Role of modulation on the effect of microwaves on ornithine decarboxylase activity in L929 cells

The effect of 835 MHz microwaves on the activity of ornithine decarboxylase (ODC) in L929 murine cells was investigated at an SAR of ∼2.5 W/kg. The results depended upon the type of modulation employed. AM frequencies of 16 Hz and 60 Hz produced a transient increase in ODC activity that reached a peak at 8 h of exposure and returned to control levels after 24 h of exposure. In this case, ODC was increased by a maximum of 90% relative to control levels. A 40% increase in ODC activity was also observed after 8 h of exposure with a typical signal from a TDMA digital cellular telephone operating in the middle of its transmission frequency range (∼840 MHz). This signal was burst modulated at 50 Hz, with approximately 30% duty cycle. By contrast, 8 h exposure with 835 MHz microwaves amplitude modulated with speech produced no significant change in ODC activity. Further investigations, with 8 h of exposure to AM microwaves, as a function of modulation frequency, revealed that the response is frequency dependent, decreasing sharply at 6 Hz and 600 Hz. Exposure with 835 MHz microwaves, frequency modulated with a 60 Hz sinusoid, yielded no significant enhancement in ODC activity for exposure times ranging between 2 and 24 h. Similarly, exposure with a typical signal from an AMPS analog cellular telephone, which uses a form of frequency modulation, produced no significant enhancement in ODC activity. Exposure with 835 MHz continuous wave microwaves produced no effects for exposure times between 2 and 24 h, except for a small but statistically significant enhancement in ODC activity after 6 h of exposure. Comparison of these results suggests that effects are much more robust when the modulation causes low-frequency periodic changes in the amplitude of the microwave carrier. Bioelectromagnetics 18:132–141, 1997. © 1997 Wiley-Liss, Inc.     

Antibody responses of mice exposed to low-power microwaves under combined, pulse-and-amplitude modulation

Irradiation by pulsed microwaves (9.4 GHz, 1 microsecond pulses at 1,000/s), both with and without concurrent amplitude modulation (AM) by a sinusoid at discrete frequencies between 14 and 41 MHz, was assessed for effects on the immune system of Balb/C mice. The mice were immunized either by sheep red blood cells (SRBC) or by glutaric-anhydride conjugated bovine serum albumin (GA-BSA), then exposed to the microwaves at a low rms power density (30 microW/cm2; whole-body-averaged SAR approximately 0.015 W/kg). Sham exposure or microwave irradiation took place during each of five contiguous days, 10 h/day. The antibody response was evaluated by the plaque-forming cell assay (SRBC experiment) or by the titration of IgM and IgG antibodies (GA-BSA experiment). In the absence of AM, the pulsed field did not greatly alter immune responsiveness. In contrast, exposure to the field under the combined-modulation condition resulted in significant, AM-frequency-dependent augmentation or weakening of immune responses.     

A dosimeter for assessment of exposures to ELF fields

There are currently no biological indicators of electromagnetic field exposure to aid investigation of the biological effects of ELF fields, but this hiatus is partly compensated for by the ease with which the external fields can be measured. The subject of this report is a small electronic instrument that can log electric-, magnetic-, and transient-field intensities of individual subjects for a duration of two weeks, thus allowing exposure to be assessed in considerable detail. A preliminary survey conducted with the instrument has shown clear differences in domestic and occupational exposures of specific groups. Simple analytical models can be fitted to the field-exposure data by adjusting a few parameters such as current, intensity, and distance. Such modeling may be increasingly useful as the data base on exposures expands.     

Exposure system to study hypotheses of ELF and RF electromagnetic field interactions of mobile phones with the central nervous system

A novel exposure system for double-blind human electromagnetic provocation studies has been developed that satisfies the precision, control of fields and potential artifacts, and provides the flexibility to investigate the response of hypotheses-driven electromagnetic field exposure schemes on brain function, ranging from extremely low frequency (ELF) to radio frequency (RF) fields. The system can provide the same exposure of the lateral cerebral cortex at two different RF frequencies (900 and 2140 MHz) but with different exposure levels at subcortical structures, and also allows uniform ELF magnetic field exposure of the brain. The RF modulation and ELF signal are obtained by a freely programmable arbitrary signal generator allowing a wide range of worst-case exposure scenarios to be simulated, including those caused by wireless devices. The maximum achievable RF exposure is larger than 60 W/kg peak spatial specific absorption rate averaged over 10 g of tissue. The maximum ELF magnetic field exposure of the brain is 800 A/m at 50 Hz with a deviation from uniformity of 8% (SD).     

No effects of short‐term exposure to mobile phone electromagnetic fields on human cognitive performance: A meta‐analysis

During recent years, a large number of studies on the effects of electromagnetic fields emitted by cellular mobile phones on human cognitive performance have been carried out. However, the results have been ambiguous. We carried out the current meta‐analysis in order to investigate the impact of electromagnetic fields emitted by mobile phones on human cognition. Seventeen studies were included in the meta‐analysis as they fulfill several requirements such as single‐ or double‐blind experimental study design, and documentation of means and standard deviations of dependent variables. The meta‐analysis was carried out as a group comparison between exposed and non‐exposed subjects. No significant effects of electromagnetic fields emitted by Global System for Mobile Communications (GSM) and Universal Mobile Telecommunications System (UMTS) mobile phones were found. Cognitive abilities seem to be neither impaired nor facilitated. Results of the meta‐analysis suggest that a substantial short‐term impact of high frequency electromagnetic fields emitted by mobile phones on cognitive performance can essentially be ruled out. Bioelectromagnetics 33:159–165, 2012. © 2011 Wiley Periodicals, Inc.     

Inter-beat intervals of cardiac-cell aggregates during exposure to 2.45 GHz CW,pulsed, and square-wave-modulated microwaves

Inter-beat intervals of aggregated cardiac cells from chicken embryos were studied during 190 s exposures to 2.45 GHz microwaves in an open-ended coaxial device. Averaged specific-absorption rates (SARs) and modulation conditions were 1.2–86.9 W/kg continuous-wave (CW). 1.2–12.2 W/kg pulse modulation (PW, duty cycle ∽ 11%). and 12.0–43.5 W/kg square-wave modulation (duty cycle = 50%). The inter-beat interval decreased during microwave exposures at 42.0 W/kg and higher when CW or square-wave modulation was used, which is consistent with established effects of elevated temperatures. However, increases in the inter-beat interval during CW exposures at 1.2–12.2 W/kg, and decreases in the inter-beat interval after PW exposures at 8.4–12.2 W/kg. are not consistent with simple thermal effects. Analysis of variance indicated that SAR. modulation, and the modulation-SAR interaction were all significant factors in altering the interbeat interval. The latter two factors indicated that the cardiac cells were affected by athermal as well as thermal effects of microwave exposure. © 1993 Wiley-Liss. Inc.     

Suppression of T-lymphocyte cytotoxicity following exposure to sinusoidally amplitude-modulated fields

Significant inhibition of allogeneic cytotoxicity of the target cell MPC-11 by the murine cytotoxic T-lymphocyte line CTLL-1 was observed when the 4-h cytotoxicity assay was conducted in the presence of a 450-MHz field sinusoidally amplitude-modulated at 60 Hz. Exposure of the effector cells to the field prior to adding them to the target cells in the cytolytic assay resulted in a similar inhibition, suggesting a direct interaction of the field with the cytolytic T lymphocyte. The inhibition was preferentially expressed during the early allogeneic recognition phase. Field-exposed cytolytic cells recovered their full cytolytic capacity in 12.5 h. A differential susceptibility was observed with modulation frequencies from 0 to 100 Hz. Peak suppression occurred at 60 Hz modulation, with progressively smaller effects at 40, 16, and 3 Hz. The unmodulated carrier wave did not affect the cytotoxicity. Effects with 80- and 100-Hz modulation were smaller than at 60 Hz. These results demonstrate an inhibitory but recoverable effect by certain amplitude modulations of weak nonionizing radiation upon the cell-mediated cytolytic immune response.     

Frequency-dependent alterations in enolase activity in Escherichia coli caused by exposure to electric and magnetic fields

Some neurochemical effects of low-intensity electric and magnetic fields have been shown to be nonlinear functions of exposure parameters. These effects occurred within narrow ranges of frequency and intensity. Previous studies on membrane-associated endpoints in cell culture preparations demonstrated changes in calcium efflux and in acetylcholinesterase activity following exposure to radiofrequency radiation, amplitude modulated (AM) at 16 and at 60 Hz, at a specific absorption rate of 0.05 W/kg. In this study, these modulation frequencies were tested for their influence on the activity of a cytoplasmic enzyme, enolase, which is being tested clinically for detection of neoplasia. Escherichia coli cultures containing a plasmid with a mammalian gene for enolase were exposed for 30 min, and cell extracts were assayed for enolase activity by measuring absorbance at 240 nm. The enolase activity in exposed cultures was compared to the activity in paired control cultures. Exposure to 147 MHz carrier waves at 0.05 W/kg, AM at 16 Hz showed enolase activity enhanced by 62%, and AM at 60 Hz showed enolase activity reduced by 28%. Similarly, exposure to 16 Hz fields alone, at 21.2 V/m_rms (electric) and 97 nT_rms (magnetic), showed enhancement in enolase activity by 59%, whereas exposure to 60 Hz fields alone, at 14.1 V/m_rms (electric) and 65 nT_rms (magnetic), showed reduction in activity by 24%. Sham exposures as well as exposure to continuous-wave 147 MHz radiation at 0.05 W/kg showed no change in enolase activity. Although the underlying basis for these field effects in the cytoplasmic compartment has not been established, differential sensitivities to 16 Hz and to 60 Hz signals provide a clear focus for additional research to determine the responsible mechanism. © 1994 Wiley-Liss, Inc.     

Whole body exposure of rats to microwaves emitted from a cell phone does not affect the testes

The objective of this study was to investigate the effects of radiofrequency radiation emitted from cellular phones on the lipid composition, malondialdehyde concentration, p53 immune reactivity, sperm count, morphology, histological structure of testes, and on rectal temperature of rats exposed to microwave radiation emitted from cellular phones. Sixteen Spraque-Dawley rats were separated into two groups of eight, sham exposed (control) and experimental. The rats were confined in plexiglas cages specially designed for this study, and cellular phones were placed 0.5 cm under the cages. For the experimental group, cellular phones were activated 20 min per day (7 days a week) for 1 month. For the control group, the cellular phones were placed beneath the cages for 20 min a day, but the phones were turned off. Rectal temperatures were measured weekly. For 250 mW radiated power, the whole body average SAR (rms) is 0.52 W/kg and 1 g averaged peak SAR (rms) is 3.13 W/kg. The Mann-Whitney U-test was used for statistical comparisons of groups. No statistically significant alteration in any of the endpoints was noted. This study found no evidence suggesting an adverse effect of cell phone exposure on measures of testicular function or structure.     

Alterations in protein kinase activity following exposure of cultured human lymphocytes to modulated microwave fields

Cultures of human tonsil lymphocytes were exposed in a Crawford cell to a 450-MHz field (peak envelope intensity 1.0 mW/cm2), sinusoidally amplitude modulated (depth 80%) at frequencies between 3 and 100 Hz for periods up to 60 min. The Crawford cell was housed in a temperature-controlled chamber (35 degrees C) and control cultures were placed in the same chamber. Activity of cAMP-dependent protein kinase relative to controls remained unaltered by fields modulated at 16 or 60 Hz with exposures of 15, 30, and 60 min. By contrast, total non-cAMP-dependent kinase activity fell to less than 50% of unexposed control levels after 15 and 30 min exposures, but, despite continuing field exposure, returned to control or preexposure levels by 45 and 60 min. A smaller reduction (20-25%) also occurred with 60-Hz modulation and was also restricted to exposure durations of 15 and 30 min. CW 450-MHz fields were without effect. Reduced enzyme activity occurred with 16-, 40-, and 60-Hz modulation frequencies, but not with 3-, 6-, 80-, or 100-Hz modulation. The specific identity of this kinase is unknown. This rapid but transient reduction in lymphocyte protein kinase activity restricted to modulation frequencies between 16 and 60 Hz and to less than 30 min exposure is consistent with "windowing" with respect to modulation frequency and exposure duration.     

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.     

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.     

Calcium-ion movement and contractility in atrial strips of frog heart are not affected by low-frequency-modulated, 1 GHz electromagnetic radiation

Calcium efflux from electrically stimulated, ⁴⁵Ca²⁺-preloaded atrial strips of the frog heart was measured from samples of the rinsing perfusate collected at 2-min intervals for 32 min in a continuous perfusion chamber. Contractile force was simultaneously monitored. The specimen chamber was located in a stripline apparatus in which the atrial strips were exposed for 32 min to constant (CW) or amplitude-modulated (AM), 1 GHz electromagnetic (EM) fields at specific absorption rates (SAR) ranging from 3.2 μW/kg to 1.6 W/kg. Amplitude modulation was either at 0.5 Hz, in synchrony with the electrical stimulus applied to the preparation, or at 16 Hz. Neither unmodulated nor 0.5 Hz or 16 Hz modulated 1 GHz waves affected the movement of calcium ions or the contractile force in isolated atrial strips of the frog heart. © 1993 Wiley-Liss, Inc.     

Molecular change signal-to-noise criteria for interpreting experiments involving exposure of biological systems to weakly interacting electromagnetic fields

We describe an approach to aiding the design and interpretation of experiments involving biological effects of weakly interacting electromagnetic fields that range from steady (dc) to microwave frequencies. We propose that if known biophysical mechanisms cannot account for an inferred, underlying molecular change signal-to-noise ratio, (S/N)gen, of a observed result, then there are two interpretation choices: (1) there is an unknown biophysical mechanism with stronger coupling between the field exposure and the ongoing biochemical process, or (2) the experiment is responding to something other than the field exposure. Our approach is based on classical detection theory, the recognition that weakly interacting fields cannot break chemical bonds, and the consequence that such fields can only alter rates of ongoing, metabolically driven biochemical reactions, and transport processes. The approach includes both fundamental chemical noise (molecular shot noise) and other sources of competing chemical change, to be compared quantitatively to the field induced change for the basic case that the field alters a single step in a biochemical network. Consistent with pharmacology and toxicology, we estimate the molecular dose (mass associated with field induced molecular change per mass tissue) resulting from illustrative low frequency field exposures for the biophysical mechanism of voltage gated channels. For perspective, we then consider electric field-mediated delivery of small molecules across human skin and into individual cells. Specifically, we consider the examples of iontophoretic and electroporative delivery of fentanyl through skin and electroporative delivery of bleomycin into individual cells. The total delivered amount corresponds to a molecular change signal and the delivery variability corresponds to generalized chemical noise. Viewed broadly, biological effects due to nonionizing fields may include animal navigation, medical applications, and environmental hazards. Understanding necessary conditions for such effects can be based on a unified approach: quantitative comparison of the estimated chemical change due to a particular electromagnetic field exposure to that due to competing influences, with both estimates based on a biophysical mechanism model within the context of a model of a biological system.     

A replication study of human exposure to 60-Hz fields: effects on neurobehavioral measures.

The purpose of this study was to reproduce and extend an earlier investigation of the effects of human exposure to combined, 60-Hz electric and magnetic fields. This paper presents the neurobehavioral results. Thirty men participated in one training session and four testing sessions. Subjects were randomly assigned to two groups. The 18 subjects in Group I were exposed (9 kV/m, 20 microT) and sham exposed in two counterbalanced orders. In Group II, half of 12 subjects were exposed (9 kV/m, 20 microT) every session, and the remaining half were sham exposed every session. The study was doubly blinded. Measures of cardiac interbeat interval, event-related brain potentials, and performance were obtained before, during, and after exposures. As in the earlier study, exposure to the combined field resulted in a statistically significant slowing of heart rate, in changes in late components of event-related brain potentials, and in decreased errors on a choice reaction-time task. In addition, field effects on several other measures approached statistical significance. The physiological measures obtained during exposure indicated that effects were greatest soon after the field was switched on, and again when it was switched off. The data indicate that changes in exposure level may be more important than duration of exposure for producing effects in human beings.     

Differentiation of Y79 cells induced by prolonged exposure to insulin

Y79 human retinoblastoma cells are known to contain receptors for both insulin and insulin-like growth factors (IGFs), to produce these cytokines and release them in the culture medium. Previously we have demonstrated that IGFs and insulin stimulate Y79 cell proliferation through the involvement of type I IGF receptor and Insulin Receptor Substrate 1 (IRS-1). This paper studies the effect of prolonged exposure to insulin on Y79 cells. Cells grown for 10 days in the presence of insulin were reseeded and incubated once more with insulin. In the reseeded cells proliferation lowered and morphological changes appeared. After 10 days of reseeding, cells stopped proliferating and showed long ramifying neurite processes and varicosities consistent with neuronal differentiation. Morphological differentiation was accompanied by a marked increase in the content of total protein and in that of tubulin, the major protein constituent of microtubules, a marked increase in the content of specialized protein markers of dopaminergic and cholinergic differentiation (dopamine -hydroxylase and choline acetyltransferase activities, respectively); a contemporaneous decrease in the content of glial fibrillary acidic protein (GFAP), a specific marker of glial cells, was also observed. Our results demonstrate that prolonged exposure to insulin induces Y79 cells to differentiate into a neuronal-like phenotype. At this moment it is not possible to establish the mechanism by which insulin induces this differentiative effect.     

Response of cyclic AMP by DMSO differentiated HL-60 cells exposed to electric interferential current after prestimulation

The action of interferential current (IFC) upon intracellular content of cyclic adenosine monophosphate (cAMP) after prestimulation with the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP) has been investigated. Human promyelocytes (HL-60) differentiated to granulocytes by dimethylsulphoxide (DMSO) have been treated with different concentrations of fMLP. This enhances their cAMP content. The half maximal effective concentration (EC50) was about 4 nM. Exposure to IFC with modulation frequencies of 35 and 125 Hz (5 min, 250 microA/cm2) after prestimulation with various concentrations of fMLP had no enhancing effect at low or high concentrations of fMLP. In contrast, at medium concentrations in the range of about 100 pM fMLP, a significant enhancement of cAMP could be observed. This synergistic effect of fMLP and IFC has been examined in detail by varying the modulation frequency, current density, and exposure time. Enhancement of cAMP content could be observed at certain modulation frequencies and exposure times suggesting a window effect, whereas for the current density in the range measured (8.5 microA/cm2-2.5 mA/cm2) a constant enhancement could be observed. The synergistic effect of fMLP and IFC could only be observed in the treatment sequence of fMLP followed by IFC; an inverse sequence had no effect on the cAMP content. .     

Carry‐over effects of multiple stressors on benthic embryos are mediated by larval exposure to elevated UVB and temperature

Damaging effects of UVB in conjunction with other stressors associated with global change are well‐established, with many studies focused on vulnerable early life stages and immediate effects (e.g., mortality, developmental abnormalities). However, for organisms with complex life cycles, experiences at one life stage can have carry‐over effects on later life stages, such that sublethal effects may mediate later vulnerability to further stress. Here, we exposed embryos in benthic egg masses of the New Zealand intertidal gastropod Siphonaria australis to treatments of either periodic stress (e.g., elevated UVB, salinity, and water temperature mimicking tidepool conditions in which egg masses are commonly found during summer) or control conditions (low UVB, ambient salinity, and water temperatures). Although there was high mortality from stressed egg masses, 24% of larvae hatched successfully. We then exposed the hatching larvae from both egg mass treatments to different combinations of water temperature (15 or 20 °C) and light (high UVB or shade) 12 h per day for 10 days. The most stressful larval conditions of 20 °C/high UVB resulted in low survival and stunted growth. Carry‐over effects on survival were apparent for shaded larvae exposed to elevated temperature, where those from stressed egg masses had 1.8× higher mortality than those from control egg masses. Shaded larvae were also larger and had longer velar cilia if they were from control egg masses, independent of larval temperature. These results demonstrate that previous experience of environmental stress can influence vulnerability of later life stages to further stress, and that focus on a single life stage will underestimate cumulative effects of agents of global change.     

Structural changes in the innercortex cells of soybean root nodules are induced by short-term exposure to high salt or oxygen concentrations

After a 2 h exposure of intact soybean nodules to high concentrations of NaCl (100mol m_?3) or oxygen (8OkPa O₂), morphometric computations carried out using an image analysis technique on semi-thin sections showed that both treatments induced a decrease in the area of the inner-cortex cells, which were then characterized by a tangential elongation. In contrast, no significant change in area occurred in the middle-cortex cells although their elongation decreased. Electron microscopic observations showed that in the inner-cortex cells changes included the presence of wall infoldings, an enlarged periplasmic space and a lobate nucleus whose chromatin distribution differed from that of the control. Structural changes also occurred in the endoplasmic reticulum, microbodies, mitochondria and plastids. From several of these changes, which are similar to those noted in osmocontractil cells in response to external stimuli, it can be hypothesized that the inner cortex may provide a potential mechanism for the control of oxygen diffusion through the nodules.     

An SR-protein induced by HSVI binding to cells functioning as a splicing inhibitor of viral pre-mRNA

The interaction between a virus and its specific receptor on the membrane of the host cell mimics the physiological combination of signal ligand and its receptor, and initiates the specific signal transduction from this activated receptor to induce a relative gene response. During the investigation of the interaction between Herpes simplex virus I (HSVI) and human fibroblast via the virus binding to its receptor complex on the cellular membrane, a new gene of cellular response against the specific stimulation of HSVI binding to fibroblasts was cloned from a cDNA library established from mRNA of an early gene response. This gene encoded a protein of 14.9kDa with the structural characteristics of Arg-rich and RS repeats. The analysis of the role of this protein in the infection by HSVI indicated that this protein, expressed only in G(1)/S phase and phosphorylated, functioned as a splicing inhibitor of HSVI pre-mRNA. The details of the mechanism of this inhibition of HSVI pre-mRNA splicing is still unclear.