proof of principle tests confirm genotoxic potential of...

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V E R U MSTIFTUNG FÜR VERHALTEN UND UMWELT Genotoxic potential of RF-EMF - NIR and Health Workshop, Porto Alegre - May 18-19, 2009 - page 1

Proof of principle testsconfirm

genotoxic potential of RF-EMF

Franz Adlkofer

VERUM - Foundation for Behaviour and EnvironmentMunich, Germany


Scientific approach

Exposure Search for Effects Investigations

50 HzExtremely low

frequency (ELF) fields

900 MHz1800 MHz

Radiofrequency(RF) fields

UMTS

Epidemiological studies

Animal studies

Studies on cell and

tissue culture

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Risk Evaluation of Potential Environmental Hazards From Low Energy Electromagnetic Field Exposure Using

Sensitive in vitro MethodsA 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-1999-01574

STUK, Helsinki

Free University of BerlinInstitute for Plant Genetics,Gatersleben

University of Hannover

RZPD, Heidelberg

Medical University of ViennaVERUM Foundation, München

University of Bordeaux

INSALUD, Madrid University of Bologna

University of Milan

ETH Zürich


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 actionvast differences in exposure and experimental conditionsthe complete lack of agreement on biological endpoints appropriate forstudy

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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 currentsafety limits cause cellular, sub-cellular or molecular alterations in isolated mammalian cells that are relevant for the development of diseases.


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.

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Working hypothesis

The hypothesis that was to be tested in the REFLEX project assumed that no biological effects of relevance for the pathogenesis of chronicdiseases will be detected and that the present safety limits reliablyprotect EMF-exposed people from any health risk.


Exposure chamber

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© Clinical Chemistry, Free University of Berlin

sham exposition gamma-radiation; 0,5 Gy

RF-EMF: 1800 MHz continuously; SAR 1.3 W/kg; 24h

Comet assay


Micronucleus test

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ELF-EMF _ Age-dependent increase of DNA strand breaksin human fibroblasts

50 Hz sinus; 1000 µT; 5 min on / 10 min off

0

2

4

6

8

10

12

14

16

18

20

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24hours of exposure

com

et ta

ilfac

tor %

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

© Division of Occupational Health, Medical University of Vienna


ELF-EMF _ Flux density-dependent increase of DNA strand breaks in human fibroblasts

50 Hz sinus; 15 h; 5 min on / 10 min off

dose dependency (15 h, intermittent)

µT magnetic flux density

com

et ta

ilfac

tor

%

0

2

4

6

8

10

12

14

16

18

20

10 20 30 40 50 60 70 80 90 100

200

300

400

500

600

700

800

900

1000

AlkalineNeutral

35 µT


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0123456789

10

sham exp. sham exp. sham exp. sham exp.

continuous intermittent 5´/10´ Pulse-modulation

intermittent 5´/10´

Talk-modulationcontinuous, SAR

1.2

Com

et ta

ilfac

tor %

4 h

16 h

24 h

GSM _ Exposure- and signal pattern-dependent increase of DNA strand breaks in human fibroblasts

1800 MHz; SAR 2 W/kg


GSM-RF-EMF _ Dose-dependent increase of DNA strand breaks

GSM Basic; 1950 MHz; 24 h; 5 min on / 10 min off

8,07,98,07,9

5,9

4,7

4,1

4,04,04,14,04,14,04,1

0

1

2

3

4

5

6

7

8

9

0,1 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

Com

et-T

ailfa

ctor

%

exp.

sham


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UMTS 1950 MHznormal human fibroblasts (cell line ES-1)exposure duration: 24 hoursexposure pattern: continuousSAR values: 0.05 W/kg, 0.1 W/kg, 0.5 W/kg, 1.0 W/kg and 2.0 W/kgendpoints: 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

UMTS _ Dose-dependency


Dose-dependent increase in DNA strand breaks after UMTS exposure

4,8 4,9 4,7 4,5 4,4

7,9

13,5

10,08,3 8,5

4,8 4,9 4,8 4,9 4,8

32,032,032,131,632,2

0,0

5,0

10,0

15,0

20,0

25,0

30,0

35,0

0,05 W/kg 0,1 W/kg 0,5 W/kg 1,0 W/kg 2,0 W/kg

Com

etAs

say

Tailf

acto

rin

%

scheinexponiert

positive Kontrolle

sham exposedexposednegative controlpositive control


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Dose-dependent increase of micronuclei after UMTS exposure

5,4 5,7 5,4 5,7 5,4

7,7

10,9 10,7 11,1 11,0

5,35,1

5,55,1

5,3

17,4 17,518,0

17,518,0

0,0

2,0

4,0

6,0

8,0

10,0

12,0

14,0

16,0

18,0

20,0

0,05 W/kg 0,1 W/kg 0,5 W/kg 1,0 W/kg 2,0 W/kg

Mic

ronu

cleu

s/50

0 B

NC

´sschein-exponiertexponiert

positive Kontrolle




UMTS 1950 MHznormal human fibroblasts (cell line ES-1)SAR value: 0.1 W/kgexposure pattern: continuousexposure duration: 4, 8, 12, 16, 20, 24 and 48 hoursendpoints: 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

UMTS _ Time-dependency

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Time-dependent increase of DNA strand breaks after UMTS exposure

4,8 4,7 4,7 4,7 4,9 4,9 4,94,9

6,88,3

10,6

14,2 13,6 13,5

4,8 4,8 4,8 4,8 4,7 4,8 4,9

32,1 32,032,132,231,932,031,7

0,0

5,0

10,0

15,0

20,0

25,0

30,0

35,0

4 h 8 h 12 h 16 h 20 h 24 h 48 h

Com

etAs

say

Tailf

acto

rin

%scheinexponiertnegative Kontrollepositive Kontrolle




Time-dependent increase of micronuclei after UMTS exposure

5,3 5,1 5,3 5,2 5,25,7 5,35,3 5,1

7,9

11,0 10,9 10,9 11,1

5,2 5,1 5,2 5,0 5,35,1 5,3

17,017,6

17,2 17,1 17,4 17,5 17,2

0,0

2,0

4,0

6,0

8,0

10,0

12,0

14,0

16,0

18,0

20,0

4 h 8 h 12 h 16 h 20 h 24 h 48 h

MN

/500

BN

C´s

scheinexponiertnegative Kontrollepositive Kontrolle



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UMTS 1950 MHzSAR value: 0.1 W/kgexposure duration: 16 hoursexposure pattern: 5 mon on / 10 min offcells: 3 different cell lines from human fibroblasts

UMTS _ Repetition with 3 different cell lines


Increase of DNA strand breaks in fibroblasts from threedifferent persons after UMTS exposure

4,6 4,9 5,2

13,1 13,8 14,1

4,6 4,9 5,3

35,2 35,938,0

0,0

5,0

10,0

15,0

20,0

25,0

30,0

35,0

40,0

45,0

ES-1 IH-9 HW-2

Com

etAs

say

Tailf

acto

rin

%

scheinexponiertnegative Kontrollepositive Kontrolle



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Increase of micronuclei in fibroblasts from threedifferent persons after UMTS exposure

5,45,7

4,4

12,4

10,910,4

6,1

4,94,6

18,017,517,6

0,0

2,0

4,0

6,0

8,0

10,0

12,0

14,0

16,0

18,0

20,0

ES-1 IH-9 HW-2

Mic

ronu

cleu

s/50

0 B

NC

´sscheinexponiert

positive Kontrolle




RF-EMF _ Effects on gene and protein expressionGSM; 900 MHz; SAR 2 W/kg; 1h

from: Dariusz Leszczynski, Radiation and Nuclear Safety Authority (STUK), Finland

RF-EMF activatesgene expression (MAPK p38, PKC)protein expression (HSP27)metabolism (phosphorylation of HSP27)

Cells react to exposure with RF-EMF with stress the physiologicalrelevance of which has not been sufficiently investigated yet.

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RF-EMF _ Contribution to pathogenesis 875 MHz; SAR < 2 W/kg; minutes

Friedman et al. (2007) Mechanisms of short-term ERK activation by electromagnetic fields at mobile phone frequencies. Biochem J 405: 559-568 (The Weizman Institute of Science, Israel)

RF-EMF activates

NADH oxidase ROS gene damages?

matrix metalloproteinase Hb-EGF

ERK cascades signal transductioncell differentiationcell proliferationcoping with stressmetabolism


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

A significant increase of DNA strand breaks was observed in human fibro-blasts after GSM exposure at a SAR of 0.3 W/kg and after UMTS exposure at a SAR of 0.05 W/kg.

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

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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. ..."

Swisscom patent

International Publication Number: WO 2004/075583 A1


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.

A risk to our health?

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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 asignificantly increased risk for brain tumours in long-term users of mobilephones.

Causal or not? We do not know yet

Lennard Hardell recently reported in young people below 20 years of age amore than 5-fold increase in the risk for brain tumours at the side of thehead at which they used the mobile phone.

Causal or not? We do not know yet

Worst case scenario


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.

Conclusions

proof of principle tests confirm genotoxic potential of...

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