ABSTRACTS â ORAL PRESENTATIONS - AMCA, spol. s r.o.

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P50. NON-INVASIVE ANALYSIS OF BEATING PARAMETERS IN IN VITRO CARDIOMYOCYTE CULTURE Dominika Sýkorová 1,3 , Pavel Karas 2 , Jana Navrátilová 1,3 , Lucia Binó 1,3 , Lukáš Kohút 4 , Lukáš Kubala 1,3,5 , Jiří Pacherník 1,5 1 Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic; 2 Centre for Biomedical Image Analysis, Faculty of Informatics Masaryk University, Brno, Czech Republic; 3 Institute of Biophysics ASCR v.v.i., Brno, Czech Republic; 4 Research Center for Toxic Compounds in the Environment, Faculty of Science, Masaryk University, Brno, Czech Republic; 5 International Clinical Research Center – Centre of Biomolecular and Cellular Engineering, St. Anne‘s University Hospital Brno, Czech Republic. Studying of tissue and cellular functions via non-invasive methods represents a powerful tool for monitoring of various experimental and pathological conditions with minimal effect on observed processes. Cardiomyocytes are highly specified cells with unique ability of spontaneous beating. The beating characteristics such as beating frequencies and chronotropic changes are related to cardiomyocytes differentiation/maturation status and are modulated by cell culture conditions. Beating characteristic is modulated in response to various stimuli. Response of cardiomyocyte beating to tested compound is suggested to be valuable marker for testing cardiotoxicity. However, the detection of beating patter of cardiomyocytes in cell culture conditions is highly challenging. Here we introduce the Cardio Analyser software developed for digital movie analysis of beating properties of cardiomyocytes in vitro culture. Potential and capabilities of the software were tested on cardiomyocytes in vitro culture under various experimental conditions. Based on our results, this non-invasive method gives promising results in studies of the response of cells to various external stimulations, such as differentiation promoting agents or treatments. Cardio Analyser can be also useful in preliminary drug screening studies as a quick and inexpensive tool which doesn’t require extensive expertise. Acknowledgements This work was supported by grants from MEYS CR MSM0021622430 and Cost CZ LD11015, from GAČR 13-29358S, and from the European Social Fund - projects OganoNET (CZ.1.07/2.4.00/31.0245) and HistoPARK (CZ.1.07/2.3.00/20.0185). LK was supported by the European Regional Development Fund - Project FNUSA-ICRC (No. CZ.1.05/1.1.00/02.0123). Analytical Cytometry VII 147
P51. FLUORESCENCE-LABELED FERROMAGNETIC NANOPARTICLES FOR MRI-GUIDED MAGNETIC FLUID HYPERTHERMIA-BASED GLIOBLASTOMA TREATMENT AND MR IMAGING Karolina Turnovcova 1 , Vit Herynek 2 , Emil Pollert 3 , Pavel Veverka 3 , Pavel Zvatora 4 , Magda Vosmanska 4 , Daniel Jirak 2 , Milan Hajek 2 , Eva Sykova 1 , Pavla Jendelova 1 1 Institute of Experimental Medicine ASCR, Prague, Czech Republic, karolina. turnovcova@biomed.cas.cz 2 MR-Unit, Department of Radiodiagnostic and Interventional Radiology, Institute for Clinical and Experimental Medicine, Prague, Czech Republic 3 Institute of Physics, ASCR, Prague, Czech Republic 4 The Institute of Chemical Technology, Prague, Czech Republic Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor occuring in humans, and its cells are resistant to conventional therapy. Fluorescencelabeled ferromagnetic nanoparticles may serve for both diagnostic and therapeutic purposes. They represent intracellular labeling using endocytosis uptake, are a suitable contrast agent for magnetic resonance imaging (MRI) in vivo, and can also be used for guided temperature-controlled thermoablation. The method is based on the deposition of stable and nontoxic suspensions of the magnetic nanoparticles inside the tumor followed by exposure to a high frequency (HF) electromagnetic field. Magnetic hysteresis losses result in local heating by the particles and consequently to the apoptosis of the cells. Perovskite nanoparticles (La 1-x Sr x MnO 3 ), coated by SiO 2 with FITC, were synthesized, their surface was modificated with different materials [chitosan, 2-[methoxy(polyethyleneoxy) propyl]trimethoxysilane (PEET)], and they were tested in vitro and in vivo. Rat and human mesenchymal stromal cells (rMSC, hMSC) and GBM cell lines (C6, A172 and GaMG) were cultivated with nanoparticles for 48 hours using the xCELLigence System; the real-time growth curves were recorded, and the culture viability was analysed by trypan blue staining. The nanoparticle uptake into the cells was then analysed using FACS-based fluorescence intensity measurements and directly by ICP-MS spectrometry. The viability of cells incubated with the nanoparticles was in the range of 60 – 95%, whereas a control sample reached 98%. The amount of incorporated nanoparticles was up to 10-fold higher in the GBM cell lines and was affected by surface modification. Two million C6 cells were injected intradermally into rats (n=10). In 2 weeks, 50 ul of a 4 mM Mn suspension of perovskite nanoparticles was injected into the glioblastoma tissue. MRI images were obtained to confirm the distribution of the nanoparticles in the tumor. The rats were exposed to a HF electromagnetic field (480 kHz, 11 mT) for 30 minutes. Four control animals with tumors underwent HF field exposure without nanoparticles, or the application of nanoparticles without exposure to the HF field. The temperature in the tumor with nanoparticles increased to 41.7°C, while control animals without nanoparticles reached 40°C. Immunohistochemistry (staining for caspase3 and TUNEL) revealed massive apoptosis in the tumors of the animals with injected nanoparticles after exposure to the HF field. 148 Analytical Cytometry VII
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ABSTRACTS - ORAL PRESENTATIONS 14.
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and (iii) siRNA-mediated knockdown
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25. PROFILING OF CHILDHOOD ACUTE LE
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more dominantly than Th1 and CD4 ce
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29. INFLUENCE OF THE LEVEL OF CD133
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36. B-CELL IMMUNOPHENOTYPING OF LAR
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possible mechanism behind risk alle
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isotype controls and FMO for CD14 a
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NEP developmental stages were disti
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50. THE PROGRESSION OF HIV INFECTIO
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52. PRŮKAZ REGULAČNÍCH T LYMFOCY
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values when analysing paediatric ly
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42,0) vs. 1,5 (0,0-28), p
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for application of proton therapy,
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effects will be also characterized
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Although multiple signalling pathwa
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an ability to recreate the tumour f
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incubated with non-filtered superna
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approaches how to detect and isolat
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progress in development of automate
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tissues is characteristic for a num
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kappaB. Beside that, we found OANO
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therapy in patients (Calderwood et
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difference in the effect caused eit
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4 CIC bioGUNE Technology Park of Bi
Page 51 and 52: P8. PROADIFEN (SKF-525A) ATTENUATES
Page 53 and 54: models. As manifested by compromise
Page 55 and 56: P11. NEW BIOACTIVE AND FLUORESCENT
Page 57 and 58: Acetylation of histones, along with
Page 59 and 60: stimulated cells. The physiological
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Page 63 and 64: chemotherapeutic drug LA-12. These
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Page 67 and 68: P21. ASSESSMENT OF MITOCHONDRIAL FU
Page 69 and 70: P23. PTEROSTILBENE: COMPARISON OF A
Page 71 and 72: Fam123b. Amer1 has been shown very
Page 73 and 74: In summary, we described different
Page 75 and 76: this process. Combined treatment wi
Page 77 and 78: Homolog 1) gene; previously identif
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Page 87 and 88: P38. EVIDENCE OF ABNORMAL PERIPHERA
Page 89 and 90: acid to 5-lipoxygenase. Similarly t
Page 91 and 92: P41. WHY CAN WE SEE DIFFERENT RADIO
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Page 109 and 110: cells (SFC) using IFN-γ Elispot in
Page 111 and 112: FACSCantoII flow cytometer (Becton
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Page 115 and 116: cross-reacting group (CREG) and sha
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Page 121 and 122: P65. OVULATION STIMULATION PROTOCOL
Page 123 and 124: Acknowledgements This work was supp
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Page 127 and 128: triggers a process of normal blood
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Page 135 and 136: We conclude that decrease in the po
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Page 141 and 142: LD11015, from GAČR 13-29358S, and
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Page 145 and 146: given cell type are missing and the
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for chemokines in attracting “inf
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in immune organs (Bursa of Fabricii
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