Effect of amplitude-modulated 147 MHz radiofrequency radiation on calcium ion efflux from avian brain tissue

Cerebral cortex tissue slices and cerebral hemispheres prepared from Gallus domesticus chicks were exposed to 147 MHz radiofrequency radiation, amplitude modulated at 16 Hz and applied at a power density of 0.75 mW/cm2, to determine the effect of such exposure of 45Ca2+ efflux from the avian brain tissue. Statistical analysis of these data demonstrates that such exposure has no significant effect on 45Ca2+ efflux.     

Effect of amplitude modulated RF radiation on calcium ion efflux and ODC activity in chronically exposed rat brain

The effect of exposing rats to amplitude modulated radiofrequency radiation (112 MHz modulated to 16 Hz) during development and growth has been examined. Wistar rats (35 days old) when exposed at above frequency at the power level 1.0 mW/cm2 (SAR, 0.75 W/kg) for 35 days showed enhanced ornithine decarboxylase activity and Ca2+ efflux in brain indicating potential health hazards due to exposure.     

Frequency-dependent alterations in enolase activity in Escherichia coli caused by exposure to electric and magnetic fields

Some neurochemical effects of low-intensity electric and magnetic fields have been shown to be nonlinear functions of exposure parameters. These effects occurred within narrow ranges of frequency and intensity. Previous studies on membrane-associated endpoints in cell culture preparations demonstrated changes in calcium efflux and in acetylcholinesterase activity following exposure to radiofrequency radiation, amplitude modulated (AM) at 16 and at 60 Hz, at a specific absorption rate of 0.05 W/kg. In this study, these modulation frequencies were tested for their influence on the activity of a cytoplasmic enzyme, enolase, which is being tested clinically for detection of neoplasia. Escherichia coli cultures containing a plasmid with a mammalian gene for enolase were exposed for 30 min, and cell extracts were assayed for enolase activity by measuring absorbance at 240 nm. The enolase activity in exposed cultures was compared to the activity in paired control cultures. Exposure to 147 MHz carrier waves at 0.05 W/kg, AM at 16 Hz showed enolase activity enhanced by 62%, and AM at 60 Hz showed enolase activity reduced by 28%. Similarly, exposure to 16 Hz fields alone, at 21.2 V/m_rms (electric) and 97 nT_rms (magnetic), showed enhancement in enolase activity by 59%, whereas exposure to 60 Hz fields alone, at 14.1 V/m_rms (electric) and 65 nT_rms (magnetic), showed reduction in activity by 24%. Sham exposures as well as exposure to continuous-wave 147 MHz radiation at 0.05 W/kg showed no change in enolase activity. Although the underlying basis for these field effects in the cytoplasmic compartment has not been established, differential sensitivities to 16 Hz and to 60 Hz signals provide a clear focus for additional research to determine the responsible mechanism. © 1994 Wiley-Liss, Inc.     

Calcium-ion movement and contractility in atrial strips of frog heart are not affected by low-frequency-modulated, 1 GHz electromagnetic radiation

Calcium efflux from electrically stimulated, ⁴⁵Ca²⁺-preloaded atrial strips of the frog heart was measured from samples of the rinsing perfusate collected at 2-min intervals for 32 min in a continuous perfusion chamber. Contractile force was simultaneously monitored. The specimen chamber was located in a stripline apparatus in which the atrial strips were exposed for 32 min to constant (CW) or amplitude-modulated (AM), 1 GHz electromagnetic (EM) fields at specific absorption rates (SAR) ranging from 3.2 μW/kg to 1.6 W/kg. Amplitude modulation was either at 0.5 Hz, in synchrony with the electrical stimulus applied to the preparation, or at 16 Hz. Neither unmodulated nor 0.5 Hz or 16 Hz modulated 1 GHz waves affected the movement of calcium ions or the contractile force in isolated atrial strips of the frog heart. © 1993 Wiley-Liss, Inc.     

Microwave radiation-induced calcium ion efflux from human neuroblastoma cells in culture

Monolayer cultures of human neuroblastoma cells were exposed to 915-MHz radiation, with or without sinusoidal amplitude modulation (80%) at 16 Hz, at specific absorption rates (SAR) for the culture medium and cells of 0.00, 0.01, 0.05, 0.075, 0.1, 0.5, 0.75, 1.0, 1.5, 2, or 5 mW/g. A significant increase in the efflux of calcium ions (⁴⁵Ca²⁺) as compared to unexposed control cultures occurred at two SAR values: 0.05 and 1 mW/g. Increased efflux at 0.05 mW/g was dependent on the presence of amplitude modulation at 16 Hz but at the higher value it was not. These results indicate that human neuroblastoma cells are sensitive to extremely low levels of microwave radiation at certain narrow ranges of SAR.     

Multiple power-density windows and their possible origin

We have previously reported that in vitro exposure of chick forebrain tissue to 50-MHz radiofrequency (RF) electromagnetic radiation, amplitude modulated (AM) at 16 Hz, would enhance the efflux of calcium ions within only two power-density ranges: one from 1.44 to 1.67 mW/cm2, and the other including 3.64 mW/cm2. No effect on efflux occurred at 0.37, 0.72, 2.17, and 4.32 mW/cm2. We confirmed and extended these results by testing at another set of power densities, which included the range of the previous study. Forebrain tissue from 1-7-day-old chickens was labeled in vitro with radioactive calcium ions (30 min, at 37 degrees C), rinsed, placed in a physiological salt solution, and then exposed for 20 min to 50-MHz radiation, AM at 16 Hz, in a transverse electric and magnetic field (TEM) cell maintained at 37 degrees C. The solution was then assayed for radioactive calcium activity. A power-density series was tested. An enhanced efflux of calcium ions was found at 1.75, 3.85, 5.57, 6.82, 7.65, 7.77, and 8.82 mW/cm2; no change was observed at 0.75, 2.30, 4.50, 5.85, 7.08, 8.19, 8.66, 10.6, and 14.7 mW/cm2. Power density is converted to specific absorption rate (SAR) by 0.36 mW/kg per mW/cm2. Even the highest SAR tested (0.005 W/kg) is much too low to result in generalized heating of the sample and thus to be the underlying cause of the enhanced response. A hypothetical mechanism is proposed involving dynamic systems that may account for the power-density dependency as well as for part of the frequency dependency observed with both modulated RF radiation and extremely-low-frequency (ELF) fields.     

Dose dependence of acetylcholinesterase activity in neuroblastoma cells exposed to modulated radio-frequency electromagnetic radiation.

Radio-frequency electromagnetic radiation (RFR) at 915 and 147 MHz, when sinusoidally amplitude modulated (AM) at 16 Hz, has been shown to enhance release of calcium ions from neuroblastoma cells in culture. The dose-response relation is unusual, consisting of two power-density "windows" in which enhanced efflux occurs, separated by power-density regions in which no effect is observed. To explore the physiological importance of these findings, we have examined the impact of RFR exposure on a membrane-bound enzyme, acetylcholinesterase (AChE), which is intimately involved with the acetylcholine (ACh) neurotransmitter system. Neuroblastoma cells (NG108), exposed for 30 min to 147-MHz radiation, AM at 16 Hz, demonstrated enhanced AChE activity, as assayed by a procedure using 14C-labeled ACh. Enhanced activity was observed within a time window between 7.0 and 7.5 h after the cells were plated and only when the exposure occurred at power densities identified in a previous report as being effective for altering the release of calcium ions. Thus RFR affects both calcium-ion release and AChE activity in nervous system-derived cells in culture in a common dose-dependent manner.     

Absorption of microwave radiation by the anesthetized rat: electromagnetic and thermal hotspots in body and tail

Anatomic variability in the deposition of radiofrequency electromagnetic energy in mammals has been well documented. A recent study [D'Andrea et al., 1985] reported specific absorption rate (SAR) hotspots in the brain, rectum and tail of rat carcasses exposed to 360- and to 2,450-MHz microwave radiation. Regions of intense energy absorption are generally thought to be of little consequence when predicting thermal effects of microwave irradiation because it is presumed that heat transfer via the circulatory system promptly redistributes localized heat to equilibrate tissue temperature within the body. Experiments on anesthetized, male Long-Evans rats (200-260 g) irradiated for 10 or 16 min with 2,450, 700, or 360 MHz radiation at SARs of 2 W/kg, 6 W/kg, or 10 W/kg indicated that postirradiation localized temperatures in regions previously shown to exhibit high SARs were appreciably above temperatures at body sites with lower SARs. The postirradiation temperatures in the rectum and tail were significantly higher in rats irradiated at 360 MHz and higher in the tail at 2,450 MHz than temperatures resulting from exposure to 700 MHz. This effect was found for whole-body-averaged SARs as low as 6 W/kg at 360 MHz and 10 W/kg at 2,450 MHz. In contrast, brain temperatures in the anesthetized rats were not different from those measured in the rest of the body following microwave exposure.     

Effect of low-frequency pulse-modulated 460 MHz electromagnetic irradiation on Drosophila embryos

Effect of electromagnetic radiation 460 MHz with 2.5-40 Hz pulse modulation rate on Drosophila embryos of 15 h 10 m age was studied. It was demonstrated that a 5-min irradiation with 0.12 W/kg average SAR (3 W/kg pulsed SAR) alters the Drosophila percentage of interrupted development. The effect strength depended on the modulation rate with a pronounced decrease at 10 and 16 Hz. A hypothesis about the presence of thermal and non-thermal mechanisms of action of pulse-modulated microwave radiation diversely effecting the embryos has been put forward and grounded.     

Effects of low level microwave radiation on carcinogenesis in Swiss Albino mice

This study concerns with the multiple treatment of the target site to potent carcinogen and the super imposition of low level radiofrequency and microwave radiation. Swiss albino mice (male) were used for this investigation. The study has been divided in two parts, part A: a single dose of 7,12-dimethylbenz(a)anthracene (DMBA) 100 μg/animal was applied topically on the skin of mice and were exposed to 112 MHz amplitude modulated (AM) at 16 Hz (power density 1.0 mW/cm(2), specific absorption rate (SAR) 0.75 W/kg). Similarly after a single dose of DMBA, mice were exposed to 2.45 GHz radiation (power density of 0.34 mW/cm(2), SAR, 0.1 W/kg), 2 h/day, 3 days a week for a period of 16 weeks. The two sets of experiments were carried out separately. Part B: mice were transplanted intraperitoneally (ip) with ascites 8 × 10(8) (Ehrlich-Lettre ascites, strain E) carcinoma cells per mouse. These mice were exposed to 112 MHz amplitude modulated at 16 Hz and 2.45 GHz radiation separately for a period of 14 days. There was no tumor development in mice exposed to RF and MW. Similarly a topical application of single dose of DMBA followed by RF/MW exposure also did not produce any visible extra tumor on the skin of mice. On the other hand mice were transplanted intraperitoneally with ascites (8 × 10(8) cell/ml) and subsequently exposed to above mentioned fields for 14 days showed a slight increase in the cell numbers as compared to the control group. However, the increase is insignificant. There were insignificant differences either in the mortality or cell proliferation among the control and exposed group. This results show that low level RF or MW do not alter tumor growth and development as evidenced by no observable change in tumor size.     

Attempts to alter 45Ca2+ binding to brain tissue with pulse-modulated microwave energy

Rat brain tissue, loaded with ⁴⁵Ca²⁺ by intraventricular injection was exposed in vitro to pulsemodulated 1-GHz (SAR of 0.29 or 2.9 W/kg) or 2.45-GHz radiation (SAR = 0.3 W/kg), and in vivo to 2.06-GHz radiation (SAR of 0.12 to 2.4 W/kg). There were no significant differences in efflux of ⁴⁵Ca²⁺ between the microwave- and sham-irradiated groups.     

Calcium-ion efflux from brain tissue: Power-density versus internal field-intensity dependencies at 50-MHz RF radiation

In previous experiments changes were found in calcium-ion efflux from chickbrain tissue that had been exposed in vitro to 147-MHz radiation across a specific range of power densities when the field was amplitude modulated at 16 Hz. In the present study, 50-MHz radiation, similarly modulated as a sinusoid, was found to produce changes in calcium-ion efflux from chick brains exposed in vitro in a Crawford cell. Exposure conditions were optimized to broaden any power-density window and to enhance the opportunity to detect changes in the calcium-ion efflux. The results of a power-density series demonstrated two effective ranges: One spanning a range from 1.44 to 1.67 mW/cm², and the other including 3.64 mW/cm², which were bracketed by no-effect results at 0.72, 2.17, and 4.32 mW/cm². Peaks of positive findings are associated with near-identical rates of energy absorption: 1.4 μW/g at 147 MHz, and 1.3 μW/g at 50 MHz, which indicates that the enhanced-efflux phenomenon is more dependent on the intensity of fields in the brain than on the power density of incident radiation. In addition, the phenomenon appears to occur at multiples of some, as yet unknown, rate of radiofrequency (RF) energy absorption. Because of the extremely small increments of temperature associated with positive findings (< 4 × 10^?4°C), and the existence of more than one productive absorption rate, a solely thermal explanation appears extremely unlikely.     

Effects of ELF (1-120 Hz) and modulated (50 Hz) RF fields on the efflux of calcium ions from brain tissue in vitro

We have previously shown that 16-Hz, sinusoidal electromagnetic fields can cause enhanced efflux of calcium ions from chick brain tissue, in vitro, in two intensity regions centered on 6 and 40 Vp-p/m. Alternatively, 1-Hz and 30-Hz fields at 40 Vp-p/m did not cause enhanced efflux. We now demonstrate that although there is no enhanced efflux associated with a 42-Hz field at 30, 40, 50, or 60 Vp-p/m, a 45-Hz field causes enhanced efflux in an intensity range around 40 Vp-p/m that is essentially identical to the response observed for 16-Hz fields. Fields at 50 Hz induce enhanced efflux in a narrower intensity region between 45 and 50 Vp-p/m, while radiofrequency carrier waves, amplitude modulated at 50 Hz, also display enhanced efflux over a narrow power density range. Electromagnetic fields at 60 Hz cause enhanced efflux only at 35 and 40 Vp-p/m, intensities slightly lower than those that are effective at 50 Hz. Finally, exposures over a series of frequencies at 42.5 Vp-p/m reveal two frequency regions that elicit enhanced efflux--one centered on 15 Hz, the other extending from 45 to 105 Hz.     

Radio frequency radiation effects on protein kinase C activity in rats' brain

The present work describes the effect of amplitude modulated radio frequency (rf) radiation (112 MHz amplitude-modulated at 16 Hz) on calcium-dependent protein kinase C (PKC) activity on developing rat brain. Thirty-five days old Wistar rats were used for this study. The rats were exposed 2 h per day for 35 days at a power density of 1.0 mW/cm2 (SAR = 1.48 W/kg). After exposure, rats were sacrificed and PKC was determined in whole brain, hippocampus and whole brain minus hippocampus separately. A significant decrease in the enzyme level was observed in the exposed group as compared to the sham exposed group. These results indicate that this type of radiation could affect membrane bound enzymes associated with cell signaling, proliferation and differentiation. This may also suggest an affect on the behavior of chronically exposed rats.     

Carcinogenicity study of 217 Hz pulsed 900 MHz electromagnetic fields in Pim1 transgenic mice

In an 18-month carcinogenicity study, Pim1 transgenic mice were exposed to pulsed 900 MHz (pulse width: 0.577 ms; pulse repetition rate: 217 Hz) radiofrequency (RF) radiation at a whole-body specific absorption rate (SAR) of 0.5, 1.4 or 4.0 W/kg [uncertainty (k = 2): 2.6 dB; lifetime variation (k = 1): 1.2 dB]. A total of 500 mice, 50 per sex per group, were exposed, sham-exposed or used as cage controls. The experiment was an extension of a previously published study in female Pim1 transgenic mice conducted by Repacholi et al. (Radiat. Res. 147, 631-640, 1997) that reported a significant increase in lymphomas after exposure to the same 900 MHz RF signal. Animals were exposed for 1 h/day, 7 days/week in plastic tubes similar to those used in inhalation studies to obtain well-defined uniform exposure. The study was conducted blind. The highest exposure level (4 W/kg) used in this study resulted in organ-averaged SARs that are above the peak spatial SAR limits allowed by the ICNIRP (International Commission on Non-ionizing Radiation Protection) standard for environmental exposures. The whole-body average was about three times greater than the highest average SAR reported in the earlier study by Repacholi et al. The results of this study do not suggest any effect of 217 Hz-pulsed RF-radiation exposure (pulse width: 0.577 ms) on the incidence of tumors at any site, and thus the findings of Repacholi et al. were not confirmed. Overall, the study shows no effect of RF radiation under the conditions used on the incidence of any neoplastic or non-neoplastic lesion, and thus the study does not provide evidence that RF radiation possesses carcinogenic potential.     

Exposure of frog hearts to CW or amplitude-modulated VHF fields: selective efflux of calcium ions at 16 Hz

Isolated frog hearts were exposed for 30-min periods in a Crawford cell to a 240-MHz electromagnetic field, either continuous-wave or sinusoidally modulated at 0.5 or 16 Hz. Radiolabeled with calcium (45Ca), the hearts were observed for movement of Ca2+ at calculated SARs of 0.15, 0.24, 0.30, 0.36, 1.50, or 3.00 mW/kg. Neither CW radiation nor radiation at 0.5 Hz, which is close to the beating frequency of the frog's heart, affected movement of calcium ions. When the VHF field was modulated at 16 Hz, a field-intensity-dependent change in the efflux of calcium ions was observed. Relative to control values, ionic effluxes increased by about 18% at 0.3 mW/kg (P less than .01) and by 21% at 0.15 mW/kg (P less than .05), but movement of ions did not change significantly at other rates of energy deposition. These data indicate that the intact myocardium of the frog, akin to brain tissue of neonatal chicken, exhibits movement of calcium ions in response to a weak VHF field that is modulated at 16 Hz.     

Effects of weak amplitude-modulated microwave fields on calcium efflux from awake cat cerebral cortex

Calcium (45Ca2+) efflux was studied from preloaded cortex in cats immobilized under local anesthesia, and exposed to a 3.0-mW/cm2 450-MHz field, sinusoidally amplitude modulated at 16 Hz modulation depth 85%). Tissue dosimetry showed a field of 33 V/m in the interhemispheric fissure (rate of energy deposition 0.29 W/kg). Field exposure lasted 60 min. By comparison with controls, efflux curves from field exposed brains were disrupted by waves of increased 45Ca2+ efflux. These waves were irregular in amplitude and duration, but many exhibited periods of 20-30 min. They continued into the postexposure period. Binomial probability analysis indicates that the field-exposed efflux curves constitute a different population from controls at a confidence level of 0.96. In about 70% of cases, initiation of field exposure was followed by increased end-tidal CO2 excretion for about 5 min. However, hypercapnea induced by hypoventilation did not elicit increased 45Ca2+ efflux. Thus this increase with exposure does not appear to arise as a secondary effect of raised cerebral CO2 levels. Radioactivity measurements in cortical samples after superfusion showed 45Ca2+ penetration at about 1.7 mm/hr, consistent with diffusion of the ion in free solution.     

Stress proteins are not induced in mammalian cells exposed to radiofrequency or microwave radiation

The induction of stress proteins in HeLa and CHO cells was investigated following a 2 h exposure to radiofrequency (RF) or microwave radiation. Cells were exposed or sham exposed in vitro under isothermal (37 ± 0.2 °C) conditions. HeLa cells were exposed to 27- or 2450 MHz continuous wave (CW) radiation at a specific absorption rate (SAR) of 25 W/kg. CHO cells were exposed to CW 27 MHz radiation at a SAR of 100 W/kg. Parallel positive control studies included 2 h exposure of HeLa or CHO cells to 40 °C or to 45 μM cadmium sulfate. Stress protein induction was assayed 24 h after treatment by electrophoresis of whole-cell extracted protein labeled with [³⁵S]-methionine. Both cell types exhibited well-characterized responses to the positive control stresses. Under these exposure conditions, neither microwave nor RF radiation had a detectable effect on stress protein induction as determined by either comparison of RF-exposed cells with sham-exposed cells or comparison with heat-stressed or Cd⁺⁺ positive control cells. Bioelectromagnetics 18:499–505, 1997. © 1997 Wiley-Liss, Inc.     

In vitro study of the stress response of human skin cells to GSM-1800 mobile phone signals compared to UVB radiation and heat shock

The evolution of mobile phone technology is toward an increase of the carrier frequency up to 2.45 GHz. Absorption of radiofrequency (RF) radiation becomes more superficial as the frequency increases. This increasingly superficial absorption of RF radiation by the skin, which is the first organ exposed to RF radiation, may lead to stress responses in skin cells. We thus investigated the expression of three heat-shock proteins (HSP70, HSC70, HSP27) using immunohistochemistry and induction of apoptosis by flow cytometry on human primary keratinocytes and fibroblasts. A well-characterized exposure system, SXC 1800, built by the IT'IS foundation was used at 1800 MHz, with a 217 Hz modulation. We tested a 48-h exposure at an SAR of 2 W/kg (ICNIRP local exposure limit). Skin cells were also irradiated with a 600 mJ/cm2 single dose of UVB radiation and subjected to heat shock (45 degrees C, 20 min) as positive controls for apoptosis and HSP expression, respectively. The results showed no effect of a 48-h GSM-1800 exposure at 2 W/kg on either keratinocytes or fibroblasts, in contrast to UVB-radiation or heat-shock treatments, which injured cells. We thus conclude that the GSM-1800 signal does not act as a stress factor on human primary skin cells in vitro.     

Nonthermal effects of radiofrequency-field exposure on calcium dynamics in stem cell-derived neuronal cells: elucidation of calcium pathways

Intracellular Ca(2+) spikes trigger cell proliferation, differentiation and cytoskeletal reorganization. In addition to Ca(2+) spiking that can be initiated by a ligand binding to its receptor, exposure to electromagnetic stimuli has also been shown to alter Ca(2+) dynamics. Using neuronal cells differentiated from a mouse embryonic stem cell line and a custom-built, frequency-tunable applicator, we examined in real time the altered Ca(2+) dynamics and observed increases in the cytosolic Ca(2+) in response to nonthermal radiofrequency (RF)-radiation exposure of cells from 700 to 1100 MHz. While about 60% of control cells (not exposed to RF radiation) were observed to exhibit about five spontaneous Ca(2+) spikes per cell in 60 min, exposure of cells to an 800 MHz, 0.5 W/kg RF radiation, for example, significantly increased the number of Ca(2+) spikes to 15.7+/-0.8 (P<0.05). The increase in the Ca(2+) spiking activities was dependent on the frequency but not on the SAR between 0.5 to 5 W/kg. Using pharmacological agents, it was found that both the N-type Ca(2+) channels and phospholipase C enzymes appear to be involved in mediating increased Ca(2+) spiking. Interestingly, microfilament disruption also prevented the Ca(2+) spikes. Regulation of Ca(2+) dynamics by external physical stimulation such as RF radiation may provide a noninvasive and useful tool for modulating the Ca(2+)-dependent cellular and molecular activities of cells seeded in a 3D environment for which only a few techniques are currently available to influence the cells.