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.

Can Low-Level Exposure to Radiofrequency Fields Effect Cognitive Behaviour in Laboratory Animals? A Systematic Review of the Literature Related to Spatial Learning and Place Memory.

This review considers whether exposure to low-level radiofrequency (RF) fields, mostly associated with mobile phone technology, can influence cognitive behaviour of laboratory animals. Studies were nominated for inclusion using an a priori defined protocol with preselected criteria, and studies were excluded from analysis if they did not include sufficient details about the exposure, dosimetry or experimental protocol, or if they lacked a sham-exposed group. Overall, 62 studies were identified that have investigated the effects of RF fields on spatial memory and place learning and have been published since 1993. Of these, 17 studies were excluded, 20 studies reported no significant field-related effects, 21 studies reported significant impairments or deficits, and four studies reported beneficial consequences. The data do not suggest whether these outcomes are related to specific differences in exposure or testing conditions, or simply represent chance. However, some studies have suggested possible molecular mechanisms for the observed effects, but none of these has been substantiated through independent replication. Further behavioural studies could prove useful to resolve this situation, and it is suggested that these studies should use a consistent animal model with standardized exposure and testing protocols, and with detailed dosimetry provided by heterogeneous, anatomically-realistic animal models.

Effects of 50 Hz magnetic fields on circadian rhythm control in mice.

Artificial light and power frequency magnetic fields are ubiquitous in the built environment. Light is a potent zeitgeber but it is unclear whether power frequency magnetic fields can influence circadian rhythm control. To study this possibility, 8-12-week-old male C57BL/6J mice were exposed for 30 min starting at zeitgeber time 14 (ZT14, 2 h into the dark period of the day) to 50 Hz magnetic fields at 580 µT using a pair of Helmholtz coils and/or a blue LED light at 700 lux or neither. Our experiments revealed an acute adrenal response to blue light, in terms of increased adrenal per1 gene expression, increased serum corticosterone levels, increased time spent sleeping, and decreased locomotor activity (in all cases, P < 0.0001) compared to an unexposed control group. There appeared to be no modulating effect of the magnetic fields on the response to light, and there was also no effect of the magnetic fields alone (in both cases, P > 0.05) except for a decrease in locomotor activity (P < 0.03). Gene expression of the cryptochromes cry1 and cry2 in the adrenals, liver, and hippocampus was also not affected by exposures (in all cases, P > 0.05). In conclusion, these results suggest that 50 Hz magnetic fields do not significantly affect the acute light response to a degree that can be detected in the adrenal response. Bioelectromagnetics. 2019;9999:XX-XX. © 2019 Bioelectromagnetics Society.

Are Exposures to Multiple Frequencies the Key to Future Radiofrequency Research?

There is an extensive literature investigating possible effects of exposure to radiofrequency (RF) electromagnetic fields associated with mobile phone technologies. This has not identified any public health risks with any degree of certainty. Some epidemiological studies have observed associations between heavy users of mobile phones and some types of cancer, but animal studies do not support this association, although a few studies have reported increased tumor yields. However, there is a crucial difference between epidemiology studies and laboratory work in terms of signals investigated: most people are exposed to a complex mixture of frequencies and signals at varying intensities, whereas the majority of animal studies have been performed using a single frequency or intensity. Whether this might explain the differences in outcome will be discussed, and whether there is a need for additional laboratory investigations that reproduce more accurately realistic exposure conditions will be considered.

A Closer Look at the Thresholds of Thermal Damage: Workshop Report by an ICNIRP Task Group.

The International Commission on Non-Ionizing Radiation Protection issued guidelines in 1998 for limiting public and occupational exposure to radiofrequency electromagnetic fields (100 kHz to 300 GHz). As part of the process of updating this advice, a 2-d workshop titled "A closer look at the thresholds of thermal damage" was held from 26-28 May 2015 in Istanbul to re-examine the thermal basis of the guidelines and to provide further information on heat-related effects and thresholds of thermal damage. Overall, the workshop provided much useful information relevant to revision of the guidelines. Participants indicated that the effects of heating from radiofrequency fields are consistent with those from other sources, and that the information derived from those studies can be applied to radiofrequency-induced heating. Another conclusion was that absolute temperature of tissues was more important for thermal damage than temperature change. The discussion suggested that the 6-min averaging time used in international guidelines was valid for whole-body exposures but with a large uncertainty: 30 min may be a more appropriate averaging time for localized exposures, and less than 1 min for implanted medical devices. The duration of whole-body radiofrequency exposure is a critical parameter that often determines the effect threshold, but this will be affected by other, ongoing thermoregulation, which is dependant on many factors. The thresholds for localized radiofrequency exposure were difficult to determine because of the potential range of exposure conditions and the possibility of radiofrequency-induced local hotspots. Suggestions for future dose metrics and further research were discussed and are included in this report.

The rate of X-ray-induced DNA double-strand break repair in the embryonic mouse brain is unaffected by exposure to 50 Hz magnetic fields.

PURPOSE: Following in utero exposure to low dose radiation (10-200 mGy), we recently observed a linear induction of DNA double-strand breaks (DSB) and activation of apoptosis in the embryonic neuronal stem/progenitor cell compartment. No significant induction of DSB or apoptosis was observed following exposure to magnetic fields (MF). In the present study, we exploited this in vivo system to examine whether exposure to MF before and after exposure to 100 mGy X-rays impacts upon DSB repair rates. MATERIALS AND METHODS: 53BP1 foci were quantified following combined exposure to radiation and MF in the embryonic neuronal stem/progenitor cell compartment. Embryos were exposed in utero to 50 Hz MF at 300 µT for 3 h before and up to 9 h after exposure to 100 mGy X-rays. Controls included embryos exposed to MF or X-rays alone plus sham exposures. RESULTS: Exposure to MF before and after 100 mGy X-rays did not impact upon the rate of DSB repair in the embryonic neuronal stem cell compartment compared to repair rates following radiation exposure alone. CONCLUSIONS: We conclude that in this sensitive system MF do not exert any significant level of DNA damage and do not impede the repair of X-ray induced damage.

The effects of extremely low frequency magnetic fields on mutation induction in mice.

The growing human exposure to extremely low frequency (ELF) magnetic fields has raised a considerable concern regarding their genotoxic effects. The aim of this study was to evaluate the in vivo effects of ELF magnetic fields irradiation on mutation induction in the germline and somatic tissues of male mice. Seven week old BALB/c×CBA/Ca F1 hybrid males were exposed to 10, 100 or 300µT of 50Hz magnetic fields for 2 or 15h. Using single-molecule PCR, the frequency of mutation at the mouse Expanded Simple Tandem Repeat (ESTR) locus Ms6-hm was established in sperm and blood samples of exposed and matched sham-treated males. ESTR mutation frequency was also established in sperm and blood samples taken from male mice exposed to 1Gy of acute X-rays. The frequency of ESTR mutation in DNA samples extracted from blood of mice exposed to magnetic fields did not significantly differ from that in sham-treated controls. However, there was a marginally significant increase in mutation frequency in sperm but this was not dose-dependent. In contrast, acute exposure X-rays led to significant increases in mutation frequency in sperm and blood of exposed males. The results of our study suggest that, within the range of doses analyzed here, the in vivo mutagenic effects of ELF magnetic fields are likely to be minor if not negligible.

Increased apoptosis and DNA double-strand breaks in the embryonic mouse brain in response to very low-dose X-rays but not 50 Hz magnetic fields.

The use of X-rays for medical diagnosis is enhancing exposure to low radiation doses. Exposure to extremely low-frequency electromagnetic or magnetic fields is also increasing. Epidemiological studies show consistent associations of childhood leukaemia with exposure to magnetic fields but any causal relationship is unclear. A limitation in assessing the consequence of such exposure is the availability of sensitive assays. The embryonic neuronal stem and progenitor cell compartments are radiosensitive tissues. Using sensitive assays, we report a statistically significant increase in DNA double-strand break (DSB) formation and apoptosis in the embryonic neuronal stem cell compartment following in utero exposure to 10-200 mGy X-rays. Both endpoints show a linear response. We also show that DSB repair is delayed following exposure to doses below 50 mGy compared with 100 mGy. Thus, we demonstrate in vivo consequences of low-dose radiation. In contrast to these impacts, we did not observe any significant induction of DSBs or apoptosis following exposure to 50 Hz magnetic fields (100 or 300 µT). We conclude that any DSB induction by treatment with magnetic fields is lower than following exposure to 10 mGy X-rays. For comparison, certain procedures involving computed tomography scanning are equivalent to 1-5 mGy X-rays.

International workshop on non-ionizing radiation protection in medicine.

An international workshop brought together a range of stakeholders to consider protection from non-ionizing radiation used in medicine, research and cosmetics. Presentations on specific topics were followed by a general discussion on possible improvements in protection. Participants considered that adherence to science-based, harmonized exposure guidelines to limit exposures for clinical staff and other workers was a key prerequisite to safety in all situations. In addition, to engender an awareness of the risks involved to both the patient as well as the operator, equipment should be operated only by suitably qualified persons who have received appropriate training in the safe use of that device. This training should be carried out under the auspices of an accredited safety provider, and preferably offer a recognized qualification. Specific advice included the necessity for correct eye protection with higher power optical radiation sources, and avoiding the use of ultrasound for all exposures without medical benefit. Finally, the possibility of a harmonized approach to safety for both non-ionizing and ionizing radiation was considered worthy of further discussion.

Investigation of transcriptional responses of juvenile mouse bone marrow to power frequency magnetic fields.

To seek alterations in gene transcription in bone marrow cells following in vivo exposure of juvenile mice to power frequency magnetic fields, young (21-24-day old) C57BL/6 mice were exposed to a 100µT 50Hz magnetic field for 2h. Transcription was analysed by three methods, High Coverage Expression Profiling (HiCEP), Illumina microarrays and quantitative real-time polymerase chain reaction (QRT-PCR). A pilot HiCEP experiment with 6 exposed (E) and 6 non-exposed (NE) mice identified four candidate responsive transcripts (two unknown transcripts (AK152075 and F10-NED), phosphatidylinositol binding clathrin assembly protein (Picalm) and exportin 7 (Xpo7)). A larger experiment compared 19 E and 15 NE mice using two independent QRT-PCR assays and repeated microarray assays. No significant field-dependent changes were seen, although Picalm showed a trend to significance in one QRT-PCR assay (E/NE=0.91; P=0.06). However, the study was underpowered to detect an effect of this magnitude (52% power at P=0.05). These data indicate the current experimental constraints in detecting small changes in transcription that may occur in response to magnetic fields. These constraints result from technical limitations in the accuracy of assays and biological variation, which together were sufficient to account statistically for the number of differentially expressed transcripts identified in the pilot experiment.

Systematic review of wireless phone use and brain cancer and other head tumors.

We conducted a systematic review of scientific studies to evaluate whether the use of wireless phones is linked to an increased incidence of the brain cancer glioma or other tumors of the head (meningioma, acoustic neuroma, and parotid gland), originating in the areas of the head that most absorb radiofrequency (RF) energy from wireless phones. Epidemiology and in vivo studies were evaluated according to an agreed protocol; quality criteria were used to evaluate the studies for narrative synthesis but not for meta-analyses or pooling of results. The epidemiology study results were heterogeneous, with sparse data on long-term use (≥ 10 years). Meta-analyses of the epidemiology studies showed no statistically significant increase in risk (defined as P < 0.05) for adult brain cancer or other head tumors from wireless phone use. Analyses of the in vivo oncogenicity, tumor promotion, and genotoxicity studies also showed no statistically significant relationship between exposure to RF fields and genotoxic damage to brain cells, or the incidence of brain cancers or other tumors of the head. Assessment of the review results using the Hill criteria did not support a causal relationship between wireless phone use and the incidence of adult cancers in the areas of the head that most absorb RF energy from the use of wireless phones. There are insufficient data to make any determinations about longer-term use (≥ 10 years).

Are the young more sensitive than adults to the effects of radiofrequency fields? An examination of relevant data from cellular and animal studies.

It has sometimes been assumed that children are more sensitive than adults to the effects of radiofrequency (RF) fields associated with cellular wireless telephones. However, relatively few in vitro or animal models have examined this possibility. In vitro studies have used several cell types, from both humans and rodents, including primary cells, embryonic cell lines, undifferentiated cancer cell lines, and stem cells. Overall, the balance of evidence does not suggest that field-related effects occur in any cell type: gene and protein expression were not significantly changed by exposure in nine out of 15 studies; genotoxicity was evaluated in 13 papers and in most, of these studies, no damage to DNA was detected; eight studies failed to demonstrate induction of apoptosis; and three studies reported lack of oxidative stress induction by RF-exposures. Five of eight studies investigating the effects of combined exposures to RF fields and chemical or physical agents reported a lack of field-related effects. In addition, few papers have been published on the effects of low level exposure of immature animals. The available results are very limited, both in terms of signals used and biological endpoints investigated, but the evidence does not indicate that prenatal or early postnatal exposures are associated with acute adverse responses or the development of detrimental changes in the long-term. Overall, this suggests that young animals may not be significantly more sensitive than adults, but there is clearly a need for further studies to be carried out.

A precautionary public health protection strategy for the possible risk of childhood leukaemia from exposure to power frequency magnetic fields.

BACKGROUND: Epidemiological evidence showing a consistent association between the risk of childhood leukaemia and exposure to power frequency magnetic fields has been accumulating. This debate considers the additional precautionary intervention needed to manage this risk, when it exceeds the protection afforded by the exposure guidelines as recommended by the International Commission on Non-Ionizing Radiation Protection. METHODS: The Bradford-Hill Criteria are guidelines for evaluating the scientific evidence that low frequency magnetic fields cause childhood leukaemia. The criteria are used for assessing the strength of scientific evidence and here have been applied to considering the strength of evidence that exposures to extremely low frequency magnetic fields may increase the risk of childhood leukaemia. The applicability of precaution is considered using the risk management framework outlined in a European Commission (EC) communication on the Precautionary Principle. That communication advises that measures should be proportionate, non-discriminatory, consistent with similar measures already taken, based on an examination of the benefits and costs of action and inaction, and subject to review in the light of new scientific findings. RESULTS: The main evidence for a risk is an epidemiological association observed in several studies and meta-analyses; however, the number of highly exposed children is small and the association could be due to a combination of selection bias, confounding and chance. Corroborating experimental evidence is limited insofar as there is no clear indication of harm at the field levels implicated; however, the aetiology of childhood leukaemia is poorly understood. Taking a precautionary approach suggests that low-cost intervention to reduce exposure is appropriate. This assumes that if the risk is real, its impact is likely to be small. It also recognises the consequential cost of any major intervention. The recommendation is controversial in that other interpretations of the data are possible, and low-cost intervention may not fully alleviate the risk. CONCLUSIONS: The debate shows how the EC risk management framework can be used to apply the Precautionary Principle to small and uncertain public health risks. However, despite the need for evidence-based policy making, many of the decisions remain value driven and therefore subjective.

Absence of nonlinear responses in cells and tissues exposed to RF energy at mobile phone frequencies using a doubly resonant cavity.

A doubly resonant cavity was used to search for nonlinear radiofrequency (RF) energy conversion in a range of biological preparations, thereby testing the hypothesis that living tissue can demodulate RF carriers and generate baseband signals. The samples comprised high-density cell suspensions (human lymphocytes and mouse bone marrow cells); adherent cells (IMR-32 human neuroblastoma, G361 human melanoma, HF-19 human fibroblasts, N2a murine neuroblastoma (differentiated and non-differentiated) and Chinese hamster ovary (CHO) cells) and thin sections or slices of mouse tissues (brain, kidney, muscle, liver, spleen, testis, heart and diaphragm). Viable and non-viable (heat killed or metabolically impaired) samples were tested. Over 500 cell and tissue samples were placed within the cavity, exposed to continuous wave (CW) fields at the resonant frequency (f) of the loaded cavity (near 883 MHz) using input powers of 0.1 or 1 mW, and monitored for second harmonic generation by inspection of the output at 2f. Unwanted signals were minimised using low pass filters (≤ 1 GHz) at the input to, and high pass filters (≥ 1 GHz) at the output from, the cavity. A tuned low noise amplifier allowed detection of second harmonic signals above a noise floor as low as -169 dBm. No consistent second harmonic of the incident CW signals was detected. Therefore, these results do not support the hypothesis that living cells can demodulate RF energy, since second harmonic generation is the necessary and sufficient condition for demodulation.

Rapporteur report: Roundup, discussion and recommendations.

There is a relative paucity of recent information regarding the long-term health effects associated with exposure to ultrasound, and to infrasound and low-frequency noise (LFN). For ultrasound, further epidemiological studies are recommended, and priority should be given to studies investigating the effects of handedness and to studies assessing possible subtle effects on brain function. These studies should reflect contemporary practises in diagnostic ultrasound and have sufficiently long follow-up periods to examine the possibility of effects into late adolescence or beyond. In the absence of a non-exposed control group, it would be advisable to make comparisons between a highly exposed group with a less exposed group, and to compare groups exposed at differing gestational stages. The effects associated with ultrasound contrast agents should also be studied, and the appropriateness of the thermal index (TI) and mechanical index (MI) should be reviewed. It is recommended that animal models should be used to investigate the effects of exposure at differing gestational ages on development, and modern cellular and molecular techniques used to investigate potential mechanisms of interaction. Although explicit morphological changes have been reported following occupational and experimental exposures to infrasound and low LFN, it was recommended that a thorough review of the relevant biological and health effects literature was necessary before specific proposals could be made. Uncertainties about the characterisation of these low frequencies also indicated the need to develop appropriate measurement techniques and protocols.

Neurobehavioural effects of electromagnetic fields.

Very few laboratory studies in children have explored the effects of exposure to low level electromagnetic fields (EMFs) on neurobehavioural function. Studies investigating effect on neurotransmitters, cognitive function and brain activity in adults and animals indicate that acute exposure to EMFs does not appear to engender any consistent physiological or behavioural impairment although a few subtle effects may occur. This suggests that exposure of children to low level EMFs may not cause significant detrimental effects on brain function. However the available evidence is not sufficient to draw any definite conclusions, and further laboratory studies are required. In particular, experiments investigating the effects of radiofrequency (RF) fields on the performance of well-characterised cognitive and behavioural tasks by immature and developing animals are recommended, if studies with children cannot be performed for ethical and practical reasons.

Rapporteur report: implications for exposure guidelines.

There is a paucity of information regarding the long-term health effects associated with exposure to static magnetic fields. Perceptual and other acute effects have been demonstrated above a threshold of about 2 T, and these form the basis for human exposure standards at present. Exposures well above this threshold are increasingly becoming more common as the technology associated with magnetic resonance imaging advances. Therefore, priority should be given to assessing the health risks associated with exposures to such fields. Studies should include a prospective cohort study investigating cancer risks of workers and patients exposed to fields in excess of 2 T, a study investigating effects on human cognitive performance from repeated exposures, and a molecular biology study investigating acute changes in genomic responses in volunteers exposed to fields of up to 8 T. Studies investigating the effects of long-term exposure on cancer, and on neurobehavioural development are also recommended using animals, where the use of transgenic models is encouraged. In addition, dosimetric studies should be conducted using high-resolution male, female and pregnant voxel phantoms, as should theoretical studies investigating the local currents induced in the eye and in the heart by movement during exposure. Finally, studies are recommended to investigate further the ability of static magnetic fields to significantly affect radical pair reactions in biological systems.

Hunger Modulates the Responses to Gustatory Stimuli of Single Neurons in the Caudolateral Orbitofrontal Cortex of the Macaque Monkey.

1. In order to determine whether the responsiveness of neurons in the caudolateral orbitofrontal cortex (a secondary cortical gustatory area) is influenced by hunger, the activity evoked by prototypical taste stimuli (glucose, NaCl, HCl, and quinine hydrochloride) and fruit juice was recorded in single neurons in this cortical area before, while, and after cynomolgous macaque monkeys were fed to satiety with glucose or fruit juice. 2. It was found that the responses of the neurons to the taste of the glucose decreased to zero while the monkey ate it to satiety during the course of which his behaviour turned from avid acceptance to active rejection. 3. This modulation of responsiveness of the gustatory responses of the neurons to satiety was not due to peripheral adaptation in the gustatory system or to altered efficacy of gustatory stimulation after satiety was reached, because modulation of neuronal responsiveness by satiety was not seen at earlier stages of the gustatory system, including the nucleus of the solitary tract, the frontal opercular taste cortex, and the insular taste cortex. 4. The decreases in the responsiveness of the neurons were relatively specific to the food with which the monkey had been fed to satiety. For example, in seven experiments in which the monkey was fed glucose solution, neuronal responsiveness decreased to the taste of the glucose but not to the taste of blackcurrant juice. Conversely, in two experiments in which the monkey was fed to satiety with fruit juice, the responses of the neurons decreased to fruit juice but not to glucose. 5. These and earlier findings lead to a proposed neurophysiological mechanism for sensory-specific satiety in which the information coded by single neurons in the gustatory system becomes more specific through the processing stages consisting of the nucleus of the solitary tract, the taste thalamus, and the frontal opercular and insular taste primary taste cortices, until neuronal responses become relatively specific for the food tasted in the caudolateral orbitofrontal cortex (secondary) taste area. Then sensory-specific satiety occurs because in this caudolateral orbitofrontal cortex taste area (but not earlier in the taste system) it is a property of the synapses that repeated stimulation results in a decreased neuronal response. 6. Evidence was obtained that gustatory processing involved in thirst also becomes interfaced to motivation in the caudolateral orbitofrontal cortex taste projection area, in that neuronal responses here to water were decreased to zero while water was drunk until satiety was produced.