Graduated master students

The thesis is reviewing PCR-based methods of mutation detection in NRAS gene while experimental part is focused on pyrosequencing.

SVOČ 2015

The thesis is focused into critical evaluation of three different RGD peptides coupled with chelators to enable their radiolabelling with gallium-68. RGD peptides are accumulated in tumors expressing αvβ3 integrin (marker of angiogenesis). Such radiolabelled RGD peptides could be succesfully used to image tumors in vivo. Our study will include all appropriate stages of radiopharmaceutical testing i.e. optimalisation of lablelling conditions, radiochemical purity tests (HPLC, TLC), plasma stability assays, testing of stability in various chemical enviroments, octanol-water partition coefficient determination. Suitable derivatives will be passed to in vitro tests employing cell line U-87 MG (glioblastoma multiforme) to assess their ability to bind to tumor cells. Finally the appropriate candidates will be used for in vivo imaging of gliomas in mouse model. The imaging will be carried out in microPET/CT system and its results will be compared with ex vivo biodistribution study.

SVOČ

The aim of the study is to find mechanisms of action of the plant steroids in human prostate cancer cells using cellular and molecular techniques indicated the possible involvement of steroid receptors in their action.

Master’s thesis is focused on cytogenetic changes in colorectal cancer patients. The student will propose and construct series of DNA probes suitable for fluorescence in situ hybridizations (FISH). By using this set will be possible to describe degree of deletions on q-arm of chromosome 18. Deletions of 18q involve DCC gene and are often find in colorectal cancer patients. The impact of this mutation is still discussed and the data are not consitent. As was newly found, in this region other 3 genes responsible for chomosome instability are located. Therfore is possible that for clinical imact would be important the the degree of the deletion.

The theoretical part of the diploma thesis summarizes information about nucleolus, ribosomal biogenesis, Mdm2 and p53 dependent and independent ribosomal stress. In the experimental part, vectors expressing nucleolin (NCL) and fibrillarin (FBL) fused with green fluorescent protein at its N-terminus were created. Transfection of human U2OS cells with plasmids expressing GFP-NCL and subsequent selection resulted in a stable fluorescent- labeled nucleoline (U2OS-pEGFP-NCL) cell line. By transfecting the U2OS cell line with plasmids expressing GFP-FBL and subsequent selection, fluorescent-labeled fibrillarin cell line (U2OS-pEGFP-FBL) was created. For the validation of FBL and NCL as possible markers of ribosomal stress, selected cytostatics known to disturb ribosomal biogenesis have been applied to U2OS-pEGFP-FBL and U2OS-pEGFP-NCL cell lines. Double RNA fluorescent in situ hybridization on specific rRNA locus was performed after application of cytostatic. Cisplatin, oxaliplatin and actinomycin-D caused the breakdown of the nucleus integrity and the release of FBL and NCL from the nucleolus to the nucleus, whereas 5-fluorouracil did not affect the integrity of the nucleus and the localization of FBL and NCL. From the results of the double rRNA FISH was evident that cisplatin, oxaliplatin and actinomycin-D blocked transcription of 47S rRNA, but did not block processing of pre-rRNA. Application of 5-fluorouracil caused stopping pre-rRNA processing, but transcription of 47S rRNA was not affected. The U2OS-pEGFP-NCL cell line was transfected with siRNA-RPL11 to silence expression of RPL11. The RPL11 depletion did not cause the loss of integrity of the nucleus and did not trigger p53 dependent ribosomal stress. Cells transfected with siRNA-RPL11 have been identified to accumulate pre-rRNA precursors. The combination of localization of these fluorescently labeled proteins with the double rRNA FISH method for pre-rRNA specific regions has proven to be a suitable system for identifying substances that may cause ribosomal stress.

The aim of the study is to identify possible molecular targets of anti-tumour indazole derivatives by immuno-affinity purification and MS analysis.

The aim of the study is to find specific biomarkers for prediction of successful in vitro fertilization in cervical mucus.

The aim of the study is to prepare resistant cell lines to 5-azacytidine and characterise multidrug resistance to epigenetic therapeutics.

The aim of the thesis is to mapp the prevalence, phenotype and genotype of vancomycin-resistant enterococci (VRE) isolated from clinical materials of patients hospitalized in Faculty Hospital Olomouc, Czech Republic and from poultry samples obtained from the Olomouc and South Moravian Region. Vysoká škola zdravotníctva a sociálnej práce sv. Alžbety v Bratislave (Obor Laboratórne vyšetrovacie metódy v zdravotníctve)

The main task in this thesis is to introduce and optimize methods used in non covalent and intact protein mass spectrometry. It is expected that student will get a deep knowledge about this problematics and will get used to operate with premium grade mass spectrometers.

This work is dealing with GC×GC TOF MS metabolomic analysis of urine sample. Silylation agent is used for derivatization. Urine sample used for optimization is analyzed by different column sets with varied polarity, width of stationary phase and internal diameter of column. BPX5/SolGel-WAX, BPX5/SLB-IL59, BPX50/SolGel-WAX, BPX50/BPX5, SolGel-WAX/BPX5, SLB-IL59/BPX50 are columns used for analysis in this work and compared. Separation success is characterized by orthogonality. Column combination BPX5/SolGel-WAX was determined as the best. This combination is used for urine sample analysis of healthy children and patients with glutaric aciduria.