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].     

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.     

Correlation of magnetic AC field on cardiac myocyte Ca2+ transients at different magnetic DC levels

The purpose of this study was to determine the effect of extremely low frequency and weak magnetic fields (WMF) on cardiac myocyte Ca²⁺ transients, and to explore the involvement of potassium channels under the WMF effect. In addition, we aimed to find a physical explanation for the effect of WMF on cardiac myocyte Ca²⁺ transients. Indo‐1 loaded cells, which were exposed to a WMF at 16 Hz and 40 nT, demonstrated a 75 ± 4% reduction in cytosolic Ca²⁺ transients versus control. Treatment with the K_ATP channel blocker, glibenclamide, followed by WMF at 16 Hz exposure, blocked the reduction in cytosolic calcium transients while treatment with pinacidil, a K_ATP channel opener, or chromanol 293B, a selective potassium channel blocker of the delayed rectifier K⁺ channels, did not inhibit the effect. Based on these finding and the ion cyclotron resonance frequency theory, we further investigated the effect of WMF by changing the direct current (DC) magnetic field (B₀). When operating different DC magnetic fields we showed that the WMF value changed correspondingly: for B₀ = 44.5 µT, the effect was observed at 17.05 Hz; for B₀ = 46.5 µT, the effect was observed at 18.15 Hz; and for B₀ = 49 µT the effect was observed at 19.1 Hz. We can conclude that the effect of WMF on Ca²⁺ transients depends on the DC magnetic field level. Bioelectromagnetics 33:634–640, 2012. © 2012 Wiley Periodicals, Inc.     

A Lorentz model for weak magnetic field bioeffects: Part I—Thermal noise is an essential component of AC/DC effects on bound ion trajectory

We have previously employed the Lorentz–Langevin model to describe the effects of weak exogenous magnetic fields via the classical Lorentz force on a charged ion bound in a harmonic oscillator potential, in the presence of thermal noise forces. Previous analyses predicted that µT‐range fields give rise to a rotation of the oscillator orientation at the Larmor frequency and bioeffects were based upon the assumption that the classical trajectory of the bound charge itself could modulate a biochemical process. Here, it is shown that the thermal component of the motion follows the Larmor trajectory. The results show that the Larmor frequency is independent of the thermal noise strength, and the motion retains the form of a coherent oscillator throughout the binding lifetime, rather than devolving into a random walk. Thermal equilibration results in a continual increase in the vibrational amplitude of the rotating oscillator towards the steady‐state amplitude, but does not affect the Larmor orbit. Thus, thermal noise contributes to, rather than inhibits, the effect of the magnetic field upon reactivity. Expressions are derived for the ensemble average of position and the velocity of the thermal component of the oscillator motion. The projection of position and velocity onto a Cartesian axis measures the nonuniformity of the Larmor trajectory and is illustrated for AC and combined AC/DC magnetic fields, suggesting a means of interpreting resonance phenomena. It is noted that the specific location and height of resonances are dependent upon binding lifetime and initial AC phase. Bioelectromagnetics 30:462–475, 2009. © 2009 Wiley‐Liss, Inc.     

Comment: analyses of models of ion actions under the combined action of AC and DC magnetic fields

I show that the interaction of weak DC and ELF magnetic fields with contained ions cannot generate significant biological effects through changing the character of the ion orbits.     

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.     

On the cyclotron resonance mechanism for magnetic field effects on transmembrane ion conductivity

The cyclotron resonance model, recently proposed to account for physiological response to weak environmental magnetic fields, is shown to violate the laws of classical mechanics. Further, it is argued that the ubiquitous presence of dynamic friction in fluid media precludes significant magnetic effects on membrane ion transport.     

Calcium binding to metallochromic dyes and calmodulin in the presence of combined, AC-DC magnetic fields.

The possibility that weak, ac and dc magnetic fields in combination may affect binding equilibria of calcium-ions (Ca2+) was investigated with two metallochromic dyes as calcium-binding molecules: murexide and arsenazo III. Calcium-dye equilibria were followed by measuring solution absorbances with a fiber-optic spectrophotometer. A Ca(2+)-arsenazo solution was also used indirectly to monitor the binding of Ca2+ to calmodulin. Parallel, ac and dc magnetic fields were applied to each preparation. The ac magnetic field was held constant during each of a series of experiments at a frequency in the range between 50 and 120 Hz (sine wave) or at 50 pps (square wave) and at an rms flux density in the range between 65 and 156 microT. The dc magnetic field was then varied from 0 to 299 microT at 1.3 microT increments. The magnetic fields did not measurably affect equilibria in the binding of metallochromic dyes or calmodulin to Ca2+.     

Frequency and amplitude windows in the combined action of DC and low frequency AC magnetic fields on ion thermal motion in a macromolecule: theoretical analysis

We solved the differential equation describing combined action of DC and AC magnetic fields on thermal motion of ions in a biological macromolecule. The solution showed the occurrence of a new set of resonant peaks for ion oscillations under the influence of magnetic fields. After establishment of steady ion oscillations in the macromolecule interior that is well shielded from the action of small particles of the medium surrounding this molecule, the change in energy of ion thermal motion could be sufficient to alter the conformation state of the macromolecule. On this basis, a diversity of biological phenomena can be explained, including the appearance of the known "frequency" and "amplitude" windows, without any resort to the ideas of participation of cyclotron or parametric resonances in these effects.     

Using planar lipid-bilayers to study plant ion channels

Ion channels are found in most plant membranes. They catalyse the rapid passive uniport of particular ions with varying selectivity. Planar lipid-bilayer (PLB) techniques have been developed to study the electrical activities of single ion channels in well-defined lipid and aqueous environments. They greatly facilitate both the biophysical and biochemical characterisation of ion channels and complement both conventional impaling electrode and membrane-patch voltage-clamping (patch-clamping) electrophysiological techniques applied in vivo. Bilayers can be formed across the end of patch-clamp pipettes or across apertures in specifically designed chambers. Ion channels in native membranes and purified, genetically altered or synthetic ion channels, proteins and peptides can all be studied in PLBs. The main applications of PLBs are (1) to study ion channels in membranes inaccessible to patch-clamp electrodes, (2) to provide a functional assay system during channel-protein purification and (3) to investigate the relationship between the molecular structure of ion channels and their conductance properties. In the present article we describe the techniques available for reconstitution and analysis of ion channels in PLBs and discuss how the PLB technique has been, and may be, useful to the study of plant ion channels.     

Porous membranes for reconstitution of ion channels

Functional biological synthetic composite (BSC) membranes were made using phospholipids, biological membrane proteins and permeable synthetic supports or membranes. Lipid bilayers were formed on porous polycarbonate (PC), polyethylene terephthalate (PETE) and poly (l-lactic acid) (PLLA) membranes and in 10-100 μm laser-drilled pores in a 96-well plastic plate as measured by increased resistance or decreased currents. Bilayers in 50 μm and smaller pores were stable for up to 4 h as measured by resistance changes or a current after gramicidin D reconstitution. Biological membrane transport reconstitution was then carried out. Using vesicles containing Kv1.5 K⁺ channels, K⁺ currents and decreased resistance were measured across bilayers in 50 μm pores in the plastic plate and PLLA membranes, respectively, which were inhibited by compound B, a Kv1.5 K⁺ channel inhibitor. Functional reconstitution of Kv1.5 K⁺ channels was successful. Incorporation of membrane proteins in functional form in stable permeable membrane-supported lipid bilayers is a simple technology to create BSC membranes that mimic biological function which is readily adaptable for high throughput screening.     

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.     

Weak extremely-low-frequency magnetic fields and regeneration in the planarian Dugesia tigrina

Extremely-low-frequency (ELF), low-intensity magnetic fields have been shown to influence cell signaling processes in a variety of systems, both in vivo and in vitro. Similar effects have been demonstrated for nervous system development and neurite outgrowth. We report that regeneration in planaria, which incorporates many of these processes, is also affected by ELF magnetic fields. The rate of cephalic regeneration, reflected by the mean regeneration time (MRT), for planaria populations regenerating under continuous exposure to combined DC (78.4 μT) and AC (60.0 Hz at 10.0 μT _peak) magnetic fields applied in parallel was found to be significantly delayed (P ? 0.001) by 48 ± 1 h relative to two different types of control populations (MRT ? 140 ± 12 h). One control population was exposed to only the AC component of this field combination, while the other experienced only the ambient geomagnetic field. All measurements were conducted in a low-gradient, low-noise magnetics laboratory under well-maintained temperature conditions. This delay in regeneration was shown to be dependent on the planaria having a fixed orientation with respect to the magnetic field vectors. Results also indicate that this orientation-dependent transduction process does not result from Faraday induction but is consistent with a Ca²⁺ cyclotron resonance mechanism. Data interpretation also permits the tentative conclusion that the effect results from an inhibition of events at an early stage in the regeneration process before the onset of proliferation and differentiation. © 1995 Wiley-Liss, Inc.     

Effect of magnetic field exposure on calcium channel currents using patch clamp technique

Calcium influxes through the membrane of PC-12D cells were measured under exposure to DC biased AC magnetic fields in resonant conditions of the ion cyclotron and the ion parametric resonance hypotheses and compared with influxes in cells without exposure to the magnetic field. After cancellation of the geomagnetic field, the cells were exposed to the horizontal fields generated by a current sheet, a planar sheet of conductor which generated a satisfactorily homogeneous horizontal magnetic field on the stage of a microscope without hindering treatment of a cell under observation. At or near any resonant conditions, no change in calcium influx could be detected under standard patch clamp conditions.     

Exposure to ELF magnetic field tuned to Zn inhibits growth of cancer cells

The effects of ELF alternating magnetic fields tuned to Zn(2+) on the growth of cancer cells with different status of p53 were investigated using a cell proliferation assay. Human cancer cells HeLa (cervix cancer, p53(+/+)), Saos-2 and Saos-2-His-273 (osteosarcoma, p53(-/-) and p53 His-273 mutant, respectively), H1299tTA and H1299tTA-His175 (lung carcinoma, p53(-/-) and p53 His-175 mutant), and normal human fibroblasts VH-10 (p53(+/+)) were used. Exposure parameters were calculated for the first harmonic of Zn(2+) based either on the magnetic parametric resonance (MPR) model of Lednev or the ion parametric resonance (IPR) model of Blanchard and Blackman. ELF exposure was for 72 and 96 h. The vertical alternating field was 20 Hz at amplitudes of either 38.7 or 77.4 microT (peaks, IPR or MPR, respectively). The vertical static magnetic field was 43 microT, and the horizontal static magnetic field was zeroed. Treatments of cells with PRIMA-1 and gamma-rays were used as positive controls. Growth inhibition was observed in cells after exposure to ELF at 38.7 microT. Inhibition of HeLa, VH-10, and Saos-2-His-273 cells was statistically significant, P=0.0003, 0.02, and 0.006, respectively. No consistent ELF effects following exposure 77.4 microT were seen. PRIMA-1 inhibited the growth of all cell lines with the strongest effect in mutant p53-carrying cell line H1299tTA-His175. The effects of gamma-rays were relatively weak, suggesting that the cell proliferation assay under conditions employed in this study is not very sensitive to apoptosis. In conclusion, ELF under conditions of exposure tuned to Zn(2+) according to the IPR model inhibited the growth of cancer and normal cells. No clear relationship of the observed growth inhibition to p53 status was found. Further experiments, using complementary techniques, are required to test whether p53 reactivation by ELF is feasible.     

A few remarks on 'combined action of DC and AC magnetic fields on ion motion in a macromolecule'

Zhadin and Barnes [2005:26:323-330] concluded that they solved the differential equation describing combined action of DC and AC magnetic fields on thermal motion of ions in a biological macromolecule and, as a result, a diversity of biological phenomena could be explained. It is shown here that biological phenomena cannot be explained based on this model. Adair [2006:27:332-334] gave several arguments for the statement that the interaction of weak magnetic fields with ions trapped in protein cavities cannot produce detectable biological effects through changing the character of the ion orbits. The arguments are analyzed here and some are shown to be questionable or unjustified. We stress the difference between the conclusion made by Adair and that stated in this article.     

Waveform magnetic field survey in Russian DC and Swiss AC powered trains: a basis for biologically relevant exposure assessment

Recent epidemiological studies suggest a link between transport magnetic fields (MF) and certain adverse health effects. We performed measurements in workplaces of engineers on Russian DC and Swiss AC powered (16.67 Hz) electric trains using a computer based waveform capture system with a 200 Hz sampling rate. MF in DC and AC trains show complex combinations of static and varying components. The most probable levels of quasistatic MF (0.001-0.03 Hz) were in the range 40 microT. Maximum levels of 120 microT were found in DC powered locomotives. These levels are much higher than the geomagnetic field at the site of measurements. MF encountered both in DC and AC powered rail systems showed irregular temporal variability in frequency composition and amplitude characteristics across the whole frequency range studied (0-50 Hz); however, more than 90% of the magnetic field power was concentrated in frequencies 相似文献   

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.     

Combined action of static and alternating magnetic fields on ion motion in a macromolecule: Theoretical aspects

This is an attempt to solve the energetic problem of the primary detection of weak parallel static (DC) and alternating (AC) extremely low frequency (ELF) magnetic fields. We studied the equations of motion for an ion situated inside a macromolecule under the influence of these fields. The main concern is with the magnetic field influence on thermal motion of the ion in the macromolecule. The resonance effects are revealed at discrete frequencies of the ion thermal oscillations determined by the DC field magnitude and the AC field frequency. These phenomena result from the Larmor precession of the ion thermal motion. When the DC field or, to a greater extent, the combined DC and AC fields with the specific frequencies are turned on or cut off, changes occur in the energy of the ion thermal motion. If, inside the macromolecule, the ion is sufficiently protected against immediate impacts of particles of the medium surrounding the macromolecule, these changes may be enough to trigger alteration in the quantum state of the macromolecule. Bioelectromagnetics 19:279–292, 1998. © 1998 Wiley-Liss, Inc.