Toxicity and SOS response to ELF magnetic field and nalidixic acid in E. coli cells

Extremely low frequency (ELF) magnetic fields have previously been shown to affect conformation of chromatin and cell proliferation. Possible genotoxic and carcinogenic effects of ELF have also been discussed and tested. In this study, we analyzed the effect of ELF on chromatin conformation in E. coli GE499 cells by the anomalous viscosity time dependence (AVTD) technique. Possible genotoxic ELF effects at the specific combination of static and ELF magnetic fields, that has been proven to have effects on chromatin conformation, were investigated by clonogenic assay, cell growth kinetics, and analysis of SOS-response using inducible recA-lacZ fusion and the β-galactosidase assay. Genotoxic agent nalidixic acid (NAL) was used as positive control and in combination with ELF. Nalidixic acid at 3-30μg/ml decreased the AVTD peaks and induced cytotoxic effect. In contrast to NAL, ELF increased AVTD, stimulated cell growth, and increased cloning efficiency. These effects depended on frequency within the frequency range of 7-11Hz. While NAL induced SOS response, ELF exposure did not induce the recA-lacZ fusion. Exposure to ELF did not modify the genotoxic effects of NAL either. All together, the data show that ELF, under specific conditions of exposure, acted as nontoxic but cell growth stimulating agent.     

Cell density dependent response of E. Coli cells to weak ELF magnetic fields

The effects of weak magnetic fields of extremely low frequency (ELF) on E. coli K12 AB1157 cells were studied by the method of anomalous viscosity time dependencies (AVTD). E. coli cells at different densities within a range of 5 × 10⁵–10⁹ cell/ml were exposed to ELF (sinusoidal, 30 μT peak, 15 min) at a frequency of 9 Hz. A transient effect with maximum 40–120 min after exposure was observed. Kinetics of the per-cell-normalised ELF effects fitted well to a Gaussian distribution for all densities during exposure. A maximum value of these kinetics and a time for this maximum were strongly dependent on the cell density during exposure. These data suggest a cell-to-cell interaction during response to ELF. Both dependencies had three regions close to a plateau within the ranges of 3 × 10⁵ − 2 × 10⁷ cell/ml, 4 × 10⁷ − 2 × 10⁸ cell/ml and 4 × 10⁸–10⁹ cell/ml and two rather sharp transitions between these plateaus. The effect reached a maximum value at a density of 4 × 10⁸ cell/ml. Practically no effect was observed at the lowest density of 3 × 10⁵ cell/ml. The data suggested that the ELF effect was mainly caused by a secondary rather than a primary reaction. The filtrates from exposed cells neither induced significant AVTD changes in unexposed cells nor increased the ELF effect when were added to cells before exposure. The data did not provide evidence for significant contribution of stable chemical messengers, but some unstable compounds such as radicals could be involved in the mechanism of cell-to-cell interaction during response to ELF. The results obtained were also in accordance with a model based on an re-emission of secondary photons during resonance fluorescence. Bioelectromagnetics 19:300–309, 1998. © 1998 Wiley-Liss, Inc.     

Factors confounding cytosolic calcium measurements in Jurkat E6.1 cells during exposure to ELF magnetic fields

Reported changes in the cytosolic calcium concentration ([Ca2+](c)) as a result of exposure to extremely low frequency (ELF) magnetic fields (MF) have been equivocal. In this study, we examine the possibility that some of these differences are attributable to variability associated with the cell cycle, pH of the suspension medium, and response to a calcium agonist. We used a custom designed spectrofluorimeter to measure [Ca2+](c) in Indo 1-AM loaded Jurkat E6.1 cells suspended in conditioned RPMI 1640 medium containing 10% fetal bovine serum. Four exposures were examined: zero static MF (Null), 60 Hz 100 microT(peak) sinusoidal MF (AC), 78 microT static MF (DC), and the combination of the 60 Hz and the 78 microT static MF (AD + DC). A significant decrease in normalized [Ca2+](c) values between 375-495 s for the DC and AC + DC groups was found in comparison to the Null group. However, statistical analysis indicated that cell cycle and quality of the alpha-CD3 monoclonal antibody response were significant covariates, while pH was not a significant covariate. When the effect of these covariates was taken into account, all exposure groups were significantly different from the control. Our results suggest that ELF MF effects may not be seen unless correction is made for biological variability of each cell preparation with respect to cell cycle and [Ca2+](c) response to antigen stimulation.     

Increased antibiotic resistance of E. coli exposed to static magnetic fields

The present study demonstrates that exposure of bacteria to medium strength static magnetic fields can significantly alter antibiotic sensitivity. Cultures of Escherichia coli were exposed to fields produced by permanent magnets. Samples of bacterial cultures continuously growing in the presence and in the absence of static magnetic fields were left untreated or were treated with an antibiotic and measured at 45 min intervals for cell growth and survival. It was found that exposure of E. coli to the static fields significantly increased antibiotic resistance. Bioelectromagnetics 22:129-137, 2001. Published 2001 Wiley-Liss, Inc.     

The effect of nalidixic acid on expression from related E. coli promoters

The effect of the DNA gyrase inhibitor, nalidixic acid, on expression from E. coli promoters was studied using the pKO-1, galactokinase expression vector system. Expression from a series of related hybrid promoters, tet promoter variants and the trp promoter flanked by oligonucleotide blocks was measured after incubation with nalidixic acid. Expression from the pBR322 tet promoter and tet promoter mutants within the -10 region was reduced after the drug treatment. The lacUV5, trp, and tettrp promoters were essentially unaffected while the trplac and the trptet promoters were stimulated. Studies of the trp promoter flanked by upstream or downstream oligonucleotide blocks revealed similar responses to the trp promoter parent control plasmids.     

Effect of ELF magnetic fields on protein synthesis in Escherichia coli K12

Escherichia coli K12 was used as a model system to determine whether ELF magnetic fields (MFs) are a general stress factor. The cells were exposed to ELF MFs (5-100 Hz) at a maximum intensity of 14 mT r. m.s. for circularly polarized MFs and 10 mT r.m.s. for vertically polarized MFs. The response of the cells to the MFs was estimated from the change in protein synthesis by using 2D PAGE. Approximately 1,000 proteins were separated on the 2D gels. The stress-responsive proteins such as CH10, DNAK, CH60, RECA, USPA, K6P1 and SODM were identified from the SWISS-2DPAGE database on the 2D gels. These proteins respond to most stress factors, including temperature change, chemical compounds, heavy metals, and nutrients. When the bacterial cells were exposed to each MF at 5-100 Hz under aerobic conditions (6.5 h) or at 50 Hz under anaerobic conditions (16 h) at the maximum intensity (7.8 to 14 mT r.m.s.), no reproducible changes were observed in the 2D gels. Changes in protein synthesis were detected by 2D PAGE with exposure to heat shock (50 degrees C for 30 min) or under anaerobic conditions (no bubbling for 16 h). Increases in the levels of synthesis of the stress proteins were observed in heat-shocked cells (CH60, CH10, HTPG, DNAK, HSLV, IBPA and some unidentified proteins) and in cells grown under anaerobic conditions (DNAK, PFLB, RECA, USPA and many unidentified proteins). These results suggest that 2D PAGE is sufficient to detect cell responses to environmental stress. The high-intensity ELF MFs (14 mT at power frequency) did not act as a general stress factor.     

ELF magnetic fields in a city environment

Some epidemiological studies indicate an association between extremely low frequency electromagnetic field (ELF-EMF) exposure and cancer risks. These studies have mainly taken residential and occupational exposure into consideration. Outdoor environments are often considered as low level areas, but in this paper we show that this is not true in a city environment. We have mapped the ELF magnetic flux densities along certain stretches of sidewalk in central G?teborg City, Sweden. About 50% of the investigated street length shows flux densities of the same order of magnitude (0.2 microT and above) as those associated with increased risks of cancer in epidemiological studies. We conclude that the outdoor exposures in a city environment also should be considered in exposure assessments and risk evaluations. These elevated flux densities are probably due to stray currents. We also found strong magnetic flux densities (> 1.0 microT) close to ordinary distribution pillars, power substations, shoplifting alarms, and other electrical devices.     

Behavioral changes in adult rats exposed to ELF magnetic fields

In two experiments, male rats that had been exposed to an ELF (0.5 Hz), 3–50 gauss rotating magnetic field (RMF) for 21–30 days, displayed significantly (p < 0.05) greater ambulatory behavior (activity) than the control group in an open field test, when removed from the RMF. In a third experiment rats were exposed to a different RMF apparatus (3–30 gauss), and tested in a different open field for longer duration. Again the RMF-exposed group displayed greater activity (p < 0.10) than the control group.

---

Zusammenfassung In zwei Untersuchungen zeigten männliche Ratten,die einem ELF (0,5 Hz) 3–50 Gauss rotierenden magnetischen Feld (RMF) für 21–30 Tage ausgesetzt wurden, in einem Open-Field Test signifikant (p < 0,05) mehr Bewegungsaktivität als die Kontrollgruppe,nachdem sie von dem RMF entfernt wurden. In einer dritten Untersuchung wurden Ratten in einem anderen RMF Gerät gehalten und in einem anderen Open-Field für längere Zeit getestet. Wieder zeigte die Gruppe, die dem RMF ausgesetzt war, mehr Bewegungsaktivität (p < 0,10) als die Kontrollgruppe.

---

Resume Lors de deux essais, on a placé des rats mâles dans un champ magnétique tournant (CMT) de 3 à 50 gauss et des très basse fréquence (FTB) (0,5 Hz). Après 21 à 30 jours de ce traitement, on les a remis dans des conditions normales. Leur activité (marche) était plus élevée que celle d'un groupe de contrôle et cela de façon significative (p < 0,05). Dans un troisième essai, d'autres rats furent placés dans un autre appareil (3 à 30 gauss) et examinés ensuite dans un autre champ ouvert sur une plus longue période. Le groupe traité a déployé dans ce cas également une plus grande activité qu'un groupe de contrôle (p < 0,10).

     

Electric fields and surface charges induced by ELF magnetic fields

A method is described for evaluating electric fields induced by ELF magnetic fields into electrically inhomogeneous, low-conductivity (less than 5 S/m) structures. It is applied to cylinders and spheres, and numerical results are given for electrical properties that are representative of some tissues, or of cells embedded either in saline solution or a tissue matrix. Surface currents on spherical cell boundaries are estimated and compared with thermal noise due to ion motion.     

Fluctuation analysis of mutations to nalidixic acid resistance in Escherichia coli.

Mutations of Escherichia coli from sensitivity to nalidixic acid resistance were studied by fluctuation analysis. The mutant distributions in replicate cultures were not significantly affected either by the age of the carbon-starved preculture used for inocula or by the inoculum size. The data from 23 fluctuation tests (48 cultures each) were pooled. The mean number of mutations per culture was estimated to be 0.71 from the fraction of cultures without mutants or 0.74 and 0.77 by maximum-likelihood estimation based on the two models under consideration. When the pooled data were compared with the theoretical expectations, the fits were unsatisfactory (P < 0.005). The lack of fit was caused mainly by too high a frequency of cultures with between 17 and 32 mutants and too high a frequency of cultures with more than 128 mutants. Possible reasons for the lack of fit and its implications with respect to estimation of mutation rates from fluctuation tests are discussed.     

Penetration of nalidixic acid into Escherichia coli K-12 cells

Transport of nalidixic acid (NAL) into Escherichia coli cells subjected to osmotic shock, permeabilised with toluene or treated with DNP, CCCP or EDTA, was studied. It was found that osmotic shock and protonophores do not inhibit the transport of [3H]NAL, however, the transport of [3H]DAP and [3H]glucose is reduced. EDTA and toluene enhance penetration of [3H]NAL. This effect is, however, abolished in the presence of Mg++ ions. It is suggested that NAL penetrates into the cell by simple or facilitated diffusion and that the outer membrane of E. coli is the penetration barrier for the drug.     

Facility for chronic exposure of rats to ELF magnetic fields

The facility consists of a 12 × 11.5 × 2.4 m high room containing six sets of exposure apparatus and the other equipment necessary to maintain a pathogen-free system. The apparatus sets produced 5 mT (rms), 0.5 mT, or a sham exposure. The apparatus was arranged in the room to minimize the fringing field of the 5 mT set at the sham position. Each set was 3.85 × 1.80 × 0.66 m in outside dimension, containing 24 cages in the magnetically homogeneous region. The apparatus was designed using Harvey's figure-eight-configuration and generated a horizontal sinusoidal alternating field. In order to save electric power, the coil of the apparatus constituted a 50 Hz LC resonance circuit with a condensor bank to which electric power was supplied to compensate losses. Magnetic flux density was kept constant by controlling the coil current. Although mild steel was used in the skeleton of the building, the fringing flux at the sham was as low as 0.1 to 1 μT. Stainless steel was used for ventilating ducts, racks for the cages, cage covers, feeder baskets, and watering nozzles. The homogeneity of the field was measured to be ± 10% in the animal residence area, and food and water consumption was found to be unaffected by the field. At 5 mT, the coil current was 370 A, and the hollow coil was cooled by a stream of 20°C water to prevent both heat and dew on the coil surface. Vibration and acoustic noise was prevented by fiber reinforced plastic framework of the coil. High harmonic distortion was not observed at the output terminal of the coil driver. The facility has operated without trouble for 2 years. © 1993 Wiley-Liss, Inc.     

Multigeneration exposure test of Drosophila melanogaster to ELF magnetic fields

Mutations, other than dominant lethals, were accumulated on wild type second chromosomes (+) of Drosophila melanogaster during exposure to 50 Hz sinusoidal alternating magnetic fields of 0.5 or 5 mT (rms) for 40 generations by the Curly/Plum(Cy/Pm) accumulation method. We maintained, for 40 generations under continuous exposure, each (+) chromosome as a heterozygote with (Cy) chromosome. Viability of the (+) chromosome was tested by sib-mating of (Cy/+) male and (Cy/+) female in a culture every 10th generation to obtain the homozygote. Viability indices, defined as twice the ratio of number of (+/+) flies to that of (Cy/+) flies plus 1 in the progeny of the test mating, also were calculated, which equaled 1.00 at the starting point. For the control and 0.5 and 5 mT exposed groups, percent frequencies of recessive lethal lines, defined as a line with (+/+) flies less than 0.3% in the test mating, were, respectively, 1.9, 0.9, and 2.9% (10th), 9.0, 4.9, and 9.5% (20th), 30.3, 22.9, and 30.4% (30th), and 39.9, 32.4, and 43.3% (40th generation). For the control and 0.5 and 5 mT groups, average viability indices, excluding lethals and markedly deleterious, were, respectively, 0.778, 0.796, and 0.752 (20th), 0.704, 0.698, and 0.694 (30th), and 0.669, 0.678, and 0.595 (40th generation). Their decreasing rates were 0.0054, 0.0059, and 0.0078 per generation. No significant difference was detected among the exposure levels in either the recessive lethal mutation frequency or the viability index. Bioelectromagnetics 19:335–340, 1998. © 1998 Wiley-Liss, Inc.     

Exposure of welders and other metal workers to ELF magnetic fields

This study assessed exposure to extremely low frequency (ELF) magnetic fields of welders and other metal workers and compared exposure from different welding processes. Exposure to ELF magnetic fields was measured for 50 workers selected from a nationwide cohort of metal workers and 15 nonrandomly selected full-time welders in a shipyard. The measurements were carried out with personal exposure meters during 3 days of work for the metal workers and 1 day of work for the shipyard welders. To record a large dynamic range of ELF magnetic field values, the measurements were carried out with “high/low” pairs of personal exposure meters. Additional measurements of static magnetic fields at fixed positions close to welding installations were done with a Hall-effect fluxmeter. The total time of measurement was 1273 hours. The metal workers reported welding activity for 5.8% of the time, and the median of the work-period mean exposure to ELF magnetic fields was 0.18 μT. DC metal inert or active gas welding (MIG/MAG) was used 80% of the time for welding, and AC manual metal arc welding (MMA) was used 10% of the time. The shipyard welders reported welding activity for 56% of the time, and the median and maximum of the workday mean exposure to ELF magnetic fields was 4.70 and 27.5 μT, respectively. For full-shift welders the average workday mean was 21.2 μT for MMA welders and 2.3 μT for MIG/MAG welders. The average exposure during the effective time of welding was estimated to be 65 μT for the MMA welding process and 7 μT for the MIG/MAG welding process. The time of exposure above 1 μT was found to be a useful measure of the effective time of welding. Large differences in exposure to ELF magnetic fields were found between different groups of welders, depending on the welding process and effective time of welding. MMA (AC) welding caused roughly 10 times higher exposure to ELF magnetic fields compared with MIG/MAG (DC) welding. The measurements of static fields suggest that the combined exposure to static and ELF fields of MIG/MAG (DC) welders and the exposure to ELF fields of MMA (AC) welders are roughly of the same level. Bioelectromagnetics 18:470–477, 1997. © 1997 Wiley-Liss, Inc.     

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.     

Vertical circularly polarized ELF magnetic fields and induced electric fields in culture media

Some properties of induced electric fields in cell culture media produced by vertical circularly polarized magnetic fields are examined. The described geometry is not advantageous for determining effects that may be attributable to induced electric fields or currents. Bioelectromagnetics 18:524–526, 1997. Published 1997 Wiley-Liss, Inc.     

Frequency-dependent effects of ELF magnetic field on chromatin conformation in Escherichia coli cells and human lymphocytes.

The effects of magnetic fields of extremely low frequency (ELF, 21 microT r.m.s.) on cells of different Escherichia coli K12 strains and human lymphocytes were studied by the method of anomalous viscosity time dependence (AVTD). Within the frequency range of 6-24 Hz, two resonance-type frequency windows with maximal effects at 9 Hz and 16 Hz were observed in response of GE499 strain. Only one frequency window with maximum effect at 8.5 Hz was found for GE500 cells. These data along with previously obtained for two other E. coli strains, AB1157 and EMG2, indicate that frequency windows are dependent on genotype of cells exposed to ELF. Resonance-type effects of ELF were also observed in human lymphocytes in frequency windows around 8 and 58 Hz. These ELF effects differed significantly between studied donors, but were well reproducible in independent experiments with lymphocytes from the same donors. The frequency windows in response of E. coli strains and human lymphocytes to ELF significantly overlapped suggesting that the same targets may be involved in this response. We compared the frequency windows with predictions based on the ion cyclotron resonance (ICR) model and the magnetic parametric resonance model. These models predicted effects of ELF magnetic fields at the 'cyclotron' frequencies of some ions of biological relevance. According to the ICR model, ELF effects should be also observed at harmonics of cyclotron frequencies and, contrary, parametric resonance model predicted effects at subharmonics. While we observed coincidence of each experimental resonance frequency with predictions of one of these two models, all experimentally defined effective frequency windows were in good agreement with relatively narrow frequency ranges of both harmonics and subharmonics for natural isotopes of Na, K, Ca, Mg, and Zn ions. The experimental data support idea that both harmonics and subharmonics of several biologically important ions are involved in frequency-dependent ELF effects in cells of different types.     

In vitro exposure parameters with linearly and circularly polarized ELF magnetic fields.

A comparison is made of induced current densities, electric fields, and rates of energy deposition during in vitro studies with linearly and circularly polarized, extremely low frequency magnetic fields for a cylindrical volume of culture medium.     

ELF magnetic fields in electric and gasoline‐powered vehicles

We conducted a pilot study to assess magnetic field levels in electric compared to gasoline‐powered vehicles, and established a methodology that would provide valid data for further assessments. The sample consisted of 14 vehicles, all manufactured between January 2000 and April 2009; 6 were gasoline‐powered vehicles and 8 were electric vehicles of various types. Of the eight models available, three were represented by a gasoline‐powered vehicle and at least one electric vehicle, enabling intra‐model comparisons. Vehicles were driven over a 16.3 km test route. Each vehicle was equipped with six EMDEX Lite broadband meters with a 40–1,000 Hz bandwidth programmed to sample every 4 s. Standard statistical testing was based on the fact that the autocorrelation statistic damped quickly with time. For seven electric cars, the geometric mean (GM) of all measurements (N = 18,318) was 0.095 µT with a geometric standard deviation (GSD) of 2.66, compared to 0.051 µT (N = 9,301; GSD = 2.11) for four gasoline‐powered cars (P < 0.0001). Using the data from a previous exposure assessment of residential exposure in eight geographic regions in the United States as a basis for comparison (N = 218), the broadband magnetic fields in electric vehicles covered the same range as personal exposure levels recorded in that study. All fields measured in all vehicles were much less than the exposure limits published by the International Commission on Non‐Ionizing Radiation Protection (ICNIRP) and the Institute of Electrical and Electronics Engineers (IEEE). Future studies should include larger sample sizes representative of a greater cross‐section of electric‐type vehicles. Bioelectromagnetics 34:156–161, 2013. © 2012 Wiley Periodicals, Inc.