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

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

3. 1. Introduction. 2. Materials and methods. 3. Results. Outline:

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

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

6. DNA double- strand break Apoptosis Mutations Chromosomeaberrations Neoplastictransformation of a cell 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. Consequences of DSB induction in the cell genome 1. Introduction Studying of regularity of programmed cell death after influence of demages factors

7. Figure 1. Major diffrences between apoptosis and necrosis. 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). Differences between necrosis and apoptosis 1. Introduction Studying of regularity of programmed cell death after influence of demages factors

8. Materials and methods Figure 2. Fluorescent microscope (Carl Zeiss). 1. Seperate lymphocytes from whole human blood. 2. Add Hydrogen Peroxide. 3. Do preparations stained with mix of dyes (ethidium bromide and arcidine orange). 4. Use fluorescent microscope for tracing morphological changes in preparations. 5. Master the morphological method of detecting various forms of cell death. Studying of regularity of programmed cell death after influence of demages factors

9. Apoptosis visualization Clasification criteria: 1. Bright green cells (irregularly shaped nucleus with heterogeneously poited chromatin) → living cells 2. Green or red cells (intact chromatine structure) → necrotic cells 3. Green or red cells (characteristic nuclear morphology: a rounded nucleus with collapsed chromatin) → apoptotic cells Figure 3. Different forms of cell death Studying of regularity of programmed cell death after influence of demages factors 2. Materials and methods

10. Apoptotic cell induction after adding H 2 O 2 Studying of regularity of programmed cell death after influence of demages factors Results

11.  -rays 0 h24 h 48 h Studying of regularity of programmed cell death after influence of demages factors 3. Results Apoptotic cell induction after  iradiation

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