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Optical speed sensor of surface
Vaněk, Ondřej ; Hadaš, Zdeněk (referee) ; Sedlák, Petr (advisor)
The aim of this work is development of an optical sensor for surface-velocity measurement based on ordinary optical mouse sensor. The proposed sensor should have find its usage in industrial applications for low-cost contactless measurement of conveyor belts or semi-finished products, thus some other gauges could check quality of products. Theoretical part of this thesis includes introduction to optical sensors and their usage. Rest of the work describes practical part consisting of programming of communication between sensor and PC in MATLAB and Arduino environments, development of simple device based on Arduino hardware, the set-up of experimental workplace and providing speed tests.
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Sanitation installation and gas installation in multifunctional building
Vaněk, Ondřej ; Bárta,, Ladislav (referee) ; Vrána, Jakub (advisor)
Bachelor Thesis ,,Technical and gas installations inmulti-building‘‘ is processed in the form of project documentationplumbing and gas installations for building construction in accordance with applicable regulations. Two-storey building is designed, besement. The building is used on the first floor of the municipal office and on the second floor of meeting andbanqueting facilities. Project part focuses on the design and solvingplumbing and gas installations. Part of the thesis are the total property assessmentsand literature review focused on the issue of hot water.
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Sanitation installation and gas installation in residential building
Vaněk, Ondřej ; Vaščáková, Alena (referee) ; Vrána, Jakub (advisor)
The thesis deals with sanitary engineering and gas installation in an apartment building with a healthcare facility. The theoretical part is focused on the installation systems, which are addressed to the suitability of the system in the given project, and the advantages and disadvantages of these systems. Computational and project part addresses the sewer, water mains and gas in the specified object. Designed building is without a basement and eight storeys. On the first floor is located technical facilities around the building and three residential units. All other floors are located on each floor of four residential units. This thesis is done on the basis of current Czech and European regulations.
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Comparative study of body temperature measurement using various technologies
Vaněk, Ondřej ; Sekora, Jiří (referee) ; Provazník, Ivo (advisor)
The aim of this work is to compare the body temperature measurement with different contact thermometers. The theoretical part includes studying the topic, familiarization with the situation at the Faculty Hospital Brno, Bohunice and suggestion of method for comparison. The practical part includes thermometers measurement in the laboratory to verify the accuracy, clinical measurement on patients in different parts of the body and under various conditions and statistical evaluation of results.
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Activating antitumor immune response using bispecific fusion proteins
Chytrá, Gabriela ; Vaněk, Ondřej (advisor) ; Černá, Věra (referee)
Natural killer (NK) cells are lymphocytes of the innate immune system that recognize and eliminate transformed and potentially harmful cells in a mechanism termed immunosurveillance. Malignant cells strive to escape immunosurveillance, and if successful, oncological disease develops. To restore immune recognition, immunotherapy utilizing NK cell-directed therapeutic fusion proteins can be employed. Therapeutic fusion proteins target tumour markers expressed on the surface of malignant cells and, at the same time, stimulate immune response through binding to NK cell activating receptors, for example receptor NKG2D or NKp30. A relevant example of a tumour marker is the HER2 receptor, which is often overexpressed in several types of cancer, most notably breast carcinoma. This thesis describes the preparation of several bispecific fusion proteins with potential use in immunotherapy. Bispecific fusion proteins consist of an NK cell activating ligand (ligand MICA or B7-H6) and nanobody targeting selected tumour marker (receptor HER2), which are connected by flexible glycine-serine linker. The constructs of fusion proteins were prepared in two configurations - with nanobody located on the N-terminus and the ligand on the C-terminus and vice versa. In addition, bispecific fusion proteins introducing...
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Effect of selected compounds on NMPylation of nsp9 SARS-CoV-2 catalyzed by NiRAN domain of nsp12 SARS-CoV-2
Fedák, Michal ; Šilhán, Jan (advisor) ; Vaněk, Ondřej (referee)
SARS-CoV-2 pandemic that begun in the end of 2019 caused over 770 milion confirmed cases of infection and almost 7 milion deaths worliwide. Despite pandemic being suppressed thanks to vaccines and restriction measures, the research of new medication targeting this virus continues. One of the possible targets of SARS-CoV-2 is nonstructural protein 9 (nsp9), which is essential for virus repliation. Just recently it was revealed that this protein can covalently bind nukleoside monophosphate (NMPylation) or RNA chain of various length (RNAylation). These reactions are catalyzed by nidovirus RNA-dependent RNA polymerase (RdRp) asociated nucleotidyltransferase (NiRAN) domain of nonstuctural protein 12 (nsp12). RNAylation of nsp9 showed to be crucial step in synthesis of RNA cap. The function of NMPylation was not jet revealed, but it is believed that this modification of nsp9 could play a role in priming of RNA synthesis. Some studies have already investigated inhibition of NMPylation. In this diploma thesis recombinant nsp9 was prepared and used in NMPylation reactions. Negative effect of DMSO on nsp9 NMPylation was observed. Three compounds (saquinavir, darunavir and conivaptan) selected based on the results of in silico studies did not show ability to inhibit NMPylation of nsp9. Next, it was confirmed...
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Cancer Immunotherapy exploiting engineered antibody fragments against prostate-specific membrane antigen
Das, Gargi ; Bařinka, Cyril (advisor) ; Vaněk, Ondřej (referee) ; Ormsby, Tereza (referee)
Prostate cancer (PCa) remains a leading cause of male cancer-related mortality, necessitating thus the development of novel therapeutic approaches as conventional treatments have limited efficacy. Prostate-specific membrane antigen (PSMA) is an established biomarker for both imaging and therapy of PCa, as it is highly upregulated in neoplastic PCa tissues and metastatic castration- resistant prostate cancer. Consequently, immunological targeting of PSMA has gained significant attention as a therapeutic platform for the management of the disease. The thesis is focused on engineering of antibody fragments and fusion proteins derived from the high affinity anti-PSMA 5D3 monoclonal antibody that can be used as immune cell engagers to target and eliminate PSMA-positive cells. To this end, we engineered 5D3 single chain variable fragments (scFv) that were subsequently fused to anti-CD3 scFv and CP33 sequences, creating thus immune cell engagers targeting T-cells (BiTE) and monocytes (5D3-CP33), respectively. The engagers were expressed in insect cells, purified to homogeneity and their biophysical and functional characteristics evaluated using size exclusion chromatography, differential scanning fluorimetry, ELISA and flow cytometry. Ensuing cell-based assays revealed that both BiTE and 5D3-CP33 can...
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Preparation of growth factor TGF-β3 with coiled-coil tag
Otépka, Tomáš ; Vaněk, Ondřej (advisor) ; Kubíčková, Božena (referee)
Growth factors represent a group of significant substances in the metabolism of organisms. They are signaling molecules that control cell activity at the endocrine, paracrine, or autocrine levels. They act as key mediators binding to cell receptors, triggering a cascade of reactions leading to the regulation of genetic transcription in the cell nucleus and stimulation of cellular response. Growth factors influence various physiological functions such as cell proliferation, cell differentiation, and tissue healing. The utilization of growth factors is evident, for example, in regenerative medicine. For a similar purpose, research has been initiated to prepare the growth factor TGF-β3 with the possibility of attaching it to a polymeric carrier using a coiled-coil tag. This work focuses on the recombinant production of TGF-β3 - its analog with a latency-associated peptide (LAP) - and the application of techniques applicable to the intention of this work, specifically, attaching the protein to a polymeric carrier based on amino acids. Given the structural complexity with which growth factors are physiologically released from cells, the preparation of growth factors with a coiled-coil tag in vitro represents an unexplored challenge in the field of recombinant protein expression. In our HEK293T cell line...
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Preparation of a polymeric therapeutic targeting CD20 positive B-lymphomas and NK cells
Hejl, Maxmilián ; Vaněk, Ondřej (advisor) ; Bělonožníková, Kateřina (referee)
Malignant transformation of B-cells is manifested by a marked increase in the number of surface markers 20 (CD20, cluster of differentiation 20). In studying this trend, chimeric monoclonal antibodies targeting CD20 were introduced to induce apoptosis in B-lymphomas. Since the introduction of the first therapeutic monoclonal antibody rituximab, many others have been developed, with some still used to treat B-lymphomas today. Unfortunately, in many cases, resistance to these drugs is developing, and therefore the development of new types of therapeutics is still relevant. This work aims to develop a polymer-protein macromolecular conjugate capable of inducing apoptosis in CD20 positive leukemia cell lines. For this purpose, we work with biologically active vectors, so-called anti-CD20 nanobodies. This is a variable binding domain derived from the "heavy chain only" antibodies found in, e.g., llamas or camels. Compared to conventional antibodies, nanobodies are approximately ten times smaller, but their binding affinity for the antigen is not altered. For this reason, nanobodies are ideal candidates for attachment to a polymeric carrier, where poly-N-[2-(hydroxypropyl)methacrylamide] (pHPMA) was chosen in this work. The transpeptidase reaction catalyzed by recombinant sortase A, which recognizes and...
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