Effects of protons and HZE particles on glutamate transport in astrocytes, neurons and mixed cultures

Radiation-induced neurotoxicity is a well-characterized phenomenon. However, the underlying mechanism of this toxicity is poorly understood. In the central nervous system (CNS), excitotoxic mechanisms are implicated in many neurodegenerative disease processes. Pivotal to the excitotoxic pathway is dysfunction of glutamate signaling. We reported previously that exposure to low-LET γ radiation results in altered glutamate transport in neurons and astrocytes. In the present study, we sought to investigate the effects of various particle radiations of differing LET on glutamate transport as a measure of the neurochemical vulnerability of the CNS. NTera2-derived neurons and astrocytes isolated as pure and mixed cultures were exposed to doses of 10 cGy, 50 cGy or 2 Gy of 250 MeV protons, 290 MeV/nucleon carbon ions, or 1000 MeV/nucleon iron ions. Transporter function was assessed at 3 h, 2 days and 7days after exposure. Functional assessment of glutamate transport revealed that neurons and astrocytes respond in a reciprocal manner after exposure to particle radiation. Uptake activity in neurons increased after particle irradiation. This effect was evident as late as our last time (7 days) after exposure (P < 0.05). In astrocytes, transporter activity decreased after exposure. The decrease in uptake observed in astrocytes was evident 7 days after exposure to carbon and iron ions. Uptake in mixed cultures after exposure to all three forms of radiation revealed a muted interactive response suggestive of the individual responses of each cellular phenotype acting in opposition.     

Impact of Microwave at X-Band in the aetiology of male infertility

Reports of declining male fertility have renewed interest in assessing the role of environmental and occupational exposures to electromagnetic fields (EMFs) in the aetiology of human infertility. Testicular functions are particularly susceptible to electromagnetic fields. The aim of the present work was to investigate the effect of 10-GHz EMF on male albino rat's reproductive system and to investigate the possible causative factor for such effect of exposure. The study was carried out in two groups of 70-day old adult male albino rats: a sham-exposed and a 10-GHz-exposed group (2?h a day for 45 days). Immediately after completion of the exposure, animals were sacrificed and sperms were extracted from the cauda and caput part of testis for the analysis of MDA, melatonin, and creatine kinase. Creatine kinase results revealed an increased level of phosphorylation that converts creatine to creatine phosphate in sperms after EMF exposure. EMF exposure also reduced the level of melatonin and MDA. It is concluded that microwave exposure could adversely affect male fertility by reducing availability of the above parameters. These results are indications of deleterious effects of these radiations on reproductive pattern of male rats.     

Impact of Microwave at X-Band in the aetiology of male infertility

Reports of declining male fertility have renewed interest in assessing the role of environmental and occupational exposures to electromagnetic fields (EMFs) in the aetiology of human infertility. Testicular functions are particularly susceptible to electromagnetic fields. The aim of the present work was to investigate the effect of 10-GHz EMF on male albino rat's reproductive system and to investigate the possible causative factor for such effect of exposure. The study was carried out in two groups of 70-day old adult male albino rats: a sham-exposed and a 10-GHz-exposed group (2 h a day for 45 days). Immediately after completion of the exposure, animals were sacrificed and sperms were extracted from the cauda and caput part of testis for the analysis of MDA, melatonin, and creatine kinase. Creatine kinase results revealed an increased level of phosphorylation that converts creatine to creatine phosphate in sperms after EMF exposure. EMF exposure also reduced the level of melatonin and MDA. It is concluded that microwave exposure could adversely affect male fertility by reducing availability of the above parameters. These results are indications of deleterious effects of these radiations on reproductive pattern of male rats.     

Calcium Ion Cyclotron Resonance (ICR), 7.0 Hz, 9.2 μT Magnetic Field Exposure Initiates Differentiation of Pituitary Corticotrope-Derived AtT20 D16V Cells

The aim of this work is the study of the effect of electromagnetic radiations (ELF-EMF) tuned to the calcium cyclotron resonance condition of 7.0?Hz, 9.2?μT on the differentiation process of pituitary corticotrope-derived AtT20 D16V cells. These cells respond to nerve growth factor by extending neurite-like processes. To establish whether exposure to the field could influence the molecular biology of the pituitary gland, a corticotrope-derived cells line (AtT20 D16V) was exposed to ELF-EMF at a frequency of 7.0?Hz, 9.2?μT electromagnetic field by a Vega Select 719 power supply. Significant evidence was obtained to conclude that as little as 36?h exposure to the Ca²⁺ ICR condition results in enhanced neurite outgrowth, with early expression and aggregation of the neuronal differentiation protein NF-200 into neurite structures.     

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.     

Long-term exposure to a continuous 900 MHz electromagnetic field disrupts cerebellar morphology in young adult male rats

The pathological effects of exposure to an electromagnetic field (EMF) during childhood and adolescence may be greater than those from exposure during adulthood. We investigated possible pathological changes in the cerebellum of adolescent rats exposed to 900 MHz EMF daily for 25 days. We used three groups of six 21-day-old male rats as follows: unexposed control group (Non-EG), sham-exposed group (Sham-EG) and an EMF-exposed group (EMF-EG). EMF-EG rats were exposed to EMF in an EMF cage for 1 h daily from postnatal days 21 through 46. Sham-EG rats were placed in the EMF cage for 1 h daily, but were not subjected to EMF. No procedures were performed on the Non-EG rats. The cerebellums of all animals were removed on postnatal day 47, sectioned and stained with cresyl violet for histopathological and stereological analyses. We found significantly fewer Purkinje cells in the EMF-EG group than in the Non-EG and Sham-EG groups. Histopathological evaluation revealed alteration of normal Purkinje cell arrangement and pathological changes including intense staining of neuron cytoplasm in the EMF-EG group. We found that exposure to continuous 900 MHz EMF for 1 h/day during adolescence can disrupt cerebellar morphology and reduce the number of Purkinje cells in adolescent rats.     

Effect of high SARs produced by cell phone like radiofrequency fields on mollusk single neuron

During exposure to the cell phone electromagnetic field (EMF), some neurons in the brain at areas of peak specific absorption rate (SAR) absorb more electromagnetic energy than is permitted by existing guidelines. The goal of the present work was to investigate the influence of cell phone-like EMF signal on excitability and memory processes in single neurons. A Transverse Electromagnetic Cell (TEM Cell) was used to expose single neurons of mollusk to the EMF.

Finite-Difference Time-Domain (FDTD) method was used for modeling the TEM Cell and the EMF interactions with living nerve ganglion and neurons. Neuron electrophysiology was investigated using standard microelectrode technique. SAR deposited into the single neuron was calculated to be 8.2 W/kg with a temperature increment of 1.21°C. After acute exposure, the threshold of firing of action potentials (AP) was significantly decreased (p ≈ 0.001). Time of habituation to stimulation with the intracellular current injection was increased (p ≈ 0.003). These results indicate that acute exposure to EMF at high SARs impairs the ability of neurons to store information.     

Effects of radiation emanating from base transceiver station and mobile phone on sleep,circadian rhythm and cognition in humans – a review

The electromagnetic fields (EMF) are ubiquitous. The base transceiver station (BTS) and mobile phones (MPs) contribute to the generation of EMF around their locations and are regarded as important sources of non-ionizing radiations. The use of mobile phone has increased dramatically in recent years so also the skepticism regarding its effects. In this review, we have made an attempt to scan the key research papers those aimed at elucidating the effects of EMF starting from extreme low frequency (ELF) to radio frequency (RF) through low frequency (LF). We have selected papers that dealt with the effects of radiations emanating from the BTS and MPs on human sleep, circadian rhythm, and cognition. Mostly, we have concentrated on papers published in the last 15 years. We came across conflicting reports. The findings reported in many papers suggest that the exposure to EMF has potentiality to compromise parameters related to sleep quality; in contrast, there are several reports those have given a clean sheet to the EMF exposure. The effects of EMF on circadian rhythms also remain inconclusive. The EMF exposure while did not produce any effect on circadian rhythm of heart rate and blood chemistry, it modulated the rhythms in cortisol and melatonin characterized by a decline in their 24-h circulating levels. The effects of exposure to EMF on cognitive parameters, like performance and memory, are also equivocal. The existing contradictory findings could be attributed to inter-individual variability in tolerance, gender-, and age-dependent differences in response, latitudinal differences in efficacy, variability among employed methodologies and differences in specific absorption rate, frequency of the mobile phone usage, and interaction of EMF with other physiological and environmental factors, among others. The future research should be carried out with added focus on elucidating the modulatory effects of these factors to put an end to the existing controversies on the biological effects of low/RF EMF radiations.     

Ceftriaxone Preserves Glutamate Transporters and Prevents Intermittent Hypoxia-Induced Vulnerability to Brain Excitotoxic Injury

Hypoxia alters cellular metabolism and although the effects of sustained hypoxia (SH) have been extensively studied, less is known about chronic intermittent hypoxia (IH), commonly associated with cardiovascular morbidity and stroke. We hypothesize that impaired glutamate homeostasis after chronic IH may underlie vulnerability to stroke-induced excitotoxicity. P16 organotypic hippocampal slices, cultured for 7 days were exposed for 7 days to IH (alternating 2 min 5% O₂ - 15 min 21% O₂), SH (5% O₂) or RA (21% O₂), then 3 glutamate challenges. The first and last exposures were intended as a metabolic stimulus (200 µM glutamate, 15 min); the second emulated excitotoxicity (10 mM glutamate, 10 min). GFAP, MAP2, and EAAT1, EAAT2 glutamate transporters expression were assessed after exposure to each hypoxic protocol. Additionally, cell viability was determined at baseline and after each glutamate challenge, in presence or absence of ceftriaxone that increases glutamate transporter expression. GFAP and MAP2 decreased after 7 days IH and SH. Long-term IH but not SH decreased EAAT1 and EAAT2. Excitotoxic glutamate challenge decreased cell viability and the following 200 µM exposure further increased cell death, particularly in IH-exposed slices. Ceftriaxone prevented glutamate transporter decrease and improved cell viability after IH and excitotoxicity. We conclude that IH is more detrimental to cell survival and glutamate homeostasis than SH. These findings suggest that impaired regulation of extracellular glutamate levels is implicated in the increased brain susceptibility to excitotoxic insult after long-term IH.     

Spindle disturbances in human-hamster hybrid (A(L) ) cells induced by the electrical component of the mobile communication frequency range signal

The production of spindle disturbances in a human–hamster hybrid (A_L) cell line by an electromagnetic field (EMF) with field strength of 90 V/m at a frequency of 900 MHz was studied in greater detail. The experimental setup presented allows investigating whether either the electrical (E) and/or the magnetic (H) field component of EMF can be associated with the effectiveness of the spindle‐disturbing potential. Therefore, both field components of a transversal electromagnetic field (TEM) wave have been separated during exposure of the biological system. This procedure should give more insight on understanding the underlying mechanisms of non‐thermal effects of EMF. A statistical comparison of the proportions of the fractions of ana‐ and telophases with spindle disturbances, obtained for five different exposure conditions with respect to unexposed controls (sham condition), showed that only cells exposed to the H‐field component of the EMF were not different from the control. Therefore, the results of the present study indicate that an exposure of cells to EMF at E‐field strengths of 45 and 90 V/m, as well as to the separated E component of the EMF, induces significant spindle disturbances in ana‐ and telophases of the cell cycle. Bioelectromagnetics 32:291–301, 2011. © 2010 Wiley‐Liss, Inc.     

Alterations of thymic morphology and antioxidant biomarkers in 60-day-old male rats following exposure to a continuous 900 MHz electromagnetic field during adolescence

We investigated changes in thymic tissue of male rats exposed to a 900 megahertz (MHz) electromagnetic field (EMF) on postnatal days 22–59. Three groups of six 21-day-old male Sprague-Dawley rats were allocated as: control (CG), sham (SG) and EMF (EMFG) groups. No procedure was performed on the CG rats. SG rats were placed in a Plexiglas cage for 1 h every day between postnatal days 22 and 59 without exposure to EMF. EMFG rats were placed in the same cage for the same periods as the SG rats and were exposed to 900 MHz EMF. Rats were sacrificed on postnatal day 60. Sections of thymus were stained for histological assessment. Oxidant/antioxidant parameters were investigated biochemically. Malondialdehyde (MDA) levels in EMFG increased compared to the other groups. Extravascular erythrocytes were observed in the medullary/corticomedullary regions in EMFG sections. We found that 900 MHz EMF applied for 1 h/day on postnatal days 22–59 can increase tissue MDA and histopathological changes in male rat thymic tissue.     

Autoimmune process after long-term low-level exposure to electromagnetic field (experimental results). Part I. Mobile communications and changes in electromagnetic conditions for the population. Need for additional substantiation of existing hygienic standards

Mobile communications provides a new source of electromagnetic exposure for almost the whole population of Russia. For the first time in the history of civilization, the brain of mobile phone users is exposed to localized radiofrequency (RF) electromagnetic fields (EMF). Base stations are a factor in the exposure of the population. Existing standards for limiting exposure do not account for the role of base stations as a source of EMF and cannot guarantee the absence of adverse health effects. It has become necessary to obtain reliable information to expand databases for the development of new standards. As recommended by the World Health Organization, an additional experiment is performed under the supervision of foreign experts, which shows changes in autoimmune status in rats after long-term low-level RF EMF exposure with an incident power of 500 μW/cm².     

No effects of UMTS exposure on the function of rat outer hair cells

UMTS communication devices are becoming common in everyday use. This could raise public concern about their possible detrimental effects on human health. The aim of this study, in the framework of the EMF nEAR Project, was to evaluate possible influence of UMTS electromagnetic fields (EMF) exposure on cochlear outer hair cells' (OHCs) functionality in laboratory animals. Forty‐eight male Sprague–Dawley rats were locally exposed (right ear) or sham‐exposed to a controlled UMTS EMF, frequency of 1946 MHz, at SAR level of 10 W/kg, 2 h a day, 5 days a week, for 4 weeks. A group of 12 rats treated with kanamycin (KM) was also included as positive control. Rats were tested by recording Distortion Product Otaoacoustic Emissions (DPOAEs), a non‐invasive test capable of assessing the status of the OHCs in the inner ear. DPOAEs were performed before, during (one or three times a week) and after (1‐week) exposure to the EMF. The analysis of the data shows that no statistically significant differences were found between the audiological signals recorded from the different experimental groups. The ototoxic effect of KM has been confirmed. Bioelectromagnetics 30:385–392, 2009. © 2009 Wiley‐Liss, Inc.     

Modulation of MCP-1 and iNOS by 50-Hz sinusoidal electromagnetic field.

The purpose of this study was to investigate whether overnight exposure to 1 mT-50 Hz extremely low-frequency sinusoidal electromagnetic field (EMF) affects the expression and production of inducible nitric oxide synthase (iNOS) and monocyte chemotactic protein-1 (MCP-1) in human monocytes. RT-PCR and Western blot analysis demonstrate that EMF exposure affects the expression of iNOS and MCP-1 in cultured human mononuclear cells at the mRNA level and protein synthesis. Interestingly, the effects of EMF exposure clearly differed with respect to the potentiation and inhibition of iNOS and MCP-1 expression. Whereas iNOS was down-regulated both at the mRNA level and at the protein level, MCP-1 was up-regulated. These results provide helpful information regarding the EMF-mediated modulation of the inflammatory response in vivo. However, additional studies are necessary to demonstrate that EMF acts as a nonpharmacological inhibitor of NO and inducer of MCP-1 in some diseases where the balance of MCP-1 and NO may be important.     

Extended exposure of adult and fetal mice to 50 Hz magnetic field does not increase the incidence of micronuclei in erythrocytes.

The flow cytometer-based micronucleus assay was used to study the effects on chromosomes in erythroid cells of CBA/Ca mice after extended exposure to 50 Hz magnetic field (MF), 14 microT, peak-to-peak (p-p). The study included two different experiments: (a) mice exposed in utero during 18 days of their prenatal stage, and (b) adult mice exposed for 18 days. In experiment (a) 35 days after exposure was terminated, peripheral blood was drawn from the mice exposed in utero to determine whether the exposure had a genotoxic effect on the pluripotent erythroid stem cells. About 200000 polychromatic erythrocytes (PCE) and 200000 normochromatic erythrocytes (NCE) were analysed from each of 20 exposed mice. The EMF exposure did not significantly change the frequency of micronucleated PCE or NCE in comparison with 20 sham-irradiated mice. There was no difference in the proportion of PCE between exposed and unexposed animals. Similarly, in experiment (b) no differences were seen between EMF exposed and unexposed adult mice when samples of peripheral blood were taken at the end of exposure and analyzed for micronuclei in PCE and NCE. The proportion of PCE was the same in both groups. The results indicate that exposure to EMF does not induce direct or indirect effects on chromosomes in erythroid cells expressed as increased levels of micronucleated erythrocytes of mice. No indications of delayed genetic effects were found.     

Impairment of chondrocyte biosynthetic activity by exposure to 3-tesla high-field magnetic resonance imaging is temporary

The influence of magnetic resonance imaging (MRI) devices at high field strengths on living tissues is unknown. We investigated the effects of a 3-tesla electromagnetic field (EMF) on the biosynthetic activity of bovine articular cartilage. Bovine articular cartilage was obtained from juvenile and adult animals. Whole joints or cartilage explants were subjected to a pulsed 3-tesla EMF; controls were left unexposed. Synthesis of sulfated glycosaminoglycans (sGAGs) was measured by using [³⁵S]sulfate incorporation; mRNA encoding the cartilage markers aggrecan and type II collagen, as well as IL-1β, were analyzed by RT–PCR. Furthermore, effects of the 3-tesla EMF were determined over the course of time directly after exposure (day 0) and at days 3 and 6. In addition, the influence of a 1.5-tesla EMF on cartilage sGAG synthesis was evaluated. Chondrocyte cell death was assessed by staining with Annexin V and TdT-mediated dUTP nick end labelling (TUNEL). Exposure to the EMF resulted in a significant decrease in cartilage macromolecule synthesis. Gene expression of both aggrecan and IL-1β, but not of collagen type II, was reduced in comparison with controls. Staining with Annexin V and TUNEL revealed no evidence of cell death. Interestingly, chondrocytes regained their biosynthetic activity within 3 days after exposure, as shown by proteoglycan synthesis rate and mRNA expression levels. Cartilage samples exposed to a 1.5-tesla EMF remained unaffected. Although MRI devices with a field strength of more than 1.5 T provide a better signal-to-noise ratio and thereby higher spatial resolution, their high field strength impairs the biosynthetic activity of articular chondrocytes in vitro. Although this decrease in biosynthetic activity seems to be transient, articular cartilage exposed to high-energy EMF may become vulnerable to damage.     

Ubiquitin-mediated proteasome activity is required for agonist-induced endocytosis of GluRs

Recent studies documenting a role for local protein synthesis in synaptic plasticity have lead to interest in the opposing process, protein degradation, as a potential regulator of synaptic function. The ubiquitin-conjugation system identifies, modifies, and delivers proteins to the proteasome for degradation. We found that both the proteasome and ubiquitin are present in the soma and dendrites of hippocampal neurons. As the trafficking of glutamate receptors (GluRs) is thought to underlie some forms of synaptic plasticity, we examined whether blocking proteasome activity affects the agonist-induced internalization of GluRs in cultured hippocampal neurons. Treatment with the glutamate agonist AMPA induced a robust internalization of GluRs. In contrast, brief pretreatment with proteasome inhibitors completely prevented the internalization of GluRs. To distinguish between a role for the proteasome and a possible diminution of the free ubiquitin pool, we expressed a chain elongation defective ubiquitin mutant (UbK48R), which causes premature termination of polyubiquitin chains but, importantly, can serve as a substrate for mono-ubiquitin-dependent processes. Expression of K48R in neurons severely diminished AMPA-induced internalization establishing a role for the proteasome. These data demonstrate the acute (e.g., minutes) regulation of synaptic function by the ubiquitin-proteasome pathway in mammalian neurons.     

Effects of mobile phone electromagnetic fields on an auditory order threshold task

The effect of acute exposure to radio frequency electromagnetic fields (RF EMF) generated by mobile phones on an auditory threshold task was investigated. 168 participants performed the task while exposed to RF EMF in one testing session (either global system for mobile communication (GSM) or unmodulated signals) while in a separate session participants were exposed to sham signals. Lateralization effects were tested by exposing participants either on the left side or on the right side of the head. No significant effect of exposure to RF EMF was detected, suggesting that acute exposure to RF EMFs does not affect performance in the order threshold task.     

Idiopathic environmental intolerance attributed to electromagnetic fields (formerly ‘electromagnetic hypersensitivity’): An updated systematic review of provocation studies

Idiopathic Environmental Intolerance attributed to electromagnetic fields (IEI‐EMF; formerly ‘electromagetic hypersensitivity’) is a medically unexplained illness in which subjective symptoms are reported following exposure to electrical devices. In an earlier systematic review, we reported data from 31 blind provocation studies which had exposed IEI‐EMF volunteers to active or sham electromagnetic fields and assessed whether volunteers could detect these fields or whether they reported worse symptoms when exposed to them. In this article, we report an update to that review. An extensive literature search identified 15 new experiments. Including studies reported in our earlier review, 46 blind or double‐blind provocation studies in all, involving 1175 IEI‐EMF volunteers, have tested whether exposure to electromagnetic fields is responsible for triggering symptoms in IEI‐EMF. No robust evidence could be found to support this theory. However, the studies included in the review did support the role of the nocebo effect in triggering acute symptoms in IEI‐EMF sufferers. Despite the conviction of IEI‐EMF sufferers that their symptoms are triggered by exposure to electromagnetic fields, repeated experiments have been unable to replicate this phenomenon under controlled conditions. A narrow focus by clinicians or policy makers on bioelectromagnetic mechanisms is therefore, unlikely to help IEI‐EMF patients in the long‐term. Bioelectromagnetics 31:1–11, 2010. © 2009 Wiley‐Liss, Inc.     

Cellular neoplastic transformation induced by 916 MHz microwave radiation

There has been growing concern about the possibility of adverse health effects resulting from exposure to microwave radiations, such as those emitted by mobile phones. The purpose of this study was to investigate the cellular neoplastic transformation effects of electromagnetic fields. 916 MHz continuous microwave was employed in our study to simulate the electromagnetic radiation of mobile phone. NIH/3T3 cells were adopted in our experiment due to their sensitivity to carcinogen or cancer promoter in environment. They were divided randomly into one control group and three microwave groups. The three microwave groups were exposed to 916 MHz EMF for 2 h per day with power density of 10, 50, and 90 w/m(2), respectively, in which 10 w/m(2) was close to intensity near the antenna of mobile phone. The morphology and proliferation of NIH/3T3 cells were examined and furthermore soft agar culture and animal carcinogenesis assay were carried out to determine the neoplastic promotion. Our experiments showed NIH/3T3 cells changed in morphology and proliferation after 5-8 weeks exposure and formed clone in soft agar culture after another 3-4 weeks depending on the exposure intensity. In the animal carcinogenesis study, lumps developed on the back of SCID mice after being inoculated into exposed NIH/3T3 cells for more than 4 weeks. The results indicate that microwave radiation can promote neoplastic transformation of NIH/3T3cells.