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Macrophages selectively sense mechanical cell death in CLASP-depleted melanoma cells to trigger CXCL10 response
Otruba, Matúš ; Labzin, Larisa (advisor) ; Filipp, Dominik (referee)
Melanoma represents a significant and often fatal form of cancer, with metastasis being a primary cause of cancer-related deaths. Immunotherapy aims to stimulate the immune system to eliminate tumors, but even when used together with traditional chemotherapy, still only has <50% success rate. Chemotherapeutic drugs induce tumor cell death, primarily through apoptosis, which is 'silent' and does not incite inflammation or immune cell infiltration into the tumor. During metastasis, tumor cells migrate through confined spaces. CLASP proteins protect organelles and cellular integrity during tumor cell metastasis; when CLASP proteins are depleted, tumor cells migrating through tissue die through a mechanical cell death. The immunological impact of CLASP depletion-induced cell death remains unknown. This study aims to explore whether macrophages, innate immune cells that sense neighboring dying cells, trigger inflammatory cytokine responses specifically to mechanical, as opposed to chemotherapeutic, melanoma cell death. To model mechanical cell death during metastatic migration, CLASP1, a microtubule stabilizing protein, was depleted, and cells were physically compressed to simulate forces encountered during metastasis. Chemotherapy was simulated using a B-Raf inhibitor and the apoptosis inducer...
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Manufacturing of clasp
Polách, Libor ; Řiháček, Jan (referee) ; Podaný, Kamil (advisor)
The project is focused on proposal of clasp technology production. The clasp is a part of central lock in parachute harness. The part is made of galvanized steel sheet metal, sheet metal thickness 3 mm. The part has a spatial shape with holes. Expected manufacture series is 25 000 per year. Bassed on literary studies and manufacture requirements on the clasp, the method of cutting combinated with stamping in combined tool was found the most optimal. The final shape of clasp will be done in three steps. The nobel steel 12 050, which performs required mechanical properties, was chosen as the main material for clasp manufacture. The manufacture will be made by eccentric press LEXN 100C. The expected price of the clasp is 9,7 CZK per piece with counting the procurement price of metallurgical semi-finished product and the tool.
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Searching for mechanisms and functions of microtubular interactions with other plant cell structures
Krtková, Jana ; Schwarzerová, Kateřina (advisor) ; Vaňková, Radomíra (referee) ; Ovečka, Miroslav (referee)
Microtubular cytoskeleton is involved in many processes in plant cells, including cell division, growth and development. Other proteins enable its functions by modulation of its dynamics and organization and by mediation of functional and structural interaction with other cell structures. Identification of the mediating proteins and the functions of these interactions under specific conditions were the main aims of the thesis. Membrane proteins interacting with microtubules were identified using biochemical methods. Surprisingly, the identified proteins co-sedimenting with microtubules were not members of the "classical" microtubule associated proteins (MAPs). There were enzymes, chaperones and plant specific proteins among them. For further studies, the identified microtubule-associated heat-shock protein 90 (Hsp90_MT) was chosen. Recombinant Hsp90_MT binds directly to microtubules and tubulin dimers in vitro. The ATP-binding pocket is not responsible for this association. In BY-2, Hsp90_MT co-localizes with phragmoplast and cortical microtubules and is involved in microtubule recovery after their depolymerization during cold treatment. In plants, Hsp90 is involved in cell cycle progression, its inhibition causes cell-cycle arrest in G1 phase. Based on literature search for animal proteins...
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Searching for mechanisms and functions of microtubular interactions with other plant cell structures
Krtková, Jana ; Schwarzerová, Kateřina (advisor) ; Vaňková, Radomíra (referee) ; Ovečka, Miroslav (referee)
Microtubular cytoskeleton is involved in many processes in plant cells, including cell division, growth and development. Other proteins enable its functions by modulation of its dynamics and organization and by mediation of functional and structural interaction with other cell structures. Identification of the mediating proteins and the functions of these interactions under specific conditions were the main aims of the thesis. Membrane proteins interacting with microtubules were identified using biochemical methods. Surprisingly, the identified proteins co-sedimenting with microtubules were not members of the "classical" microtubule associated proteins (MAPs). There were enzymes, chaperones and plant specific proteins among them. For further studies, the identified microtubule-associated heat-shock protein 90 (Hsp90_MT) was chosen. Recombinant Hsp90_MT binds directly to microtubules and tubulin dimers in vitro. The ATP-binding pocket is not responsible for this association. In BY-2, Hsp90_MT co-localizes with phragmoplast and cortical microtubules and is involved in microtubule recovery after their depolymerization during cold treatment. In plants, Hsp90 is involved in cell cycle progression, its inhibition causes cell-cycle arrest in G1 phase. Based on literature search for animal proteins...
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Manufacturing of clasp
Polách, Libor ; Řiháček, Jan (referee) ; Podaný, Kamil (advisor)
The project is focused on proposal of clasp technology production. The clasp is a part of central lock in parachute harness. The part is made of galvanized steel sheet metal, sheet metal thickness 3 mm. The part has a spatial shape with holes. Expected manufacture series is 25 000 per year. Bassed on literary studies and manufacture requirements on the clasp, the method of cutting combinated with stamping in combined tool was found the most optimal. The final shape of clasp will be done in three steps. The nobel steel 12 050, which performs required mechanical properties, was chosen as the main material for clasp manufacture. The manufacture will be made by eccentric press LEXN 100C. The expected price of the clasp is 9,7 CZK per piece with counting the procurement price of metallurgical semi-finished product and the tool.
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