Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Petra Mottlová Supervisor: Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of valencene on adhesion of breast cancer cell line MDA-MB-231 Valencene belongs to sesquiterpenes and is constituent of essential oils of many plants. It is the main citrus flavoring component. Valencene is mainly used in the food and cosmetic industries, but recent studies have confirmed its biological activity. Its antitumor, antiinflammatory, antioxidant and antiallergic effects have been already proven. The objective of this study was to determine the cytotoxic effect and influence on the adhesion of sesquiterpene valencene on breast cancer cell line MDA-MB-231. Another objective was to study the mechanism of its effect from the perspective of the selected adhesion molecules, which have an important role in tumorigenesis and metastasis. Valencene's cytotoxic effect was tested with use of neutral red (NRU test). Valencene influence on cell adhesion was continuously monitored by means of the X-Celligence device and expression of selected adhesion molecules was studied by Western blot and qPCR methods. The results showed a slight cytotoxic effect of valencene. Cell viability was over 70% at a...

Although adhesion of bacteria and yeast have been extensively studied by a wide range of experimental and theoretical approaches, significantly less attention has been focused on microalgal adhesion to solid materials. Therefore this doctoral thesis is focused on physico-chemical aspects of microalgal adhesion. The results are based on experimental characterization of surface properties of both microalgae and solids by contact angle and zeta potential measurements. These data were subsequently used in modeling the surface interactions (thermodynamic, classical and extended DLVO models) resulting in quantitative prediction of the interaction intensities. Finally, the model predictions are compared with experimental adhesion tests of microalgae onto model solids in order to identify the physico-chemical forces governing the microalgae-solid interaction. The model solids were prepared in order to cover a wide range of properties (hydrophobicity, surface charge). The results revealed that in low ionic strength environment the adhesion was influenced mostly by electrostatic attraction/repulsion between surfaces, while with increasing ionic strength grew the importance of apolar (hydrophobic) interactions. Among tested materials the highest adhesion of Chlorella vulgaris cells was observed to (3-aminopropyl)triethoxysilane modified glass (APTES glass) mimicking the surface properties of stainless steel. The impact of solid surface properties on the degree of colonization by microlagae was statistically more significant than the influence of culture medium composition on cell surface of Chlorella vulgaris.

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Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Markéta Marešová Supervisor: Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of alpha-humulene on adhesion of breast cancer cell line MDA-MB-231 α-humulene is a sesquiterpene contained in the essential oil of Chinese Bayberry (Myrica rubra), which has various biological effects. The aim of this thesis was to study the cell adhesion of tumor cell line MDA-MB-231, and the effects of α-humulene on expression of cell adhesion molecules. Furthermore, the cytotoxic effect of α-humulene on this cell line was verified. Effect of α-humulene on cell proliferation was evaluated by neutral red uptake test (NRU test) and by xCelligence system. Cell adhesion was also continuously monitored by xCelligence. Expression ganges of cell adhesion molecules upon α-humulene treatment were determined by Western blotting and by quantitative polymerase chain reaction on the protein and mRNA levels, respectively. The proliferation of the cells was significantly affected by α-humulene after 24 hour treatment (IC50 13.6 µg/ml). α-humulene alone caused slightly increased cell adhesion. Adhesion molecules EpCAM and β-cathenin were almost unaffected. Level of ICAM1 mRNA was increased after 12 hours and...

Cardiovascular diseases are the major causes of death worldwide. Studying factors leading to initiation and progression of atherosclerosis and its complications leads to a better understanding of underlying mechanisms of this disease and to development of novel treatments. Adhesion of monocytes on the endothelial surface is the initial step of atherosclerosis. The main aim of this study was to establish and test an in vitro model of monocyte adhesion on the endothelial cells and to evaluate the results by means of two methods - measuring the fluorescence signal intensity and counting adhered cells. Because of its well known effects on endothelial cells activation and adhesion molecules expression TNF-α was chosen for endothelial cells stimulation. The lowest concentration of TNF-α affecting the percentage of adhered monocytes in comparision with negative control was 1 ng TNF-α/ml. The optimal concentration of TNF-α increasing the percentage of adhered monocytes was 10 ng TNF-α/ml. The influence of TNF-α on the adhesion was observed already after 5 minutes of coincubation of THP-1 monocytes with HUVEC. Using the optimal concentration of 10 ng/ml led to the highest percentage of adhered monocytes after 30 - 40 minutes of coincubation with HUVEC. Other factor affecting the percentage of adhered...

The protein p130Cas (human ortholog BCAR1) is a major substrate for phosphorylation by the Src family kinase and plays a central role in oncogenic transformation. Increased level of BCAR1 correlates with primary tumour growth and cancer progression. Localized to focal adhesion, p130Cas serves as a mechanosensor and mediates key interactions with the extracellular environment. The structure of p130Cas is crucial for its function, mainly the anchoring domains SH3 and CCH, together with the substrate domain which is extended when under tension. This Master's thesis presents a newly developer FRET mechanosensor based on the structure of p130Cas. The sensor utilizes the anchoring domains of p130Cas for proper localization to focal adhesions, where it can detect tension in living cells. Key words: p130CAS, FRET, focal adhesions, mechanosensing

This master thesis deal with the glued joint. Teoretical part is focused on basic facts and an advanteges or disadvanteges of this technique. A large part is devoted to theory of gluid process, adhesion, cohesion wettability of glued surfaces. Last part of the theoretical part is searches in the field of tension glued joints and methods to quality assessment of made joint. Experimental testing glued surfaces was made as bend test. During testing was monitored the acoustic emissions. The emissions characterize violation of glued joints. Results of this experiment are evaluated in the end of this thesis.

The protein p130Cas (human ortholog BCAR1) is a major substrate for phosphorylation by the Src family kinase and plays a central role in oncogenic transformation. Increased level of BCAR1 correlates with primary tumour growth and cancer progression. Localized to focal adhesion, p130Cas serves as a mechanosensor and mediates key interactions with the extracellular environment. The structure of p130Cas is crucial for its function, mainly the anchoring domains SH3 and CCH, together with the substrate domain which is extended when under tension. This Master's thesis presents a newly developer FRET mechanosensor based on the structure of p130Cas. The sensor utilizes the anchoring domains of p130Cas for proper localization to focal adhesions, where it can detect tension in living cells. Key words: p130CAS, FRET, focal adhesions, mechanosensing

Focal adhesions are important subcellular structures that are composed of many signaling and scaffolding proteins. They serve not only for anchoring the cell to the substratum but they are also important signaling centers that regulate various cellular behavior such as migration, invasiveness, proliferation and survival. Focal adhesion signaling needs to be strictly regulated because alteration in activity or expression of many focal adhesion proteins leads to tumorogenesis and metastasis formation. One of the most important scaffolding protein associated with focal adhesion is p130Cas. The importance of p130Cas in regulation of cell migration and invasiveness has been well established. P130Cas also plays important role in regulation of cell survival and proliferation. Moreover, high protein levels of human ortholog of p130Cas - BCAR1, has been linked to more aggressive breast tumors and poor prognosis. During my doctoral studies, I focused on the role of p130Cas in integrin signaling. At the beginning we characterized the role of tyrosine 12 phosphorylation within its SH3 domain. We confirmed that this phosphorylation is increased in Src527F transformed mouse embryonic fibroblasts compared to non-transformed counterparts and also in some human cancer cell lines. We showed that this phosphorylation...

This thesis deals with the measuring of adhesive and cohesive characteristics of composite systems filled with microparticles of glass powder. The theoretical part describes in detail composites from their historical development to the present, also their characteristics, general properties, distribution and types of fibers and matrices. Emphasis is placed on composites filled with particulate filler and current researches in the field of composites with glass filler. Furthermore, the adhesion and cohesion of filled systems is described. The experimental part of this thesis includes a methodology, which describes in detail the used materials, methods of preparation of mechanical testing and statistical analysis. The results of mechanical tests for all three composite mixtures and evaluation of cost savings are appended to this thesis.