Proof of principle tests confirm genotoxic potential of RF-EMF

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1 Proof of principle tests confirm genotoxic potential of RF-EMF Franz Adlkofer VERUM - Foundation for Behaviour and Environment Munich, Germany Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 1 Scientific approach Exposure Search for Effects Investigations 50 Hz Extremely low frequency (ELF) fields Epidemiological studies 900 MHz 1800 MHz Radiofrequency (RF) fields UMTS Animal studies Studies on cell and tissue culture Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 2

2 Risk Evaluation of Potential Environmental Hazards From Low Energy Electromagnetic Field Exposure Using Sensitive in vitro Methods A project funded by the European Union under the programme "Quality of Life and Management of Living Resources", Key Action 4 "Environment and Health": QLK4-CT STUK, Helsinki University of Hannover RZPD, Heidelberg VERUM Foundation, München University of Bordeaux INSALUD, Madrid Free University of Berlin Institute for Plant Genetics, Gatersleben Medical University of Vienna University of Milan University of Bologna ETH Zürich Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 3 State of the art in 1999 Even though the biological effects of EMF exposure have been studied for the past 80 years, no consensus has been reached with respect to either findings or their interpretation. The reasons for this are numerous: difficulties in measuring EMF exposure at the putative sites of action vast differences in exposure and experimental conditions the complete lack of agreement on biological endpoints appropriate for study Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 4

3 Assumption and aim of the study REFLEX was based on the assumption that a health risk due to EMF below the current safety limits can only exist if scientific proof of biological effects, relevant for the development of diseases, is obtained. The aim of REFLEX was to find out whether or not EMF below the current safety limits cause cellular, sub-cellular or molecular alterations in isolated mammalian cells that are relevant for the development of diseases. Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 5 Pathogenesis of chronic diseases All chronic diseases such as cancer and neurodegenerative disorders are of extremely diverse and heterogeneous origin and this variability is generated by alterations of the structure and function of genes, leading to changes in gene and protein expression. This may result in a number of critical cellular events for example deregulated cell proliferation or cell differentiation and suppressed or exaggerated programmed cell death (apoptosis). Their convergence is required for the development of chronic diseases. Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 6

4 Working hypothesis ¾ The hypothesis that was to be tested in the REFLEX project assumed that no biological effects of relevance for the pathogenesis of chronic diseases will be detected and that the present safety limits reliably protect EMF-exposed people from any health risk. VERUM Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 7 Exposure chamber VERUM Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 8 4

5 Comet assay sham exposition gamma-radiation; 0,5 Gy RF-EMF: 1800 MHz continuously; SAR 1.3 W/kg; 24h Clinical Chemistry, Free University of Berlin Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 9 Micronucleus test Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 10

6 ELF-EMF _ Age-dependent increase of DNA strand breaks in human fibroblasts 50 Hz sinus; 1000 µt; 5 min on / 10 min off comet tailfactor % ES1, male, 6 years of age AN2, female, 14 years of age IH9, female, 28 years of age KE1, male, 43 years of age HN3, female, 56 years of age WW3, male, 81 years of age hours of exposure Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 11 ELF-EMF _ Flux density-dependent increase of DNA strand breaks in human fibroblasts 50 Hz sinus; 15 h; 5 min on / 10 min off comet tailfactor % Alkaline Neutral µt dose dependency (15 h, intermittent) µt magnetic flux density Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 12

7 GSM _ Exposure- and signal pattern-dependent increase of DNA strand breaks in human fibroblasts 1800 MHz; SAR 2 W/kg Comet tailfactor % h 16 h 24 h sham exp. sham exp. sham exp. sham exp. continuous intermittent 5 /10 Pulsemodulation Talk-modulation continuous, SAR intermittent 5 / Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 13 GSM-RF-EMF _ Dose-dependent increase of DNA strand breaks GSM Basic; 1950 MHz; 24 h; 5 min on / 10 min off 9 8 exp. 7,9 8,0 7,9 8,0 7 sham 6 5,9 Comet-Tailfactor % ,1 4,1 4,7 4,0 4,1 4,0 4,1 4,0 4, ,1 W/kg 0,3 W/kg 0,6 W/kg 1,0 W/kg 1,3 W/kg 1,6 W/kg 2,0 W/kg SAR in W/kg Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 14

8 UMTS _ Dose-dependency UMTS 1950 MHz normal human fibroblasts (cell line ES-1) exposure duration: 24 hours exposure pattern: continuous SAR values: 0.05 W/kg, 0.1 W/kg, 0.5 W/kg, 1.0 W/kg and 2.0 W/kg endpoints: comet assay, micronucleus test positive controls in the comet assay: UV light, 254 nm, 3 min, 800 µw/cm² positive control in the micronucleus test: 25 nm Vincristin Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 15 Dose-dependent increase in DNA strand breaks after UMTS exposure 35,0 32,2 31,6 32,1 32,0 32,0 schein sham exposed Comet Assay Tailfactor in % 30,0 25,0 20,0 15,0 10,0 7,9 13,5 10,0 8,3 8,5 exponiert exposed negative control positive Kontrolle control 5,0 4,8 4,8 4,9 4,9 4,7 4,8 4,5 4,9 4,4 4,8 0,0 0,05 W/kg 0,1 W/kg 0,5 W/kg 1,0 W/kg 2,0 W/kg Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 16

9 Dose-dependent increase of micronuclei after UMTS exposure 20,0 18,0 17,4 17,5 18,0 17,5 18,0 schein-exponiert sham exposed exponiert exposed negative control 16,0 positive Kontrolle control Micronucleus/500 BNC s 14,0 12,0 10,0 8,0 6,0 4,0 10,9 10,7 7,7 5,4 5,3 5,7 5,4 5,5 5,1 11,1 11,0 5,7 5,4 5,3 5,1 2,0 0,0 0,05 W/kg 0,1 W/kg 0,5 W/kg 1,0 W/kg 2,0 W/kg Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 17 UMTS _ Time-dependency UMTS 1950 MHz normal human fibroblasts (cell line ES-1) SAR value: 0.1 W/kg exposure pattern: continuous exposure duration: 4, 8, 12, 16, 20, 24 and 48 hours endpoints: comet assay, micronucleus test positive controls in the comet assay: UV light, 254 nm, 3 min, 800 µw/cm² positive control in the micronucleus test: 25 nm Vincristin Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 18

10 Time-dependent increase of DNA strand breaks after UMTS exposure Comet Assay Tailfactor in % 35,0 30,0 25,0 20,0 15,0 10,0 5,0 32,2 32,1 31,7 32,0 31,9 32,1 32,0 14,2 13,6 13,5 10,6 8,3 6,8 4,8 4,94,8 4,7 4,8 4,7 4,8 4,7 4,8 4,9 4,7 4,9 4,8 4,9 4,9 schein sham exposed exponiert exposed negative Kontrolle control positive Kontrolle control 0,0 4 h 8 h 12 h 16 h 20 h 24 h 48 h Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 19 Time-dependent increase of micronuclei after UMTS exposure 20,0 18,0 16,0 17,0 17,6 17,2 17,1 17,4 17,5 17,2 schein sham exposed exponiert exposed negative Kontrolle control positive Kontrolle control 14,0 MN/500 BNC s 12,0 10,0 8,0 6,0 11,0 10,9 10,9 11,1 7,9 5,3 5,1 5,3 5,7 5,35,2 5,1 5,1 5,2 5,2 5,0 5,2 5,3 5,3 5,1 5,3 4,0 2,0 0,0 4 h 8 h 12 h 16 h 20 h 24 h 48 h Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 20

11 UMTS _ Repetition with 3 different cell lines UMTS 1950 MHz SAR value: 0.1 W/kg exposure duration: 16 hours exposure pattern: 5 mon on / 10 min off cells: 3 different cell lines from human fibroblasts Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 21 Increase of DNA strand breaks in fibroblasts from three different persons after UMTS exposure 45,0 schein sham exposed Comet Assay Tailfactor in % 40,0 35,0 30,0 25,0 20,0 15,0 10,0 13,1 35,2 35,9 13,8 14,1 38,0 exponiert exposed negative Kontrolle control positive Kontrolle control 5,0 4,6 4,6 4,9 4,9 5,2 5,3 0,0 ES-1 IH-9 HW-2 Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 22

12 Increase of micronuclei in fibroblasts from three different persons after UMTS exposure 20,0 18,0 16,0 17,6 17,5 18,0 schein sham exposed exponiert exposed negative control positive Kontrolle control Micronucleus/500 BNC s 14,0 12,0 10,0 8,0 6,0 4,0 4,4 10,4 4,6 5,7 10,9 4,9 5,4 12,4 6,1 2,0 0,0 ES-1 IH-9 HW-2 Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 23 RF-EMF _ Effects on gene and protein expression GSM; 900 MHz; SAR 2 W/kg; 1h RF-EMF activates gene expression (MAPK p38, PKC) protein expression (HSP27) metabolism (phosphorylation of HSP27) Cells react to exposure with RF-EMF with stress the physiological relevance of which has not been sufficiently investigated yet. from: Dariusz Leszczynski, Radiation and Nuclear Safety Authority (STUK), Finland Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 24

13 RF-EMF _ Contribution to pathogenesis 875 MHz; SAR < 2 W/kg; minutes RF-EMF activates NADH oxidase ROS gene damages? matrix metalloproteinase Hb-EGF ERK cascades signal transduction cell differentiation cell proliferation coping with stress metabolism Friedman et al. (2007) Mechanisms of short-term ERK activation by electromagnetic fields at mobile phone frequencies. Biochem J 405: (The Weizman Institute of Science, Israel) Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 25 Summary of the REFLEX results The results of the REFLEX study did not confirm our hypothesis which assumed that no biological effects of relevance for the pathogenesis of chronic diseases will be found below the existing safety limits. However, ELF-EMF below 100 µt and RF-EMF below 2 W/kg alter the structure and function of genes in different animal and human cells. We saw an increase of single and double DNA strand breaks in human fibroblasts, HL60 cells and granulosa cells of rats, but not in human lymphocytes an increase of micronuclei and chromosome aberrations in human fibroblasts alterations in gene and protein expression of several cell types, especially in human fibroblasts, human endothelial cells and embryonic stem cells from mice A significant increase of DNA strand breaks was observed in human fibroblasts after GSM exposure at a SAR of 0.3 W/kg and after UMTS exposure at a SAR of 0.05 W/kg. Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 26

14 Swisscom patent on the "reduction of electrosmog in wireless local networks": "Thus it has been possible to show that mobile radio radiation can cause damage to genetic material, in particular in human white blood cells, whereby both the DNA itself is damaged and the number of chromosomes changed. This mutation can consequently lead to increased cancer risk. In particular, it could also be shown that this destruction is not dependent upon temperature increases, i.e. is non-thermal...." International Publication Number: WO 2004/ A1 Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 27 A risk to our health? None of the single approaches in basic research, animal experiments and epidemiological studies is at present in the position to provide proof of a health risk caused by EMF with sufficient certainty. However, as the results from the three approaches complement each other, they strongly speak in favour of the assumption that a health risk could arise from ELF-EMF as well as from RF-EMF. Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 28

15 Worst case scenario Genotoxic effects caused by mobile phone radiation in living cells in vitro speak in favour of a causality between the long-term use of mobile phones and the increase of a brain tumour risk. This in mind, we have to consider: The Interphone study, although not published yet in a final report, found a significantly increased risk for brain tumours in long-term users of mobile phones. Causal or not? We do not know yet Lennard Hardell recently reported in young people below 20 years of age a more than 5-fold increase in the risk for brain tumours at the side of the head at which they used the mobile phone. Causal or not? We do not know yet Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 29 Conclusions Available scientific data clearly demonstrate that biological effects of electromagnetic fields below the current safety limits are athermal by nature. Since current safety limits are based upon the assumption that athermal biological effects do not exist at all, the only possible conclusion is that they are invalid and do not protect people. Lowering the safety limits by a factor between 10-1 and 10-5 may be a first and easily to accomplish step to adjust the radiation intensity to the conditions of living organisms. In the long range, the development of adequate safety limits away from physical EMF effects towards biological EMF effects by independent researchers is a top priority. All-clear signals by the industry and their scientific advisers that deny possible health risks of people exposed to RF-EMF below the current safety limits do not have a sound basis. Therefore, national governments which are responsible for the health protection of their citizens are badly advised, if they back the safety limits and the all-clear signals. Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, page 30

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