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Measurement of tissue embryos by MR imaging methods
Vrba, Jan ; Mikulka, Jan (referee) ; Bartušek, Karel (advisor)
The aim of this work is to determine the volume of water in tissue cultures of spruce MR imaging methods and changes during growth. Was measured set of samples, where was found contain of cluster in each cuts. It describes spin-echo method, which was used for measurement. In this work is given a description of the acquisition, calculation, evaluation and graphical representation of measured values.
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Ecotoxicity tests on cellular level
Procházková, Petra ; Vávrová, Milada (referee) ; Zlámalová Gargošová, Helena (advisor)
Endocrine disrupting compounds among significant environmental pollutants. They are dangerous both for humans and other organisms by affect their hormonal system. They enter the environment during production, use and disposal some chemicals, but they may be of natural origin. The aim of this work is to give a brief overview of possible methods of determination of the presence of ecotoxic substances in the environment, using the in vitro toxicity assays. These tests use cell cultures instead of test organisms. The work describes principles of ecotoxicity tests at the cellular level, such as cell viability assays, cell proliferation assays, assays based on the metabolic activity of cells or DNA microarrays. A significant part of this work is focused on the possibilities of determination of endocrine disruptors by selected assays. They are ligand binding assays, reporter gene assays (recombinant yeast assay, mammalian-based reporter gene assays) and cell proliferation assay called E-screen. Significance and rationale use of tests to prove the endocrine activities in environmental matrice is given through examples of studies in which the tests were used.
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MR imaging of somatic embryos
Chovanec, Ján ; Kubásek, Radek (referee) ; Bartušek, Karel (advisor)
The aim of this bachelor thesis is measuring of tissue cultures of spruce MR imaging methods. Was measured set of samples, where was found contain of cluster in each cuts. To obtain the image, the magnetic resonance was used, specifically spin echo sequence. Method called spin echo is based on two pulses 90° and 180°. Selected method from the beginning of the work is than used to measure the diffusion and processing of cultures. In last part the work is devoted to data processing in Matlab and Marevisi.
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Optimisation of rapeseed in vitro cultures used for CRISPR/Cas9 technology
HARENČÁK, Jan
Plant transformation using Agrobacterium tumefaciens bacteria (Rhizobium radiobacter, according to new nomenclature) is widely used method for plant genome editing. However, different oilseed rape varieties do not always have the same in vitro regeneration capacity, which is crucial for transformation. This work aims to optimize regeneration protocol for selected varieties of oilseed rape. Of the six varieties tested, only two regenerated successfully - Arabella and Obelix. The created protocol can be used for regeneration without transformation, but it is also possible to implement specific steps in it that enable transformation using A. tumefaciens.
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The comparison of properties of cell lines resistant to ellipticine, doxorubicin, and cisplatin
Černá, Tereza ; Poljaková, Jitka (advisor) ; Eckschlager, Tomáš (referee)
7 Abstract Neuroblastoma is the most common extracranial solid tumor of childhood. Despite advances in cancer diagnosis and therapy, the treatment of some forms of neuroblastoma is still complicated. One of the major complications of the chemotherapy is a developed drug resistance. This master thesis deals with the effect of cytostatics on protein and gene expression of selected proteins, which may contribute to chemoresistance of the human neuroblastoma cell line UKF-NB-4. The sensitive line UKF-NB-4 and the resistant line UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI were exposed to cisplatin, doxorubicin, ellipticine for 24, 48 and 72 hours. The Western blot analysis showed that cytostatic agents cisplatin, doxorubicin or ellipticine added to the sensitive neuroblastoma cell line UKF-NB-4 in amounts which are added to resistant neuroblastoma cell lines in order to maintain resistance induced expression of p53 and reduced expression of retinoblastoma protein pRb after 72 hours of cultivation. Differences in the expression of RAS protein, cytochrome P450 1A1, 3A4 and cytochrome b5 has not been shown. Changes in the expression of the studied proteins in resistant lines UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI cultured with and without cytostatic agents were not detected by the Western blot analysis....
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The expression of macrophage migration inhibitory factor in neuroblastoma cell lines
Polatová, Daniela ; Poljaková, Jitka (advisor) ; Groh, Tomáš (referee)
A macrophage migration inhibitory factor (MIF) is a cytokine which participates in immune responses induced by outer stimuli such as lipopolysaccharides of bacteria or glucocorticoids. It is produced mainly by macrophages. It activates other macrophages and T-lymphocytes and allows anti-inflammatory cytokines to be released. It contributes to angiogenesis - endogenously produced MIF is crucial for the proliferation of endothelial cells. The formation of new blood vessels stimulated by MIF was also described in tumors and it leads to tumor growth. The natural receptor for MIF is an integral membrane protein CD74, which could be presented on the cell surface. If it is presented on the surface of T-lymphocytes, it could be modified by chondroitin- sulphate and interact with a protein CD44. This modification and interaction is important for an effective activation of T-cells. MIF then binds to the complex CD74-CD44. The presence of CD74 in human neuroblastoma cell lines UKF-NB-3 and UKF-NB-4 was studied in this bachelor thesis. Using electrophoresis and Western blot CD74 was detected in the UKF-NB-4 neuroblastoma cell line but it wasn't detected in the UKF-NB-3 neuroblastoma cell line. Nevertheless, the presence of this receptor in neuroblastoma cells indicates, that even neuroblastomas could be...
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The expression of macrophage migration inhibitory factor in neuroblastoma cell lines
Polatová, Daniela ; Poljaková, Jitka (advisor) ; Groh, Tomáš (referee)
A macrophage migration inhibitory factor (MIF) is a cytokine which participates in immune responses induced by outer stimuli such as lipopolysaccharides of bacteria or glucocorticoids. It is produced mainly by macrophages. It activates other macrophages and T-lymphocytes and allows anti-inflammatory cytokines to be released. It contributes to angiogenesis - endogenously produced MIF is crucial for the proliferation of endothelial cells. The formation of new blood vessels stimulated by MIF was also described in tumors and it leads to tumor growth. The natural receptor for MIF is an integral membrane protein CD74, which could be presented on the cell surface. If it is presented on the surface of T-lymphocytes, it could be modified by chondroitin- sulphate and interact with a protein CD44. This modification and interaction is important for an effective activation of T-cells. MIF then binds to the complex CD74-CD44. The presence of CD74 in human neuroblastoma cell lines UKF-NB-3 and UKF-NB-4 was studied in this bachelor thesis. Using electrophoresis and Western blot CD74 was detected in the UKF-NB-4 neuroblastoma cell line but it wasn't detected in the UKF-NB-3 neuroblastoma cell line. Nevertheless, the presence of this receptor in neuroblastoma cells indicates, that even neuroblastomas could be...
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The comparison of properties of cell lines resistant to ellipticine, doxorubicin, and cisplatin
Černá, Tereza ; Poljaková, Jitka (advisor) ; Eckschlager, Tomáš (referee)
7 Abstract Neuroblastoma is the most common extracranial solid tumor of childhood. Despite advances in cancer diagnosis and therapy, the treatment of some forms of neuroblastoma is still complicated. One of the major complications of the chemotherapy is a developed drug resistance. This master thesis deals with the effect of cytostatics on protein and gene expression of selected proteins, which may contribute to chemoresistance of the human neuroblastoma cell line UKF-NB-4. The sensitive line UKF-NB-4 and the resistant line UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI were exposed to cisplatin, doxorubicin, ellipticine for 24, 48 and 72 hours. The Western blot analysis showed that cytostatic agents cisplatin, doxorubicin or ellipticine added to the sensitive neuroblastoma cell line UKF-NB-4 in amounts which are added to resistant neuroblastoma cell lines in order to maintain resistance induced expression of p53 and reduced expression of retinoblastoma protein pRb after 72 hours of cultivation. Differences in the expression of RAS protein, cytochrome P450 1A1, 3A4 and cytochrome b5 has not been shown. Changes in the expression of the studied proteins in resistant lines UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI cultured with and without cytostatic agents were not detected by the Western blot analysis....
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Somatic embryogenesis of conifers: description of structural development
Kadlecová, Marie ; Albrechtová, Jana (advisor) ; Konrádová, Hana (referee)
Somatic embryogenesis is a very important tool of in vitro cultivation, therefore, it has recently devoted adequate attention. Stages of somatic embryogenesis involve several consecutive steps: induction of embryogenic culture and its proliferation, maturation of embryo, desiccation, germination and subsequent transfer of embryos into young seedlings. All of these steps are directly influenced by many factors, such as a choice of the original explant, time of cultivation, physical conditions or the composition of the culture medium. There are still emerging new studies describing how the culture conditions and, thus, the quality of embryos could be improved. An important prerequisite for further improving the quality of somatic embryos is good knowledge of aculture protocol, i.e. right concentrations of amended substances in the culture medium, the timing of each step, using the suitable explants etc. Another very important thing is good knowledge of development of all anatomical structures established during embryogenesis. Only then it will get closer to the point when somatic embryos will not arise only in the laboratory, but could be successfully used in practice. The object of this work is to summarize existing knowledge about development of all anatomical structures during somatic...
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Ecotoxicity tests on cellular level
Procházková, Petra ; Vávrová, Milada (referee) ; Zlámalová Gargošová, Helena (advisor)
Endocrine disrupting compounds among significant environmental pollutants. They are dangerous both for humans and other organisms by affect their hormonal system. They enter the environment during production, use and disposal some chemicals, but they may be of natural origin. The aim of this work is to give a brief overview of possible methods of determination of the presence of ecotoxic substances in the environment, using the in vitro toxicity assays. These tests use cell cultures instead of test organisms. The work describes principles of ecotoxicity tests at the cellular level, such as cell viability assays, cell proliferation assays, assays based on the metabolic activity of cells or DNA microarrays. A significant part of this work is focused on the possibilities of determination of endocrine disruptors by selected assays. They are ligand binding assays, reporter gene assays (recombinant yeast assay, mammalian-based reporter gene assays) and cell proliferation assay called E-screen. Significance and rationale use of tests to prove the endocrine activities in environmental matrice is given through examples of studies in which the tests were used.
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