Dielectric measurements on live biological materials under magnetic resonance conditions

In the course of work on the interactions of electric and magnetic fields with both living and dead biological materials, it was noticed that certain published dielectrophoretic yield curves for biological cells showed unexplained deviations in the region of 2 kHz. Dielectrophoretic measurements made at frequencies and magnetic fields which satisfied the nuclear magnetic resonance conditions showed sharply resonant features. Dielectric measurements showed small, but sharp, resonances most easily seen in the dielectric loss curves which had a bandwidth of the order of one Hertz and presented at the frequencies which satisfied the magnetic resonance conditions for the ambient magnetic field. Resonances were found corresponding to the frequencies for electron spin resonance and nuclear magnetic resonance for¹H,³¹P,²³Na,³⁷Cl and³⁹K. The onset of these resonances occurs at the value of the steady magnetic field strength so that one quantum of magnetic flux (2.07×10^?15wb) would link a single biological cell or pair of cells, approximately 1 G (100μT) in the case of a 5-μm yeast cell. The effects of these magnetic resonance conditions on the mean generation time ofE. coli and on the reaction of the enzyme lysozyme with the substrateM. lysodeikticus cells are also shown.     

Nuclear magnetic resonance studies of amino acids and proteins. Rotational correlation times of proteins by deuterium nuclear magnetic resonance spectroscopy

We show that measurement of the spin-lattice (T1) and spin-spin (T2) relaxation times (or line widths) of irrotationally bound 2H nuclei in macromolecules undergoing isotropic rotational motion outside of the extreme narrowing limit (i.e., for the case omega 02 tau R2 much greater than 1) permits determination of both the rotational correlation time (tau R) of the macromolecule and the electric quadrupole coupling constant (e2qQ/h) of the 2H label. The technique has the advantage over 13C nuclear magnetic resonance (NMR) that no assumptions about bond lengths (which appear to the sixth power in 13C relaxation studies) or relaxation mechanisms need to be made, since relaxation will always be quadrupolar, even for aromatic residues at high field. Asymmetry parameter (eta) uncertainties are shown to cause negligible effects on tau R determinations, and in any case it is shown that both e2qQ/h and eta may readily be determined in separate solid-state experiments. By way of example, we report 2H NMR results on aqueous lysozyme (EC 3.2.1.17) at 5.2 and 8.5 T (corresponding to 2H-resonance frequencies of 34 and 55 MHz). Interpretation of the results in terms of the isotropic rigid-rotor model yields e2qQ/h values of approximately equal to 170 or approximately equal to 190 kHz, respectively, for the imidazolium and free-base forms of [epsilon 1-2H] His-15 lysozyme in solution, in excellent agreement with e2qQ/h values of approximately 167 and approximately 190 kHz obtained for the free amino acids in the solid state. In principle, the method may in suitable cases permit comparison between the dynamic structures of proteins in solution and in the crystalline solid state.     

Analysis of electric and magnetic fields leaking from induction heaters

Results are presented of an investigation on electric and magnetic fields leaking from inductive (magnetic) heaters that are used for thermal processing of high-power electron tubes and lasers in an industrial plant. Measurements of electric and magnetic fields were done using both commercially available and laboratory-developed instrumentation. Isotropic H-field sensors were developed to allow quantitative evaluation of high-intensity magnetic fields. Ten induction heaters with nominal A.C. power ranging from 2.5 kW to 15 kW and operating at frequencies between 300 kHz and 790 kHz were surveyed. Electric field strengths up to 8 kV/m and magnetic field strengths up to 20 A/m were measured.     

Comparison of coupling of humans to electric and magnetic fields with frequencies between 100 Hz and 100 kHz

Recent laboratory and epidemiological results have stimulated interest in the hypothesis that human beings may exhibit biological responses to magnetic and/or electric field transients with frequencies in the range between 100 Hz and 100 kHz. Much can be learned about the response of a system to a transient stimulation by understanding its response to sinusoidal disturbances over the entire frequency range of interest. Thus, the main effort of this paper was to compare the strengths of the electric fields induced in homogeneous ellipsoidal models by uniform 100 Hz through 100 kHz electric and magnetic fields. Over this frequency range, external electric fields of about 25–2000 V/m (depending primarily on the orientation of the body relative to the field) are required to induce electric fields inside models of adults and children that are similar in strength to those induced by an external 1 μT magnetic field. Additional analysis indicates that electric fields induced by uniform external electric and magnetic fields and by the nonuniform electric and magnetic fields produced by idealized point sources will not differ by more than a factor of two until the sources are brought close to the body. Published data on electric and magnetic field transients in residential environments indicate that, for most field orientations, the magnetic component will induce stronger electric fields inside adults and children than the electric component. This conclusion is also true for the currents induced in humans by typical levels of 60 Hz electric and magnetic fields in U.S. residences. Bioelectromagnetics 18:67–76, 1997. © 1997 Wiley-Liss, Inc.     

Experimental investigation on neural cell survival after dielectrophoretic trapping

Negative dielectrophoretic forces can effectively be used to trap cortical rat neurons. The creation of dielectrophoretic forces requires electric fields of high non-uniformity. High electric field strengths, however, can cause excessive membrane potentials by which cells may unrecoverably be changed or it may lead to cell death. In a previous study it was found that cells trapped at 3 Vtt/14 MHz did not change morphologically as compared to cells that were not exposed to the electric field. This study investigates the viability of fetal cortical rat neurons after being trapped by negative dielectrophoretic forces at frequencies up to 1 MHz. A planar quadrupole micro-electrode structure was used for the creation of a non-uniform electric field. The sinusoidal input signal was varied in amplitude (3 and 5 Vtt) and frequency (10 kHz-1 MHz). The results presented in this paper show that the viability of dielectrophoretically trapped postnatal cortical rat cells was greatly frequency dependent. To preserve viability frequencies above 100 kHz (at 3 Vtt) or 1 MHz (5 Vtt) must be used.     

Development and evaluation of intermediate frequency magnetic field exposure system for studies of in vitro biological effects

We have developed an intermediate frequency (IF) magnetic field exposure system for in vitro studies. Since there are no previous studies on exposure to heating-frequency magnetic fields generated from an induction heating (IH) cook top, there is a strong need for such an exposure system and for biological studies of IF magnetic fields. This system mainly consists of a magnetic-field-generating coil housed inside an incubator, inside which cultured cells can be exposed to magnetic field. Two systems were prepared to allow the experiment to be conducted in a double-blind manner. The level of the generated magnetic field was set to 532 microT rms in the exposure space, 23 kHz, 80 times the value in the International Commission on Non-ionizing Radiation Protection (ICNIRP) guidelines, with a spatial field uniformity better than 3.8%. The waveforms were nearly sinusoidal. It was also confirmed that the parasitic electric field was 157 V/m rms and the induced electric field was 1.9 V/m rms. The temperature was maintained at 36.5 +/- 0.5 degrees C for 2 h. Furthermore, leaked magnetic flux density was 0.7 microT rms or lower at extremely low frequency (ELF) and IF in the stopped system when the other system was being operated, and the environmental magnetic flux density was 0.1 microT rms or lower at the center of the coils. As a result, it was confirmed that this system could be successfully used to evaluate the biological effects of exposure to IF magnetic fields.     

A scheme for incorporating DC magnetic fields into epidemiological studies of EMF exposure

Experimental data on calcium-ion release in chicken brain tissue suggest that biological effects of electric and magnetic fields (EMFs) are concentrated near certain “active combinations” of DC magnetic field strength and “effective” AC magnetic field frequencies. We hypothesize that active AC/DC combinations may exist and suggest that epidemiologic data, coupled with DC magnetic field measurements, may be used to identify critical exposure conditions. An empirical model is used to calculate these multiple active combinations at any given DC magnetic field strength and to define a rating system that incorporates the proximity of AC magnetic field frequencies generated by electric power lines to the new, computed effective frequencies. Such an exposure score may be useful in investigating correlations of EMF exposure with disease incidence. For 60 Hz and 50 Hz, the highest EMF exposure scores occurred at DC field strengths of 506 mG and 422 mG, respectively. The exposure score contains a factor which may be adjusted to reflect the importance of harmonics of the AC magnetic field as well as of the fundamental frequency. Using this factor, we consider two important special cases consistent with chick brain data: 1) we consider active pairs associated with all detectable harmonics (up to 660 Hz) without regard to relative intensity of the harmonics, and 2) we use the relative intensities of the AC field frequencies to adjust their contribution to the exposure score. © 1993 Wiley-Liss. Inc.     

Evaluation of Radiofrequency and Extremely Low‐Frequency Field Levels at Children's Playground Sites in Greece From 2013 to 2018

From 2013 to 2018, in‐situ measurements of radiofrequency (RF) electromagnetic fields (EMF) and extremely low‐frequency (ELF) electric and magnetic fields in 317 existing and under‐construction children's playground facilities, in 16 municipalities all over Greece, were carried out by the Greek Atomic Energy Commission (EEAE). These measurements were conducted following legislative framework established in 2009, which requires that compliance with the established exposure limits for EMFs should be verified in playground areas. The results are presented by the value of the electric field (E) and exposure ratio (Λ) for the RF EMF, as well as the value of the electric field (E) and magnetic flux density (B) for the ELF electric and magnetic fields. Statistical analysis tools were applied on measurement data and conclusions have been made, taking into consideration: (i) environment type (urban/suburban), and (ii) vicinity to any transmitting installations. Measurement results correspond to the typical EMF background levels for each environment type. Concerning the environment type, RF EMF, and ELF electric/magnetic field measurements reveal no differentiation between urban and suburban environments. Bioelectromagnetics. 2019;40:602–605. © 2019 Bioelectromagnetics Society.     

RF Exposure During Use of Electrosurgical Units

Electrosurgical units (ESUs) commonly used in operating suites employ radiofrequency (RF) energy for cutting and coagulation, and operate at different frequencies in the range 0.3–5 MHz. Around the electrode and cables, electric and magnetic fields at similar frequencies will be generated, and the surgeon using the ESU will therefore be exposed to these electromagnetic fields. In this study we have measured the levels of RF fields near the lead wires of two electrosurgical units, BARD 3000 operating at a fixed frequency of 0.5 MHz, and ERBE ICC 350 with a frequency range from 0.3 to 1 MHz. Electric fields were measured at distances from 5–30 cm from the lead wire. Measurements were done with the ESU both cutting and coagulating, and power levels ranging from 10–100 W. The magnetic field outside the lead wire was calculated from the measured current through the leads using standard theory. Using those measurements as a base, the calculated local exposure of the surgeon's hand was estimated to exceed 15 kV/m for the electric field and the corresponding value for the magnetic field was 16 µT. These calculations exceed the suggested international reference levels at 0.5 MHz (610 V/m and 4 µT, respectively).     

New model for the avian magnetic compass

It is proposed that the avian magnetic compass depends on the angle between the horizontal component B(h) of the geomagnetic field (GMF) and E(r), the radial electric field distribution generated by gamma-oscillations within the optic tectum (TeO). We hypothesize that the orientation of the brain relative to B(h) is perceived as a set of electric field ion cyclotron resonance (ICR) frequencies that are distributed in spatially recognizeable regions within the TeO. For typical GMF intensities, the expected ICR frequencies fall within the 20-50 Hz range of gamma-oscillation frequencies observed during visual stimulation. The model builds on the fact that the superficial lamina of the TeO receive signals from the retina that spatially map the visual field. The ICR frequencies are recruited from the local wide-band gamma-oscillations and are superposed on the tectum for interpretation along with other sensory data. As a first approximation, our analysis is restricted to the medial horizontal plane of the TeO. For the bird to fly in a preferred, previously mapped direction relative to B(h), it hunts for that orientation that positions the frequency maxima at appropriate locations on the TeO. This condition can be maintained even as B(h) varies with geomagnetic latitude during the course of long-distance flights. The magnetovisual coordinate system (straight phi, omega) overlaying the two halves of the tectal surface in a nonsymmetric way may imply an additional orienting function for the TeO over and above that of a simple compass (e.g., homing navigation as distinct from migrational navigation).     

Calculation of electric fields and currents induced in a millimeter-resolution human model at 60 Hz using the FDTD method

The finite-difference time-domain (FDTD) method has previously been used to calculate induced currents in anatomically based models of the human body at frequencies ranging from 20 to 915 MHz and resolutions down to about 1.25 cm. Calculations at lower frequencies and higher resolutions have been precluded by the huge number of time steps that would be needed in these simulations. This paper describes a method used to overcome this problem and efficiently calculate induced currents in an MRI-based, 6-mm-resolution model of the human under a high-voltage transmission line. This model is significantly higher resolution than the 1.31-cm-resolution model previously used; therefore, it can be used to pinpoint locations of peak current densities in the body. Proposed safety guidelines would allow external electric fields of 10 kV/m and 25 kV/m for exposure to 60 Hz fields of the general public and workers, respectively. For this external electric field exposure of 10 kV/m, local induced current densities as high as 20 mA/m² are found in the head and trunk with even higher values (above 150 mA/m²) in the legs. These currents are considerably higher than the 4 or even 10 mA/m² that have been suggested in the various safety guidelines, thus indicating an inconsistency in the proposed guidelines. In addition, several ratios of E/H typical of power line exposures were examined, and it was found that the vertical electric field couples strongly to the body, whereas the horizontal magnetic field does not. Bioelectromagnetics 19:293–299, 1998. © 1998 Wiley-Liss, Inc.     

Local measurements of viscoelastic moduli of entangled actin networks using an oscillating magnetic bead micro-rheometer.

A magnetically driven bead micro-rheometer for local quantitative measurements of the viscoelastic moduli in soft macromolecular networks such as an entangled F-actin solution is described. The viscoelastic response of paramagnetic latex beads to external magnetic forces is analyzed by optical particle tracking and fast image processing. Several modes of operation are possible, including analysis of bead motion after pulse-like or oscillatory excitations, or after application of a constant force. The frequency dependencies of the storage modulus, G'(omega), and the loss modulus, G'(omega), were measured for frequencies from 10(-1) Hz to 5 Hz. For low actin concentrations (mesh sizes epsilon > 0.1 micron) we found that both G'(omega) and G'(omega) scale with omega 1/2. This scaling law and the absolute values of G' and G' agree with conventional rheological measurements, demonstrating that the magnetic bead micro-rheometer allows quantitative measurements of the viscoelastic moduli. Local variations of the viscoelastic moduli (and thus of the network density and mesh size) can be probed in several ways: 1) by measurement of G' and G' at different sites within the network; 2) by the simultaneous analysis of several embedded beads; and 3) by evaluation of the bead trajectories over macroscopic distances. The latter mode yields absolute values and local fluctuations of the apparent viscosity eta(x) of the network.     

The direct influence of electromagnetic fields on nerve- and muscle cells of man within the frequency range of 1 Hz to 30 MHz

Summary By using several biophysical approximations and considering man as free space model limiting order-of-magnitude values for external electric and magnetic field strengths which may be hazardous for human beings were calculated. Danger may occur by excitation processes below 30 kHz for field strengths exceeding these limiting values; for frequencies larger than 30 kHz, thermal effects are predominant before excitation occurs. The external electric field strength necessary for causing action potentials in the central nervous system exceeds by far the corona forming level. But excitation is possible by strong alternating magnetic fields.Furthermore, by comparing the electrically and magnetically induced currents with the naturally flowing currents in man caused by the brain's and heart's electrical activity, a lower boundary-line was estimated. Regarding electric or magnetic field strengths undercutting this boundary-line, direct effects on the central nervous system may be excluded. Other mechanisms should be responsible for demonstrated biological effects.     

TEDOR with adiabatic inversion pulses: Resonance assignments of <Superscript>13</Superscript>C/<Superscript>15</Superscript>N labelled RNAs

We have examined via numerical simulations the performance characteristics of different 15N RF pulse schemes employed in the transferred echo double resonance (TEDOR) experimental protocol for generating 13C-15N dipolar chemical shift correlation spectra of isotopically labelled biological systems at moderate MAS frequencies (omega(r) approximately 10 kHz). With an 15N field strength of approximately 30-35 kHz that is typically available in 5 mm triple resonance MAS NMR probes, it is shown that a robust TEDOR sequence with significant tolerance to experimental imperfections sa as H1 inhomogeneity and resonance offsets can be effectively implemented using adiabatic heteronuclear dipolar recoupling pulse schemes. TEDOR-based 15N-13C and 15N-13C-13C chemical shift correlation experiments were carried out for obtaining 13C and 15N resonance assignments of an RNA composed of 97 (CUG) repeats which has been implicated in the neuromuscular disease myotonic dystrophy.     

Extremely low frequency (ELF) electric and magnetic field exposure limits: rationale for basic restrictions used in the development of an Australian standard

There are large disparities between basic restrictions for exposure to extremely low-frequency (0-3 kHz) Electric and Magnetic Fields set by two major international bodies. Both bodies agree that these basic restrictions should prevent neuro-stimulatory effects: the retinal phosphene at frequencies up to a few hundred Hertz and peripheral nervous stimulation (PNS) at higher frequencies. The disparity arises from differences in estimated thresholds and frequency dependence, and whether restrictions should be of tissue induced current density or electric field. This paper argues that the latter metric more directly relates to neurostimulatory processes. By analysing available literature, a threshold for retinal phosphenes occurrence is found to be 56 mV/m (95% Confidence Interval 2-1330 mV/m), with a characteristic frequency of 20 Hz. Similarly, the smallest PNS sensation threshold is identified at 2 V/m (characteristic frequency above 3 kHz). In the case of the former, the large range of uncertainty suggests a 'power of ten' value of 100 mV/m. For the latter, because of the small margin between sensation and pain threshold, and because of the large individual variation, the smallest estimate of sensation threshold (2 V/m) represents a basic restriction with precaution incorporated.     

Kinetics of channelized membrane ions in magnetic fields

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

Deformability and stability of erythrocytes in high-frequency electric fields down to subzero temperatures.

High-frequency electric fields can be used to induce deformation of red blood cells. In the temperature domain T = 0 degrees to -15 degrees C (supercooled suspension) and for 25 degrees C this paper examines for human erythrocytes (discocytes, young cell population suspended in a low ionic strength solution with conductivity sigma(25 degrees) = 154 microS/cm) in a sinusoidal electric field (nu = 1 MHz, E0 = 0-18 kV/cm) the following properties and effects as a function of field strength and temperature: 1) viscoelastic response, 2) (shear) deformation (steady-state value obtained from the viscoelastic response time), 3) stability (by experimentally observed breakdown of cell polarization and hemolysis), 4) electrical membrane breakdown and field-induced hemolysis (theoretical calculations for ellipsoidal particles), and 5) mechanical hemolysis. The items 2-4 were also examined for the frequency nu = 100 kHz and for a nonionic solution of very low conductivity (sigma(25 degrees) = 10 microS/cm) to support our interpretations of the results for 1 MHz. Below 0 degrees C with decreasing temperature the viscoelastic response time tau(res)(T) for the cells to reach steady-state deformation values d(infinity,E) increases and the deformation d(infinity,E)(T) decreases strongly. Both effects are especially high for low field strengths. The longest response time of approximately 30 s was obtained for -15 degrees C and small deformations. For 1 MHz the cells can be highly elongated up to 2.3 times their initial diameter a0 for 25 degrees and 0 degrees C, 2.1a0 for -10 degrees C and still 1.95a0 for -15 degrees C. For T > or = 0 degrees C the deformation is limited by hemolysis of the cells, which sets in for E0(lysis)(25 degrees) approximately 8 kV/cm and E0(lysis)(0 degrees) approximately 14 kV/cm. These values are approximately three times higher than the corresponding calculated critical field strengths for electrically induced pore formation. Nevertheless, the observed depolarization and hemolysis of the cells is provoked by electrical membrane breakdown rather than by mechanical forces due to the high deformation. For the nonionic solution, where no electrical breakdown is expected in the whole range for E0, the cells can indeed be deformed to even higher values with a low hemolytic rate. Below 0 degrees C we observe no hemolysis at all, not even for the frequency 100 kHz, where the cells hemolyze at 25 degrees C for the much lower field strength E0(lysis) approximately 2.5 kV/cm. Obviously, pore formation and growth are weak for subzero temperatures.     

Electro-rotation of mouse oocytes: single-cell measurements of zona-intact and zona-free cells and of the isolated zona pellucida

Passive electrical properties of oocytes and of zonae pellucidae, and the mechanical coupling between them, can be elucidated by means of rotating-field-induced rotation. In low-conductivity media (25-100 microS/cm) rotation of mouse oocytes (with or without their zonae) requires fields in the 1-100 kHz frequency range. However, an isolated zona shows weak rotation in the opposite direction to that of a cell, and in response to much higher field frequencies (approx. 1 MHz). In zona-intact mouse oocytes, the rotation of cell and zona are not rigidly coupled: thus rotation of the cell can still be induced when the zona is held stationary. However, rotation of freely suspended zona-intact cells is much slower than that of zona-free cells and requires an optimum field frequency that is approximately 1.5 kHz higher. These observations show that the electrical properties of the oocyte that are measured by rotation are altered by the presence of the zona pellucida, even though no such influence has been detected using micro-electrodes. The data are consistent with the zona acting as a porous shell with a conductivity of 40 microS/cm (preliminary estimate made at a single medium conductivity of 26 microS/cm). Measurements on cells from which the zonae had been removed gave values for the membrane capacity and resistivity of 1.2-1.3 microF/cm2 and 400 omega.cm2, respectively. These values may reflect the presence of plasmalemma microvilli. The results strongly suggest that the technique may be useful for studies of cell maturation and for in vitro fertilization, because the cells may be further cultured after measurement.     

Reduction of electrode polarization capacitance in low-frequency impedance spectroscopy by using mesh electrodes

Dielectric measurements of biological samples are obscured by electrode polarization, which at low frequencies dominates over the actual sample response. Reduction of this artifact is especially necessary in studying interactions of electric field with biological systems in the α-dispersion range. We developed a method to reduce the influence of electrode polarization by employing mesh instead of solid electrodes as sensing probes, thereby reducing the area of the double layer. The design decreases the electrode-electrolyte contact area by almost 40% while keeping the bulk sample capacitance the same. Interrogation electric fields away from the electrode surface and sensitivity are unaffected. Electrodes were microfabricated (600μm×50μm, spacing of 100μm) with and without mesh holes 7.5μm×7.5μm in size. Simulations of electric field performed using Comsol Multiphysics showed non-uniformity of the electric field within less than 1.5μm from the electrode surface, which encompasses the double layer region, but at greater distance the solid and mesh electrodes gave the same results. Mesh electrodes reduced capacitance measurements for water and KCl solutions of different concentrations at low frequencies (<10kHz), while higher frequency capacitance remained the same for both electrode types, confirming our hypothesis that this design leaves the electric field mainly unaffected. Impedance measurements at low frequencies for water and mice heart mitochondrial suspension were lower for mesh than for solid electrodes. Comsol simulations confirmed these results by showing that mesh electrodes have a greater charge density than solid electrodes, which affects conductance. These electrodes are being used for mitochondrial membrane potential studies.