Colorectal cancer (CRC) is a common malignancy which develops due to old age and lifestyle factors, low percent of patients afflicted by a genetic disorders. Half of all colorectal cancer patients are diagnosed after metastasis. The high rate of the late detection, emphasizes on the requirement of convenient and inexpensive diagnostic methods for comprehensive screening programs. The aim of this study was to discover proto-oncogenes mutation and assessment of tumor suppressor genes expression. Formalin fixed paraffin embedded (FFPE) histologically verified colorectal cancer samples were used. APC, KRAS and BRAF mutations were investigated using polymerase chain reaction (PCR) fragments and direct sequencing. Gene expression assessment of MLH1, MGMT and CDKN2A were achieved via quantitative polymerase chain reaction (qPCR). In the present study we could detect a novel transversion heterozygous mutation in APC gene codon 1365 in three patients. BRAF codon 600 mutation were detected in one patient. KRAS codon 12 mutation was discovered in one sample and also a novel transition mutation in codon 15 was detected in 6 patients. In 80% of cases, MLH1 and MGMT expression were undetectable, in remaining 20%, MLH1 expression were reduced, but MGMT showed both reduced and increased expression compared to control. In 100% of patients CDKN2A expression was undetectable. The rate of mutations in predetermined hotspot codons and amount of uncommon mutations into APC, BRAF and KRAS in Nepalese patients indicates the requirement of further investigation in CRC patients from that part of the world. Also, the expression rate of MLH1, MGMT, CDKN2A and deficiency of an information source emphasizes the necessity of whole genome CRC expression profiling data to comparison and conclusion. / <p>-</p> / -

Vegetation sampling was carried out to determine changes on the limestone quarries after 30 years. These semi-permanent plots are located in the Czech Karst Protected Landscape Area in the Czech Republic.

Selected quarries and adjacent areas of xerophilous grassland in Bohemian karst were examined for the presence of velvet spider burrows. Selected microhabitat valuables were collected and their significance for ladybird spider analyzed.

Based on light trapping carried out in 2010, I studied moth communities of the Koda National nature reserve, part of Český kras landscape protected area, in order to compare moth communities inhabiting major biotope types within the reserve. The obtained material consisted of 295 species in 4455 individuals. Steppe enclaves hosted more species than overgrown coppices and beech-dominated high forests. Ordination analyses distinguished between steppe and forest biotopes. Steppe catches differed from forest catches in habitat associations of constituent species (more species of grassland habitats), whereas the two forest types did not differ in this. Identical patterns applied for all moths analysed together and for analyses restricted to Geometridae and Noctuidae moths. Steppic enclaves represent the most valuable sites within the Koda reserve.

Features typical for karst, such as fast groundwater flow and existence of open conduits were observed in various parts of Bohemian Cretaceous Basin (BCB). So far, vertical and areal extent of karst rocks is not known. In case of concerning conduits in quartz sandstones the observation of their evolution in profiles perpendicular to flow is missing. The thesis deals with above mentioned issues. Concerning potential to karstification it is limited to Jizera facial area of BCB. For this purpose, leaching of samples in hydrochloric acid were carried out. The hydrochloric acid was used as an accelerated simulation of natural processes of dissolution of rock by acidic solutions. The reaction of samples on leaching in acid, disintegration and content of the CaCO3 component were evaluated. CaCO3 content was determined also by calcimetry. Rocks which disintegrated can be a suitable for the evolution of karst conduits. According to the lithostratigraphic sections, the examined profiles belong to the TUR5 and TUR6 units - thus these units contain rocks suitable for the evolution of karst conduits. In Předměřice site about 23 % of samples disintegrated, in Kosmonosy site 8 % and in Turnov site 36 % disintegrated. Thus about 1/10 to 1/3 of tested profiles is prone to karstification and evolution of karst...

Head and neck cancers (HNSCC) are highly heterogeneous disease, results from two major carcinogens - tobacco and/or alcohol, or HR HPV infection. This thesis was based on 60 biopsies of head and neck tumours embedded into paraffin after histological verification. HPV infection, including particular types was monitored in different HNSCC regions by multiplex qPCR. Subsequent IHC demonstrated expression of p16INK4A and p53 as a possible diagnostic biomarker. Based on the information, patients with HNSCC can benefit from antiEGFR therapy by Cetuximab, but so far without defined predictors, the analysis of point mutations of Ras gene family (Kras, Nras) and Braf gene was performed. These mutations were monitored as potential predictive biomarkers, in correlation with gender, age and other risk factors. For all statistical processing the Chi-x2 test was used. Key words: Head and neck cancers, biopsy, HPV types, PCR, p16INK4A, p53, molecular predictors, Kras, Nras, Braf

Tumors are one of the leading causes of death worldwide. Generally, the prognosis is better if the treatment begins at an early stage. Nowadays, the conventional chemotherapy treatment of cancer, known for its limited efficacy and side effects, is being gradually replaced by targeted biological treatment, which is used when specific genetic mutations are found. A part of the treatment is a detection of a potential progression, which is mainly based on the tumor biomarkers monitoring. Currently, further investigation of a so-called liquid biopsy method are ongoing, on which this thesis is focused. The main aim of this work was the experimental development and validation of the method for detection of the ctDNA in the plasma samples based on the somatic mutations presence. For the development and optimization of the system on the principle of denaturation capillary electrophoresis, the samples of cancer patients with KRAS mutation were used. Subsequently, a clinical part of the research was performed on a pilot set of 21 plasma samples. Finally, the method was optimized for the detection of BRAF and EGFR markers. A partial objective was to improve the detection sensitivity and increase the capture of the ctDNA in patients with advanced stage of the disease. The results of this work suggest the...

The subject of the diploma thesis is monitoring of rocks blocks movements near sinking stream Bílá voda situated near the village Holštejn in Moravian Karst. The aim of thesis is to measure of the 21th stage and 22th stage in year 2017. The thesis follows on the results obtained in years 2004-2015. Achieved results are mutually compared and analysed towards the results of previous stages. The final results are shown numerically and graphically

The diploma thesis deals with the design of a new building for a children's sanatorium with speleotherapy in Ostrov u Macochy. Site is located in the valuable landscape of the Moravský kras nearby Císařská jeskyně, which is used in the healing process. The sloping plot is oriented to the southeast and offers optimal sun conditions. The designed building enters the existing terrain with respect and uses its natural slope. The sanatorium has three receding floors, their green roofs smoothly merge into the terrain. The building includes all required facilities - clinic, education, catering, speleology locker room, accommodation, administration, technical facilities, club rooms and games rooms. The construction system of the building is a combined brick and monolithic skeleton. The basic idea was playfulness and dynamism, simplicity of shapes, easy orientation in space and a terraced solution with ecological aspects of the design.

The thesis deals with monitoring of rock blocks movements near sinking stream Bílá voda situated near the village Holštejn in Moravian Karst. The thesis follows up results, which were gained in years 2004-2011. The aim of thesis is survey of the 16th stage in summer 2012 and 17th stage in autumn 2012, following processing of surveying data and evaluation of space movements. Achieved results are compared mutually and they are analysed towards results of previous periods. The final results are documented numerically and graphicallyl.