INDIRECT EVIDENCE OF NON-THERMAL BIOLOGICAL EFFECTS INDUCED BY MOBILE PHONE RADIATION IN VITRO. Dariusz Leszczynski

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1 COST 281Seminar "Subtle Temperature Effects of RF-EMF November 12-13, 2002, London UK INDIRECT EVIDENCE OF NON-THERMAL BIOLOGICAL EFFECTS INDUCED BY MOBILE PHONE RADIATION IN VITRO Dariusz Leszczynski Bio-NIR Research Group Radiation and Nuclear Safety Authority, Helsinki, Finland

2 Jokela s chamber Kuster s chamber Heat stress What are thermal/non-thermal effects?

3 Jokela s chamber

4 Jokela s chamber RF-EMF feed post Field distribution at average 2W/kg cell culture dishes circulating cooling water

5 hsp27 phosphorylation in SAR dose-dependent manner 32 P-labeling + immunoprecipitation sham 1.2SAR sham 1.8SAR sham 2.4SAR <2-folds up >2-folds up 3-4-folds up hsp27 expression in time-dependent manner sham 2.4SAR sham 2.4SAR sham 2.4SAR sham 1h 2h 6h 8h 2.4SAR h 2h 6h 8h

6 2.4SAR 2.4SAR 2.4SAR 2.4SAR expression p38 MAP kinase activity sham sham sham sham inhibitor prevents 32 P-hsp27 exposed + SB exposed sham h 2h 6h 8h 1h 2h 6h 8h RF-EMF causes activation sham 2.4SAR <2-folds up

7 1h sham hsp27 & stress fibers 1h 2.4 SAR 1h 2.4 SAR + 1h sham

cells with high expression of hsp27 (red

the cells combined effect of the increased

8 cells with high expression of hsp27 (red color) have prominent stress fibers-network (green color) and stress fiber components are present also in the ruffles (edges) of the cells combined effect of the increased expression and increased phosphorylation of hsp27

9 Active hsp27 regulates stability of stress fibers in EA.hy926 SB inhibitor of p38map kinase 1h 2SAR without SB h 2SAR with SB203580

10 Jacques Landry (Laval University, Quebec, Canada) Chinese hamster cell lines (fibroblasts): CCL39 - control cell line (hamster hsp27) CCL39 - over-expressing human wild-hsp27 3 phosphorylation sites are active CCL39 - over-expressing human mutant-hsp27 3 phosphorylation sites are inactivated Cells were exposed to 900GSM for 1h at 2SAR

11 hamster cell line CCL39 over-expressing human wild-hsp27 sham 1h 2SAR

12 hamster cell line CCL39 over-expressing human mutant-hsp27 sham 1h 2SAR

13 Kuster s chamber

17 Kuster s chamber 1800 GSM talk signal 1h exposure at 2SAR EA.hy926 EA.hy926v1 phorbol ester 2 SAR sham 2 SAR sham 3.0x 2.5x 2.8x 2.3x 1.7x 2.0x

18 hsp27 & stress fibers Kuster s chamber 1h sham 1h 2.0 W/kg

19 hsp27 & stress fibers Jokela s chamber 1h sham 1h 2.4 SAR

20 Summary of hsp27-effects Jokela s 1.2 SAR - <2 fold Kuster s 2.0 SAR fold Jokela s 2.4 SAR - >3-fold

21 Molecular event: activation of p38mapk/hsp27 pathway Physiological event: hsp27-p alters stability and distribution of stress fibers Observed in: human endothelial cell line hamster cell line hamster cell line transfected with human hsp27

22 modified from Leszczynski et al. Differentiation 70, 2002, Stress Rescue hsp 70 up-stream cellular receptors up-stream stress kinases p38 MAP kinase MAPKAP kinase-2/3 PRAK kinase Mobile phone radiation is absorbed by brain cells (nervous & endothelial) chaperone function oligomer hsp27 monomer hsp27-p cell-cell contact pinocytosis cell shape & size generation of growth factors non-apoptotic cell surface blebbing BLOOD-BRAIN BARRIER actin polymerization and stabilization of stress fibers fascin α-actinin overdrive hsp27-p CANCER & OTHER DISEASES APOPTOSOME cytochrome c Apaf-1 pro-caspase-9 apoptosis-related cell surface blebbing pro-caspase-3 caspase-9 caspase-3 APOPTOSIS

23 We see effects but are these effects thermal or non thermal? Temperature increases are <0.3 o C in Jokela s chamber <0.1 o C in Kuster s chamber

24 Thermal regulation of hsp27

25 Heat stress effect (water bath) 60 min., 37 o C 2.4 W/kg, 60 min., 37 o C 20 min., 43 o C 40 min., 43 o C 60 min., 43 o C

26 Cells exposed in cell culture incubator on copper plate for 60 min h, 2SAR

Thermal induction of hsp27 & hsp27-p hsp27 hsp27-p 36 37 38 39 40 43 180000 160000 140000

27 Thermal induction of hsp27 & hsp27-p hsp27 hsp27-p sham RF-EMF hsp27 hsp27-p

28 Thermal effects term used in safety standards average SAR vs. real distribution assumption that 4 SAR is cut-off point thermal effects are taken care of in safety guidelines

29 Health Physics 74, 1998,

30 Niels Kuster s presentation at COST 281 meeting in Rome 2002 on Children and Mobile Phones

31 Thermal effects - questions what temperature activates stress kinases? how rapid temperature increase? how big increase? timing of the increase? other concomitant insults? is it followed by cell physiological response? can it affect physiology of organ/body?

32 Do we know what we are talking when referring to: thermal vs. non-thermal effects? We do not have precise definition of what is the thermal effect

33 Should we rather talk about effects at SAR levels allowed by safety standards because we do not know what temperature elevation causes activation of cellular stress kinases or other cellular responses?

Bio-effects Reetta Kuokka Sakari Joenväärä Jukka Reivinen Hanna Tammio Pia Kontturi Teemu Kallonen Funding support: Dosimetry Kari Jokela Tim Toivo Ari Pekka Sihvonen Niels Kuster Juergen Schuderer

34 Bio-effects Reetta Kuokka Sakari Joenväärä Jukka Reivinen Hanna Tammio Pia Kontturi Teemu Kallonen Funding support: Dosimetry Kari Jokela Tim Toivo Ari Pekka Sihvonen Niels Kuster Juergen Schuderer Tekes - The National Technology Agency, Finland 5th Framework Programme of the European Union (REFLEX project) The Academy of Finland STUK-Radiation and Nuclear Safety Authority Cells Cora-Jean Edgell Jacques Landry

CURRENT STATUS OF THE RESEARCH ON BIOLOGICAL AND HEALTH EFFECTS OF RADIATION EMITTED BY WIRELESS COMMUNICATION DEVICES

CURRENT STATUS OF THE RESEARCH ON BIOLOGICAL AND HEALTH EFFECTS OF RADIATION EMITTED BY WIRELESS COMMUNICATION DEVICES Dariusz Leszczynski, PhD, DSc Adjunct Professor of Biochemistry, University of Helsinki,