- David Větvička, M.Sc . Many researchers have, in the past, focused on pathophysiological features of tumor tissue, various tumor-nonmalignant cell interactions, and secretion of active molecules within the tumor mass. All these aspects of tumor structure are known as tumor microenvironment. The composition of particular tumor ecosystem is highly variable, with differences between various tumor types, even between patients with the same diagnosis, and in separate areas of the same tumor. Moreover, further changes in tumor microenvironment often occur during the progression of the disease. Studies of tumor microenvironment have revealed both novel targets for therapy and new prognostic markers. New therapy modalities are being developed to target these discovered features, including drugs functioning to boost anti-malignancy immunity, to block pro-metastatic potential, or to utilize the unique features of this pathological environment established by the tumor. These are obviously of great interest and harbor high potential for better management of malignant diseases. The focus of this thesis is to study the interactions of polymeric drug delivery systems within the tumor microenvironment and to utilize various features of this specific niche for drug delivery research. We have followed three...

Cancer cell invasion and metastasis are hallmarks of cancer. It is becoming apparent that the interaction between cancer cells and the surrounding microenvironment are involved in their ability to invade and metastasise. In general, cancer cells can either migrate individually, in an amoeboid or mesenchymal manner, or collectively. The first aim of this thesis was to analyse the role of NG2 in amoeboid to mesenchymal transition (AMT) and Rho/ROCK signalling. We found that NG2 promotes an amoeboid morphology, and increased invasiveness, in a Rho-dependent manner. Secondly, we analysed the role of the major tumour microenvironment (TME) component, cancer-associated fibroblasts (CAFs), on melanoma cell invasiveness. We found the CAF interaction with melanoma cells leads to increased levels of interleukin-6 (IL-6) and IL-8, and this leads to increased invasiveness. Simultaneous blocking of IL-6 and IL-8, using neutralising antibodies, inhibits CAF-dependent invasion. Further analysis of another major component in the melanoma TME, keratinocytes, has highlighted the importance of the tumour cell niche in invasion. Our results indicate that cancer cells have the ability to change morphology, and that the TME plays an important role in melanoma cell invasiveness. Metastatic melanoma treatment has proven...

This Master's Thesis aims to review current wine grape production in the Czech Republic. External and competitive environment analyses and other methods from financial analysis are used here. A theoretical part of the thesis contains definition of strategy and strategic management. Beside that there are defined base tools used for macro-environment and micro-environment analyses. A practical part of the thesis is focused on PEST analysis, a delimitation of current wine grape production and competitive market. The practical part also covers main obstructions in entering to the wine industry. Last two parts of the thesis assess situation of wine grape production in the Czech Republic. SWOT analysis for the future evolution of this industry are included.

Modulation of the tumor microenvironment represents a possible way to inhibit cancer growth and enhance anti-cancer immune responses. In the presented work we employ two strategies for tumor microenvironment modulation. Firstly, we have constructed rVACV co-expressing the tumor suppressor gene insulin-like growth factor-binding protein-3 (IGFBP- 3) and the fusion gene encoding the immunogen SigE7LAMP. The expression of IGFBP-3 was regulated either by the early vaccinia virus H5 promoter or by the synthetic early/late (E/L) promoter. We have shown that expression of IGFBP-3 regulated by the H5 promoter yielded higher amounts of IGFBP-3 protein when compared with the E/L promoter. Immunization with P13-SigE7LAMP-H5-IGFBP-3 was more effective in inhibiting the growth of TC-1 tumors in mice and elicited a higher T-cell response against VACV-encoded antigens than the control virus P13-SigE7LAMP-TK- . We found that high-level production of IGFBP-3 enhanced virus replication both in vitro and in vivo, resulting in profound antigen stimulation. Production of IGFBP-3 was associated with a higher adsorption rate of P13-SigE7LAMP-H5-IGFBP-3 to CV-1 cells when compared with P13-SigE7LAMP-TK- . We have identified two structural differences between the IMVs of the IGFBP-3 expressing virus P13-SigE7LAMP-H5-IGFBP-3...

The aim of this diploma thesis was to analyze the most important points of the marketing plan of the company ZOO Prague and evaluate the results and lessons learned during the analysis. The work also contains proposals of new marketing objectives, which could enhance current marketing activities of the company. The first two chapters focus on the theoretical definitions and the following chapters deal with partial analyzes. For example, the analysis of macro and micro (environment). The thesis also includes an own survey, supported by mystery shopping.

Cancer diseases represent second most frequent cause of death after cardiovascular diseases in Europe. Nowadays used medical treatments like chemotherapy and radiotherapy are nonspecific and cause huge side effects. Various systems to deliver therapy directly inside the tumour microenvironment and reduce side effects are under development. Protein nanoparticles seem to be very promising strategy to achieve that goal. Our group in cooperation with CNR in Rome tested nanoparticles based on heavy chain of human ferritine. These constructs, modified to expose the tumor targeting molecule, were able to be specifically internalised by B16F10 melanoma cells in vitro. They also specifically target and localise at the sites of primary melanoma and lung metastases of different size in mouse in vivo model. These nanoparticles can carry either therapeutic or diagnostic molecules. Thus they represent a suitable candidate for further studies for potential use in clinical praxis as a diagnostic and/or therapeutic agents (theranostics). Powered by TCPDF (www.tcpdf.org)

TRAIL ligand can trigger apoptosis of permissive human cells via engagement of its two pro- apoptotic receptors TRAIL-R1 (DR4) and TRAIL-R2 (DR5). Its ability to induce apoptosis independently on p53 status and to selectively kill cancer cells in vitro and in vivo made this ligand an attractive target in cancer research. However, acquired resistance of primary cancer cells, unsatisfactory outcome of clinical trials and recent studies arguing that TRAIL might under specific conditions promote cancer progression, opened new plethora of questions, which need to be addressed. Though both receptors DR4 and DR5 are ubiquitously expressed, different types of tumours show preference for either of the receptors. The relative participation of DR4 and DR5 in TRAIL- induced signalling is still largely unknown. To analyse TRAIL receptor-specific signalling, I prepared Strep-tagged, trimerised variants of recombinant human TRAIL ligands with high affinity for either DR4 or DR5 receptor. Using these receptor-specific ligands, I examined a contribution of individual pro-apoptotic receptors to TRAIL-induced signalling pathways. I found that in TRAIL resistant colorectal HT-29 cells but not in pancreatic PANC-1 cancer cells, DISC formation and initial caspase-8 processing proceeded comparably in both DR4- and...

Cancer development and progression vary depending on tumor type, localization, invasion, immunogenicity and the ability of immune system to become activated. There are frequent interactions between tumor cells and immune cells, occuring locally at the site of primary tumor or distally through paracrine signalling of various mediators and cytokines. The main subject of this PhD thesis is to study key factors and aspects of immune response in cancer patients. In the first part, we analyzed immune cells infiltrating tumor tissues of ovarian cancer patients at different stages of disease. We focused on the dynamics of immune response, primarily on frequency of individual T lymphocyte populations in peripheral blood and tumor infiltrating T lymphocytes in tumors of early and advanced stages of ovarian cancer. We found that during disease progression there is a gradual decrease of proinflammatory Th17 and Th1 immune responses and a specific recruitment of regulatory T cells to the tumor site, which results in a significant immune suppression in the tumor microenvironment. In the second part, we demonstrated that the character of immune response in HPV-positive head and neck cancer patients is very different from the patients with tumors not associated with HPV infection. In HPV-positive patients, significantly...

The dissertation is focused on the study of intercellular interactions in skin tumors. It is based on 5 original publications that cover several topics. We studied the origin of tumor-associated fibroblasts concerning the primary tumor population. We demonstrated using a mouse model that tumor-associated fibroblasts are produced from the host organism and thus did not arise from transformation directly from tumor cells. We also investigated the relationship between tumor-associated fibroblasts and keratinocytes. We have shown that tumor-associated melanoma fibroblasts affect keratinocytes which, under their influence, acquire the features typically observed in migrating cells and cells undergoing epithelial-mesenchymal transition. We studied the interactions between healthy fibroblasts and tumor cells. We have demonstrated that fibroblasts acquired from healthy skin from a patient suffering from melanoma are significantly different from control fibroblasts of healthy donors in the expression profile. Changes in distal fibroblasts support the view of melanoma as a systemic disease. We have further demonstrated that melanoma-associated fibroblasts do not carry a BRAF mutation, in contrast to BRAF positivity of melanoma cells. And therefore, they did not arise from the transition from melanoma. The...

This diploma thesis is tasked to evaluate the company Kordárna Inc. with the view of its sale within the frame of reorganization as a nonliquidating solution to its insolvency. The evaluation of the company Kordárna Inc. is performed to the date of January 1st, 2009. By then the company, due to many unfavorable conditions, falls into existential problems that have to be immediately solved. The reorganization plan is allowed and processed within the framework of higher creditors' satisfaction. An integral part of this plan is also the desirable evaluation of the company Kordárna Inc. with the view of its sale to a new investor. Within these issues, both the detailed external and internal analysis of the company and the financial analysis of the 5 preceding fiscal periods are processed (years 2004-2008). Consequently, based on many assumptions, the financial plan for the 4 following fiscal periods (2009-2012) is conducted, and the evaluation of the company Kordárna using 4 evaluation methods -- FCFF, FCFE, EVA and Liquidation value methods -- is processed.