Application of a temperature‐dependent fluorescent dye (Rhodamine B) to the measurement of radiofrequency radiation‐induced temperature changes in biological samples

We have applied a non‐contact method for studying the temperature changes produced by radiofrequency (RF) radiation specifically to small biological samples. A temperature‐dependent fluorescent dye, Rhodamine B, as imaged by laser scanning confocal microscopy (LSCM) was used to do this. The results were calibrated against real‐time temperature measurements from fiber optic probes, with a calibration factor of 3.4% intensity change °C^?1 and a reproducibility of ±6%. This non‐contact method provided two‐dimensional and three‐dimensional images of temperature change and distributions in biological samples, at a spatial resolution of a few micrometers and with an estimated absolute precision of around 1.5 °C, with a differential precision of 0.4 °C. Temperature rise within tissue was found to be non‐uniform. Estimates of specific absorption rate (SAR) from absorbed power measurements were greater than those estimated from rate of temperature rise, measured at 1 min intervals, probably because this interval is too long to permit accurate estimation of initial temperature rise following start of RF exposure. Future experiments will aim to explore this. Bioelectromagnetics 30:583–590, 2009. © 2009 Wiley‐Liss, Inc.     

Proposal of 28 GHz In Vitro Exposure System Based on Field Uniformity for Three‐Dimensional Cell Culture Experiments

This paper proposes a novel in vitro exposure system operating at millimeter‐wave (mmWave) 28 GHz, one of the frequency bands under consideration for fifth generation (5G) communication. We employed the field uniformity concept along cross‐sectional observation planes at shorter distances from the radiation antenna for better efficiency and a small‐size system. A choke‐ring antenna was designed for this purpose in consideration of a wider beamwidth (BW) and a symmetric far‐field pattern across three principal planes. The permittivity of Dulbecco's modified Eagle's medium solution was measured to examine the specific absorption rate (SAR) of the skin cell layer inside a Petri dish model for a three‐dimensional (3D) cell culture in vitro experiment. The best deployment of Petri dishes, taking into account a geometrical field symmetry, was proposed. Local SAR values within the cell layer among the Petri dishes were determined with different polarization angles. It was determined that this polarization effect should be considered when the actual exposure and deployment were conducted. We finally proposed an in vitro exposure system based on the field uniformity including downward exposure from an antenna for 3D cell culture experiments. A small‐size chamber system was obtained, and the size was estimated using the planar near‐field chamber design rule. Bioelectromagnetics. 2019;40:445–457. © 2019 Bioelectromagnetics Society     

Head exposure system for a human provocation study to assess the possible influence of UMTS‐like electromagnetic fields on cerebral blood circulation using near‐infrared imaging

A head exposure setup for efficient and precisely defined exposure of human subjects equipped with a near‐infrared imaging (NIRI) sensor is presented. In a partially shielded anechoic chamber the subjects were exposed to Universal Mobile Telecommunications System (UMTS)‐like electromagnetic fields (EMF) by using a patch antenna at a distance of 4 cm from the head. The non‐contact design of the exposure setup enabled NIRI sensors to easily attach to the head. Moreover, different regions of the head were chosen for localised exposure and simultaneous NIRI investigation. The control software enabled the simple adaptation of the test parameters during exploratory testing as well as the performance of controlled, randomised, crossover and double‐blind provocation studies. Four different signals with a carrier frequency of 1900 MHz were chosen for the exposure: a simple continuous wave signal and three different UMTS signals. Furthermore, three exposure doses were available: sham, low (spatial peak specific absorption rate (SAR) = 0.18 W/kg averaged over 10 g) and high (spatial peak SAR = 1.8 W/kg averaged over 10 g). The SAR assessment was performed by measurement and simulation. Direct comparison of measurement and numerical results showed good agreement in terms of spatial peak SAR and SAR distribution. The variability analysis of the spatial peak SAR over 10 g was assessed by numerical simulations. Maximal deviations of ?22% and +32% from the nominal situation were observed. Compared to other exposure setups, the present setup allows for low exposure uncertainty, combined with high SAR efficiency, easy access for the NIRI sensor and minimal impairment of test subjects. Bioelectromagnetics 33:124–133, 2012. © 2011 Wiley Periodicals, Inc.     

Early‐Life Exposure to Pulsed LTE Radiofrequency Fields Causes Persistent Changes in Activity and Behavior in C57BL/6 J Mice

Despite much research, gaps remain in knowledge about the potential health effects of exposure to radiofrequency (RF) fields. This study investigated the effects of early‐life exposure to pulsed long term evolution (LTE) 1,846 MHz downlink signals on innate mouse behavior. Animals were exposed for 30 min/day, 5 days/week at a whole‐body average specific energy absorption rate (SAR) of 0.5 or 1 W/kg from late pregnancy (gestation day 13.5) to weaning (postnatal day 21). A behavioral tracking system measured locomotor, drinking, and feeding behavior in the home cage from 12 to 28 weeks of age. The exposure caused significant effects on both appetitive behaviors and activity of offspring that depended on the SAR. Compared with sham‐exposed controls, exposure at 0.5 W/kg significantly decreased drinking frequency (P ≤ 0.000) and significantly decreased distance moved (P ≤ 0.001). In contrast, exposure at 1 W/kg significantly increased drinking frequency (P ≤ 0.001) and significantly increased moving duration (P ≤ 0.005). In the absence of other plausible explanations, it is concluded that repeated exposure to low‐level RF fields in early life may have a persistent and long‐term effect on adult behavior. Bioelectromagnetics. 2019;40:498–511. © 2019 The Authors. Bioelectromagnetics Published by Wiley Periodicals, Inc.     

Effects of 171 MHz Low‐Intensity Electromagnetic Field on Glucocorticoid and Mineral Corticoid Activity of the Adrenal Glands of Rats

A sub‐acute electromagnetic field (EMF) biological effect study was carried out on rats exposed in the Transverse ElectroMagnetic exposure chamber at 171 MHz Continuous Wave (CW). The experiments involved three exposure levels (15, 25, and 35 V/m) for 15 days with triplicate parallel sham‐exposed controls in each series. All exposure conditions were simulated for the evaluation of the electromagnetic energy distribution and specific absorption rate (SAR) in the rat phantoms. Studies have shown a biphasic biological response depending on time and absorbed electromagnetic energy. Under low SAR, approximately 0.006 W/kg, EMF exposure leads to the stimulation of adrenal gland activity. This process is accompanied by an initial increase of daily excretion of corticosterone and Na⁺, which is seen as a higher Na⁺/K⁺ ratio, followed by a decrease of these parameters over time. It is possible that EMF exposure causes a stress response in animals, which is seen as an increased adrenal activity. Bioelectromagnetics. 2019;40:578–587. © 2019 Bioelectromagnetics Society.     

Electromagnetic and thermal evaluation of an applicator specialized to permit high‐resolution non‐perturbing optical evaluation of cells being irradiated in the W‐band

To permit epi‐illuminated, high‐resolution optical microscopy of cells in monolayer culture during unperturbed W‐band (75–110 GHz) irradiation, a new class of applicator has been developed based upon WR10 rectangular waveguide components: the cells are normally plated onto the underside of a coverslip which is then placed against the under side of a waveguide flange and receives a roughly circular exposure pattern, with the ±1 dB central spot roughly 1 mm in diameter. Constructed and tested with 94 GHz millimeter waves, water‐immersion optics, and free‐convection cooling, the applicator works robustly and permits SARs at the cell layer as high as 4500 W/kg before the steady‐state temperature rise at the cell layer exceeds 0.5 K. Bioelectromagnetics 31:140–149, 2010. © 2009 Wiley‐Liss, Inc.     

Numerical modeling of heat and mass transfer in the human eye under millimeter wave exposure

Human exposure to millimeter wave (MMW) radiation is expected to increase in the next several years. In this work, we present a thermal model of the human eye under MMW illumination. The model takes into account the fluid dynamics of the aqueous humor and predicts a frequency‐dependent reversal of its flow that also depends on the incident power density. The calculated maximum fluid velocity in the anterior chamber and the temperature rise at the corneal apex are reported for frequencies from 40 to 100 GHz and different values of incident power density. Bioelectromagnetics 34:291–299, 2013. © 2012 Wiley Periodicals, Inc.     

Statistical multi‐path exposure method for assessing the whole‐body SAR in a heterogeneous human body model in a realistic environment

Assessing the whole‐body absorption in a human in a realistic environment requires a statistical approach covering all possible exposure situations. This article describes the development of a statistical multi‐path exposure method for heterogeneous realistic human body models. The method is applied for the 6‐year‐old Virtual Family boy (VFB) exposed to the GSM downlink at 950 MHz. It is shown that the whole‐body SAR does not differ significantly over the different environments at an operating frequency of 950 MHz. Furthermore, the whole‐body SAR in the VFB for multi‐path exposure exceeds the whole‐body SAR for worst‐case single‐incident plane wave exposure by 3.6%. Moreover, the ICNIRP reference levels are not conservative with the basic restrictions in 0.3% of the exposure samples for the VFB at the GSM downlink of 950 MHz. The homogeneous spheroid with the dielectric properties of the head suggested by the IEC underestimates the absorption compared to realistic human body models. Moreover, the variation in the whole‐body SAR for realistic human body models is larger than for homogeneous spheroid models. This is mainly due to the heterogeneity of the tissues and the irregular shape of the realistic human body model compared to homogeneous spheroid human body models. Bioelectromagnetics 34:240–251, 2013. © 2012 Wiley Periodicals, Inc.     

Morphological Changes Induced By Extremely Low‐Frequency Electric Fields

In this paper, morphological effects of electric fields on avian erythrocytes (nucleated red blood cells) have been studied in detail. Morphological changes include rounding and cytoplasm transparency. It has been shown that the effect is non‐thermal. Careful imaging and image analyses have been carried out to show that the degree of this effect is frequency‐dependent, and has a higher conversion rate at higher temperatures. Furthermore, to better understand the mechanisms behind the morphological changes, we investigated the dedifferentiation hypothesis and performed a series of tests on avian erythrocytes including fluorescence spectroscopy for hemoglobin, and tests on human umbilical cord blood, mesenchymal stem cells, and bone marrow mesenchymal stem cells including flow‐cytometry analysis for expression of certain markers and calcium staining. Bioelectromagnetics. 2019;40:375–390. © 2019 Bioelectromagnetics Society.     

The effect of a 94 GHz electromagnetic field on neuronal microtubules

Hardware that generates electromagnetic waves with wavelengths from 1 to 10 mm (millimeter waves, “MMW”) is being used in a variety of applications, including high‐speed data communication and medical devices. This raises both practical and fundamental issues concerning the interaction of MMW electromagnetic fields (EMF) with biological tissues. A 94 GHz EMF is of particular interest because a number of applications, such as active denial systems, rely on this specific frequency. Most of the energy associated with MMW radiation is absorbed in the skin and, for a 94 GHz field, the power penetration depth is shallow (≈0.4 mm). At sufficiently high energies, skin heating is expected to activate thermal pain receptors, leading to the perception of pain. In addition to this “thermal” mechanism of action, a number of “non‐thermal” effects of MMW fields have been previously reported. Here, we investigated the influence of a 94 GHz EMF on the assembly/disassembly of neuronal microtubules in Xenopus spinal cord neurons. We reasoned that since microtubule array is regulated by a large number of intracellular signaling cascades, it may serve as an exquisitely sensitive reporter for the biochemical status of neuronal cytoplasm. We found that exposure to 94 GHz radiation increases the rate of microtubule assembly and that this effect can be entirely accounted for by the rapid EMF‐elicited temperature jump. Our data are consistent with the notion that the cellular effects of a 94 GHz EMF are mediated entirely by cell heating. Bioelectromagnetics 34:133–144, 2013. © 2012 Wiley Periodicals, Inc.     

Effects of gestational exposure to 1.95‐GHz W‐CDMA signals for IMT‐2000 cellular phones: Lack of embryotoxicity and teratogenicity in rats

The present study was designed to evaluate whether gestational exposure to an EMF targeting the head region, similar to that from cellular phones, might affect embryogenesis in rats. A 1.95‐GHz wide‐band code division multiple access (W‐CDMA) signal, which is one applied for the International Mobile Telecommunication 2000 (IMT‐2000) system and used for the freedom of mobile multimedia access (FOMA), was employed for exposure to the heads of four groups of pregnant CD(SD) IGS rats (20 per group) for gestational days 7–17. The exposure was performed for 90 min/day in the morning. The spatial average specific absorption rate (SAR) for individual brains was designed to be 0.67 and 2.0 W/kg with peak brain SARs of 3.1 and 7.0 W/kg for low (group 3) and high (group 4) exposures, respectively, and a whole‐body average SAR less than 0.4 W/kg so as not to cause thermal effects due to temperature elevation. Control and sham exposure groups were also included. At gestational day 20, all dams were killed and fetuses were taken out by cesarean section. There were no differences in maternal body weight gain. No adverse effects of EMF exposure were observed on any reproductive and embryotoxic parameters such as number of live (243–271 fetuses), dead or resorbed embryos, placental weights, sex ratios, weights or external, visceral or skeletal abnormalities of live fetuses. Bioelectromagnetics 30:205–212, 2009. © 2008 Wiley‐Liss, Inc.     

Near-field dosimetry for in vitro exposure of human cells at 60 GHz

Due to the expected mass deployment of millimeter‐wave wireless technologies, thresholds of potential millimeter‐wave‐induced biological and health effects should be carefully assessed. The main purpose of this study is to propose, optimize, and characterize a near‐field exposure configuration allowing illumination of cells in vitro at 60 GHz with power densities up to several tens of mW/cm². Positioning of a tissue culture plate containing cells has been optimized in the near‐field of a standard horn antenna operating at 60 GHz. The optimal position corresponds to the maximal mean‐to‐peak specific absorption rate (SAR) ratio over the cell monolayer, allowing the achievement of power densities up to 50 mW/cm² at least. Three complementary parameters have been determined and analyzed for the exposed cells, namely the power density, SAR, and temperature dynamics. The incident power density and SAR have been computed using the finite‐difference time‐domain (FDTD) method. The temperature dynamics at different locations inside the culture medium are measured and analyzed for various power densities. Local SAR, determined based on the initial rate of temperature rise, is in a good agreement with the computed SAR (maximal difference of 5%). For the optimized exposure setup configuration, 73% of cells are located within the ±3 dB region with respect to the average SAR. It is shown that under the considered exposure conditions, the maximal power density, local SAR, and temperature increments equal 57 mW/cm², 1.4 kW/kg, and 6 °C, respectively, for the radiated power of 425 mW. Bioelectromagnetics 33:55–64, 2012. © 2011 Wiley Periodicals, Inc.     

Novel ETV6‐RUNX1 Mouse Model to Study the Role of ELF‐MF in Childhood B‐Acute Lymphoblastic Leukemia: a Pilot Study

Exposure to extremely low‐frequency magnetic fields (ELF‐MFs) has been classified by the International Agency for Research on Cancer (IARC) as “possibly carcinogenic to humans,” based on limited scientific evidence concerning childhood leukemia. This assessment emphasized the lack of appropriate animal models recapitulating the natural history of this disease. Childhood B‐cell acute lymphoblastic leukemia (B‐ALL) is the result of complex interactions between genetic susceptibility and exposure to exogenous agents. The most common chromosomal alteration is the ETV6‐RUNX1 fusion gene, which confers a low risk of developing the malignancy by originating a preleukemic clone requiring secondary hits for full‐blown disease to appear. To develop potential prophylactic interventions, we need to identify the environmental triggers of the second hit. Recently, we generated a B‐ALL mouse model of the human ETV6‐RUNX1⁺ preleukemic state. Here, we present the results from the ARIMMORA pilot study, obtained by exposing 34 Sca1‐ETV6‐RUNX1 mice (vs. 27 unexposed) to a 50 Hz magnetic field of 1.5 mT with both fundamental and harmonic content, with an on/off cycle of 10 min/5 min, for 20 h/day, from conception until 3 months of age. Mice were monitored until 2 years of age and peripheral blood was periodically analyzed by flow cytometry. One of the exposed mice developed B‐ALL while none of the non‐exposed did. Although the results are statistically non‐significant due to the limited number of mice used in this pilot experiment, overall, the results show that the newly developed Sca1‐ETV6‐RUNX1 mouse can be successfully used for ELF‐MF exposure studies about the etiology of childhood B‐ALL. Bioelectromagnetics. 2019;40:343–353. © 2019 Bioelectromagnetics Society.     

Effect of Exposure to a Radiofrequency Electromagnetic Field on Body Temperature in Anesthetized and Non-Anesthetized Rats

Exposure to a radiofrequency (RF) signal at a specific absorption rate (SAR) of 4 W/kg can increase the body temperature by more than 1 °C. In this study, we investigated the effect of anesthesia on the body temperature of rats after exposure to an RF electromagnetic field at 4 W/kg SAR. We also evaluated the influence of body mass on rats’ body temperature. Rats weighing 225 and 339 g were divided into sham- and RF-exposure groups. Each of the resulting four groups was subdivided into anesthetized and non-anesthetized groups. The free-moving rats in the four RF-exposure groups were subjected to a 915 MHz RF identification signal at 4 W/kg whole-body SAR for 8 h. The rectal temperature was measured at 1-h intervals during RF exposure using a small-animal temperature probe. The body temperatures of non-anesthetized, mobile 225 and 339 g rats were not significantly affected by exposure to an RF signal. However, the body temperatures of anesthetized 225 and 339 g rats increased by 1.9 °C and 3.3 °C from baseline at 5 and 6 h of RF exposure, respectively. Three of the five 339 g anesthetized and exposed rats died after 6 h of RF exposure. Thus, anesthesia and body mass influenced RF exposure-induced changes in the body temperature of rats. Bioelectromagnetics. 2020;41:104–112. © 2019 Bioelectromagnetics Society.     

Design,implementation, and dosimetry analysis of an S‐band waveguide in vitro system for the exposure of cell culture samples to pulsed fields

The interaction between electromagnetic fields and biological media, particularly regarding very high power, short pulses as in radar signals, is not a fully understood phenomenon. In the past few years, many in vitro, cellular communications‐oriented exposure studies have been carried out. This article presents a high‐power waveguide exposure system capable of dealing with monochromatic, multicarrier or pulsed signals between 1.8 and 3.2 GHz (L‐ and S‐band) with a pulse duration as low as 90 ns, minimum pulse repetition of 100 Hz, and maximum instantaneous power of 100 W. The setup is currently being used with a 2.2 GHz carrier modulated by 5 µs pulses with a 100 Hz repetition period and approximately 30 W of instantaneous power. After a worst‐case temperature analysis, which does not account for conduction and convection thermal effects, the experiment's exposure is considered sub‐thermal. Evaluation of the results through the specific absorption rate distribution is not considered sufficient enough in these cases. An electromagnetic field distribution analysis is needed. For monochromatic signals, the representation of the modulus of the electric and magnetic field components is proposed as a suitable method of assessment. Bioelectromagnetics 31:479–487, 2010. © 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.     

The relation between the specific absorption rate and electromagnetic field intensity for heterogeneous exposure conditions at mobile communications frequencies

The relation between the incident electromagnetic field strength and both the whole‐body and the local specific absorption rate (SAR) was investigated for typical heterogeneous exposure scenarios for frequencies relevant for mobile communication. The results were compared to results from plane wave exposure. Heterogeneous exposure arises from multiple path propagation of the electromagnetic waves to the location of interest. It is shown that plane wave exposure does not represent worst‐case exposure conditions. When the electric field strength arising at plane wave exposure is compared to the electric field strength averaged over the volume of the human body occurring during multipath exposure, 12% of all heterogeneous cases examined represent worse exposure conditions than plane wave exposure for whole‐body exposure at 946 MHz, 15% at 1840 MHz, and 22% at 2140 MHz. The deviation between plane wave and heterogeneous whole‐body SAR ranges from ?54% to 54%. For partial‐body SAR averaged over 10 g of tissue, a range from ?93% to 209% was found when comparing multiple wave exposure to single incoming plane waves. The investigations performed using the Visible Human as phantom showed that the basic restrictions are met as long as the reference levels are not exceeded. However, this must not be necessarily the case when different phantoms are used to perform similar investigations because recent studies demonstrated that reference levels might not be conservative when phantoms of children are used. Therefore, the results of this work indicate the need to extend the investigations to numerical simulations with additional human phantoms representing parts of the human population having different anatomy and morphology compared to the phantom used within the frame of this project. This also applies to phantoms of children. Bioelectromagnetics 30:651–662, 2009. © 2009 Wiley‐Liss, Inc.     

Melatonin Levels and Low‐Frequency Magnetic Fields in Humans and Rats: New Insights From a Bayesian Logistic Regression

The present analysis revisits the impact of extremely low‐frequency magnetic fields (ELF‐MF) on melatonin (MLT) levels in human and rat subjects using both a parametric and non‐parametric approach. In this analysis, we use 62 studies from review articles. The parametric approach consists of a Bayesian logistic regression (LR) analysis and the non‐parametric approach consists of a Support Vector analysis, both of which are robust against spurious/false results. Both approaches reveal a unique well‐ordered pattern, and show that human and rat studies are consistent with each other once the MF strength is restricted to cover the same range (with B ? 50 μT). In addition, the data reveal that chronic exposure (longer than ～22 days) to ELF‐MF appears to decrease MLT levels only when the MF strength is below a threshold of ~30 μT (), i.e., when the man‐made ELF‐MF intensity is below that of the static geomagnetic field. Studies reporting an association between ELF‐MF and changes to MLT levels and the opposite (no association with ELF‐MF) can be reconciled under a single framework. Bioelectromagnetics. 2019;40:539–552. © 2019 Bioelectromagnetics Society.