is responsible for oxaliplatin treatment. These changes has been previously analyzed by quantitative proteomics. The main techniques used in this bachelory thesis are western blot and classical microscopy.

Identification of genetic changes typical for particular subtypes of glioma is crucial for the improvement of diagnostics and choosing personalised therapy. This thesis describes the detection of BRAF:KIAA1549 fusion, which is considered a possible diagnostic marker for pilocytic astrocytoma. Fusion was detected by polymerase chain reaction (PCR) in 25 % of examined brain tumours (2 out of 8), in 40% of pilocytic astrocytoma (2 out of 5) specifically. Fusion was also detected by fluorescent in situ hybridization (FISH) using a specifically labelled fluorescence DNA probe. Amplification of the PDGFRA gene is common in high-grade gliomas and is considered a possible prognostic factor. Amplification of PDGFRA was detected by FISH using specifically labelled fluorescence DNA probe in 5.6 % of brain tumours (2 out of 36). From the examined subset of brain tumours FFPE samples, only 57 % (36 out of 63) provided countable signals. The significant connection between FFPE sample age and quality of the provided signal was found.

E2Fs are transcription factors which regulate the cell cycle by different mechanism. E2F family contains eight members E2F1 – E2F8. Some are able to activate and some repress gene expression. Disruption E2Fs signaling pathways is one of the reason cancer diagnosis. Theoretical part summarizes basic information about E2Fs, the cell cycle with its control by cyclin dependent kinases. Also is focused on retinoblastoma protein pRb and drugs used for high throughput screening assay. In experimental part were tested three types of cell clone 293H which were kept in 10% EMEM medium. I chose one clone with the best properties and used it for testing anti cancer drugs by high throughput screening.

The main target of this work was preparation of the precursors leading to the new type of 3-hydroxy-4-(1H)-quinolinone derivatives, compounds with potential biological importance. In this regard, we first focused our attention on the synthesis of intermediates resulting in the connection of the heterocycle and hydroxyquinoline skelet in position 7 via spacer with amide group. For this purpose we prepared a model intermediate methyl 4-((3-hydroxypropyl)carbamoyl)-2-nitrobenzoate using the solid-phase synthesis. Later, we worked on the successful conversion of hydroxy group of model intermediate to primary or secondary amino group. After synthesis of target intermediate, we tried to prepare a model bisheterocycle of hydroxyquinoline-thiazole. Unfortunately, the attempt was not successful and it will be necessary to focus on optimisation of the final reaction step.

This work is focused on preclinical testing of selected peptides labelled with 68Ga as potential radiodiagnostics for glioblastoma multiforme. The main goal of the study is to determine in vitro characteristics of studied peptides, their in vivo behaviour in mice and to evaluate the potential of 68Ga labelled peptides for diagnosis of glioblastoma multiforme using positron emisson tomography.

The main aim is to prepare a small set of new heterocyclic triterpenoid derivatives using condensation reactions of triterpenoid precursors with ethylene diamine, urea derivatives, hydrazine and other small molecules. Our previous research of triterpenoid heterocycles resulted in several compounds with high in vitro cytotoxic activity and this work should further explore this topic. New derivatives prepared within this work will be tested for their in vivo cytotoxic activity and the best candidates will be selected for more biological examinations.

The aim of the thesis is the detection and characterization of bacteria producing beta-lactamases isolated from patients with leukemia. All isolates of enterobacteria will be tested for the presence of blaTEM, blaSHV and blaCTX-M genes (responsible for extended-spectrum beta-lactamase production) using PCR method and the eventual occurrence of genes enabling carbapenemases production (KPC, SME, IMI, VIM, IMP, OXA enzyme types etc.) will be assessed. For direct detection of extended-spectrum beta-lactamase (ESBL) producers, new real-time PCR assay will be performed.

Cardiomyopathies are illnesses of myocardium connected to myocardial dysfunction. Some are connected to high risk of life treating arrhythmia and already first manifestation could end in sudden death. The most frequent type of cardiomyopathy is dilatation cardiomyopathy (DM). In many cases the causative of disease is unknown. In part of the patients, the inheritance of genetic variation or mutation could play a role in pathogenesis. The aim of this works is the study and identification of inherited forms of DM using high-throughput sequencing on Illumina MiSeq platform.