W. Ross Adey

Magnetic field-induced changes in specific gene transcription

Magnetic fields are physical, environmental agents that have been shown to produce a variety of responses in cellular and animal studies, including general changes in gene transcription. In this study, the nuclear run-off assay has been employed to assess alterations in specific gene transcription in CEM-CM3 T-lymphoblastoid cells exposed for 15-120 min to a 1 gauss sinusoidal magnetic field at 60 Hz. Time-dependent and cell density-dependent changes in the transcription of c-fos, c-jun, c-myc and protein kinase C (beta-form) have been observed and quantitated. Additionally, changes in transcript levels, assessed by slot-blot analysis, have been found to parallel the changes in gene transcription. These data suggest an important role for magnetic field exposure in altering cellular processes.

Electrophysiological studies of hippocampal connections and excitability

Abstract 1. 1. An electrophysiological study of the afferent and efferent pathways of the hippocampus has been made. 2. 2. The excitability of the hippocampus has been studied before and after isolation from the rest of the brain. 3. 3. Evidence for afferent projections to the hippocampus through the fornix has been adduced. 4. 4. Evidence that the dentate gyrus may present and intermediary relay station is advanced. 5. 5. Evidence is presented for connections between the hippocampus and various cortical and subcortical areas, including limbic cortex, amygdala, diecephalic areas and the midbrain tegmentum. 6. 6. The physiological role of the hippocampus and its significance in seizure discharge are discussed.

Exposure of nerve growth factor-treated PC12 rat pheochromocytoma cells to a modulated radiofrequency field at 836.55 MHz: Effects on c-jun and c-fos expression

Rat PC12 pheochromocytoma cells have been treated with nerve growth factor and then exposed to athermal levels of a packet-modulated radiofrequency field at 836.55 MHz. This signal was produced by a prototype time-domain multiple-access (TDMA) transmitter that conforms to the North American digital cellular telephone standard. Three slot average power densities were used: 0.09, 0.9, and 9 mW/cm2. Exposures were for 20, 40, and 60 min and included an intermittent exposure regimen (20 min on/20 min off), resulting in total incubation times of 20, 60, and 100 min, respectively. Concurrent controls were sham exposed. After extracting total cellular RNA, Northern blot analysis was used to assess the expression of the immediate early genes, c-fos and c-jun, in all cell populations. No change in c-fos transcript levels were detected after 20 min exposure at each field intensity (20 min was the only time period at which c-fos message could be detected consistently). Transcript levels for c-jun were altered only after 20 min exposure to 9 mW/cm2 (average 38% decrease).

DNA synthesis and cell proliferation in C6 glioma and primary glial cells exposed to a 836.55 MHz modulated radiofrequency field.

We have tested the hypothesis that modulated radiofrequency (RF) fields may act as a tumor-promoting agent by altering DNA synthesis, leading to increased cell proliferation. In vitro tissue cultures of transformed and normal rat glial cells were exposed to an 836.55 MHz, packet-modulated RF field at three power densities: 0.09, 0.9, and 9 mW/cm2, resulting in specific absorption rates (SARs) ranging from 0.15 to 59 muW/g. TEM-mode transmission-line cells were powered by a prototype time-domain multiple-access (TDMA) transmitter that conforms to the North American digital cellular telephone standard. One sham and one energized TEM cell were placed in standard incubators maintained at 37 degrees C and 5% CO2. DNA synthesis experiments at 0.59-59 muW/g SAR were performed on log-phase and serum-starved semiquiescent cultures after 24 h exposure. Cell growth at 0.15-15 muW/g SAR was determined by cell counts of log-phase cultures on days 0, 1, 5, 7, 9, 12, and 14 of a 2 week protocol. Results from the DNA synthesis assays differed for the two cell types. Sham-exposed and RF-exposed cultures of primary rat glial cells showed no significant differences for either log-phase or serum-starved condition. C6 glioma cells exposed to RF at 5.9 muW/g SAR (0.9 mW/cm2) exhibited small (20-40%) significant increases in 38% of [3H]thymidine incorporation experiments. Growth curves of sham and RF-exposed cultures showed no differences in either normal or transformed glial cells at any of the power densities tested. Cell doubling times of C6 glioma cells [sham (21.9 +/- 1.4 h) vs. field (22.7 +/- 3.2 h)] also demonstrated no significant differences that could be attributed to altered DNA synthesis rates. Under these conditions, this modulated RF field did not increase cell proliferation of normal or transformed cultures of glial origin.

Nonlinear wave mechanisms in interactions between excitable tissue and electromagnetic fields.

It is now well established that intrinsic electromagnetic fields play a key role in a broad range of tissue functions, including embryonic morphogenesis, wound healing, and information transmission in the nervous system. These same processes may be profoundly influenced by eletromagnetic fields induced by an external force. Tissue exposure to extremely low frequency (ELF) and ELF-modulated microwave fields at levels below those inducing significant thermal effects has revealed highly nonlinear mechanisms as a basis for observed effects. Interactions of phonons and excitons along linear molecules may produce nonlinear molecular vibrations in the form of soliton waves. Solitons exist in a minimal energy state and are extremely long-lived in comparison to linear oscillations. Solitons may convey energy released by chemical reactions from one site to another in enzymes of other long-chain proteins. These nonlinear waves may also couple reaction-diffusion processes in the intracellular and extracellular domains. A model is proposed for interaction between excitable tissue and electromagnetic fields, based on nonlinear waves in the cell membrane, with ionic interactions as an essential step. Calcium fluxes in the extracellular space of the central system are modeled by a nonlinear reaction-diffusion system. Membrane molecular solitons may exist in long-chain molecules (Davydov type) and play a significant role in charge transfer; or they may exist as nonlinear waves conveying energy along gel-lipid domains from one protein site to another (Sine-Gordon soliton). Soliton movements occur at subsonic velocities.

On the role of subthalamic areas in the maintenance of brain-stem reticular excitability

Abstract The responsiveness of midbrain tegmental zones constituting the reticular formation to various sensory inputs has long been recognized. The influence of cortical and subcortical influxes on the excitability of these reticular zones is less clear. The influence of various diencephalic areas, and particularly of the subthalamic region, on midbrain responses to peripheral volleys has been tested here in cat and monkey. In animals with chronic subthalamic lesions, spontaneous unit firing and unit responsiveness to peripheral stimuli in dorsal tegmental zones of the rostral midbrain were much reduced. In acute experiments, bilateral subthalamic coagulation was followed by marked reduction in the amplitude of the midbrain reticular responses to sciatic stimulation. Brief tetanization of the midbrain recording site restored the responses to their original amplitude, but they then decayed in a fashion resembling declining post-tetanic potentiation. Behavior changes in cat and monkey following these lesions included altered feeding habits, “catatonic” postures, reduced spontaneous movement, and a failure to make avoidance responses in previously trained animals. It is concluded that the subthalamic areas may exercise a tonic excitatory influence on midbrain reticular neurons, which may not be inherently available to peripheral influxes in the absence of this tonic excitation. Evidence is also presented for a phasic inhibitory subthalamic influence occurring briefly after a peripheral influx, and which is lost following subthalamic lesions. The possible relationship of these findings to mechanisms of sleep and wakefulness is discussed.

Effect of immobilization and concurrent exposure to a pulse-modulated microwave field on core body temperature, plasma ACTH and corticosteroid, and brain ornithine decarboxylase, Fos and Jun mRNA.

Abstract Stagg, R. B., Hawel, L. H., III, Pastorian, K., Cain, C., Adey, W. R. and Byus, C. V. Effect of Immobilization and Concurrent Exposure to a Pulse-Modulated Microwave Field on Core Body Temperature, Plasma ACTH and Corticosteroid, and Brain Ornithine Decarboxylase, Fos and Jun mRNA. Exposure of humans and rodents to radiofrequency (RF) cell phone fields has been reported to alter a number of stress- related parameters. To study this potential relationship in more detail, tube-restrained immobilized Fischer 344 rats were exposed in the near field in a dose-dependent manner to pulse-modulated (11 packets/s) digital cell phone microwave fields at 1.6 GHz in accordance with the Iridium protocol. Core body temperatures, plasma levels of the stress-induced hormones adrenocorticotrophic hormone (ACTH) and corticosterone, and brain levels of ornithine decarboxylase (Odc), Fos and Jun mRNAs were measured as potential markers of stress responses mediated by RF radiation. We tested the effects of the loose-tube immobilization with and without prior conditioning throughout a 2-h period (required for near-field head exposure to RF fields), on core body temperature, plasma ACTH and corticosteroids. Core body temperature increased transiently (±0.3°C) during the initial 30 min of loose- tube restraint in conditioned animals. When conditioned/tube- trained animals were followed as a function of time after immobilization, both the ACTH and corticosterone levels were increased by nearly 10-fold. For example, within 2–3 min, ACTH increased to 83.2 ± 31.0 pg/dl, compared to 28.1 ± 7.7 pg/dl for cage controls, reaching a maximum at 15–30 min (254.6 ± 46.8 pg/dl) before returning to near resting levels by 120 min (31.2 ± 10.2 pg/dl). However, when non-tube-trained animals were submitted to loose-tube immobilization, these animals demonstrated significantly higher (3–10-fold greater) hormone levels at 120 min than their tube-trained counterparts (313.5 ± 54.8 compared to 31.2 ± 10.2 pg/dl; corticosterone, 12.2 ± 6.2 μg/dl compared to 37.1 ± 6.4 μg/dl). Hormone levels in exposed animals were also compared to those in swim-stressed animals. Swimming stress also resulted in marked elevation in both ACTH and corticosterone levels, which were 10–20 fold higher (541.8 compared to 27.2–59.1 pg/dl for ACTH) and 2–5 fold higher (45.7 compared to 8.4– 20.0 μg/dl for corticosteroids) than the cage control animals. Three time-averaged brain SAR levels of 0.16, 1.6 and 5 W/ kg were tested in a single 2-h RF-field exposure to the Iridium cell phone field. When RF-exposed and sham-exposed (immobilized) animals were compared, no differences were seen in core body temperature, corticosterone or ACTH that could be attributed to near-field RF radiation. Levels of Odc, Fos and Jun mRNA were also monitored in brains of animals exposed to the RF field for 2 h, and they showed no differences from sham-exposed (loose-tube immobilized) animals that were due to RF-field exposure. These data suggest that a significant stress response, indicated by a transient increase in core body temperature, ACTH and corticosterone, occurred in animals placed in even the mild loose-tube immobilization required for near-field RF exposure employed here and in our other studies. Failure to adequately characterize and control this immobilization response with appropriate cage control animals, as described previously, could significantly mask any potential effects mediated by the RF field on these and other stress-related parameters. We conclude that the pulse-modulated digital Iridium RF field at SARs up to 5 W/kg is incapable of altering these stress-related responses. This conclusion is further supported by our use of an RF-field exposure apparatus that minimized immobilization stress; the use of conditioned/tube-trained animals and the measurement of hormonal and molecular markers after 2 h RF-field exposure when the stress-mediated effects were complete further support our conclusion.

Focus formation of C3H/10T1/2 cells and exposure to a 836.55 MHz modulated radiofrequency field

Disruption of communication between transformed cells and normal cells is involved in tumor promotion. We have tested the hypothesis that exposures to radiofrequency (RF) fields using a form of digital modulation (TDMA) and a chemical tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), are copromoters that enhance focus formation of transformed cells in coculture with parental C3H/10T1/2 murine fibroblasts. RF field exposures did not influence TPAs dose-dependent promotion of focus formation in coculture. Cell cultures were exposed to an 836.55 MHz TDMA-modulated field in TEM transmission line chambers, with incident energies that simulated field intensities at a users head. Specific absorption rates (SARs) of 0.15, 1.5, and 15 muW/g were used during each digital packet, and the packet frequency was 50/s. The TEM chambers were placed in a commercial incubator at 37 degrees C and 95% humidity/5% CO2. The RF field exposures were in a repeating cycle, 20 min on, 20 min off, 24 h/day for 28 days. At 1.5 muW/g, TPA-induced focus formation (at 10, 30, and 50 ng/ml) was not significantly different in RF-exposed cultures compared to parallel sham-exposed cultures in ten independent experiments in terms of the number, density, and area of foci. Similarly, at 0.15 and 15.0 muW/g, in two and four experiments, respectively, RF exposure did not alter TPA-induced focus formation. The findings support a conclusion that repeated exposures to this RF field do not influence tumor promotion in vitro, based on the RF fields inability to enhance TPA-induced focus formation.

The Extracellular Space and Energetic Hierarchies in Electrochemical Signaling Between Cells

It is a reasonable assumption that the first living organisms existed as single cells floating or swimming at the surface of primitive oceans. This concept of a cell emphasizes the role of a bounding membrane that delimits an organized interior composed of biomolecules that may have first existed in the absence of cells, perhaps simply as an unconstrained molecular “soup” at the surface of primordial seas. Within cells, these molecular systems mediate processes essential for all terrestrial life in metabolism, reproduction and responses to environmental stimuli.

Biological Effects of Low Energy Electromagnetic Fields on the Central Nervous System

There has been extensive speculation about the possibility of enhanced sensitivities of central nervous tissue to environmental electromagnetic fields. Brain tissue possesses its own well known intrinsic oscillating field, the electroencephalogram (EEG). The functional significance of this internal field has been a matter of conjecture. Based on classical membrane electrophysiology, the majority of opinion has most frequently dismissed it as merely “the noise of the brain’s motor.” Evidence that this may not be an accurate evaluation has come from a variety of studies of effects of imposed oscillating electromagnetic fields which induce weak extracellular electrochemical oscillations in the fluid surrounding brain cells, and which mimic in varying degrees components of the natural electrochemical oscillations of the EEG in the same domain of brain tissue.