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5.1 ELF fields and calcium ion effects in the immune system:

Electromagnetic fields, through their effect on calcium ions, play a vital role in the immune system, Walleczek (1992). Walleczek (1992) quotes research relating to the role of calcium, sodium and potassium ions, including research showing that EMF could alter the activity of the membrane incorporated Ca2+_{-ATPase responsible for pumping Ca}2+ _{out of the}

cell (calcium ion efflux).

In addition, data from two laboratories demonstrate that ELF fields alter the activity of another membrane ion pump, Na+_/K+_{-ATPase with current densities as low as 50µA/cm}2

and estimated, by the authors, to also have an effect at 1µA/cm2. At 50µA/cm2, J = 0.5 A/m2; E=2.5 V/m, assuming σ=0.2 S/m. Hence S= 1.7 µW/cm2 and SAR = 0.00063 W/kg. If the extrapolation to 1µA/cm2 is confirmed then the EMR effects will be occurring at 1/2500th _{of the S and SAR levels estimated here.}

This demonstrates the extremely low induced currents, SARs and energy densities which are associated with EMR induced changes in ion pumping and calcium, sodium and potassium efflux at the cellular level.

Walleczek and Budinger (1992) report that:

“To date, at least 10 different laboratories, including our own, have reported ELF magnetic influences on lymphoid cells, and stimulatory as well as inhibitory effects on parameters related to calcium metabolism or RNA- and DNA-synthesis have been observed.”

They also state that:

“A plausible magnetic interaction mechanism based on radical pair recombination reactions which are linked to cellular signal transduction and application processes has been proposed (Grundler et al. (1992)). Magnetic field intensities similar to the intensities used in most experiments (e.g. 1-30 mT) are known from magnetochemistry to be able to influence non-thermally

the kinetics and product yields from radical pair reactions in vitro, Steiner et al. (1989). The underlying reaction scheme is well known and is described by the radical pair mechanism.

For this mechanism to be applicable to the data reported here, a pathway by which magnetically-sensitive radical-dependent processes could influence mitogen-induced lymphocyte Ca2+ signaling must be postulated. There is new evidence that such pathways exist.

For example, Con A-induced Ca2+ uptake in rat thymic lymphocytes has been shown to depend on the generation of reactive oxygen radical species. There is also evidence from inhibition studies that cytochrome P-450 activity may be

involved in Ca2+ uptake regulation in rat thymic lymphocytes, Alvarez et al. (1992), and it is known that P-450 function proceeds via radical pair recombination steps, Hollenberg (1992). Thus it is plausible to investigate if externally applied magnetic fields may interfere with radical pair reactions and as a consequence, may alter lymphocyte Ca2+ regulation.”

Calcium ion influx has been shown to play a role in the transcript levels of proto-oncogenes c-myc and c-fos which alters in the presence of electromagnetic fields, Karabakhtsian et al. (1994). (Proto-oncogenes: altered genes which become carcinogenic.) Lin et al. (1994) identified a specific part of the c-myc promoter which is responsive to electric and magnetic fields. Phillips (1993) exposed T-lymphoblastoid cells to 60 Hz magnetic field and found alterations to the transcription of genes encoding for c-fos, c-jun, c-myc and protein Kinase- C.

Modulated electric and magnetic fields alter key genetic characteristics and therefore are mutagenic. This is further confirmed with studies showing EMR/EMF exposure is associated with chromosome aberrations and DNA damage, e.g. Vijayalaxmi et al. (1997) and Lai and Singh (1997). Lindstrom et al. (1995) replicated and extended the research of Walleczek (1992), using the T-cell line (lymphocytes) for human leukaemia cells, and show that oscillating low-level magnetic fields produce the same calcium ion reaction as does an antibody. They show that weak magnetic fields initiate calcium ion oscillations with a threshold flux density of 40 µT, a plateau at 150 µT and a frequency range from 5 to 100 Hz, with a fairly broad peak at 50 Hz. Galvanovskis et al. (1996) report significant 30% reductions in the calcium ion oscillation amplitude in human leukaemia T-cells when exposed to 50 Hz magnetic fields. The key role of modulation frequency in the alteration of calcium ions was recognized early.

5.2 Calcium ion efflux and modulation frequency

The very early research on brain cells efflux and influx of calcium ions using external ELF frequency fields in the same range of EEG frequencies was carried out by Dr Ross Adey and his research team, e.g. Figure 24a.

Figure 24a: ELF induced calcium ion efflux in chick brain cells from (A) an ELF modulated 147 MHz signal and (B) an ELF signal, Adey (1988).

Table 2 shows a concentration on using a modulation frequency of 16 Hz which was identified early to be an ELF frequency associated with strong calcium ion efflux compared with frequencies near it. A leading researcher in this area, Dr Carl Blackman of the U.S.E.P.A. has shown that research has identified modulation frequencies which significantly alter calcium ion efflux out to 510 Hz, Figure 24b.

Figure 24b: The effect of 15 V/m electromagnetic fields on the efflux of calcium ions from chicken brain tissue as a function of modulation frequency. The relative efflux is the difference between exposed and unexposed samples. The data from 1 to 120 Hz are taken from Blackman et al. (1985). Blackman et al. (1988).

Their research further shows the involvement of polypeptide molecules, specifically poly-L- lysine, which the authors postulate may explain the intracellular calcium ion EMR effects on cell membrane surfaces, through the polylysine causing strong deformations on the cell surface which could trigger the release of stored calcium cations from intracellular pools, thus starting the oscillations.

Through replicating and extending the experiments of other laboratories, Dr Carl Blackman and his team at the U.S. Environmental Protection Agency have become the world leaders in calcium ion efflux research. That is why he was well qualified to review the research results and conclude, Blackman (1990) that

"Taken together, the evidence overwhelmingly indicates that electric and magnetic fields can alter normal calcium ion homeostasis and lead to changes in the response of biological systems to their environment".

There is extremely strong evidence that both ELF and ELF modulated RF/MW radiation causes calcium ion efflux from cells which significantly alters the intracellular calcium concentrations, reducing the efficacy of lymphocytes in the immune system, participating in the alteration of transformation of pineal serotonin to melatonin and altering the damaged cells likelihood of becoming neoplastic or dying by apoptosis.

5.3 The melatonin - calcium ion efflux link:

The interdependence of cyclic AMP and calcium ions is outlined in section 3.3 above. The following outlines an hypothesis for modulated RF/MW effects on melatonin.

Since it has been shown:

• That ELF electric fields do reduce melatonin production in living rats brains; Wilson et al. (1986).

• That RF/MW signals produce tissue level electric fields about a million times higher than imposed ELF signals, Adey (1981).

• That RF/MW signals are resonantly absorbed at the cell membrane, Liu and Cleary (1995).

• That altering the electric and thermal fields on the surface of the cell membrane change the binding characteristics of H+ and Ca2+ ions on the outer surface of the membrane, Adey (1990).

• That modulated RF/MW has been shown to induce significant calcium ion efflux from cells, Table 2 above.

• That it known that the cyclic AMP signal transduction pathway and the Calcium ion signal transduction pathway interact, Alberts et al. (1994).

• That in the pinealocyte cell the cAMP pathway is involved in regulating the transformation of serotonin to melatonin.

The calcium ion mediated responses to neurotransmitters on the membrane of the pineal cells has been discussed by Wilson et al. (1989) in relation to ELF induced melatonin reduction. Thus it is highly probable that pinealocytes exposed to modulated RF/MW will experience an outflow of calcium ions, a reduction of the cAMP signal transduction activity and a reduction in the production of melatonin. This is a highly plausible mechanism to explain why RF/MW can reduce pineal melatonin production with consequent the adverse health effects.

Table 2: Summary of Studies concerning Calcium ion efflux and ELF Modulation of RF.

RF Modn Intensity Time SAR

Effects Species (MHz) (Hz) (mW/cm2) (min) (W/kg) Reference

_______________________________________________________________________________________ Altered calcium-ion efflux in brain tissue in vitro:

Frequency specificity Chicken 147 6-20 1-2 20 0.002* Bawin et al.(1975) influence of pH and

lanthanum Chicken 450 16 0.75 20 0.0035 Bawin et al.(1978) frequency and intensity

specificity Chicken 147 16 0.83 20 0.0014 Blackman et al.(1979) intensity specificity and

sample spacing Chicken 147 9,16 0.083 20 0.0014 Blackman et al.(1980a) intensity specificity and

sample spacing Chicken 147 16 0.083 20 0.0014 Joines et al (1981) intensity specificity Chicken 450 16 0.1-1 20 0.005- Sheppard et al.(1979)

0.005

two intensity ranges Chicken 50 16 1.5 20 0.0013 Blackman et al.(1980b)

3.6 20 0.0035

theoretical analysis of Chicken 50 16 - 20 ~0.001 Joines and Blackman(1980)

RF dependence 147 Athey (1981); Joines and

450 Blackman (1981).

test of predictions Chicken 147 16 0.37 20 0.0006 Blackman et al.(1981) of theoretical analyses 0.49 0.0008

no effect for pulse Rat 1000 16,32 0.5-15 20 0.15-4.35 Shelton and Merritt (1981) modulation

no effect for pulse Rat 1000 16 1,10 20 0.29-2.9 Merritt et al. (1982) modulation 2450 8,16,32 1 0.3

change in calcium efflux Rat 450 16 0.5 10 - Lin-Liu and Adey (1982) kinetics in synaptosomes

frequency and intensity Human 915 16 - 30 0.05 Dutta et al.(1984) specificity in cultured being

neuroblastoma cells

Altered calcium ion efflux in brain tissue in vivo:

no effect for pulse mod. Rat 2060 8,16,32 0.5-10 20 0.12-2.4 Merritt et al.(1982) change in efflux kinetics Cat 450 16 3 60 0.29 Adey et al.(1982) Changes found in Rat 147 16 2 60-150 <0.075 Albert et al.(1980) pancreatic slices

Suppressed T-lymphocyte Mouse 450 16-100 1.5 120 - Lyle et al.(1983) Changes in Hearts Frog 240 0.5,16 30 0.00015 Schwartz et al (1990)