Studying of regularity of programmed cell death after influence of demages factors

1. 2011Student Practice in JINR Fields of Research Dubna, Russia July 13 – 29, 2011 Studying of regularity of programmed cell death after influence of demages factors University of Wroclaw Wroclaw Medical University Speaker: Karolina Socha

2. Our Group: Laboratory of Radiation Biology (LRB) Joint Institute for Nuclear Research Supervisor: Ivanka Ravnachka JINR Group: Elena Baranova Maria Saveleva Student Group: Tatiana Alexandru (Romania) Karolina Socha (Poland) Katarzyna Shteklova (Czech Republic)

3. Outline: Introduction. Materials and methods. Results.

4. Relation between LET(Linear Energy Transfer) and the action type Direct actionis predominant with high LET radiation, e.g. alpha particles and neutrons. 1. Introduction Studying of regularity of programmed cell death after influence of demages factors Indirect action is predominant with low LET radiation,e.g. X and gamma rays.

5. Types of radiation-induced lesions in DNA 1. Introduction Studying of regularity of programmed cell death after influence of demages factors Base damage Double-strand breaks Single-strand breaks

6. 1. Introduction Studying of regularity of programmed cell death after influence of demages factors Consequences of DSB induction in the cell genome Studying regularities in the induction of apoptosis and DNA double-strand breaks as the main initiating events in apoptotic cell death after the action of radiations with different physical properties.

7. Figure 1. Major diffrences between apoptosis and necrosis. 1. Introduction Studying of regularity of programmed cell death after influence of demages factors Differences between necrosis and apoptosis Necrosis Accidental uncontrolled cell death leads to lysis of cell, inflammatory responses and serious health problems. Apoptosis Programmed cell death occurs normally during development and aging and as a homeostatic mechanism to maintain cell populations in tissues; defense mechanism (immune reactions or when sells demaged by disease or noxious agents (Norbury and Hickson, 2001).

8. Materials and methods Figure 2.Fluorescent microscope (Carl Zeiss). Studying of regularity of programmed cell death after influence of demages factors Seperate lymphocytes from whole human blood. Add Hydrogen Peroxide. Do preparations stained with mix of dyes (ethidium bromide and arcidine orange). Use fluorescent microscope for tracing morphological changes in preparations. Master the morphological method of detecting various forms of cell death.

9. Apoptosis visualization 2. Materials and methods Studying of regularity of programmed cell death after influence of demages factors Clasification criteria: Bright green cells (irregularly shaped nucleus with heterogeneously poited chromatin)→living cells Green or red cells (intact chromatine structure) →necroticcells Green or red cells (characteristic nuclear morphology: a rounded nucleus with collapsed chromatin)→apoptoticcells Figure 3.Different forms of cell death

10. Studying of regularity of programmed cell death after influence of demages factors Results Apoptotic cell induction after adding H2O2

11. 0 h 24 h 48 h 3. Results Studying of regularity of programmed cell death after influence of demages factors Apoptotic cell induction after  iradiation  -rays

12. Studying of regularity of programmed cell death after influence of demages factors Thank you for attention!