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mTOR mediated phosphorylation of Akt at Ser473 regulates cancer cell survival after severe hypertermia in PTEN-coordinated manner
Valentová, Iva ; Sadílková, Lenka (advisor) ; Černý, Jan (referee)
Hyperthermia (HT), a term used in general for temperatures higher than 42řC, induces cell stress response in dependence on its intensity, heating duration and target cell type (this is called a thermal dose). In vitro, HT causes changes in Ca2+ homeostasis as a consequence of endoplasmic reticulum stress and permeabilisation of the outer mitochondrial membrane. The release of cytochrom c further leads to activation of caspases that can result in cell death. In parallel, the presence of denatured proteins in the cell cytosol leads to the development of the so called unfolded protein response (UPR). UPR is characterized by increased expression and activation of molecular chaperones, increase in the intensity of proteosomal degradation and enhancement in autophagy. This process is accompanied by the attenuation of translation machinery, iniciation of DNA repair and activation of cytoprotective mechanisms leading to cell survival. This highly complex stress response mechanism is regulated mainly by the signalling pathways in the PI3K/Akt/mTOR axis. Cell survival is in the end fully dependent on the amount of energetic substrates, denatured proteins accumulation and on the level of irreversibly demaged DNA. If the termal dose is too high, the cell will ultimatively die. Hyperthermia is also one of the...
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Examination of the ion levels in the blood before and after plasmapheresis
VALENTOVÁ, Iva
In my bachelor thesis I worked on the topic which is focused on the managing and measuring of the chosen ion levels (Na, K, Ca, P) before and after plasmapheresis. This thesis consists of the theoretical and practical part. The blood sample was collected from each of 25 donors who agreed with the providing their blood samples for research purposes, in cooperation with the plasmapheretic centre in České Budějovice. The first tube of blood was collected from each volunteer before plasmapheresis and the second tube was collected after plasmapheresis. The second blood sample was collected after the break of 10 minutes. Plasmapheresis is a separation method, which consists of the collection, treatment and return part of blood plasma into the blood circulation. There are two methods of obtaining plasma. There are plasmaferetic systems which work on the principle of centrifugation or membrane filtration. In my thesis, I was acquainted with the process of plasmapheresis, whose function was focused on collecting plasma for donor purposes and other plasma processing. The plasmapheretic center named Plasmafera s.r.o. based in České Budějovice works with automatic plasmapheretic separator Autopheresis-C. This system uses a plasmapheresis system , which is characterized by a discontinuous separator method. In the second part, I have measured 50 obtained blood samples from the plasma donors. The main goal was based on the collaboration with the laboratory during the measuring of minerals sodium, potassium, calcium and phosphorus. I have measured the blood samples in the cooperation with the Laboratory of clinical chemistry in the hospital in České Budějovice. Nowadays there are methods used for measuring ion levels which can be automatized. This is very useful especially for the large-scale laboratory which are these assesments matter routine. For the measurement of ion levels in the blood sample was used automatic analyzer ADVIA 1800, which examinates the serum, plasma and urine samples using the method of potentiometric measurements by ISE for the measuring of sodium, potassium and chloride concentration and by the photometric method, which works via the spectrophotometer for the determination of calcium and phosphorus levels. In the Laboratory of clinical chemismy in the hospital in České Budějovice I have gain a lot of information about the requirements during the pre-analytical phase in the laboratory and out of the laboratory. It was allowed me to participate the whole laboratory process under the supervision of the laboratory technician. The result of this process was obtaining the measured concentrations of the chosen ions.
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