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

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SPsubpopulation occurrence and its nature in human lung adenocarcinoma cells A549. We hypothesized that HYP could act as enhancer of SP phenotype and stimulate the aggressive nature of these cells, as the result of promotion of ABC-transporters expression. We demonstrated strong correlation between higher activity of ABC transporters in SP cells and HYP elimination. It supports an idea of HYP being the substrate for ABCtransporters as suggested by Jendzelovsky and colleagues (Jendzelovky et al., 2009). On the other hand, however, we found that HYP reversibly suppresses SP population in dose-dependent manner. To assess the influence of HYP on the clonogenic capacity of A549 cells, we chose two methodologies, i.e. standard assay and single cell cloning. We observed certain paradox, when HYP enforced the clonogenicity of A549 when established by standard test, in contrary, though, we saw lower clonogenic efficiency but larger colonies established by single cell cloning. Our results demonstrate the influence of non-photoactivated HYP on suppression of Side population in reversible manner. We suggest, that this effect could by caused via one of known pathways involved in effect of HYP in dark which include affection of key targets in CSC, e.g. enhanced ubiquitinylation of Hsp90 (Blank et al., 2003; Peng et al., 2007). HYP was also capable to induce more aggressive phenotype in A549 cells. We conclude that differences in observed clonogenic capability could be explained by elevated dependence of cells with more aggressive phenotype on factors produced by surrounding cells. Further studies will be necessary to confirm this suggestion and to assess the impact of “niché – CSC“ relation on their behavior. Acknowledgements This work was supported by the Slovak Research and Development Agency under the contract No. APVV-0040-10 and VVCE-0001-07 and the Scientific Grant Agency of the Ministry of Education of the Slovak Republic under contract No. VEGA 1/0626/11. References Blank, M., Mandel, M., Keisari, Y., Meruelo, D. and Lavie, G.: Enhanced ubiquitinylation of heat shock protein 90 as a potential mechanism for mitotic cell death in cancer cells induced with hypericin. – Cancer research 63: 8241-8247, 2003. Blank, M., Lavie, G., Mandel, M., Hazan, S., Orenstein, A., Meruelo, D. and Keisari, Y.: Antimetastatic activity of the photodynamic agent hypericin in the dark. – International journal of cancer 111: 596-603, 2004. Jendzelovský, R., Mikes, J., Koval‘, J., Soucek, K., Procházková, J., Kello, M., Sacková, V., Hofmanová, J., Kozubík, A. and Fedorocko, P.: Drug efflux transporters, MRP1 and BCRP, affect the outcome of hypericin-mediated photodynamic therapy in HT-29 adenocarcinoma cells. – Photochemical & Photobiological Sciences 12: 1716-1723, 2009. Peng, C., Brain, J., Hu, Y., Goodrich, A., Kong, L., Grayzel, D., Pak, R., Read, M. and Li, S.: Inhibition of heat shock protein 90 prolongs survival of mice with BCR-ABL-T315Iinduced leukemia and suppresses leukemic stem cells. – Blood 110: 678-685, 2007. Zhou, S., Schuetz, J.D., Bunting, K.D., Colapietro, A.M., Sampath, J., Morris, J.J., Lagutina, I., Grosveld, G.C., Osawa, M., Nakauchi, H. and Sorrentino, B.P.: The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. – Nature Medicine 7:1028-1034, 2001. Analytical Cytometry VII 195
P90. TOLL-LIKE RECEPTOR 2 TRACKS THE EMERGENCE OF EARLIEST MYELOID PROGENITORS IN PRECIRCULATION EMBRYO Jana Balounová, Tereza Vavrochová, Martina Benešová and Dominik Filipp Institute of Molecular Genetics, AS CR, Prague, Czech Republic; jana.balounova@img.cas.cz Toll like receptors (Tlrs) are critically important in the pathogen recognition and regulation of the innate and adaptive immune responses (Akira et al., 2001; Medzhitov, 2007). In addition, a direct pathogen sensing of bone marrow hematopoietic stem cells and hematopoietic progenitors via Tlrs seem to play a crucial role in directing the hematopoietic cell fates under inflammatory conditions (De Luca et al., 2009; Nagai et al., 2006; Sioud et al., 2006). So far the expression of Tlrs during early stages of embryonic development has not been addressed. Using a transgenic model to trace cells of embryonic origin we show that Tlrs are expressed on embryonic myeloid cells as well as hematopoietic precursors committed to the myeloid lineage. Together with the prototypic marker of hematopoietic progenitors, c-kit, TLR2 is specifically expressed on the surface of hematopoietic precursors in early gastrulation embryos (E6.5-E7.0). E7.5-E8.5 TLR2 + c-kit + cells express CD45 mRNA and gradually differentiate through an intermediate c-kit + CD45 + stage to c-kit – CD45 + myeloid cells. Moreover E6.5 TLR2 + precursors differentiate to myeloid TLR2 + ckit – CD45 + cells in vitro. Upon TLR2 ligand recognition, E8.5 TLR2 + c-kit + cells proliferate and differentiate to CD45 + CD11b + myeloid cells in a MyD88 dependent manner. Our results indicate that TLR2 could be used as one of the earliest markers of embryonic hematopoietic progenitors and suggest a functional link between embryonic hematopoiesis and inflammation. Acknowledgements This study was supported by the Grant Agency of the Academy of Sciences of the Czech Republic (grant IAA500520707) and by The Grant Agency of Charles University (grant 259109). References Akira S, Takeda K, Kaisho T. 2001. Nat Immunol 2: 675-680. De Luca K, et al. 2009. Leukemia 23: 2063-2074. Medzhitov R. 2007. Nature 449: 819-826. Nagai Y, et al. 2006. Immunity: 24(6): 801-812 Sioud M, et al. 2006. Journal of Molecular Biology 364(5): 945-954 196 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
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P8. PROADIFEN (SKF-525A) ATTENUATES
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models. As manifested by compromise
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P11. NEW BIOACTIVE AND FLUORESCENT
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Acetylation of histones, along with
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stimulated cells. The physiological
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h induced very minor manifestations
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chemotherapeutic drug LA-12. These
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aberrant expression, localization a
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P21. ASSESSMENT OF MITOCHONDRIAL FU
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P23. PTEROSTILBENE: COMPARISON OF A
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Fam123b. Amer1 has been shown very
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In summary, we described different
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this process. Combined treatment wi
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Homolog 1) gene; previously identif
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egulate expression of different gen
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was harmful neither alone nor in co
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P34. PHARMACOLOGICAL INHIBITION OF
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novel chemotherapeutics with specif
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P38. EVIDENCE OF ABNORMAL PERIPHERA
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acid to 5-lipoxygenase. Similarly t
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P41. WHY CAN WE SEE DIFFERENT RADIO
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P42. ELUCIDATION OF CROSSTALK BETWE
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P44. INSTRUMENT SETTINGS FOR EUROFL
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e used to cultivate endothelial cel
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