900 MHz modulated electromagnetic fields accelerate the clathrin‐mediated endocytosis pathway

We report new data regarding the molecular mechanisms of GSM‐induced increase of cell endocytosis rate. Even though endocytosis represents an important physical and biological event for cell physiology, studies on modulated electromagnetic fields (EMF) effects on this process are scarce. In a previous article, we showed that fluid phase endocytosis rate increases when cultured cells are exposed to 900 MHz EMF similar to mobile phones' modulated GSM signals (217 Hz repetition frequency, 576 µs pulse width) and to electric pulses similar to the GSM electrical component. Trying to distinguish the mechanisms sustaining this endocytosis stimulation, we exposed murine melanoma cells to Lucifer Yellow (LY) or to GSM–EMF/electric pulses in the presence of drugs inhibiting the clathrin‐ or the caveolin‐dependent endocytosis. Experiments were performed at a specific absorption rate (SAR) of 3.2 W/kg in a wire patch cell under homogeneously distributed EMF field and controlled temperature (in the range of 28.5–29.5 °C). Thus, the observed increase in LY uptake was not a thermal effect. Chlorpromazine and ethanol, but not Filipin, inhibited this increase. Therefore, the clathrin‐dependent endocytosis is stimulated by the GSM–EMF, suggesting that the cellular mechanism affected by the modulated EMF involves vesicles that detach from the cell membrane, mainly clathrin‐coated vesicles. Bioelectromagnetics 30:222–230, 2009. © 2008 Wiley‐Liss, Inc.     

Effects of continuous and intermittent exposure to RF fields with a wide range of SARs on cell growth, survival, and cell cycle distribution

To examine the biological effects of radio frequency (RF) electromagnetic fields in vitro, we have examined the fundamental cellular responses, such as cell growth, survival, and cell cycle distribution, following exposure to a wide range of specific absorption rates (SAR). Furthermore, we compared the effects of continuous and intermittent exposure at high SARs. An RF electromagnetic field exposure unit operating at a frequency of 2.45 GHz was used to expose cells to SARs from 0.05 to 1500 W/kg. When cells were exposed to a continuous RF field at SARs from 0.05 to 100 W/kg for 2 h, cellular growth rate, survival, and cell cycle distribution were not affected. At 200 W/kg, the cell growth rate was suppressed and cell survival decreased. When the cells were exposed to an intermittent RF field at 300 W/kg(pk), 900 W/kg(pk) and 1500 W/kg(pk) (100 W/kg(mean)), no significant differences were observed between these conditions and intermittent wave exposure at 100 W/kg. When cells were exposed to a SAR of 50 W/kg for 2 h, the temperature of the medium around cells rose to 39.1 degrees C, 100 W/kg exposure increased the temperature to 41.0 degrees C, and 200 W/kg exposure increased the temperature to 44.1 degrees C. Exposure to RF radiation results in heating of the medium, and the thermal effect depends on the mean SAR. Hence, these results suggest that the proliferation disorder is caused by the thermal effect.     

Growth characteristics of mung beans and water convolvuluses exposed to 425‐MHz electromagnetic fields

Effects of high‐frequency, continuous wave (CW) electromagnetic fields on mung beans (Vigna radiata L.) and water convovuluses (Ipomoea aquatica Forssk.) were studied at different growth stages (pre‐sown seed and early seedling). Specifically, the effects of the electromagnetic source's power and duration (defined as power‐duration level) on the growth of the two species were studied. Mung beans and water convolvuluses were exposed to electromagnetic fields inside a specially designed chamber for optimum field absorption, and the responses of the seeds to a constant frequency at various power levels and durations of exposure were monitored. The frequency used in the experiments was 425 MHz, the field strengths were 1 mW, 100 mW, and 10 W, and the exposure durations were 1, 2, and 4 h. Results show that germination enhancement is optimum for the mung beans at 100 mW/1 h power‐duration level, while for water convolvuluses the optimum germination power‐duration level was 1 mW/2 h. When both seed types were exposed at the early sprouting phase with their respective optimum power‐duration levels for optimum seed growth, water convolvuluses showed growth enhancement while mung bean sprouts showed no effects. Water content analysis of the seeds suggests thermal effects only at higher field strength. Bioelectromagnetics 31:519–527, 2010. © 2010 Wiley‐Liss, Inc.     

Cytostatic response of NB69 cells to weak pulse-modulated 2.2 GHz radar-like signals

The present study investigates the response of two human cancer cell lines to a 24‐h treatment with a 2.2‐GHz, pulse‐modulated (5 µs pulse duration, 100 Hz repetition rate) radar‐like signal at an average SAR = 0.023 W/kg, using a newly designed setup for in vitro exposure to radiofrequency (RF) fields. A complete discretized model of the setup was created for numerical dosimetry using finite‐difference time‐domain (FDTD) software, SEMCAD X. The average dose of RF radiation absorbed by the cultures was calculated to be subthermal (ΔT < 0.1 °C). The RF exposure induced a consistent, statistically significant reduction in the cell number (13.5% below controls, P < 0.001) in the neuroblastoma NB69 line. This effect was accompanied with slight but statistically significant increases in the proportions of cells in phases G0/G1 and G2/M of the cell cycle (6% and 9%, respectively; P < 0.05 over controls). By contrast, the hepatocarcinoma cell line HepG2 did not respond to the same RF treatment. These results indicate that a pulse‐modulated RF radiation with high instantaneous amplitude and low average power can induce cytostatic responses on specific, sensitive cancer cell lines. The effect would be mediated, at least in part, by alterations in the kinetics of the cell cycle. Bioelectromagnetics 32:340–350, 2011. © 2010 Wiley‐Liss, Inc.     

Dependence of microwave effect on the secondary structure of DNA on molecular weight of polynucleotide

The effect of ultralow power pulse-modulated electromagnetic radiation (average power density 60 microW/cm2, carrying frequency 1.05; 2.12; or 2.39 GHz; modulating pulses with frequency 4 Hz) on the secondary structure of DNA was investigated. It was established that the exposure of beta-alanine and formaldehyde containing aqueous DNA solution to electromagnetic radiation had activated the process of DNA despiralization under the action of beta-alanine--formaldehyde reaction product. The effect of electromagnetic radiation on the secondary structure of DNA can be removed by lowering of molecular weight of DNA to 0.46 x 10(6) (at carrying frequency 1.05 GHz), or to 0.25 x 10(3) (at carrying frequency 2.39 GHz).     

Stimulation of Burn Wound Healing in Rats by Nanosecond Repetitive Pulsed Microwave Radiation

Biology Bulletin - The effect of nanosecond repetitive pulsed microwave radiation (RPMR, 10 GHz, 100 ns pulse duration, 8 Hz pulse repetition rate, 140 and 1500 W/cm2 peak power flux density...     

New fin-line devices for radiofrequency exposure of small biological samples in vitro allowing whole-cell patch clamp recordings

The development and analysis of three waveguides for the exposure of small biological in vitro samples to mobile communication signals at 900 MHz (GSM, Global System for Mobile Communications), 1.8 GHz (GSM), and 2 GHz (UMTS, Universal Mobile Telecommunications System) is presented. The waveguides were based on a fin‐line concept and the chamber containing the samples bathed in extracellular solution was placed onto two fins with a slot in between, where the exposure field concentrates. Measures were taken to allow for patch clamp recordings during radiofrequency (RF) exposure. The necessary power for the achievement of the maximum desired specific absorption rate (SAR) of 20 W/kg (average over the mass of the solution) was approximately P_in = 50 mW, P_in = 19 mW, and P_in = 18 mW for the 900 MHz, 1800 MHz, and 2 GHz devices, respectively. At 20 W/kg, a slight RF‐induced temperature elevation in the solution of no more than 0.3 °C was detected, while no thermal offsets due to the electromagnetic exposure could be detected at the lower SAR settings (2, 0.2, and 0.02 W/kg). A deviation of 10% from the intended solution volume yielded a calculated SAR deviation of 8% from the desired value. A maximum ±10% variation in the local SAR could occur when the position of the patch clamp electrode was altered within the area where the cells to be investigated were located. Bioelectromagnetics 32:102–112, 2011. © 2010 Wiley‐Liss, Inc.     

Absence of corneal endothelium injury in non‐human primates treated with and without ophthalmologic drugs and exposed to 2.8 GHz pulsed microwaves

Microwave‐induced corneal endothelial damage was reported to have a low threshold (2.6 W/kg), and vasoactive ophthalmologic medications lowered the threshold by a factor of 10–0.26 W/kg. In an attempt to confirm these observations, four adult male Rhesus monkeys (Macaca mulatta) under propofol anesthesia were exposed to pulsed microwaves in the far field of a 2.8 GHz signal (1.43 ± 0.06 µs pulse width, 34 Hz pulse repetition frequency, 13.0 mW/cm² spatial and temporal average, and 464 W/cm² spatial and temporal peak (291 W/cm² square wave equivalent) power densities). Corneal‐specific absorption rate was 5.07 W/kg (0.39 W/kg/mW/cm²). The exposure resulted in a 1.0–1.2 °C increase in eyelid temperature. In Experiment I, exposures were 4 h/day, 3 days/week for 3 weeks (nine exposures and 36 h total). In Experiment II, these subjects were pretreated with 0.5% Timolol maleate and 0.005% Xalatan® followed by 3 or 7 4‐h pulsed microwave exposures. Under ketamine–xylazine anesthesia, a non‐contact specular microscope was used to obtain corneal endothelium images, corneal endothelial cell density, and pachymetry at the center and four peripheral areas of the cornea. Ophthalmologic measurements were done before and 7, 30, 90, and 180 days after exposures. Pulsed microwave exposure did not cause alterations in corneal endothelial cell density and corneal thickness with or without ophthalmologic drugs. Therefore, previously reported changes in the cornea exposed to pulsed microwaves were not confirmed at exposure levels that are more than an order of magnitude higher. Bioelectromagnetics 31:324–333, 2010. Published 2010 Wiley‐Liss, Inc.     

Changes in physico-chemical parameters of homeopathic remedies ferrum metallicum CH6 and ferrum metallicum CH30 after exposure to high frequency electromagnetic radiation of low intensity

It is considered the microwaves electromagnetic radiation do not affect the materials, alive or not, when used in low power. In high power, the interaction effects would be the material warming (thermal effect). However, in the last years, the studies about electromagnetic radiation with low power (non thermal effect) in the human being have been increasing. It was found out the electromagnetic radiation, even with low power, can affect the living organisms and biosubstratum. In the present work the influence of electromagnetic radiation (2.45 GHz 500 W/cm2), on physical and chemical parameters of the homeopathic pharmaceutics products in shown.     

Features of anti‐inflammatory effects of modulated extremely high‐frequency electromagnetic radiation

Using a model of acute zymosan‐induced paw edema in NMRI mice, we test the hypothesis that anti‐inflammatory effects of extremely high‐frequency electromagnetic radiation (EHF EMR) can be essentially modified by application of pulse modulation with certain frequencies. It has been revealed that a single exposure of animals to continuous EHF EMR for 20 min reduced the exudative edema of inflamed paw on average by 19% at intensities of 0.1–0.7 mW/cm² and frequencies from the range of 42.2–42.6 GHz. At fixed effective carrier frequency of 42.2 GHz, the anti‐inflammatory effect of EHF EMR did not depend on modulation frequencies, that is, application of different modulation frequencies from the range of 0.03–100 Hz did not lead to considerable changes in the effect level. On the contrary, at “ineffective” carrier frequencies of 43.0 and 61.22 GHz, the use of modulation frequencies of 0.07–0.1 and 20–30 Hz has allowed us to restore the effect up to a maximal level. The results obtained show the critical dependence of anti‐inflammatory action of low‐intensity EHF EMR on carrier and modulation frequencies. Within the framework of this study, the possibility of changing the level of expected biological effect of modulated EMR by a special selection of combination of carrier and modulation frequencies is confirmed. Bioelectromagnetics 30:454–461, 2009. © 2009 Wiley‐Liss, Inc.     

The study of the mechanisms of formation of reactive oxygen species in aqueous solutions on exposure to high peak-power pulsed electromagnetic radiation of extremely high frequencies

Using the method of enhanced chemiluminescence in a peroxidase-luminol-p-iodophenol system, we found the formation of reactive oxygen species (in equivalent of hydrogen peroxide concentration) in 1 mM phosphate buffer under the exposure to high peak-power pulsed electromagnetic radiation of extremely high frequencies (37 GHz, peak power 20 kW, pulse width 400 ns, repetition rate 500 Hz). The results obtained show that the formation of hydrogen peroxide in aqueous solutions under the action of electromagnetic radiation is the result of the summary influence of heat and thermoacoustic waves excited in the solutions.     

Sleep after mobile phone exposure in subjects with mobile phone‐related symptoms

Several studies show increases in activity for certain frequency bands (10–14 Hz) and visually scored parameters during sleep after exposure to radiofrequency electromagnetic fields. A shortened REM latency has also been reported. We investigated the effects of a double‐blind radiofrequency exposure (884 MHz, GSM signaling standard including non‐DTX and DTX mode, time‐averaged 10 g psSAR of 1.4 W/kg) on self‐evaluated sleepiness and objective EEG measures during sleep. Forty‐eight subjects (mean age 28 years) underwent 3 h of controlled exposure (7:30–10:30 PM; active or sham) prior to sleep, followed by a full‐night polysomnographic recording in a sleep laboratory. The results demonstrated that following exposure, time in Stages 3 and 4 sleep (SWS, slow‐wave sleep) decreased by 9.5 min (12%) out of a total of 78.6 min, and time in Stage 2 sleep increased by 8.3 min (4%) out of a total of 196.3 min compared to sham. The latency to Stage 3 sleep was also prolonged by 4.8 min after exposure. Power density analysis indicated an enhanced activation in the frequency ranges 0.5–1.5 and 5.75–10.5 Hz during the first 30 min of Stage 2 sleep, with 7.5–11.75 Hz being elevated within the first hour of Stage 2 sleep, and bands 4.75–8.25 Hz elevated during the second hour of Stage 2 sleep. No pronounced power changes were observed in SWS or for the third hour of scored Stage 2 sleep. No differences were found between controls and subjects with prior complaints of mobile phone‐related symptoms. The results confirm previous findings that RF exposure increased the EEG alpha range in the sleep EEG, and indicated moderate impairment of SWS. Furthermore, reported differences in sensitivity to mobile phone use were not reflected in sleep parameters. Bioelectromagnetics 32:4–14, 2011. © 2010 Wiley‐Liss, Inc.     

Confirmation studies of Soviet research on immunological effects of microwaves: Russian immunology results

This paper presents the results of a replication study performed to investigate earlier Soviet studies conducted between 1974 and 1991 that showed immunological and reproductive effects of long‐term low‐level exposure of rats to radiofrequency (RF) electromagnetic fields. The early studies were used, in part, for developing exposure standards for the USSR population and thus it was necessary to confirm the Russian findings. In the present study, the conditions of RF exposure were made as similar as possible to those in the earlier experiments: Wistar rats were exposed in the far field to 2450 MHz continuous wave RF fields with an incident power density in the cages of 5 W/m² for 7 h/day, 5 days/week for a total of 30 days, resulting in a whole‐body SAR of 0.16 W/kg. Effects of the exposure on immunological parameters in the brain and liver of rats were evaluated using the complement fixation test (CFT), as in the original studies, and an additional test, the more modern ELISA test. Our results, using CFT and ELISA, partly confirmed the findings of the early studies and indicated possible effects from non‐thermal RF exposure on autoimmune processes. The RF exposure resulted in minor increases in formation of antibodies in brain tissue extract and the exposure did not appear to be pathological. In addition, a study was conducted to replicate a previous Soviet study on effects from the injection of blood serum from RF‐exposed rats on pregnancy and foetal and offspring development of rats, using a similar animal model and protocol. Our results showed the same general trends as the earlier study, suggesting possible adverse effects of the blood serum from exposed rats on pregnancy and foetal development of intact rats, however, application of these results in developing exposure standards is limited. Bioelectromagnetics 31:589–602, 2010. © 2010 Wiley‐Liss, Inc.     

Rhesus monkey behavior during exposure to high-peak-power 5.62-GHz microwave pulses

Limits on the exposure to high-peak-power, short-duration microwave pulses have only recently been adopted. Additional data, however, are needed to understand the effects that may be produced by exposure to high-peak-power pulsed microwaves. Four male rhesus monkeys (Macaca mulatta) were trained on an operant task for food pellet reward to investigate the behavioral effects of very high-peak-power 5.62 GHz microwaves. The operant task required monkeys to pull one plastic lever on a variable interval schedule (VI-25 s) and then respond to color signals and pull a second lever to obtain food. The monkeys were conditioned to perform a color discrimination task using one of three colors displayed by a fiber-optic cable. A red signal was the discriminative stimulus for responding on the first lever. A response on the second lever when a green signal was presented (1 s duration) delivered a food pellet. If a response on the second lever was made in the presence of a white signal, a 30-s timeout occurred. While performing the behavioral task, the monkeys were exposed to microwave pulses produced by either a military radar (FPS-26A) operating at 5.62 GHz or the same radar coupled to a Stanford linear energy doubler (SLED) pulse-forming device (ITT-2972) that enhanced peak power by a factor of nine by adding a high power pulse to the radar pulse. The effects of both types of pulses were compared to sham exposure. Peak field power densities tested were 518, 1270, and 2520 W/cm² for SLED pulses and 56, 128, and 277 W/cm² for the radar pulses. The microwave pulses (radar or SLED) were delivered at 100 pps (2.8 μs radar pulse duration, ≈ 50 ns SLED pulse duration) for 20 min and produced averaged whole-body SARs of 2,4, or 6 W/kg. Compared to sham exposures, significant alterations of lever responding, reaction time, and earned food pellets occurred during microwave exposure at 4 and 6 W/kg but not 2 W/kg. There were no differences between radar or SLED pulses in producing behavioral effects. ©1994 Wiley-Liss, Inc.

1 This is a US Government work and, as such, is in the public domain in the United States of America.

     

Ultra-wide band electromagnetic radiation does not affect UV-induced recombination and mutagenesis in yeast

Cell samples of the yeast Saccharomyces cerevisiae were exposed to 100 J/m² of 254 nm ultraviolet (UV) radiation followed by a 30 min treatment with ultra-wide band (UWB) electromagnetic pulses. The UWB pulses (101–104 kV/m, 1.0 ns width, 165 ps rise time) were applied at the repetition rates of 0 Hz (sham), 16 Hz, or 600 Hz. The effect of exposures was evaluated from the colony-forming ability of the cells on complete and selective media and the number of aberrant colonies. The experiments established no effect of UWB exposure on the UV-induced reciprocal and non-reciprocal recombination, mutagenesis, or cell survival. Bioelectromagnetics 19: 128–130, 1998. © 1998 Wiley-Liss, Inc.     

Dependence of anti-inflammatory effects of high peak-power pulsed electromagnetic radiation of extremely high frequency on exposure parameters

A pronounced anti-inflammatory effect of high peak-power pulsed electromagnetic radiation of extremely high frequency was shown for the first time in a model of zymosan-induced footpad edema in mice. Exposure to radiation of specific parameters (35, 27 GHz, peak power 20 kW, pulse widths 400-600 ns, pulse repetition frequency 5-500 Hz) decreased the exudative edema and local hyperthermia by 20% compared to the control. The kinetics and the magnitude of the anti-inflammatory effect were comparable with those induced by sodium diclofenac at a dose of 3 mg/kg. It was found that the anti-inflammatory effect linearly increased with increasing pulse width at a fixed pulse repetition frequency and had threshold dependence on the average incident power density of the radiation at a fixed pulse width. When animals were whole-body exposed in the far-field zone of radiator, the optimal exposure duration was 20 min. Increasing the average incident power density upon local exposure of the inflamed paw accelerated both the development of the anti-inflammatory effect and the reactivation time. The results obtained will undoubtedly be of great importance in the hygienic standardization of pulsed electromagnetic radiation and in further studies of the mechanisms of its biological action.     

Lack of direct DNA damage in human blood leukocytes and lymphocytes after in vitro exposure to high power microwave pulses

Currently, the potential genotoxicity of high power microwave pulses (HPMP) is not clear. Using the alkaline single cell gel electrophoresis assay, also known as the alkaline comet assay, we studied the effects of HPMP (8.8 GHz, 180 ns pulse width, peak power 65 kW, pulse repetition frequency 50 Hz) on DNA of human whole-blood leukocytes and isolated lymphocytes. The cell suspensions were exposed to HPMP for 40 min in a rectangular waveguide. The average SAR calculated from the temperature kinetics was about 1.6 kW/kg (peak SAR was about 300 MW/kg). The steady-state temperature rise in the 50 microl samples exposed to HPMP was 3.5 +/- 0.1 degrees C. In independent experiments, we did not find any statistically significant DNA damage manifested immediately after in vitro HPMP exposure of human blood leukocytes or lymphocytes or after HPMP exposure of leukocytes subsequently incubated at 37 degrees C for 30 min. Our results indicate that HPMP under the given exposure conditions did not induce DNA strand breaks, alkali-labile sites, and incomplete excision repair sites, which could be detected by the alkaline comet assay.     

Evaluation of Viral Inactivation on Dry Surface by High Peak Power Microwave (HPPM) Exposure

Previous research has shown that virus infectivity can be dramatically reduced by radio frequency exposure in the gigahertz (GHz) frequency range. Given the worldwide SARS-CoV-2 pandemic, which has caused over 1 million deaths and has had a profound global economic impact, there is a need for a noninvasive technology that can reduce the transmission of virus among humans. RF is a potential wide area-of-effect viral decontamination technology that could be used in hospital rooms where patients are expelling virus, in grocery and convenience stores where local populations mix, and in first responder settings where rapid medical response spans many potentially infected locations within hours. In this study, we used bovine coronavirus (BCoV) as a surrogate of SARS-CoV-2 and exposed it to high peak power microwave (HPPM) pulses at four narrowband frequencies: 2.8, 5.6, 8.5, and 9.3 GHz. Exposures consisted of 2 µs pulses delivered at 500 Hz, with pulse counts varied by decades between 1 and 10,000. The peak field intensities (i.e. the instantaneous power density of each pulse) ranged between 0.6 and 6.5 MW/m², depending on the microwave frequency. The HPPM exposures were delivered to plastic coverslips containing BCoV dried on the surface. Hemagglutination (HA) and cytopathic effect analyses were performed 6 days after inoculation of host cells to assess viral infectivity. No change in viral infectivity was seen with increasing dose (pulse number) across the tested frequencies. Under all conditions tested, exposure did not reduce infectivity more than 1.0 log_10. For the conditions studied, high peak power pulsed RF exposures in the 2–10 GHz range appear ineffective as a virucidal approach for hard surface decontamination. © 2023 Bioelectromagnetics Society.