Calcium uptake by leukemic and normal T-lymphocytes exposed to low frequency magnetic fields

Calcium-ion uptake by normal and leukemia lymphocytes increased during a 30-min exposure to a 13.6 Hz, sinusoidal magnetic field at 20 microT peak. The time-varying field was horizontal and parallel to a 16.5 microT component of the ambient static magnetic field. The uptake of 45Ca2+ increased 102% in a line of murine, cytotoxic T-lymphocytes (C57B1/6-derived CTLL-1), increased 126% in freshly-isolated spleen lymphocytes (C57B1/6 mice), and increased 75% in a line of lymphoma cells (C57B1/6-derived EL4). In contrast, there was no effect when the same field was applied for 30 min immediately before--as opposed to during--incorporation of calcium ions. When spleen lymphocytes were exposed during incubation with 45Ca2+ to a 60 Hz magnetic field at 20 microT peak, a small but statistically significant increase (37%) in uptake of the labeled ions occurred. These results indicate that weak, alternating magnetic fields might affect calcium-dependent functions of normal and leukemic lymphocytes.     

Investigation of AC-DC magnetic field effects in planar phospholipid bilayers.

Observations recently reported by others indicate that a combination of a weak dc magnetic field and extremely-low-frequency ac magnetic field can produce resonant effects in biological systems. We report measurements of the effects of combined dc and ac magnetic fields on the dc current through channel-free planar phospholipid membranes. The combined dc-ac magnetic fields did affect the dc current through planar phospholipid membranes, but not in every membrane, and not consistently at the same values of magnetic flux density and frequency. None of our measurements showed resonant response akin to the cyclotron-like resonance reported in diatoms [Smith et al., 1987] and lymphocytes [Liboff et al., 1987].     

Criticism of Lednev's mechanism for the influence of weak magnetic fields on biological systems.

V. V. Lednev has proposed a mechanism that he suggests would allow very weak magnetic fields, at the cyclotron resonance frequency for Ca2+ ions in the earth's field, to induce biological effects. I show that for four independent reasons no such mechanism can operate.     

Ca2+ ion transport through patch-clamped cells exposed to magnetic fields

The total current of Ca²⁺ ions through patch-clamped cell membranes was measured while exposing clonal insulin-producing β-cells (RINm5F) to a combination of DC and AC magnetic fields at so-called cyclotron resonance conditions. Previous experimental evidence supports the theory that a resonant interaction between magnetic fields and organisms can exist. This experiment was designed to test one possible site of interaction: channels in the cell membrane. The transport of Ca²⁺ ions through the protein channels of the plasma membrane did not show any resonant behavior in the frequency range studied. © 1995 Wiley-Liss, Inc.     

Experimental search for combined AC and DC magnetic field effects on ion channels

The hypothesis that specific combinations of DC and low frequency AC magnetic fields at so-called cyclotron-resonance conditions could affect the transport of ions through ion channels, or alter the kinetics of ion channels (opening and closing rates), has been tested. As a model system, the ion channels formed by gramicidin A incorporated in lipid bilayer membranes were studied. No significant changes in channel conductance, average lifetime, or formation rate as a function of applied fields could be detected over a wide range of frequencies and field strengths. Experiments were carried out to measure the time-resolved single-channel events and the average conductances of many-channel events in the presence of K⁺ and H⁺ ions. The channel blocking effect of Ca⁺⁺ was also studied. © 1993 Wiley-Liss. Inc.     

Effects of extremely-law-frequency electromagnetic fields on ion transport in several mammalian cells

We have investigated the effects of sinusoidal electromagnetic fields (EMF) on ion transport (Ca²⁺, Na⁺, K⁺, and H⁺) in several cell types (red blood cells, thymocytes, Ehrlich ascites tumor cells, and HL60 and U937 human leukemia cells). The effects on the uptake of radioactive tracers as well as on the cytosolic Ca²⁺ concentration ([Ca²⁺]_i), the intracellular pH (pH_i), and the transmembrane potentsial (TMP) were studied. Exposure to EMF at 50 Hz and 100–2000 μT (rms) had no significant effects on any of these parameters. Exposure to EMF of 20–1200 μT (rms) at the estimated cyclotron magnetic resonance frequencies for the respective ions had no significant effects except for a 12–32% increase of the uptake of ⁴²K within a window at 14.5–15.5 Hz and 100–200 μT (rms), which was found in U937 and Ehrlich cells but not in the other cell types. © 1994 Wiley-Liss, Inc.     

Possible mechanism for the influence of weak magnetic fields on biological systems

A physical mechanism is suggested for a resonant interaction of weak magnetic fields with biological systems. An ion inside a Ca(2+)-binding protein is approximated by a charged oscillator. A shift in the probability of ion transition between different vibrational energy levels occurs when a combination of static and alternating magnetic fields is applied. This in turn affects the interaction of the ion with the surrounding ligands. The effect reaches its maximum when the frequency of the alternating field is equal to the cyclotron frequency of this ion or to some of its harmonics or sub-harmonics. A resonant response of the biosystem to the magnetic field results. The proposed theory permits a quantitative explanation for the main characteristics of experimentally observed effects.     

Calcium binding to calmodulin. Cooperativity of the calcium-binding sites

The effects of Mg2+ ion, pH, and KCl concentration on Ca2+ binding to calmodulin were studied by using a Ca2+ ion-sensitive electrode. The Ca2+ ion affinity of calmodulin increased with increasing pH or decreasing KCl concentration. Cooperativity between the Ca2+-binding sites was observed, and increased with decreasing pH or increasing KCl concentration. Free Ca2+ ion concentration was decreased by adding MgCl2 ion at low Mg2+ concentration and increased at higher concentrations in the presence of small amounts of Ca2+ ion. The decrease of free Ca2+ ion concentration by Mg2+ ion strongly suggests cooperativity between the Ca2+-binding sites, and it is difficult to explain the decrease in terms of the ordered binding models previously proposed. These results can be explained by a simple model which has four equivalent binding sites that bind Ca2+ and Mg2+ competitively, and showing cooperativity when either Ca2+ or Mg2+ is bound. Mg2+ ion binding to calmodulin was measured in the presence or absence of Ca2+ to confirm the validity of this model, and no Mg2+-specific site was observed.     

Electric fields induced in rat and human models by 60-Hz magnetic fields: comparison of calculated and measured values.

The calculated distribution of electric fields induced in homogeneous human and rat models by a 60-Hz magnetic field is compared with values measured in instrumented mannequins. The calculated values agree well with measured values.     

Kinetics of channelized membrane ions in magnetic fields

The cyclotron resonance model for channel ion transport in weak magnetic fields is extended to include damping losses. The conductivity tensor is obtained for different electric field configurations, including the circuital field E phi normal to the channel axis. The conductivity behavior close to the cyclotron resonance frequency omega c is compared to existing Ca2+-efflux data in the literature. A collision time of .023 s results from this comparison under the assumption that K+ ions are transiting in a 0.35 G field. We estimate a mean kinetic energy of 3.5 eV for this ion at resonance. This model leads to discrete modes of vibration (eigenfrequencies) in the ion-lattice interaction, such that omega n = n omega c. The presence of such harmonics is compatible with recent results by Blackman et al. [1985b] and McLeod et al. [1986] with the interesting exception that even modes do not appear in their observations, whereas the present model has no restriction on n. This harmonic formalism is also consistent with another reported phenomenon, that of quantized multiple conductances in single patch-clamped channels.     

Electric fields induced in chicken eggs by 60-Hz magnetic fields and the dosimetric importance of biological membranes.

Chicken eggs are convenient models for observing the effects of inhomogeneities and variations, such as those found in biological membranes and in cellular conductivities, on the distribution of internal electric fields as induced by exposure to magnetic fields. The vitelline membrane separates the yolk, which has a conductivity of 0.26 S/m, from the white, which has a conductivity of 0.85 S/m. A miniaturized probe with 2.4-mm resolution was used to measure induced fields in eggs placed in a uniform, 1-mT magnetic field at 60 Hz. The E fields induced in eggs with homogenized contents agreed with expectations based on simple theory. Results were similar to intact eggs unless the probe moved the yolk off-center, which greatly perturbed the induced fields. A more reproducible arrangement, which consisted of saline-agar filled dishes with a hole cut for test samples, was developed to enhance definition of electrical parameters. With this test system, the vitelline membrane was found to be responsible for most of the perturbation of the induced field, because it electrically isolates the yolk from the surrounding white. From a theoretical viewpoint, this dosimetry for the macroscopic egg yolk is analogous to the interaction of fields with microscopic cells. These findings may have important implications for research on biological effects of ELF electromagnetic fields, especially for studies of avian embryonic development.     

Biologic effects of prolonged exposure to ELF electromagnetic fields in rats: II. 50 Hz magnetic fields

To provide possible laboratory support to health risk evaluation associated with long-term, low-intensity magnetic field exposure, 256 male albino rats and an equal number of control animals (initial age 12 weeks) were exposed 22 h/day to a 50 Hz magnetic flux density of 5 μmT for 32 weeks (a total of about 5000 h). Hematology was studied from blood samples before exposure to the field and at 12 week intervals. Morphology and histology of liver, heart, mesenteric lymph nodes, and testes as well as brain neurotransmitters were assessed at the end of the exposure period. In two identical sets of experiments, no significant differences in the investigated variables were found between exposed and sham-exposed animals. It is concluded that continuous exposure to a 50 Hz magnetic field of 5 μT from week 12 to week 44, which makes up ?70% of the life span of the rat before sacrifice, does not cause changes in growth rate, in the morphology and histology of liver, heart, mesenteric lymph nodes, testes, and bone marrow, in hematology and hematochemistry, or in the neurotransmitters dopamine and serotonin. © 1995 Wiley-Liss, Inc.     

An analysis of associations between social class and ambient magnetic fields in metropolitan Melbourne.

In the course of a study on residential magnetic-field exposure, some incidental data were obtained that bear on the issue of confounding of magnetic field exposure by social class. We have explored the possibility that the magnetic flux density of 50 Hz fields measured in Melbourne streets is correlated with a number of variables that index the socio-economic status of the neighborhood. We have examined also for a correlation between field-intensity levels and sums of some or all of the indicators, which were weighted to provide an overall score on socio-economic status. Although some of the indexes were weakly, but significantly, correlated with environmental levels of magnetic fields, the combined indices were not. These results indicate that socio-economic status is not likely to be a confounder in epidemiological studies of residential exposure to ELF magnetic fields in Melbourne.     

Sex-linked recessive lethal test of Drosophila melanogaster after exposure to 50-Hz magnetic fields.

To determine whether a 50-Hz magnetic field will induce mutations, a sex-linked recessive lethal test of Drosophila melanogaster was performed. Adult flies were exposed at an rms flux density of 500 mu T or 5 mT to the homogeneous field of a Helmholtz coil. The ambient field to which controls were exposed was less than 1 mu T. Exposures took place continuously for 13 to 14 days, which correspond to the life cycle of Drosophila at 25 degrees C. About 10,000 X-chromosomes were tested at each flux density. Recessive lethal mutation rates of 0.13, 0.21, and 0.18 percent were observed, respectively, for control, 500-mu T, and 5-mT conditions. By the Kastenbaum-Bowman significance test, the recessive lethal mutation rates in the 500-mu T and 5-mT conditions did not differ from the mutation rate of controls.     

Dose response study of human exposure to 60 Hz electric and magnetic fields

This human exposure study examined the relationship between field strength and biological response and tested whether the exposure levels at which the greatest effects occur differ for different endpoints. Three matched groups of 18 men each participated in two 6 h exposure test sessions. All subjects were sham exposed in one session. In the other session, each group of subjects was exposed at a different level of combined electric and magnetic field strength (low group: 6 kV/m, 10 μT; medium group: 9 kV/m, 20 μT; and high group: 12 kV/m, 30 μT). The study was performed double blind, with exposure order counterbalanced. Significant slowing of heart rate, as well as alterations in the latency and amplitude of event-related brain potential measures derived from the electro encephalogram (EEG), occurred in the group exposed to the 9 kV/m, 20 μT combined field (medium group). Exposure at the other field strength levels had no influence on cardiac measures and differential effects on EEG activity. Significant decrements in reaction time and in performance accuracy on a time estimation task were observed only in the low group. These results provide support for the hypothesis that humans may be more responsive to some combinations or levels of field strength than to others and that such differences in responsivity may depend, in part, on the endpoint of interest. © 1994 Wiley-Liss, Inc.     

Effects of 60 Hz electric and magnetic fields on maturation of the rat neopallium

This study was undertaken to determine the effects of extremely low frequency (ELF; 60 Hz) electromagnetic (EM) fields on somatic growth and cortical development, as well as biochemical and morphological maturation, of the rat neopallium. On the fifth day of pregnancy, female rats were put in pairs into plastic cages that were housed in a specially constructed apparatus for irradiation under three separate sets of combination and intensity: 1) 1 kV/m and 10 gauss; 2) 100 kV/m and 1 gauss; and 3) 100 kV/m and 10 gauss. The dams were exposed for 23 h daily, from days 5 through 19 postconception after which they were returned to cages outside the exposure apparatus until they littered. The neonates were culled to eight pups per litter. At 0 (birth), 5, 12, and 19 days postnatally, they were killed for biochemical and morphological studies. Another group of pregnant rats was sham-exposed in an identical apparatus, which was not energized, and the pups were used as controls. The irradiated rats exhibited no physical abnormalities, nor did they show brain deformities such as swelling or herniation following exposure to ELF-EM fields. There was no difference in somatic growth between control and exposed rats, but a small reduction in cortical weight was observed in rats exposed at 1 kV/m and 10 gauss, and 100 kV/m and 1 gauss, respectively. Biochemical measurements of DNA. RNA, protein, and cerebroside concentrations indicated that among the three separate exposures, only the neopallium of rats exposed at 1 kV/m and 10 gauss showed a small reduction in DNA level, as well as small reductions in RNA and protein levels. No changes were noticed in cerebroside levels in any exposed animals, and there were no differences in protein/DNA and cerebroside/DNA ratios between control and exposed rats. Morphological observations did not reveal any detectable alterations in the irradiated rats. These results indicate that exposure to ELF-EM fields caused minimal or no changes in somatic growth and cerebral development of the rat. © 1993 Wiley-Liss, Inc.     

Effects of prenatal exposure to 50 Hz magnetic fields on development in mice: I. Implantation rate and fetal development

Pregnant CD1 mice were exposed or sham-exposed from day 0 to day 17 of gestation to a 50 Hz sinusoidal magnetic field at 20 mT (rms). Preimplantation and postimplantation survival were assessed and fetuses examined for the presence of gross external, internal, and skeletal abnormalities. There were no statistically significant field-dependent effects on preimplantation or postimplantation survival, sex ratio, or the incidence of fetuses with internal or skeletal abnormalities. Magnetic field exposure was, however, associated with longer and heavier fetuses at term, with fewer external abnormalities. The results lend no support to suggestions of increased rates of spontaneous abortion or congenital malformation following prenatal exposure to power frequency magnetic fields. © 1994 Wiley-Liss, Inc.