Magnetically driven separation techniques using magnetic solid carriers are one of modern methods to speed up and facilitate the previously used separation and purification procedures. The use of magnetic particles in biology imposes strict requirements on physical, and chemical properties of the particles, including low toxicity, biocompatibility and non-interference with the chemical environment in diagnostics. The aim of this study was to evaluate carboxyl-functionalised magnetic non-porous P(HEMA-co-GMA), P(HEMA-co-EDMA), PGMA, silica-coated lanthanum manganese peroskvite La0.75Sr0.25MnO3 and thermosensitive poly(N-isopropylacrylamide) microspheres – P(NIPAAm) for DNA isolation from different types of complex food and environmental samples containing PCR inhibitors. The solid-phase reversible immobilisation (SPRI) of nucleid acids on microsphere surface and the release of adsorbed DNA were optimised. DNA from real samples (milk products and probiotic food suplements, mouse faeces) was apparently adsorbed on solid particles from the aqueous phase system composed of 16% PEG 6000 and 2M NaCl. The conditions of the subsequent release absorbed DNA to the elution buffer (pH of elution buffer, temperature and time of elution) were optimized. The quality of eluted DNA and the presence of target DNA were examined by PCR and q-PCR using domain-specific Bacteria and genus-specific Lactobacillus primer set. Real-time PCR was used for an estimation of the PCR interference by comparing the amplification efficiencies of purified DNA containing solid nanoparticles with the DNA standards free of any nanoparticles

This thesis describes the study and characterization of secondary created structures in PN junction of silicon solar cells. Secondary created structure is the term which means the structures created for the purpose of suppressing the negative influence of local defects and edges of the solar cell. This means in particular laser notches used to isolate the edges. Furthermore, the secondary created structure means modification defective area using focused ion beam milling. Theoretical part of this work deals with short introduction to the topic of solar cells. There are mentioned the physical nature of the solar cell and the technology associated with the issue of solar cells. Experimental section begins with a description of the experimental methods. For diagnostic methods were used both electrical (UI characteristics, noise characteristics) and optical methods (measuring local radiation - CCD camera, thermal imager, lock-in thermography). Furthermore, there was also used a scanning electron microscope (SEM) equipped with technology using Focused Ion Beam (FIB). Sequentially there are presented individual results of characterization of created structures by laser. These partial results are incorporated into a comprehensive methodology developed for characterizing laser-created structures. The experimental part is finished by a presentation of the results of the research use of focused ion beam technology for sputtering defective areas of solar cells.

This diploma thesis solves design of the building for temporary accommodation - hotel in the city of Brno Lisen street Novolíšeňská on plot no. 5049/24. The land is gently sloping and its area is 41,546 square meters. The outer dimensions of the building are 49.0 m x 28.3 m. The hotel has four floors and a basement. The roof is designed single-layer flat. The building is of brick construction of two-way system of Porotherm blocks based on reinforced concrete footings. Ceilings consist of reinforced concrete monolithic slabs. The hotel is equipped with 27 rooms for 50 guests.

The introduction of theoretical part deals history of floor heating. Here is an overview of the technology, which was for centuries used for floor heating. Further included in the introductory part of the aggregate materials used, which are used for floor heating, and used. In the next part of the theoretical introduction we are given the best known ecological heat source. These are mainly heat pumps and solar collectors. The last part is made in the proposal which are important values calculated for the design and seamless use of central heating system.

This thesis presents a comparison of building materials for vertical bearing structures of buildings, thermal insulation materials and mutual combination of structural materials and insulators. Furthermore, there are possibilities for solving the most common insulation thermal bridges construction. The paper evaluated the pros and cons of different combinations of building materials. Teplo and Area programs were used for the calculations.

The subject of this thesis is to compare the price of a variant of the technical solution to a specific building. On introduction describes the various parts of the construction budget and their origins. In the practical part the comparison of alternative building materials on the basis of their technical characteristics. Victorious materials are inserted into the original budget, so a change was made final price of the building.

This work was focused on the detection of probiotic bacteria in four different probiotic products (probiotic cream, probiotic tampons, oral probiotics and soy beverages with probiotics). The viability of the bacteria contained in the products was verified. Complex matrices of the products were used to isolate DNA in a quality suitable for the PCR method, followed by identification of the declared bacterial genus and species. Amplification was achieved with conventional PCR and real-time PCR, genus- and species-specific primers were used. Bacteria, of the genus Lactobacillus and Bacillus and bacterial species Lactobacillus pentosus, Lactobacillus rhamnosus, Lactobacillus fermentum and Lactobacillus gasseri, were proven to be within the products. Subsequently, the DNA from mixed bacterial species in the probiotic tampon were distinguished using PCR-HRM. Five sets of primers were used to test this. Two sets of primers (primers P1V1, P2V1 and V1F-HRM, V1R-HRM) were evaluated as the most suitable for resolution.

Circulating tumor DNA (ctDNA) in peripheral blood of patients with metastatic colorectal cancer appears to be a promising molecular marker that provides various applications. ctDNA levels vary depending on the presence, alternatively on the volume of tumor mass within patient's body, which can be used primarily for early detection of disease progression or recurrence and moreover for evaluating radicality of surgical treatment, all within long-term postoperative follow-up of the patient. Due to minimal invasivity of ctDNA analysis from peripheral blood (so-called liquid biopsy), it is possible to perform it repeatedly at relatively short time intervals. On account of very low fraction of ctDNA in total cell-free DNA (cfDNA) ranging between units and hundreds of percent, the key factor is optimal methodology covering all steps from the isolation process to a sufficiently sensitive detection technology. In this thesis I focus on an optimization of isolation process and analysis of ctDNA obtained from tumor tissue and plasma of selected patients with metastatic colorectal cancer in connection with surgical radicality and correlation with a clinical status of the patients.

CHARLES UNIVERSITY IN PRAGUE FACULTY OF SCIENCE DEPARTMENT OF BIOCHEMISTRY PRAGUE 2010 Supervisor: Doc. MUDr. Jiří Gallo, PhD. Supervisor-consultant: Prof. RNDr. Gustav Entlicher, CSc. Eva Zolotarevová Summary of PhD Thesis POLYETHYLENE WEAR PARTICLES AROUND TOTAL JOINT REPLACEMENTS - THEIR PROPERTIES, DISTRIBUTION AND POSSIBLE MECHANISM OF THEIR ADVERSE BIOLOGICAL EFFECTS PRAGUE 2010 The research project was supported by the Grant project of National research program II. (no. 2B06096) of the Czech Ministry of Education, Youth and Sports named "Observation and minimization of UHMWPE wear in joint replacements". Front-page picture: Polyethylene wear particle isolated from granuloma tissue surrounding total hip replacement; Electron microscope microphotograph; particle size app. 90 x 80 µm; retouched Eva Zolotarevová Introduction Introduction For many people all over the world a replacement with an implant of irreversibly damaged joint is often the only way how to come back to normal life without pain and mobility limitation. Nowadays, greater and greater requirements are asked of quality of all replacement components, especially in terms of quality of used materials. Biomechanical properties of hip and knee replacements have been already solved. The most often used materials for manufacturing joint...

The aim of this dissertation study was to optimize the isolation and long term cultivation protocols for human dental pulp stem cells. The protocols which showed best results were used for cultivation of dental pulp stem cell isolated from exfoliated teeth (SHED). Additional aims were to characterize DPSC and SHED and prove their ability to proliferate over Hayflick's limit and differentiate into mature cell lines (osteoblasts, chondroblasts and adipocytes). In order to find optimal protocols for isolation of dental pulp from tooth, we tried three different approaches. During optimization of cultivation protocol we focused on decreasing amount of fetal calf serum (FCS) from 10 % FCS in cultivation media (most often used in literature) into 2 % and thus get closer to cultivation conditions suitable for clinical usage. We compared DPSC cultivated in three different media (medium with 10 % FCS, 2 % FCS supplemented with growth factors and media with 2 % FCS supplemented with ITS and growth factors). For characterization of DPSC and SHED we used basic biological properties (proliferation activity, viability, morphology), their phenotype and karyotype. The study demonstrated that the best protocol for isolation of dental pulp from tooth was to break the roots and extract the pulp throw this aperture. We...