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T cells labelling by bimodal contrast agent for in vivo studies of stroke
Krijt, Matyáš ; Poljaková, Jitka (advisor) ; Kříž, Jan (referee)
Stroke is a serious brain injury, which causes sudden death or terminates in permanent neurological disability. Nowadays, tissue plasminogen activator (tPA) is used as the only effective treatment of stroke. One of the potential targets for novel therapy are T cells. Even though the explicit role of T cells in the pathogenesis of brain injury, amounts and timing of all T cell subtypes infiltrating into brain during the stroke still needs further investigation. The research in this field is complicated by the lack of efficient methods for in vivo cell tracking. Therefore the aim of this thesis was to develop a method of T cells labelling by MRI contrast agent in order to investigate T cells distribution in ischemic mice model using in vivo MR imaging. T cells were isolated from C57/BL6 mice in two step isolation protocol using gradient centrifugation and magnetic separation with the efficiency of 97 %. The isolated cells were labelled with 100 μg Fe/mL of Molday ION Rhodamine B contrast agent. The labelling efficiency after 17 hours of cells incubation was higher than 99 %. The labelled cells were cultured with CD3 and CD28 antibodies resulting into the 74 % viability of labelled T cells compared to 83 % viability of non labelled T cells. The labelled T cells were visualized by fluorescent...
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New ligands for applications in medicine
Holá, Kateřina ; Kotek, Jan (advisor) ; Holzhauser, Petr (referee)
The aim of this Thesis was the preparation of macrocyclic ligands (Figure 1) for the complexation of lantanoides 3+ . Their complexes can be used in nuclear medicine (in case of ligand L1) or as MRI contrast agents (in case of ligand L2). Ligand L1 was designed to increase the rate of complexation due to the weakly binding pendant group based on iminodiacetic acid (IDA). The complexation mechanism was spectrophotometrically studied with Ce3+ . Spectrophotometric kinetic measurements proved that ligand L1 not only shown no improvement in the complexation rate but its complexation is rather noticeable slowed. Ligand L2 contains amino group whose protonization/coordination could be influenced by pH or temperature. It can be utilized in determination of physical status of different tissues. However, it is difficult to obtain the ligand L2 in sufficient purity for further studies N N NN COOHHOOC HOOC N COOH COOH P OH O L1 N N NN COOHHOOC HOOC NH2 L2 Figure 1: Ligands prepared in course of this Bc Thesis
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Macrobicyclic ligands for use in MRI
Pniok, Miroslav ; Kotek, Jan (advisor) ; Veselý, Jan (referee)
4 Title: Macrobicyclic ligands for use in MRI Author: Bc. Miroslav Pniok Department: Department of Inorganic Chemistry, Faculty of Science Supervisor: doc. RNDr. Jan Kotek, Ph.D. Supervisor's e-mail address: modrej@natur.cuni.cz Abstract: The aim of this project is to synthesize new kind of macrobicyclic ligands for Ln3+ ions, especially Gd3+ ion. Gadolinium(III) complexes are widely used in the clinical practice as contrast agents for MR imaging and an improvement of their properties is in the center of interest of many scientific teams. The macrobicyclic ligands have been designed to make possible coordination of two water molecules in the first coordination sphere with fast water exchange. The macrocyclic core (cyclen) is combined with a linking phosphinate chain in N1 ,N4 - and N1 ,N7 -positions forming a cryptand-like structure. The N1 ,N7 -isomer Et2L1 was synthesized in a small amount so only the N1 ,N4 -isomer H2L3 has been studied. The protonation constants have been determined by pH 31 P NMR titration. Keywords: cyclen, chelating agent, cryptand, MRI, phosphinate
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New ligands for magnetic resonance imaging; pH-sensitive probes
Táborská, Zdeňka ; Kotek, Jan (advisor) ; Kubíček, Vojtěch (referee)
TÁBORSKÁ, Z. New ligands for magnetic resonance imaging; pH-sensitive probes. Prague: Charles University in Prague, Faculty of Science, Department of Inorganic Chemistry, 2011, 32 pages. Supervisor: doc. RNDr. Kotek J., Ph.D. Magnetic resonance imaging is a modern method used in diagnostic for assessing morphology and pathology of soft tissues. This thesis briefly outlines the principles of magnetic resonance imaging, including instrumentation. In many cases intravenously administered contrast agents are used, which are used for diagnostic purposes. At present, the most applied are CAs based on Gd(III) complexes. The experimental part of the thesis is focused on preparation of a derivative of DTPA with aminomethylphosphinate pendant group for using as a potential ligand for CAs for the magnetic resonance imaging. KEYWORDS: magnetic resonance imaging, contrast agents, lanthanides, phosphinates
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