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"The cancer cell proteome and its changes after anti-cancer drug treatment".
Tylečková, Jiřina ; Kovářová, Hana (advisor) ; Strnad, Miroslav (referee) ; Petrák, Jiří (referee)
Cancers represent a group of unprecedented heterogeneous diseases and currently available anti-cancer therapies provide highly variable efficacy with unsatisfactory cure rates. A wide range of proteomic technologies are being used in quest for newer approaches which could significantly contribute to the discovery and development of selective and specific cancer biomarkers for monitoring the disease state and anti-cancer therapy success. Taking into consideration the above aspects, this research was undertaken to study cancer cell proteomes and their changes after anti-cancer treatment with specific focus on: (a) response to conventional anthracycline/anthracenedione drugs with respect to their different clinical efficacy and (b) identification of novel targets for therapy in cancer cells resistant to biological drugs such as inhibitors of (b1) cyclin-dependent kinases and (b2) Aurora kinases. This study identified several interesting key aspects related to the effects of daunorubicin, doxorubicin and mitoxantrone. With the main focus on early time intervals when the influence of apoptosis is minimised, changes common for all three drugs belonging mainly to metabolic and cellular processes were observed. More importantly, significant changes in proteins involved in the generation of precursor...
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Proteomic analysis of selected oncohematological diseases
Pimková, Kristýna ; Dyr, Jan (advisor) ; Kodíček, Milan (referee) ; Petrák, Jiří (referee)
Oxidative stress is an important factor in carcinogenesis of oncohematological diseases. However its role in the pathogenesis of myelodysplastic syndromes (MDS) remains unclear. In this study, we have determined the oxidative status and evaluated proteomic changes in plasma of MDS patients as a consequence of oxidative dysbalance (oxidative modifications, protein-protein interaction and complex forming). We measured the levels of total cysteine, homocysteine, cysteinyglycine, glutathione, nitrites and nitrates in the plasma from 61 MDS patients and 23 healthy donors using high performance liquid chromatography. Glutathione and nitrites levels reduced significantly while other aminothiols levels increased significantly in plasma of MDS patients. This association with oxidative stress did not correlate with iron overload. We also found enhanced levels of asymmetric dimethylarginine in serums of middle aged patients with MDS that correlate to posttranslational modifications of proteins arginyl residues. Furthermore, carbonylated proteins level was significantly elevated in MDS patients compared to healthy donors. Using mass spectrometry, 5 S-nitrosylated blood platelets proteins were identified in plasma and blood platelets of MDS patients and set of 16 plasma proteins with high probability of...
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Molecular mechanisms of Diamond-Blackfan anemia
Handrková, Helena ; Petrák, Jiří (advisor) ; Šebela, Marek (referee) ; Trka, Jan (referee)
Diamond-Blackfan anemia (DBA) is a rare congenital syndrome that presents with ane- mia and selective deficiency of erythroid precursors, while other blood lineages are usu- ally unaffected. Approximately half of the patients display additional somatic anoma- lies and growth retardation. The therapy is mostly symptomatic and is dominated by corticosteroids, other modalities include regular blood transfusions or hematopoietic stem cell transplantation. At the beginning of this work, only two DBA causal genes were known, RPS19 and RPS24, being mutated in approximately 1/4 of all DBA patients. The goals of this work were to study the consequences of the known DBA causal mutations on cellular level and to find novel DBA causal genes. To date, over a half of DBA patients have been reported to carry a mutation in one of nine known DBA causal genes, including RPS17, RPL11 and RPL5, that are reported in this dissertation. All confirmed DBA causal genes encode for ribosomal proteins (RPs) that were essential for ribosome assembly. We further hypothesized a non- ribosomal protein participating in this process might be involved in DBA pathogenesis, too. In one DBA patient, we identified a rare sequence variant in one such candidate, a protein arginine methyltransferase 3 (PRMT3). We reported that the patient PRMT3...
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Proteomics of biological fluids
Jarkovská, Karla ; Kovářová, Hana (advisor) ; Petrák, Jiří (referee) ; Ulčová-Gallová, Zdeňka (referee)
Reproductive diseases, mainly those resulting in the infertility affect the chances of human being to reproduce. On the contrary, the heart disease, cancer and degenerative diseases currently account for majority of deaths in the world. Usually, these lifestyle diseases need longer lifespan to become the cause of death. The proteins secreted by cells carry important information about the cell's well-being, as well as about the condition of the tissues formed by these cells. Once secreted, these proteins may also be transferred throughout the body by means of body fluids, many of which are easily accessible for further 'in-depth' studies. Cellular and secreted proteins are often a focus of studies using proteomic means and the revelation of protein alterations can lead us to new ideas about the molecular mechanisms of diseases as well as possible identification of proteins that may be used as new targets for pharmaceutical intervention or molecules that could be used for diagnostic or prognostic purposes. Taking into consideration the above aspects, this research was undertaken to find proteins that could: (a) characterise the human follicular fluid as microenvironment of the maturing oocyte, to increase understanding of reproductive processes to improve the techniques of assisted repro- duction;...
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Proteomic analysis of liver iron overload
Myslivcová, Denisa ; Petrák, Jiří (advisor) ; Kovářová, Hana (referee) ; Hrkal, Zbyněk (referee)
V našich experimentech jsme prokázali, že lidské jaterní buňky mění expresi proteinů v důsledku akutního toxického přetížení železem a následně v důsledku oxidativního stresu a že v játrech přetížených železem dochází ke změnám v močovinovém cyklu, metylačním cyklu, oxidaci mastných kyselin a metabolizmu cukrů. Dále jsme zjistili, že pro dědičnou hemochromatózu je typická odlišná jaterní exprese sarkozin dehydrogenázy, selenium binding proteinu 2, glutathion S-transferázy P1, karboxylesterázy 1 a thioredoxin-like proteinu 2. Identifikované proteiny v první a druhé studii přímo nebo nepřímo souvisejí s přetížením modelového organizmu železem. Jiné jsou spíše obecnou odpovědí buňky na stresové podmínky. Jsou to především PDI, enoláza, keratiny, HSP70, GST, GRP78/Bip a podjednotky proteazómu. Tyto proteiny patří mezi nejčastěji identifikované diferenciálně exprimované proteiny v proteomických studiích využívajících 2-DE (Petrak et al., 2008) a jejich expresní změny tedy nelze považovat za změny specifické pro metabolizmus železa či hemochromatózu. Výsledky naší studie nám poskytly nové informace nezbytné pro detailní molekulárních pochopení mechanizmů uplatňujících se v metabolizmu železa, které je podmínkou rozvoje nových postupů v prevenci i terapii onemocnění souvisejících s metabolizmem železa, především...
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Proteomic analysis of selected oncohematological diseases
Pimková, Kristýna ; Petrák, Jiří (referee)
Oxidative stress is an important factor in carcinogenesis of oncohematological diseases. However its role in the pathogenesis of myelodysplastic syndromes (MDS) remains unclear. In this study, we have determined the oxidative status and evaluated proteomic changes in plasma of MDS patients as a consequence of oxidative dysbalance (oxidative modifications, protein-protein interaction and complex forming). We measured the levels of total cysteine, homocysteine, cysteinyglycine, glutathione, nitrites and nitrates in the plasma from 61 MDS patients and 23 healthy donors using high performance liquid chromatography. Glutathione and nitrites levels reduced significantly while other aminothiols levels increased significantly in plasma of MDS patients. This association with oxidative stress did not correlate with iron overload. We also found enhanced levels of asymmetric dimethylarginine in serums of middle aged patients with MDS that correlate to posttranslational modifications of proteins arginyl residues. Furthermore, carbonylated proteins level was significantly elevated in MDS patients compared to healthy donors. Using mass spectrometry, 5 S-nitrosylated blood platelets proteins were identified in plasma and blood platelets of MDS patients and set of 16 plasma proteins with high probability of...
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Snížení energetické náročnosti průmyslových chladicích zařízení
E - CONSULT, Praha ; Petrák, Miroslav ; Petrák, Jiří
Zpráva obsahuje souhrn prací, které byly vykonány na projektu v roce 2005, kdy bylo řešení zaměřeno na snížení energetické náročnosti provozu průmyslových chladicích zařízení jejich začleněním do energetického hospodářství podniku. Zpráva obsahuje řadu řešení umožňujících použít kondenzační teplo k ohřevu teplé vody a vytápění. byly analyzovány tyto metody zvyšující využitelnost odpadního tepla: a) zvýšení kondenzační teploty, b) vnitřní výměna tepla a c) použití tepelného čerpadla. Základem prací byla podrobná analýza tepelného oběhu parního používaného u průmyslových chladicích zařízení a tepelných čerpadel.
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