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.     

Noise magnetic fields abolish the gap junction intercellular communication suppression induced by 50 hz magnetic fields

Previously, we have reported that exposure to 50 Hz coherent sinusoidal magnetic fields (MF) for 24 h inhibits gap junction intercellular communication (GJIC) in mammalian cells at an intensity of 0.4 mT and enhances the inhibition effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) at 0.2 mT. In the present study, we further explored the effects of incoherent noise MF on MF-induced GJIC inhibition. GJIC was determined by fluorescence recovery after photobleaching (FRAP) with a laser-scanning confocal microscope. The rate of fluorescence recovery (R) at 10 min after photobleaching was adopted as the functional index of GJIC. The R-value of NIH3T3 cells exposed to 50 Hz sinusoidal MF at 0.4 mT for 24 h was 30.85 +/- 14.70%, while the cells in sham exposure group had an R-value of 46.36 +/- 20.68%, demonstrating that the GJIC of NIH3T3 cells was significantly inhibited by MF exposure (P < .05). However, there were no significant differences in the R-values of the sham exposure, MF-plus-noise MF exposure (R: 49.58 +/- 19.38%), and noise MF exposure groups (R: 46.74 +/- 21.14%) (P > .05), indicating that the superposition of a noise MF alleviated the suppression of GJIC induced by the 50 Hz MF. In addition, although MF at an intensity of 0.2 mT synergistically enhanced TPA-induced GJIC inhibition (R: 24.90 +/- 13.50% vs. 35.82 +/- 17.18%, P < .05), further imposition of a noise MF abolished the synergistic effect of coherent MF (R: 32.51 +/- 18.37%). Overall, the present data clearly showed that although noise MF itself had no effect on GJIC of NIH3T3 cells, its superposition onto a coherent sinusoidal MF at the same intensity abolished MF-induced GJIC suppression. This is the first report showing that noise MF neutralizes 50 Hz MF-induced biological effect by using a signaling component as the test endpoint.     

A role for the magnetic field in the radiation-induced efflux of calcium ions from brain tissue in vitro

Two independent laboratories have demonstrated that electromagnetic radiation at specific frequencies can cause a change in the efflux of calcium ions from brain tissue in vitro. In a local geomagnetic field (LGF) at a density of 38 microTesla (microT), 15- and 45-Hz electromagnetic signals (40 Vp-p/m in air) have been shown to induce a change in the efflux of calcium ions from the exposed tissues, whereas 1- and 30-Hz signals do not. We now show that the effective 15-Hz signal can be rendered ineffective when the LGF is reduced to 19 microT with Helmholtz coils. In addition, the ineffective 30-Hz signal becomes effective when the LGF is changed to +/- 25.3 microT or to +/- 76 microT. These results demonstrate that the net intensity of the LGF is an important variable. The results appear to describe a resonance-like relationship in which the frequency of the electromagnetic field that can induce a change in efflux is proportional to a product of LGF density and an index, 2n + 1, where n = 0,1. These phenomenological findings may provide a basis for evaluating the apparent lack of reproducibility of biological effects caused by low-intensity extremely-low-frequency (ELF) electromagnetic signals. In future investigations of this phenomenon, the LGF vector should be explicitly described. If the underlying mechanism involves a general property of tissue, then research conducted in the ambient electromagnetic environment (50/60 Hz) may be subjected to unnoticed and uncontrolled influences, depending on the density of the LGF.     

Lack of alterations on meiotic chromosomes and morphological characteristics of male germ cells in mice exposed to a 60 Hz and 2.0 mT magnetic field

The effect of in vivo exposure of mice to a 60 Hz sinusoidal magnetic field (MF) at 2.0 mT on male germ cells was studied. The cytological endpoints measured included meiotic chromosome aberrations in spermatocytes and sperm morphology. Three independent experiments were carried out: (a) animals exposed for 72 h, (b) 10 days/8 h daily, and (c) 72 h exposure to MF plus 5 mg/kg of Mitomycin-C. No statistically significant differences indicative of MF effects were observed between MF exposed and control animals. In addition, an opposite effect between MF exposure and Mitomycin-C treatment in terms of chromosomal aberrations and sperm morphology was observed.     

Intracellular recording during magnetic field application to monitor neurotransmitter release events: methods and preliminary results

A method for simultaneous magnetic field application and intracellular recording is presented. A little used method for magnetic field application was exploited; the field generator consisted of a flat copper sheet through which current was passed to generate a magnetic field. The resultant magnetic field was relatively homogeneous, exhibiting a variation of +/- 5%. This compact, current-sheet field generator was mounted on the condensor of a microscope. The current induced in the intracellular electrode was reduced by injecting current equal and opposite to the induced current into the microelectrode. This step reduces the possibility of cellular effects and voltage artifacts due to the induced electrode current. The technique was used to conduct preliminary studies on the effects of extremely low frequency (ELF) linearly and circularly polarized magnetic fields (1.0 Gauss, 60 and 70 Hz) on miniature end plate potential (mepp) frequency (frequency of neurotransmitter release events) of rat flexor digitorum brevis muscle. The same synapse was utilized for both the sham-exposed control and the exposed experimental halves of an experiment. After 10 min of exposure to a 60-Hz linear field, mepp frequency was significantly increased by 12%, but exposure to a 60-Hz circular field did not significantly alter mepp frequency. Exposure to a 70-Hz linear field did not significantly change mepp frequency, but application of a 70-Hz circular field appears to decrease mepp frequency by 4%. These results indicate that both types of magnetic fields can alter mepp frequency, depending upon the frequency and configuration of the field.     

Addition of magnetic field capability to existing extremely-low-frequency electric field exposure systems

Magnetic field systems were added to existing electric field exposure apparatuses for exposing cell suspensions in vitro and small animals in vivo. Two horizontally oriented, rectangular coils, stacked one directly above the other, have opposite electric currents. This configuration minimizes leakage fields and allows sham- and field-exposure systems to be placed in the same room or incubator. For the in vitro system, copper plates formed the loop-pair, with up to 900 A supplied by a 180:1 transformer. Electric fields were supplied via electrodes at the ends of cell-culture tubes, eight of which can be accommodated by each exposure system. Two complete systems are situated in an incubator to allow simultaneous sham and field exposure up to 1 mT. For the in vivo system, four pairs of 0.8 x 2.7-m coils made of copper bus bar are employed. This arrangement is energized from the power grid via a 30:1 transformer; horizontal magnetic flux densities up to 1 mT can be generated. Pairs of electrode plates spaced 30.5 cm apart provide electric field exposure of up to 130 kV/m. Four systems with a capacity of 48 rats each are located in one room. For both the in vitro and in vivo systems, magnetic exposure fields are uniform to within +/- 2.5%, and sham levels are at least 2,500-fold lower than exposure levels. Potential confounding factors, such as heating and vibration, were examined and found to be minimal.     

Miniature-probe measurements of electric fields and currents induced by a 60-Hz magnetic field in rat and human models

A miniaturized probe was designed and built to provide detailed data on fields induced by a uniform 60-Hz magnetic field in homogeneous models of rat and human. The probe employed three silver wires twisted and potted in an 8-cm hypodermic needle. The exposed tips of the wires formed three sensing electrodes with a centered ground; highly sensitive voltage measurements were enabled by a lock-in amplifier. Tests were conducted in a 1-mT rms field that was uniform within +/- 5%. The models were made by casting 1.5% agar at 1-S/m conductivity into plastic-foam molds. The rat model was scaled 1:1 as an adult (22 cm length; mass about 640 g). The human model was scaled 1:4 as an adult (height = 46.5 cm; mass 1.4 kg). The probe was inserted into each model in several regions, and readings of induced fields were made under different exposure geometries. Maximal strengths of fields induced near the surface of the torso were as high as 120 microV/cm in the laterally exposed rat model. Data extrapolated from the quarter-scale human model revealed that an induced field as high as 700 microV/cm could occur at the torso of a frontally exposed human adult. An overall size-scale factor of about 5 appears to be appropriate for experimental exposures of rats that are intended to simulate currents induced in human beings by magnetic fields. The average strength of electric fields induced in the torso by a 1-mT magnetic field is comparable to that by a vertical electric-field at 60 kV/m and 28 kV/m, respectively, for the rat and human.     

Lack of an effect of static magnetic field on calcium efflux from isolated chick brains

45Ca2+ efflux from neonatal isolated chick brains was measured. The brains were exposed to uniform or nonuniform static magnetic fields. The field intensity ranged from 200-900 mT. The exposure took place during incubation and/or when efflux was being measured. No difference appeared in the 45Ca2+ efflux between controls and exposed brains.     

Increased resorptions in CBA mice exposed to low-frequency magnetic fields: An attempt to replicate earlier observations

This paper has two aims. First, it reports the findings of a study on the effects of low-frequency magnetic fields on reproduction. Second, it serves as an example of an attempt to replicate the results of an experimental study in an independent laboratory and discusses some of the problems of replication studies. To try to replicate the findings of a study reporting increased resorptions (fetal loss) in mice exposed to 20 kHz magnetic fields with sawtooth waveform and to study the possible effects of 50 Hz sinusoidal fields, pregnant mice were exposed to magnetic fields from day 0 to 18 of pregnancy, 24 h per day. The flux densities of the vertical magnetic fields were 15 μT (peak-to-peak) at 20 kHz and 13 or 130 μT (root mean square) at 50 Hz. Two strains of animals were used: CBA/S mice imported from the laboratory reporting the original observations, and a closely related strain CBA/Ca. The CBA/S mice were cleaned of pathogenic microbes and parasites before they were imported into our laboratory. The magnetic field exposures did not affect resorption rate in CBA/Ca mice. In CBA/S, the frequency of resorptions was higher in the exposed mice than in the control group. However, the increase was not significantly different from either the no-effect hypothesis or the results of the original study we were attempting to replicate. Differences between the two studies and difficulties in interpreting the results are discussed. It is concluded that the results tend more to support than argue against increased resorptions in CBA/S mice exposed to the 20 kHz magnetic field. The results demonstrate that animal strain is an important variable in bioelectromagnetics research: even closely related strains may show different responses to magnetic field exposure. Bioelectromagnetics 18:410–417, 1997. © 1997 Wiley-Liss, Inc.     

Action of extremely low frequency electric fields on the cytosolic calcium concentration of differentiated HL-60 cells: Nonactivated cells

The effect of sinusoidal electric fields on the cytosolic free [Ca²⁺]_i concentration in differentiated HL-60 cells was measured. The calcium concentration was measured in a fluorescence spectrometer using the fluorescence sample fluo-3. In the fluorescence spectrometer two samples can be measured simultaneously, one as the sham-exposed control and the other as the field-exposed sample. The effects of an external field, applied using two capacitor plates outside the cuvettes, and a field applied directly to the medium, using two platinum electrodes inside the cuvettes, were measured at selected frequencies between 0 and 100 Hz and field strengths from 1 to 2000 V_pp/m (external field) and from 0.1 to 1000 V_pp/m (in medium). No significant effects of the fields on the cytosolic free [Ca²⁺]_i concentration in HL-60 cells have been observed at the measured frequencies and field strengths. Bioelectromagnetics 19:32–40, 1998. © 1998 Wiley-Liss, Inc.     

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.     

A four coil exposure system (tetracoil) producing a highly uniform magnetic field

We propose a magnetic field exposure system (tetracoil) for in vitro and in vivo experiments, composed of two couples of circular coils satisfying a spherical constraint, whose characteristics are chosen in order to maximize the uniformity region of the magnetic field. Analytical calculations and computer simulations show that our system, as compared to the other most largely used magnetic field exposure systems, represents an optimal compromise in terms of field uniformity, accessibility for biological experiments, and ratio between overall dimension and uniformity region.     

No association between occupational exposure to ELF magnetic field and urinary 6-sulfatoximelatonin in workers

A suppression in melatonin secretion is one of the mechanisms proposed to explain the possible adverse effects of extremely low frequency magnetic fields (ELF-MF), but the results of research are inconclusive. This study investigated the effect of occupational ELF-MF exposure on 6-sulfatoximelatonin (6-OHMS). Exposure was monitored for three complete work shifts in 59 workers using personal exposure meters. Environmental exposure was also evaluated. Urinary 6-OHMS in morning samples, an indicator of night-time melatonin production, was measured. Urine was collected twice on Friday and the following Monday. Workers were classified according to ELF exposure as low exposed (0.2 microT): 6-OHMS did not differ between groups (P > .05) in either Friday or Monday urine samples. In addition, 6-OHMS was not related to exposure under multivariate analysis. The ratio between 6-OHMS in Monday versus Friday samples was also calculated to test the hypothesis of a possible variation in pineal function after 2 days, interruption of occupational ELF-MF exposure: again no exposure-related difference was observed. Our results do not support the hypothesis that occupational exposure to ELF-MF significantly influences melatonin secretion.     

Effect of extremely low frequency magnetic fields on calpain activation

The effects of low intensity, low frequency magnetic fields (MFs) on catalytic activity of the calcium dependent protease calpain was determined following the enzyme activation both in "in vitro" and "in vivo" conditions. We have observed that a 0.3 mT MF induces a significant increase in the requirement of the protease for this metal ion. This change is detectable at low [Ca(2+)] and disappears when the level of Ca(2+) is raised to saturating amounts. The observed effects are not due to transient MF(-) induced conformational changes occurring in calpain, but to direct effects of the MF on Ca(2+) ions, which become less available for the binding sites present in calpain. Altogether, these results indicate that exposure to low intensity, low frequency MFs alters the intracellular Ca(2+) "availability," thereby modifying the related cell response.     

Occupational magnetic field exposure and melatonin: interaction with light-at-night

The evidence of magnetic field (MF) effects on melatonin production in humans is limited and inconsistent. Part of the inconsistencies might be explained by findings suggesting interaction with light in pineal responses to MFs. To test this hypothesis, we reanalyzed data from a previously published study on 6-hydroxy melatonin sulfate (6-OHMS) excretion in women occupationally exposed to extremely low-frequency MFs. Based on questionnaire data on exposure to light-at-night (LAN), and measurement-based MF data, the 60 women were classified to four groups: no MF, no LAN; MF, no LAN; no MF, LAN; MF, LAN. The lowest excretion of 6-OHMS was observed in the group of women who were exposed to both MF and LAN, and the differences between the four groups were significant (P < .0001). The result is based on low numbers, but supports the hypothesis that daytime occupational exposure to MF enhances the effects of nighttime light exposure on melatonin production.     

Morphological anomalies in pollen tubes of Actinidia deliciosa (kiwi) exposed to 50 Hz magnetic field

The role of the pollen grain, with respect to the reproductive process of higher plants, is to deliver the spermatic cells to the embryo sac for egg fertilisation. Delivery occurs through the pollen tube, a self produced organ that is generated when the pollen grain reaches the stigma surface. The effect of magnetic fields on pollen tube growth was reported in a recent publication by Germanà et al. Pollen tube growth is an interesting candidate for the detailed study of the effects of electromagnetic fields on cytoplasmic structures and organelles. In this research Actinidia deliciosa (kiwifruit) pollen grains were germinated in the presence of an alternating magnetic field (50 Hz). Our results, although of preliminary nature, show that pollen tube growth is affected by magnetic fields. The analysis of the observed anomalies in the pollen tube appear to be the result of changes in the ionic charges within the pollen tube cytoplasm.     

50 Hz magnetic field effects on the performance of a spatial learning task by mice

Intense magnetic fields have been shown to affect memory-related behaviours of rodents. A series of experiments was performed to investigate further the effects of a 50 Hz magnetic field on the foraging behaviour of adult, male C57BL/6J mice performing a spatial learning task in an eight-arm radial maze. Exposure to vertical, sinusoidal magnetic fields between 7.5 μT and 7.5 mT for 45 min immediately before daily testing sessions caused transient decreases in performance that depended on the applied flux density. Exposure above a threshold of between 7.5 and 75 μT significantly increased the number of errors the animals made and reduced the rate of acquisition of the task without any effect on overall accuracy. However, the imposition of a 45-minute delay between exposure at 0.75 mT and behavioural testing resulted in the elimination of any deficit. Similarly, exposure to fields between 7.5 μT and 0.75 mT for 45 min each day for 4 days after training had no amnesic effects on the retention and subsequent performance of the task. Overall, these results provide additional evidence that 50 Hz magnetic fields may cause subtle changes in the processing of spatial information in mice. Although these effects appear dependent on field strength, even at high flux densities the field-induced deficits tend to be transient and reversible. Bioelectromagnetics 19:486–493, 1998. © 1998 Wiley-Liss, Inc.     

Measuring temporal variability in residential magnetic field exposures

Considerable interest has developed during the past ten years regarding the hypothesis that living organisms may respond to temporal variability in ELF magnetic fields to which they are exposed. Consequently, methods to measure various aspects of temporal variability are of interest. In this paper, five measures of temporal variability were examined: Arithmetic means (D(mean)) and rms values (D(rms)) of the first differences (i.e., absolute value of the difference between consecutive measurements) of magnetic field recordings; "standardized" forms of D(rms), denoted RCMS, obtained by dividing D(rms) by the standard deviations of the magnetic field data; and mean (F(mean)) and rms (F(rms)) values of fractional first differences. Theoretical investigations showed that D(mean) and D(rms) are virtually unaffected by long-term systematic trends (changes) in exposure. These measures thus provide rather specific measures of short-term temporal variability. This was also true to a lesser extent for F(mean) and F(rms). In contrast, the RCMS metric was affected by both short-term and long-term exposure variabilities. The metrics were also investigated using a data set consisting of twice-repeated two-calendar-day recordings of bedroom magnetic fields and personal exposures of 203 women residing in the western portion of Washington State. The predominant source of short-term temporal variability in magnetic field exposures arose from the movement of subjects through spatially varying magnetic fields. Spearman correlations between TWA bedroom magnetic fields or TWA personal exposures and five measures of temporal variability were relatively low. Weak to moderate levels of correlation were observed between temporal variability measured during two different sessions separated in time by 3 or 6 months. We conclude that first difference and fractional difference metrics provide specific and fairly independent measures of short-term temporal variability. The RCMS metric does not provide an easily interpreted measure of short-term or long-term temporal variability. This last result raises uncertainties about the interpretation of published studies that use the RCMS metric.     

Absence of genotoxicity in human blood cells exposed to 50 Hz magnetic fields as assessed by comet assay, chromosome aberration, micronucleus, and sister chromatid exchange analyses

In the past, epidemiological studies indicated a possible correlation between the exposure to ELF fields and cancer. Public concern over possible hazards associated with exposure to extremely low frequency magnetic fields (ELFMFs) stimulated an increased scientific research effort. More recent research and laboratory studies, however, have not been able to definitively confirm the correlation suggested by epidemiological studies. The aim of this study was to evaluate the effects of 50 Hz magnetic fields in human blood cells exposed in vitro, using several methodological approaches for the detection of genotoxicity. Whole blood samples obtained from five donors were exposed for 2 h to 50 Hz, 1 mT uniform magnetic field generated by a Helmholtz coil system. Comet assay, sister chromatid exchanges (SCE), chromosome aberrations (CA), and micronucleus (MN) tests were used to assess DNA damage, one hallmark of malignant cell transformation. The effects of a combined exposure with X-rays were also evaluated. Results obtained do not show any significant difference between ELFMFs exposed and unexposed samples. Moreover, no synergistic effect with ionizing radiation has been observed. A slight but significant decrease of cell proliferation was evident in ELFMFs treated samples and samples subjected to the combined exposure.     

Alternative magnetic field exposure metrics: relationship to TWA, appliance use, and demographic characteristics of children in a leukemia survival study.

The ongoing Childhood Leukemia Survival Study is examining the possible association between magnetic field exposure and survival of children with newly diagnosed acute lymphocytic leukemia (ALL). We report the results of the first year 24 h personal magnetic field monitoring for 356 US and Canadian children by time weighted average TWA and alternative exposure metrics. The mean TWA of 0.12 microT was similar to earlier personal exposure studies involving children. A high correlation was found between 24 h TWA and alternative metrics: 12 h day TWA, 12 night TWA, geometric mean, 95th percentile value, percentage time over 0.2 and 0.3 microT, and an estimate of field stability (Constant Field Metric). Two measures of field intermittency, rate of change metric (RCM) and standardized rate of change metric (RCMS), were not highly correlated with TWA. The strongest predictor of TWA was location of residence, with highest TWAs associated with urban areas. Residence in an apartment, lower paternal educational level, and residential mobility were also associated with higher TWAs. There were no significant differences in the appliance use patterns of children with higher TWA values. Children with the highest field intermittency (high RCM) were more likely to sit within 3 feet of a video game attached to the TV. Our results suggest that 24 h TWA is a representative metric for certain patterns of exposure, but is not highly correlated with two metrics that estimate field intermittency.