The diploma thesis deals with the analysis of the effects surface texturing on starved contact. Experimental verification was realized on apparatus simulating contact between a steel ball and glass disc using colorimetric interferometry and high-speed camera. The work deals with lubrication regimes, problems with starved contacts and surface texturing which can reduce effects of starvation on non-conformal surface contacts.

This master thesis deals with influence of topography of friction surfaces on rolling contact fatigue. This influence is examined by newly reconstructed R-MAT station, whose reconstruction and fully report about it, is part of this master thesis. Influence is examined on area with higher surface's roughness which it wasn't examined before. Further research is made on area of surfaces with topographical modification with higher surface's roughness.

The goal of the submitted Diploma Thesis is the Proposal of modifications of the horizontal tail (HT) of the plane VUT 100 by increasing the shoulder of HT and reduce of the HT surface. The Diploma Thesis consists of assessing the benefits of position adjustments, load calculation and the proposal of the basic design.

The diploma thesis deals with The study of transition between fully flooded and starved regimes in EHL Contact. The work is focused on problems of starved contact and surface texturing of rubbing surfaces. Surface texturing of rubbing surfaces is one of the adjustments that cause in a sense improved lubricity and reduce friction and wear. The aim of this study is to compare numerical models with real experimental results measured on the Experimental apparatus.

This master thesis deals with the interaction of humic acids with organic ions and effect of chemical modification of humic acids on this interaction. Rhodamine 6G was used as a model organic ion. The interaction were studied by fluorescence quenching, dialysis in diffusion cells and differential UV-VIS spectroscopy. Results showed that the bond between the reactants was not primarily dependent on carboxylic functional groups of humic acids. Fluorescence experiments indicated static quenching mechanism and differential UV-VIS spectroscopy demonstrated the existence of - interactions.

It is well known that chromatin posttranslational state, transcription and splicing influence each other. Nevertheless, the details of this coupling are not fully understood. In S. cerevisiae, it is possible to induce conditions, in which splicing is uncoupled from transcription. Such situation occurred in cells expressing a mutated splicing factor Prp45, whose human homolog has been proved to participate in transcription regulation and also in splicing reactions. Based on previously indicated interactions in high throughput two-hybrid screens, we have been looking for physical links between Prp45 and proteins involved in chromatin posttranslational modifications. Finding of such a link would provide insight into the relationships of gene expression processes. Using coimmunoprecipitation and affinity purification, we were unable to detect physical interactions between Prp45 and our candidate chromatin regulators. Alternative approaches are discussed. Using the precipitation techniques, we mapped the interaction of Prp46 with truncated variants of Prp45. This observation contributes to our knowledge of protein-protein interactions within the spliceosome.

The subject of my thesis was to genetically modify a potato for increased resistance against its pathogens and pests. In developing a resistant plant, it is quite common to use the same type of molecules that plants use themselves in their defense reactions. In this work I used the gene SPI-2 originating from a honeycomb moth (Galleria mellonella). The protein SPI-2 is a member of serine protease inhibitors. Since the previous attempts of the team to detect the protein in transformed plants haven't been successful, the basic form of the gene was modified by adding a Kozak sequence near the start codon, which should have increased the translation initiation and hence increase the level of the protein. Two constructs were prepared for the transformation: SPI-2-T a SPI-2-Y. They differ by one amino acid, which slightly changes their inhibitory activity. First, the construct SPI-2-T was used for a transient transformation of tobacco Nicotiana benthamiana by agroinfiltration of its leaves. Then both constructs were used for a stable transformation of Solanum tuberosum cv. Desireé. The detection of the protein has not been successful, although the inserted gene was transcribed and his sequence was verified by sequencing. It is therefore most likely that the protein has a low stability in the cytoplasm....

ABSTRACT Mixed function oxygenase system participates in biosynthesis of endogenous and metabolism of exogenous substances (e.g. drugs or chemical procarcinogens) in an organism. Substrates are biotransformed by terminal oxygenases - cytochromes P450 (P450). Catalytic properties of certain P450s (e.g. studied isoform 2B4) are altered in the presence of a redox partner - cytochrome b5 (cyb5). Both cytochromes are anchored by hydrophobic domains in a lipid membrane of endoplasmic reticulum whereas their catalytic domains are exposed to cytosol. Two zero-length cross-linking approaches were employed to extend present knowledge of P450 2B4 and cyb5 protein structure and protein-protein interactions: (1) interlinking of carboxylate and primary amine groups of amino acids by water soluble 1- ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), and (2) photo-initiated cross-linking by photo-labile methionine analog (pMet), which links to any amino acid after activation by UV-irradiation, either in hydrophilic or hydrophobic environment. pMet was incorporated to methionine site(s) of cyb5 during recombinant expression in E. coli, which was carried out in limit medium supplemented with amino acid analog. Optimization of experimental conditions led to ~20-30% substitution of the natural amino acid. Covalent...

This paper deals with the expression of vascular endothelial growth factor (vascular endothelial growth factor, VEGF) and its use in tissue engineering of vascular wall. During the work interaction of endothelial cells with the modified fibrin-based biomaterial into which vascular endothelial growth factor (VEGF-A121) has been incorporated was monitored. This modification supported the adhesion and growth of endothelial cells. Vascular endothelial growth factor VEGF-A121 is signal glycoprotein that activates transmembrane receptors on endothelial cells. VEGF-A121 is a key regulator in vasculogenesis, angiogenesis, proliferation, migration and survival of endothelial cells. In this work, this protein was heterologously expressed at a thioredoxin fusion partner in an expression system of E. coli Origami B (DE3). Recombinant VEGF-A121 was additionally coexpressed with bacterial chaperones GroEL/GroES for potential increase of its solubility and biological activity. In the next part of this work thin fibrin network was prepared by catalytic action of thrombin on the polystyrene-bound monolayer of fibrinogen. This network has been further enriched by vascular endothelial growth factor (VEGF-A121), which was covalently incorporated in it by enzyme activity of transglutaminase (factor XIIIa). The last...

anks to their unique properties and high biocompatibilities, fluorescent nanodiamonds are promising representatives of modern carbon nanomaterials with a broad range of applications. Nevertheless, their wider use is limited because of weak fluorescence intensity and low colloidal stability in the biological environment. e optimization of treatment procedures and development of new suitable surface designs is therefore critically needed. In this study, several key steps for fluorescent nanodiamond treatment have been optimized, leading to both a substantial increase in fluorescence intensity and to significantly lower surface damage caused by graphitization. Further, a new high-throughput irradiation technique was developed. e influence of surface chemistry on the fluorescence parameters was studied using partial fluorination of the functional groups on the nanodiamond surface. A novel method which significantly affects the interaction of nanodiamonds with biological systems by increasing of the homogeneity and circularity was developed. e potential of nanodiamonds for future medical and biological research was demonstrated on particles with complex surface architectures that enabled targeting and therapy of tumor cells. Moreover, a strong and highly selective affinity of fibroblast growth factors to diamond...