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UNIVERSITY OF ROSTOCKUNIVERSITY OF ROSTOCK
Institute for Cell Biology and Institute for Cell Biology and BiosystemBiosystem Technology Technology Division of Environmental Physiology Division of Environmental Physiology
Untersuchungen zu Wirkmechanismen an Zellen unter
Exposition mit hochfrequenten elektromagnetischen
Feldern der MobilfunktechnologieStart: September 2003
Myrtill Simkó
Simkó’s Lab
ITIS Radio frequency setupITIS Radio frequency setup
1,8 GHz: • CW • GSM 217 Hz • Hearing (GSM-DTX) • Speaking (GSM-nonDTX) • Talk (GSM-Talk)(70 : 30%)
SAR: 0.5; 1.0; 1.5; 2.0 W/kgExp. time: 10 min on/off
45 min cont.
Simkó’s Lab
CellCell systemssystems
Human primary Monocytes and Lymphocytes: isolatedfrom human umbilical cord blood within 48 h after birth
Mono Mac 6 (MM6): human acute monocytic leukemia cells
K562: human chronic myeloid leukemia
Monocytes Lymphocytes Mono Mac 6 cells K562 cells
Simkó’s Lab
1800 MHz 1800 MHz -- Free radical productionFree radical production(human Mono Mac 6 cells)(human Mono Mac 6 cells)
0.5W/kg (n=3)
1W/kg (n=5)
1.5W/kg (n=4)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
Continuous Wave
***
* ****
0.5W/kg (n=3)
1W/kg (n=5)
1.5W/kg (n=4)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
Continuous Wave
***
* ****
0.5W/kg (n=3)
1W/kg (n=5)
1.5W/kg (n=4)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
Continuous Wave
***
* ****
Simkó’s Lab
1800 MHz 1800 MHz -- Free radical productionFree radical production(human Mono Mac 6 cells)(human Mono Mac 6 cells)
0.5W/kg (n=3)1W/kg (n=4)
1.5W/kg (n=4)
2W/kg (n=3)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM-non DTX
**
**
****
0.5W/kg (n=3)1W/kg (n=4)
1.5W/kg (n=4)
2W/kg (n=3)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM-non DTX
**
**
****
0.5W/kg (n=3)1W/kg (n=4)
1.5W/kg (n=4)
2W/kg (n=3)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM-non DTX
**
**
****
Simkó’s Lab
1800 MHz 1800 MHz -- Free radical productionFree radical production(human Mono Mac 6 cells)(human Mono Mac 6 cells)
0.5W/kg (n=6)
1W/kg (n=3)
1.5W/kg (n=3)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-RF/Sham LPS/Sham LPS-RF/Sham 40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM Talk
* *
**
****
0.5W/kg (n=6)
1W/kg (n=3)
1.5W/kg (n=3)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-RF/Sham LPS/Sham LPS-RF/Sham 40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM Talk
* *
**
****
0.5W/kg (n=6)
1W/kg (n=3)
1.5W/kg (n=3)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-RF/Sham LPS/Sham LPS-RF/Sham 40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM Talk
* *
**
****
Simkó’s Lab
1800 MHz 1800 MHz -- Free radical productionFree radical production(human Mono Mac 6 cells)(human Mono Mac 6 cells)
*
0.5W/kg (n=4)
1W/kg (n=4)
1.5W/kg (n=4)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM-DTX
*
** ** **
***
0.5W/kg (n=4)
1W/kg (n=4)
1.5W/kg (n=4)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM-DTX
*
** ** **
*
0.5W/kg (n=4)
1W/kg (n=4)
1.5W/kg (n=4)
2W/kg (n=4)Sham/Cont RF/Sham RF /Cont PMA /Sham PMA-
RF/ShamLPS/Sham LPS-RF
/Sham40°C/Cont
0,0
0,5
1,0
1,5
2,0
2,5
E/C
GSM-DTX
*
** ** **
*
Simkó’s Lab
ROS ROS productionproduction ((DHRDHR--assayassay) )
GSM-DTX, 2 W/kg, for 45 min
K 562 cellsMono Mac 6 cells
0,9 1,1 1,1 1,2 1,40,0
1,0
2,0
3,0
4,0
5,0
Sham/Cont RF/Sham RF/Cont PMA/Sham PMA-RF/Sham
E/C *
3,13,20,91,40,70,0
1,0
2,0
3,0
4,0
5,0
Sham/Cont RF/Sham RF/Cont PMA/Sham PMA-RF/Sham
E/C
*
**
n ≥ 3, p< 0.05
Simkó’s Lab
ShamSham--effecteffect????
GSM-DTX- active during hearing- pulsed 1800 MHz DTX signal (2, 8 and 217 Hz modulation)- pulse maximum is variable, depending of average SAR
Cell type dependent decreased ROS production in sham:- Mono Mac 6 cells- monocytes- not K562 cells
SAR dependency: 0.5, 1.0, 1.5, 2.5 W/kg
Frequency modulation of 1800 MHz:- 2, 8, 50 or 217 Hz moduliation- fixed pulse maximum (500 W/kg) and constant average SAR
Simkó’s Lab
Hsp70 expression – DTX signal
average SAR: 2 W/kg (max. pulse: 140 W/kg)
Exposure time: 60 min
K562 cells GSM-DTX (2 W/kg)
1.2 1.1 1.20.9 0.9 1.11.1 1.0 1.31.4 1.7 1.90.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
0h 1h 2h
E/C
Sham/Cont. RF/Sham RF/Cont. Heat 42°C
* *
*
Simkó’s Lab
Hsp70 expression – DTX signal
average SAR: 2 W/kg (max. pulse: 140 W/kg)
Exposure time: 60 min
Mono Mac 6 cellsGSM-DTX (2 W/kg)
1,0 0,8 1,00,8 1,2 0,90,9 0,8 0,90,6 1,2 2,10,0
0,5
1,0
1,5
2,0
2,5
3,0
3,5
0h 1h 2h
E/C
Sham/Cont. RF/Sham RF/Cont. Heat 42°C
*
*
Simkó’s Lab
DTX signal DTX signal –– Protein profilingProtein profiling
Human monocytesHuman monocytes
Protein arraysProtein arrays: : RF: 45 min, 2 W/Kg, GSM DTX ExpositionRF: 45 min, 2 W/Kg, GSM DTX ExpositionShSh: Sham: ShamIcIc: Incubator control: Incubator control
Antibodies (Antibodies (AbAb):):512 512 AbAb dubbledubble spotted (1024 spotted (1024 spottsspotts/array)/array)Per array Per array Cy5Cy5 / / Cy3Cy3RFRF--Cy3Cy3 / Sh/ Sh--Cy5Cy5 and RFand RF--Cy5Cy5 / Sh/ Sh--Cy3Cy3
Evaluation:Evaluation:3 values of 4 must show the same direction 3 values of 4 must show the same direction of regulationof regulation
4 , 80 , 91, 30 , 7
4 , 40 ,0
1,0
2 ,0
3 ,0
4 ,0
5,0
6 ,0 *
*
*
Simkó’s Lab
SummarySummary
1800 MHz at any time and using different signal modulations do not induce free radical production or Hsp70 expression if data compared to controls
GSM-DTX signal at 2 W/kg induces a significant increase of free radicals if data compared to sham
Protein profiling showed 4-6 candidates for real-time PCR which is in progress
Simkó’s Lab
PublicationsPublications
LantowLantow M, M, SchudererSchuderer J, J, HartwigHartwig C and C and SimkSimkóó M: Free M: Free radicalradical release and release and Hsp70 expression in Hsp70 expression in twotwo human immune relevant cell human immune relevant cell lineslines after after exposureexposureto 1800 MHz to 1800 MHz radiofrequencyradiofrequency radiationradiation ((acceptedaccepted Rad. Res. 2005)Rad. Res. 2005)
SimkSimkóó M, M, HartwigHartwig C, C, LantowLantow M, M, LupkeLupke M, Mattsson MO, and M, Mattsson MO, and RollwitzRollwitz J: J: Hsp70 expression and Hsp70 expression and freefree radicalradical release after release after exposureexposure to to nonnon--thermalthermalradioradio--frequencyfrequency electromagneticelectromagnetic fieldsfields and and ultrafineultrafine particlesparticles in human in human Mono Mac 6 cells (Mono Mac 6 cells (acceptedaccepted ToxicToxic. Lett. 2005) . Lett. 2005)
LantowLantow M, M, HartwigHartwig C, C, MaerckerMaercker C and C and SimkSimkóó M: M: Free Free radicalradical ProductionProduction, , Hsp70 Hsp70 exressionexression and and proteinprotein profilingprofiling afterafter 1800 1800 mhzmhz RF RF exposureexposure in in different immune relevant different immune relevant cellscells. . 27 Annual Meeting of The BEMS, 2005, 27 Annual Meeting of The BEMS, 2005, Dublin, Ireland Abstract Book (2005).Dublin, Ireland Abstract Book (2005).
LantowLantow M and M and SimkSimkóó M: 1800 MHz RFM: 1800 MHz RF--EMF do not induce free radical EMF do not induce free radical production in different immune relevant cells. 26 Annual Meetingproduction in different immune relevant cells. 26 Annual Meeting of The of The BEMS, 2004, Washington DC, USA, Abstract Book, (2004).BEMS, 2004, Washington DC, USA, Abstract Book, (2004).
