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.     

Sinusoidal 60 Hz electromagnetic fields failed to induce changes in protein synthesis in Escherichia coli

Escherichia coli JM83 {F^? ara Δ(lac-proAB) rpsL [?80dΔ(lacZ)M15]} in midlog growth phase at 30 °C were exposed to 60 Hz sinusoidal magnetic field of 3 mT of nonuniform diverging flux, inducing a nonuniform electric field with a maximum intensity of 32 μV/cm using an inductor coil. Exposed and unexposed control cells were maintained at 30.8 ± 0.1 °C and 30.5 ± 0.1 °C, respectively. Quadruplicate samples of exposed and unexposed E. coli cells were simultaneously radiolabeled with ³⁵S-L-methionine at 10 min intervals over 2 hr. Radiochemical incorporation into proteins was analyzed via liquid scintillation counting and by denaturing 12.5% polyacrylamide gel electrophoresis. The results showed that E. coli exposed to a 60 Hz magnetic field of 3 mT exhibited no qualitative or quantitative changes in protein synthesis compared to unexposed cells. Thus small prokaryotic cells (less than 2 μm × 0.5 μm) under constant-temperature conditions do not alter their protein synthesis following exposure to 60 Hz magnetic fields at levels at 3 mT. © 1994 Wiley-Liss, Inc.     

Cancer promotion in a mouse-skin model by a 60-Hz magnetic field: I. Experimental design and exposure system.

The rationale for selection of an animal model, the experimental design, and the design and evaluation of an exposure system used in studies of 60-Hz magnetic fields are described. The studies were conceived to assay development of cancer and immune responsiveness in mice exposed to magnetic fields. The exposure system utilized a quadrupole-coil configuration to minimize stray magnetic fields. Four square-wound coil provided a uniform field within a volume occupied by 16 animal cages. The magnetic field had a mean flux density of 2 mT that varied less than +/- 10% within the volume occupied by animals' cages. The flux density decreased to less than 0.1 microT at a distance of 2 m from the coils. In each exposure system 32 animals could be housed in plastic cages.     

ELF Magnetic Field Exposure System for In Vitro Studies Based on Lee-Whiting Coils

In order to run a series of in vitro studies on the effect of extremely low-frequency magnetic fields on cell cultures, developing and characterizing an appropriate exposure system is required. The present design is based on a two-shielded Lee-Whiting coils system. The circular design was chosen because its axial symmetry allowed for both reducing simulation unknowns and measurement points during the characterization, and additionally made the machining of the parts easier. The system can generate magnetic flux densities (B fields) up to 1 mT root-mean-square amplitude (rms) with no active cooling system in the incubator, and up to 3 mT_rms with it. The double-wrapped windings with twisted pairs allow for the use of each set of coils either as exposure or control with no detectable parasitic B field in the control. The artifacts have also been analyzed; the B field in the center of the sham control chamber is about 1 µT_rms for a maximum of 3 mT_rms in the exposure chamber, the parasitic incident electric fields are less than 1 V/m, the temperature difference between sham and exposure chamber is less than or equal to 0.2 °C, and the typical vibration difference between sham and exposure is less than 0.1 m/s². © 2020 Bioelectromagnetics Society.     

Contribution of a 300 kHz alternating magnetic field on magnetic hyperthermia treatment of HepG2 cells

We investigated the relative contributions of temperature and a 300 kHz alternating magnetic field (AMF) on magnetic hyperthermia treatment (MHT). Our system consisted of an induction coil, which generated AMF by electric current flow, and a newly developed, temperature‐controlled circulating water‐jacketed glass bottle placed inside the coil. The AMF generator operated at a frequency of 300 kHz with variable field strength ranging from 0 to 11 mT. Four treatment conditions were employed: (A) control (37 °C, 0 mT), (B) AMF exposure (37 °C, 11 mT), (C) hyperthermia (46 °C, 0 mT), and (D) hyperthermia plus AMF exposure (46 °C, 11 mT) for 30 min. Cell viability and apoptotic death rate were estimated. The relative contributions or interactions of hyperthermia (46 °C) and AMF (11 mT) on MHT were evaluated using 2 × 2 factorial experiment analysis. Group A was statistically different (P < 0.05) from each of the other treatments. The observed effects on both cell viability and apoptotic cell death were influenced by temperature (97.36% and 92.15%, respectively), AMF (1.78% and 4.99%, respectively), and the interactions between temperature and AMF (0.25% and 2.36%, respectively). Thus, the effect of hyperthermia was significant. Also, AMF exposure itself might play a role in MHT, although these observations were made in vitro. These findings suggest a possible presence of an AMF effect during clinical magnetic hyperthermia. Bioelectromagnetics 34:95–103, 2013. © 2012 Wiley Periodicals, Inc.     

Mechanical vibration in “double-wound” magnetic field exposure coils

It has been suggested that “double-wound” (bifilar) exposure coils are capable of producing a sham environment in which hum and vibration will be “similar” to the field-exposed condition. We found by direct measurements in a bifilar coil system that vibration amplitude in sham and exposed conditions differed by a factor of 50 when our test system was driven at B = 10 mT. We also found that the normal laboratory environment can include vibrations of an intensity similar to that produced by the exposure system, although not necessarily of similar spectral distribution. © 1996 Wiley-Liss, Inc.     

A system for simultaneous exposure of small animals to 60-Hz electric and magnetic fields

Equipment designed for simultaneous exposure of rodents to 60-Hz electric and magnetic fields is described. Three identical systems were constructed, each capable of continuous exposure of 256 rats or 640 mice to a nominal electric field at less than 50 kV/m, and to horizontal and vertical magnetic fields at less than 1 mT. Design features, construction details, and results of various tests of the systems are described. Tests were made: of phase relations between electric and magnetic fields; of uniformity of electric and magnetic fields; of changes across time in electric-field intensity as a result of animals' soiling of cages and various washing routines; of resistance of bedding material during humid and dry conditions; and of acoustic noise due to background, to field-generation equipment, and to air conditioning equipment. The results demonstrated that fields were effectively generated but that significant and troublesome changes in electric-field intensity occurred because of cage-soiling. However, when cages were frequently cleaned, field intensities were consistent from one exposure to another.     

In vitro exposure apparatus for ELF magnetic fields

For in vitro studies on the effect of extremely low frequency (ELF) magnetic field exposures in different laboratories, a programmable, high precision exposure system enabling blinded exposures has been developed and fully characterized. It is based on two shielded 4 coil systems that fit inside a commercial incubator. The volume of uniform B field exposure with 1% field tolerance is 50% larger compared to a Merrit 4 coil system with the same coil volume. The uncertainties for the applied magnetic fields have been specified to be less than 4%. The computer controlled apparatus allows signal waveforms that are composed of several harmonics, blind protocols, monitoring of exposure and environmental conditions and the application of B fields up to 3.6 mT root-mean-square amplitude. Sources of artifacts have been characterized: sham isolation >43 dB, parasitic incident E fields <1 V/m, no recognizable temperature differences in the media for exposure or sham state, and vibrations of the mechanically decoupled dish holder <0.1 m/s(2) (= 0.01 g), which is only twice the sham acceleration background level produced by the incubator and fan vibrations.     

Electromagnetic field treatment of nerve crush injury in a rat model: effect of signal configuration on functional recovery

Electromagnetic fields (EMFs) have been demonstrated to enhance mammalian peripheral nerve regeneration in vitro and in vivo. Using an EMF signal shown to enhance neurite outgrowth in vitro, we tested this field in vivo using three different amplitudes. The rat sciatic nerve was crushed. Whole body exposure was performed for 4 h/day for 5 days in a 96-turn solenoid coil controlled by a signal generator and power amplifier. The induced electric field at the target tissue consisted of a bipolar rectangular pulse, having 1 and 0.3 ms durations in each polarity, respectively. Pulse repetition rate was 2 per second. By varying the current, the coils produced fields consisting of sham (no current) and peak magnetic fields of 0.03 mT, 0.3 mT, and 3 mT, corresponding to peak induced electric fields of 1, 10, and 100 microV/cm, respectively, at the tissue target. Walking function was assessed over 43 days using video recording and measurement of the 1-5 toe-spread, using an imaging program. Comparing injured to uninjured hind limbs, mean responses were evaluated using a linear mixed statistical model. There was no difference found in recovery of the toe-spread function between any EMF treatments compared to sham.     

Design and fabrication of well confined uniform magnetic field exposure systems

Exposure systems that provide good magnetic field uniformity, minimum stray fields, and minimal heating, vibration, and hum, as well as capability for true sham exposure in which current flows in the coils, are needed to determine rigorously the biological effects of weak magnetic fields. Designs based on acrylic polymer coil support structures and twisted pair bifilary coil windings were employed to fabricate several different systems for the exposure of laboratory animals and cell cultures to magnetic fields. These systems exhibit excellent performance characteristics in terms of exposure field uniformity, stray field containment, and exposure field cancellation in the sham exposure mode. A custom-written computer program was used to determine the best arrangement for coils with regard to field uniformity in the exposure volume and stray field containment. For in vivo exposures, modules were made up of four Merritt four-coil sets, built into a single structure and positioned to form an octapole with fields directed in the horizontal plane. For in vitro applications, two different coil configurations were selected to produce the vertical fields required. A quadrupole system, comprising modules consisting of two Merritt four-coil sets arranged side by side to limit stray fields, was built as a prototype. In the second configuration, one Merritt four-coil set was positioned inside the other to form a concentric coil set. In both in vitro systems, exposure chambers were connected to remote commercial incubators in order to reduce ambient magnetic fields in the exposure volume. An active field cancellation circuit was developed for reducing ambient AC magnetic fields in the in vitro sham exposure chamber, when necessary. These design and fabrication approaches provide systems that offer uniform field exposures and excellent stray field containment when needed and are portable, washable, and relatively inexpensive. © 1994 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

50-Hz magnetic field exposure system for small animals

The design, construction, and results of evaluation of an animal-exposure system for the study of biological effects of extremely low frequency (ELF) magnetic fields are described. The system uses a square coil arrangement based on a modification of the Helmholtz coil. Due to the cubic configuration of this exposure system, horizontal and vertical magnetic fields as high as 0.3 mT can be generated. Circularly polarized magnetic fields can also be generated by changing the current and phase difference between two sets of coils. Tests were made for uniformity of the magnetic field, stray fields, sham-exposure ratio of stray field, changes of temperature and humidity, light intensity and distribution inside the animal-housing space, and noise due to air-conditioning equipment. Variation of the magnetic field was less than 2% inside the animal housing. The stray-field level inside the sham-exposure system is less than 2% of experimental exposure levels. The system can be used for simultaneous exposure of 48 rats (2 to a cage) or 96 mice (4 to a cage). © 1993 Wiley-Liss. Inc.     

Low stray ELF magnetic field exposure system for in vitro study

An exposure facility for wide application to cell exposure to an ELF (extremely low frequency) magnetic field was developed. It is suitable for conducting experiments under a high-intensity, variable-frequency magnetic field, on the biological effects of the ELF magnetic field in an in vitro study. The exposure system consists of Merritt's 4-square coil as a basic component to generate the required magnetic field intensity of 10 mT at 50 Hz with spatial field uniformity less than +/-3% in a 400 mm cube. Concentric compensation coils are adopted to eliminate the effects of stray fields on sham (control) samples in the vicinity of the exposure system. The uniformity of the magnetic field in the exposure coil, the increase in the power supply capacity due to the existence of compensation coils, and the stray field estimation were investigated carefully. After fabricating the system, performance tests were carried out and all the characteristics were found to be satisfactory. In addition, the ideal configuration for a concentric coil system was proposed.     

A Study of the Effects of Flux Density and Frequency of Pulsed Electromagnetic Field on Neurite Outgrowth in PC12 Cells

The aim of this study was to investigate the influence of pulsed electromagnetic fields with various flux densities and frequencies on neurite outgrowth in PC12 rat pheochromocytoma cells. We have studied the percentage of neurite-bearing cells, average length of neurites and directivity of neurite outgrowth in PC12 cells cultured for 96 hours in the presence of nerve growth factor (NGF). PC12 cells were exposed to 50 Hz pulsed electromagnetic fields with a flux density of 1.37 mT, 0.19 mT and 0.016 mT respectively. The field was generated through a Helmholtz coil pair housed in one incubator and the control samples were placed in another identical incubator. It was found that exposure to both a relatively high flux density (1.37 mT) and a medium flux density (0.19 mT) inhibited the percentage of neurite-bearing cells and promoted neurite length significantly. Exposure to high flux density (1.37 mT) also resulted in nearly 20% enhancement of neurite directivity along the field direction. However, exposure to low flux density field (0.016 mT) had no detectable effect on neurite outgrowth. We also studied the effect of frequency at the constant flux density of 1.37 mT. In the range from 1 ∼ 100 Hz, only 50 and 70 Hz pulse frequencies had significant effects on neurite outgrowth. Our study has shown that neurite outgrowth in PC12 cells is sensitive to flux density and frequency of pulsed electromagnetic field.     

Influence of 50 Hz electric and magnetic fields on the pulse rate of human heart

This investigation studied the effects of 50 Hz electric and magnetic fields on the pulse rate of the human heart. The ECG (electrocardiograms) of 41 male volunteers were recorded with a Holter recorder. Twenty-six subjects were measured in and outside real fields, and 15 subjects were measured in and outside “sham” fields. The blood pressure and EEG (electroencephalogram) were also measured, but this article presents only the results of ECG recordings. The measurements took 3 h. The subjects were first sitting for 1 h outside the fields, then 1 h in the real or “sham” fields, and then, again, 1 h outside the fields. The electric field strength varied from 3.5 to 4.3 kV/m and the magnetic flux density from 1.4 to 6.6 μT. An analysis of the ECG recordings showed that the subjects' pulse rates were the same in and outside the fields. No response occurred when the subjects were exposed to real or “sham” fields. © 1994 Wiley-Liss, Inc.     

Real‐time measurement of cytosolic free calcium concentration in DEM‐treated HL‐60 cells during static magnetic field exposure and activation by ATP

This study investigated whether glutathione depletion affected the sensitivity of HL‐60 cells to static magnetic fields. The effect of Diethylmaleate (DEM) on static magnetic field induced changes in cytosolic free calcium concentration ([Ca²⁺]_c) was examined. Cells were loaded with a fluorescent dye and exposed to a uniform static magnetic field at a strength of 0 mT (sham) or 100 mT. [Ca²⁺]_c was monitored during field and sham exposure using a ratiometric fluorescence spectroscopy system. Cells were activated by the addition of ATP. Metrics extracted from the [Ca²⁺]_c time series included: average [Ca²⁺]_c during the Pre‐Field and Field Conditions, peak [Ca²⁺]_c following ATP activation and the full width at half maximum (FWHM) of the peak ATP response. Comparison of each calcium metric between the sham and 100 mT experiments revealed the following results: average [Ca²⁺]_c measured during the Field condition was 53 ± 2 nM and 58 ± 2 nM for sham and 100 mT groups, respectively. Average FWHM was 51 ± 3 s and 54 ± 3 s for sham and 100 mT groups, respectively. An effect of experimental order on the peak [Ca²⁺]_c response to ATP in sham/sham experiments complicated the statistical analysis and did not allow pooling of the first and second order experiments. No statistically significant difference between the sham and 100 mT groups was observed for any of the calcium metrics. These data suggested that manipulation of free radical buffering capacity in HL‐60 cells did not affect the sensitivity of the cells to a 100 mT static magnetic field. Bioelectromagnetics 30:213–221, 2009. © 2008 Wiley‐Liss, Inc.     

Rats are not aversive when exposed to 60-Hz magnetic fields at 3.03 mT.

Thirty-two male rats were tested in two replicates of an experiment to determine whether body currents induced by 60-Hz magnetic fields might lead to avoidance behavior comparable to that which results from exposure to strong 60-Hz electric fields. The test apparatus was a two-compartment Plexiglas shuttlebox enclosed in a sound-attenuating plywood chamber, which in turn was encompassed by two copper bus bars that, when energized, served as a source of 60-Hz magnetic fields. Location of the rat, and traverse activity in the shuttlebox were monitored by nine infra-red photo detectors equally spaced along the length of the apparatus. Rats were divided into 2 groups: 1 group of rats (n = 8 per group per replicate) was sham exposed while rats in the other group (n = 8 per group per replicate) were exposed to a 3.03 mT (30.3 G), 60-Hz magnetic field whenever they traversed to or were located on the side (L or R) predetermined as the exposed side. To control artifact incident to side preference, the side exposed (L or R) was alternated over the exposed rats. Each rat was tested individually in a 1-h session. A 2-factor ANOVA (exposed vs. control, replicate 1 vs. replicate 2) failed to reveal any significant effects due to either factor or to an interaction between factors. These data demonstrate that rats do not avoid exposure to 60-Hz magnetic fields at a flux density of 3.03 mT and further imply that the avoidance by rats of high level 60-Hz electric fields is mediated by something other than the internal body currents induced by the exposure.     

Effects of exposure to a circularly polarized 50-Hz magnetic field on plasma and pineal melatonin levels in rats

We sought to determine whether a 6-week exposure to a 50-Hz rotating magnetic field influences melatonin synthesis by 11–18 week-old Wistar-King male rats. Rats were exposed continuously to a rotating magnetic field at 1, 5, 50, or 250 μT (spatial vector rms) for 6 weeks, except for twice-weekly breaks of about 2 h for cleaning of cages and feeding. The rats were housed in exposure and sham-exposure facilities, which were located in the same room, under a 12:12 light-dark photoperiod (lights on at 06:00 h). The room was constantly illuminated by 4 small, dim red lights (< 0.07 lux in dark period). Levels of plasma and pineal gland melatonin were determined by radioimmunoassay. A significant decrease of melatonin was observed between the control group and groups exposed to a magnetic field at a flux density in excess of l μT during the night time, but no statistical differences were found among the exposed groups. These results indicate that subchronic exposure of albino rats to a 50-Hz rotating magnetic field influences melatonin production and secretion by the pineal gland. © 1993 Wiley-Liss, Inc.     

Complex magnetic field exposure system for in vitro experiments at intermediate frequencies

In occupational environments, an increasing number of electromagnetic sources emitting complex magnetic field waveforms in the range of intermediate frequencies is present, requiring an accurate exposure risk assessment with both in vitro and in vivo experiments. In this article, an in vitro exposure system able to generate complex magnetic flux density B‐fields, reproducing signals from actual intermediate frequency sources such as magnetic resonance imaging (MRI) scanners, for instance, is developed and validated. The system consists of a magnetic field generation system and an exposure apparatus realized with a couple of square coils. A wide homogeneity (99.9%) volume of 210 × 210 × 110 mm³ was obtained within the coils, with the possibility of simultaneous exposure of a large number of standard Petri dishes. The system is able to process any numerical input sequence through a filtering technique aimed at compensating the coils' impedance effect. The B‐field, measured in proximity to a 1.5 T MRI bore during a typical examination, was excellently reproduced (cross‐correlation index of 0.99). Thus, it confirms the ability of the proposed setup to accurately simulate complex waveforms in the intermediate frequency band. Suitable field levels were also attained. Moreover, a dosimetry index based on the weighted‐peak method was evaluated considering the induced E‐field on a Petri dish exposed to the reproduced complex B‐field. The weighted‐peak index was equal to 0.028 for the induced E‐field, indicating an exposure level compliant with the basic restrictions of the International Commission on Non‐Ionizing Radiation Protection. Bioelectromagnetics 34:211–219, 2013. © 2012 Wiley Periodicals, Inc.     

Intracellular calcium activity in isolated bovine adrenal chromaffin cells in the presence and absence of 60 Hz magnetic fields

This study examined whether 60 Hz magnetic field (MF) exposure alters intracellular calcium levels ([Ca(2+)](i)) in isolated bovine adrenal chromaffin cells, a classic model of neural responses. [Ca(2+)](i) was monitored by fluorescence video imaging of cells loaded with the calcium indicator fluo-4 during exposures to magnetic flux densities of 0.01, 0.1, 1.0, 1.4, or 2.0 mT. MFs generated by Helmholtz coils constructed from bifilar wire allowed both 60 Hz field and sham exposures. Following a 5 min monitoring period to establish baseline patterns, cells were subjected for 10 min to a 60 Hz MF, sham field or no field. Reference calcium responses and assessment of cell excitability were obtained by the sequential addition of the nicotinic cholinergic receptor agonist dimethylphenylpiperazinium (DMPP) and a depolarizing concentration of KCl. Throughout an 8 day culture period, cells exhibited spontaneous fluctuations in [Ca(2+)](i). Comparisons of the number of cells exhibiting transients, the number and types of calcium transients, as well as the time during monitoring when transients occurred showed no significant differences between MF exposed cells and either sham exposed or nonexposed cells. With respect to the percentage of cells responding to DMPP, differences between 1 and 2 mT exposed cells and both nonexposed and sham exposed cells reached statistical significance during the first day in culture. No statistically significant differences were observed for responses to KCl. In summary, our data indicate that [Ca(2+)](i) in chromaffin cells is unaffected by the specific 60 Hz MF intensities used in this study. On the other hand, plasma membrane nicotinic receptors may be affected in a manner that is important for ligand-receptor interactions.     

System for the exposure of cell suspensions to power-frequency electric fields

A system is described that uses an oscillating magnetic field to produce power-frequency electric fields with strengths in excess of those produced in an animal or human standing under a high-voltage electric-power transmission line. In contrast to other types of exposure systems capable of generating fields of this size, no electrodes are placed in the conducting growth media: the possibility of electrode contamination of the exposed suspension is thereby eliminated. Electric fields in the range 0.02–3.5 V/m can be produced in a cell culture with total harmonic distortions less than 1.5%. The magnetic field used to produce electric fields for exposure is largely confined within a closed ferromagnetic circuit, and experimental and control cells are exposed to leakage magnetic flux densities less than 5 μT. The temperatures of the experimental and control cell suspensions are held fixed within ±0.1°C by a water bath. Special chambers were developed to hold cell cultures during exposure and sham exposure. Chinese hamster ovary (CHO) cells incubated in these chambers grew for at least 48 h and had population doubling times of 16–17 h, approximately the same as for CHO cells grown under standard cell-culture conditions.