The effect of exposure to radiofrequency electromagnetic fields on cognitive performance in human experimental studies: Systematic review and meta-analyses.

BACKGROUND: The objective of this review is to evaluate the associations between short-term exposure to radiofrequency electromagnetic fields (RF-EMF) and cognitive performance in human experimental studies. METHODS: Online databases (PubMed, Embase, Scopus, Web of Science and EMF-Portal) were searched for studies that evaluated effects of exposure to RF-EMF on seven domains of cognitive performance in human experimental studies. The assessment of study quality was based on the Risk of Bias (RoB) tool developed by the Office of Health Assessment and Translation (OHAT). Random effects meta-analyses of Hedges's g were conducted separately for accuracy- and speed-related performance measures of various cognitive domains, for which data from at least two studies were available. Finally, the certainty of evidence for each identified outcome was assessed according to Grading of Recommendations Assessment, Development, and Evaluation (GRADE). RESULTS: 57,543 records were identified and 76 studies (80 reports) met the inclusion criteria. The included 76 studies with 3846 participants, consisting of humans of different age, sex and health status from 19 countries, were conducted between 1989 and 2021. Quantitative data from 50 studies (52 reports) with 2433 participants were included into the meta-analyses. These studies were performed in 15 countries between 2001 and 2021. The majority of the included studies used head exposure with GSM 900 uplink. None of the meta-analyses observed a statistically significant effect of RF-EMF exposure compared to sham on cognitive performance as measured by the confidence interval surrounding the Hedges's g or the significance of the z-statistic. For the domain Orientation and Attention, subclass Attention - Attentional Capacity RF-EMF exposure results in little to no difference in accuracy (Hedges's g 0.024, 95 % CI [-0.10; 0.15], I2 = 28 %, 473 participants). For the domain Orientation and Attention, subclass Attention - Concentration / Focused Attention RF-EMF exposure results in little to no difference in speed (Hedges's g 0.005, 95 % CI [-0.17; 0.18], I2 = 7 %, 132 participants) and probably results in little to no difference in accuracy; it does not reduce accuracy (Hedges's g 0.097, 95 % CI [-0.05; 0.24], I2 = 0 %, 217 participants). For the domain Orientation and Attention, subclass Attention - Vigilance RF-EMF exposure probably results in little to no difference in speed and does not reduce speed (Hedges's g 0.118, 95 % CI [-0.04; 0.28], I2 = 41 %, 247 participants) and results in little to no difference in accuracy (Hedges's g 0.042, 95 % CI, [-0.09; 0.18], I2 = 0 %, 199 participants). For the domain Orientation and Attention, subclass Attention - Selective Attention RF-EMF exposure probably results in little to no difference in speed and does not reduce speed (Hedges's g 0.080, 95 % CI [-0.09; 0.25], I2 = 63 %, 452 participants); it may result in little to no difference in accuracy, but it probably does not reduce accuracy (Hedges's g 0.178, 95 % CI [-0.02; 0.38], I2 = 68 %, 480 participants). For the domain Orientation and Attention, subclass Attention - Divided Attention RF-EMF exposure results in little to no difference in speed (Hedges's g -0.010, 95 % CI [-0.14; 0.12], I2 = 5 %, 307 participants) and may result in little to no difference in accuracy (Hedges's g -0.089, 95 % CI [-0.35; 0.18], I2 = 53 %, 167 participants). For the domain Orientation and Attention, subclass Processing Speed - Simple Reaction Time Task RF-EMF exposure results in little to no difference in speed (Hedges's g 0.069, 95 % CI [-0.02; +0.16], I2 = 29 %, 820 participants). For the domain Orientation and Attention, subclass Processing Speed - 2-Choice Reaction Time Task RF-EMF exposure results in little to no difference in speed (Hedges's g -0.023, 95 % CI [-0.13; 0.08], I2 = 0 %, 401 participants), and may result in little to no difference in accuracy (Hedges's g -0.063, 95 % CI [-0.38; 0.25], I2 = 63 %, 117 participants). For the domain Orientation and Attention, subclass Processing Speed - >2-Choice Reaction Time Task RF-EMF exposure results in little to no difference in speed (Hedges's g -0.054, 95 % CI [-0.14; 0.03], I2 = 0 %, 544 participants) and probably results in little to no difference in accuracy (Hedges's g -0.129, 95 % CI [-0.30; 0.04], I2 = 0 %, 131 participants). For the domain Orientation and Attention, subclass Processing Speed - Other Tasks RF-EMF exposure probably results in little to no difference in speed and does not reduce speed (Hedges's g 0.067, 95 % CI [-0.12; 0.26], I2 = 38 %, 249 participants); it results in little to no difference in accuracy (Hedges's g 0.036, 95 % CI [-0.08; 0.15], I2 = 0 %, 354 participants). For the domain Orientation and Attention, subclass Working Memory - n-back Task (0-3-back) we found Hedges's g ranging from -0.090, 95 % CI [-0.18; 0.01] to 0.060, 95 % CI [-0.06; 0.18], all I2 = 0 %, 237 to 474 participants, and conclude that RF-EMF exposure results in little to no difference in both speed and accuracy. For the domain Orientation and Attention, subclass Working Memory - Mental Tracking RF-EMF exposure results in little to no difference in accuracy (Hedges's g -0.047, 95 % [CI -0.15; 0.05], I2 = 0 %, 438 participants). For the domain Perception, subclass Visual and Auditory Perception RF-EMF exposure may result in little to no difference in speed (Hedges's g -0.015, 95 % CI [-0.23; 0.195], I2 = 0 %, 84 participants) and probably results in little to no difference in accuracy (Hedges's g 0.035, 95 % CI [-0.13; 0.199], I2 = 0 %, 137 participants). For the domain Memory, subclass Verbal and Visual Memory RF-EMF exposure probably results in little to no difference in speed and does not reduce speed (Hedges's g 0.042, 95 % CI [-0.15; 0.23], I2 = 0 %, 102 participants); it may result in little to no difference in accuracy (Hedges's g -0.087, 95 % CI [-0.38; 0.20], I2 = 85 %, 625 participants). For the domain Verbal Functions and Language Skills, subclass Verbal Expression, a meta-analysis was not possible because one of the two included studies did not provide numerical values. Results of both studies did not indicate statistically significant effects of RF-EMF exposure on both speed and accuracy. For the domain Construction and Motor Performance, subclass Motor Skills RF-EMF exposure may reduce speed, but the evidence is very uncertain (Hedges's g -0.919, 95 % CI [-3.09; 1.26], I2 = 96 %, 42 participants); it probably results in little to no difference in accuracy and does not reduce accuracy (Hedges's g 0.228, 95 % CI [-0.01; 0.46], I2 = 0 %, 109 participants). For the domain Concept Formation and Reasoning, subclass Reasoning RF-EMF exposure results in little to no difference in speed (Hedges's g 0.010, 95 % CI [-0.11; 0.13], I2 = 0 %, 263 participants) and probably results in little to no difference in accuracy and does not reduce accuracy (Hedges's g 0.051, 95 % CI [-0.14; 0.25], I2 = 0 %, 100 participants). For the domain Concept Formation and Reasoning, subclass Mathematical Procedures RF-EMF exposure results in little to no difference in speed (Hedges's g 0.033, 95 % CI [-0.12; 0.18], I2 = 0 %, 168 participants) and may result in little to no difference in accuracy but probably does not reduce accuracy (Hedges's g 0.232, 95 % CI [-0.12; +0.59], I2 = 86 %, 253 participants). For the domain Executive Functions there were no studies. DISCUSSION: Overall, the results from all domains and subclasses across their speed- and accuracy-related outcome measures according to GRADE provide high to low certainty of evidence that short-term RF-EMF exposure does not reduce cognitive performance in human experimental studies. For 16 out of 35 subdomains some uncertainty remains, because of limitations in the study quality, inconsistency in the results or imprecision of the combined effect size estimate. Future research should focus on construction and motor performance, elderly, and consideration of both sexes. OTHER: This review was partially funded by the WHO radioprotection programme. The protocol for this review was registered in Prospero reg. no. CRD42021236168 and published in Environment International (Pophof et al. 2021).

Biological Effects of Radiofrequency Electromagnetic Fields above 100 MHz on Fauna and Flora: Workshop Report.

ABSTRACT: This report summarizes the effects of anthropogenic radiofrequency electromagnetic fields with frequencies above 100 MHz on flora and fauna presented at an international workshop held on 5-7 November 2019 in Munich, Germany. Anthropogenic radiofrequency electromagnetic fields at these frequencies are commonplace; e.g., originating from transmitters used for terrestrial radio and TV broadcasting, mobile communication, wireless internet networks, and radar technologies. The effects of these radiofrequency fields on flora, fauna, and ecosystems are not well studied. For high frequencies exceeding 100 MHz, the only scientifically established action mechanism in organisms is the conversion of electromagnetic into thermal energy. In accordance with that, no proven scientific evidence of adverse effects in animals or plants under realistic environmental conditions has yet been identified from exposure to low-level anthropogenic radiofrequency fields in this frequency range. Because appropriate field studies are scarce, further studies on plants and animals are recommended.

Biological Effects of Electric, Magnetic, and Electromagnetic Fields from 0 to 100 MHz on Fauna and Flora: Workshop Report.

ABSTRACT: This report summarizes effects of anthropogenic electric, magnetic, and electromagnetic fields in the frequency range from 0 to 100 MHz on flora and fauna, as presented at an international workshop held on 5-7 November in 2019 in Munich, Germany. Such fields may originate from overhead powerlines, earth or sea cables, and from wireless charging systems. Animals and plants react differentially to anthropogenic fields; the mechanisms underlying these responses are still researched actively. Radical pairs and magnetite are discussed mechanisms of magnetoreception in insects, birds, and mammals. Moreover, several insects as well as marine species possess specialized electroreceptors, and behavioral reactions to anthropogenic fields have been reported. Plants react to experimental modifications of their magnetic environment by growth changes. Strong adverse effects of anthropogenic fields have not been described, but knowledge gaps were identified; further studies, aiming at the identification of the interaction mechanisms and the ecological consequences, are recommended.

The effect of radiofrequency electromagnetic fields (RF-EMF) on biomarkers of oxidative stress in vivo and in vitro: A protocol for a systematic review.

BACKGROUND: Oxidative stress is conjectured to be related to many diseases. Furthermore, it is hypothesized that radiofrequency fields may induce oxidative stress in various cell types and thereby compromise human and animal health. This systematic review (SR) aims to summarize and evaluate the literature related to this hypothesis. OBJECTIVES: The main objective of this SR is to evaluate the associations between the exposure to radiofrequency electromagnetic fields and oxidative stress in experimental models (in vivo and in vitro). METHODS: The SR framework has been developed following the guidelines established in the WHO Handbook for Guideline Development and the Handbook for Conducting a Literature-Based Health Assessment). We will include controlled in vivo and in vitro laboratory studies that assess the effects of an exposure to RF-EMF on valid markers for oxidative stress compared to no or sham exposure. The protocol is registered in PROSPERO. We will search the following databases: PubMed, Embase, Web of Science Core Collection, Scopus, and the EMF-Portal. The reference lists of included studies and retrieved review articles will also be manually searched. STUDY APPRAISAL AND SYNTHESIS METHOD: Data will be extracted according to a pre-defined set of forms developed in the DistillerSR online software and synthesized in a meta-analysis when studies are judged sufficiently similar to be combined. If a meta-analysis is not possible, we will describe the effects of the exposure in a narrative way. RISK OF BIAS: The risk of bias will be assessed with the NTP/OHAT risk of bias rating tool for human and animal studies. We will use GRADE to assess the certainty of the conclusions (high, moderate, low, or inadequate) regarding the association between radiofrequency electromagnetic fields and oxidative stress. FUNDING: This work was funded by the World Health Organization (WHO). REGISTRATION: The protocol was registered on the PROSPERO webpage on July 8, 2021.

The effect of exposure to radiofrequency electromagnetic fields on cognitive performance in human experimental studies: A protocol for a systematic review.

BACKGROUND: The World Health Organization (WHO) is currently assessing the potential health effects of exposure to radiofrequency electromagnetic fields (RF-EMFs) in the general and working population. Related to one such health effect, there is a concern that RF-EMFs may affect cognitive performance in humans. The systematic review (SR) aims to identify, summarize and synthesize the evidence base related to this question. Here, we present the protocol for the planned SR. OBJECTIVES: The main objective is to present a protocol for a SR which will evaluate the associations between short-term exposure to RF-EMFs and cognitive performance in human experimental studies. DATA SOURCES: We will search the following databases: PubMed, Embase, Web of Science, Scopus, and the EMF-Portal. The reference lists of included studies and retrieved review articles will be manually searched. STUDY ELIGIBILITY AND CRITERIA: We will include randomized human experimental studies that assess the effects of RF-EMFs on cognitive performance compared to no exposure or lower exposure. We will include peer-reviewed articles of any publication date in any language that report primary data. DATA EXTRACTION AND ANALYSIS: Data will be extracted according to a pre-defined set of forms developed and piloted by the review author team. To assess the risk of bias, we will apply the Rating Tool for Human and Animal Studies developed by NTP/OHAT, supplemented with additional questions relevant for cross-over studies. Where sufficiently similar studies are identified (e.g. the heterogeneity concerning population, exposure and outcome is low and the studies can be combined), we will conduct random-effects meta-analysis; otherwise, we will conduct a narrative synthesis. ASSESSMENT OF CERTAINTY OF EVIDENCE: The certainty of evidence for each identified outcome will be assessed according to Grading of Recommendations Assessment, Development, and Evaluation (GRADE). Performing the review according to this protocol will allow the identification of possible effects of RF-EMFs on cognitive performance in humans. The protocol has been registered in PROSPERO, an open-source protocol registration system, to foster transparency.

Unifying photocycle model for light adaptation and temporal evolution of cation conductance in channelrhodopsin-2.

Although channelrhodopsin (ChR) is a widely applied light-activated ion channel, important properties such as light adaptation, photocurrent inactivation, and alteration of the ion selectivity during continuous illumination are not well understood from a molecular perspective. Herein, we address these open questions using single-turnover electrophysiology, time-resolved step-scan FTIR, and Raman spectroscopy of fully dark-adapted ChR2. This yields a unifying parallel photocycle model integrating now all so far controversial discussed data. In dark-adapted ChR2, the protonated retinal Schiff base chromophore (RSBH+) adopts an all-trans,C=N-anti conformation only. Upon light activation, a branching reaction into either a 13-cis,C=N-anti or a 13-cis,C=N-syn retinal conformation occurs. The anti-cycle features sequential H+ and Na+ conductance in a late M-like state and an N-like open-channel state. In contrast, the 13-cis,C=N-syn isomer represents a second closed-channel state identical to the long-lived P480 state, which has been previously assigned to a late intermediate in a single-photocycle model. Light excitation of P480 induces a parallel syn-photocycle with an open-channel state of small conductance and high proton selectivity. E90 becomes deprotonated in P480 and stays deprotonated in the C=N-syn cycle. Deprotonation of E90 and successive pore hydration are crucial for late proton conductance following light adaptation. Parallel anti- and syn-photocycles now explain inactivation and ion selectivity changes of ChR2 during continuous illumination, fostering the future rational design of optogenetic tools.

Tracking Pore Hydration in Channelrhodopsin by Site-Directed Infrared-Active Azido Probes.

In recent years, gating and transient ion-pathway formation in the light-gated channelrhodopsins (ChRs) have been intensively studied. Despite these efforts, a profound understanding of the mechanistic details is still lacking. To track structural changes concomitant with the formation and subsequent collapse of the ion-conducting pore, we site-specifically introduced the artificial polarity-sensing probe p-azido-l-phenylalanine (azF) into several ChRs by amber stop codon suppression. The frequently used optogenetic actuator ReaChR (red-activatable ChR) exhibited the best expression properties of the wild type and the azF mutants. By exploiting the unique infrared spectral absorption of azF [νas(N3) ∼ 2100 cm-1] and its sensitivity to polarity changes, we monitored hydration changes at various sites of the pore region and the inner gate by stationary and time-resolved infrared spectroscopy. Our data imply that channel closure coincides with a dehydration event occurring between the interface of the central and the inner gate. In contrast, the extracellular ion pathway seems to be hydrated in the open and closed states to similar extents. Mutagenesis of sites in the inner gate suggests that it acts as an intracellular entry funnel, whose architecture and composition modulate water influx and efflux within the channel pore. Our results highlight the potential of genetic code expansion technology combined with biophysical methods to investigate channel gating, particularly hydration dynamics at specific sites, with a so far unprecedented spatial resolution.

Mitochondrial COI and nuclear RAG1 DNA sequences and analyses of specimens of the three morphologically established species in the genus Trichopsis (Perciformes: Osphronemidae) reveal new/cryptic species.

Air-breathing fish species of the genus Trichopsis have been reported in Cambodia, Lao PDR, Indonesia, Malaysia, Singapore, Thailand and Vietnam. It is only in Thailand that all three recognized species (Trichopsis vittata, Trichopsis schalleri and Trichopsis pumila), as judged by distinct external features, are found. Cambodia and Lao PDR harbor two species each. The present work involves first-time DNA sequencing and analysis based on mitochondrial (COI) and nuclear (RAG1) DNA of numerous specimens of these species and specimens of a controversial Phetchaburi (Thailand) fish population with a mixed outward appearance. In addition to confirming the morphologically clear-cut taxonomic division of the three fish species, our DNA results show that whereas the T. pumila populations form one single species, there are cryptic species in the T. vittata and T. schalleri populations and possibly a new one in the latter. Members of the putative Phetchaburi fish population have been proven to be hybrids between T. pumila and T. vittata. In addition, a new the phylogenetic tree indicating ancestral relationships is also presented. This study should generate further research to find new/cryptic species of the genus Trichopsis in all countries harboring the fish.

Early formation of the ion-conducting pore in channelrhodopsin-2.

Channelrhodopsins (ChRs) are light-gated ion channels that are widely used in optogenetics. They allow precise control of neuronal activity with light, but a detailed understanding of how the channel is gated and the ions are conducted is still lacking. The recent determination of the X-ray structural model in the closed state marks an important milestone. Herein the open state structure is presented and the early formation of the ion conducting pore is elucidated in atomic detail using time-resolved FTIR spectroscopy. Photo-isomerization of the retinal-chromophore causes a downward movement of the highly conserved E90, which opens the pore. Molecular dynamic (MD) simulations show that water molecules invade through this opened pore, Helix 2 tilts and the channel fully opens within ms. Since E90 is a highly conserved residue, the proposed E90-Helix2-tilt (EHT) model might describe a general activation mechanism and provides a new avenue for further mechanistic studies and engineering.

Southeast Asian mouth-brooding Betta fighting fish (Teleostei: Perciformes) species and their phylogenetic relationships based on mitochondrial COI and nuclear ITS1 DNA sequences and analyses.

Fighting fish species in the genus Betta are found in several Southeast Asian countries. Depending on the mode of paternal care for fertilized eggs and hatchlings, various species of the betta fish are classified as mouth brooders or nest builders whose members in turn have been grouped according to their similarities mainly in morphology. The mouth brooders as well as some nest builders involved in the present study include fishes discovered and identified subsequent to previous reports on species groupings and their positions on phylogenetic trees based on DNA sequences that differ from those used by us in this study. From the mitochondrial COI gene and nuclear ITS1 gene sequences and more accurate analyses we conclude that the following members of the mouth-brooding pairs, named differently previously, are virtually identical, viz the Betta prima-Betta pallida pair and Betta ferox-Betta apollon pair. The Betta simplex, hitherto believed to be one species, could possibly be genetically split into 2 distinct species. In addition, several other established type-locality fishes could harbor cryptic species as judged by genetic differences. Assignments of fish species to groups reported earlier may have to be altered somewhat by the present genetic findings. We propose here a new Betta fish phylogenetic tree which, albeit being similar to the previous ones, is clearly different from them. Our gene-based evidence also leads to assignments of some fishes to new species groups and alters the positions of some species on the new phylogenetic tree, thus implying different ancestral relationships.

In channelrhodopsin-2 Glu-90 is crucial for ion selectivity and is deprotonated during the photocycle.

The light-activated microbial ion channel channelrhodopsin-2 (ChR2) is a powerful tool to study cellular processes with high spatiotemporal resolution in the emerging field of optogenetics. To customize the channel properties for optogenetic experiments, a detailed understanding of its molecular reaction mechanism is essential. Here, Glu-90, a key residue involved in the gating and selectivity mechanism of the ion channel is characterized in detail. The deprotonation of Glu-90 during the photocycle is elucidated by time-resolved FTIR spectroscopy, which seems to be part of the opening mechanism of the conductive pore. Furthermore, Glu-90 is crucial to ion selectivity as also revealed by mutation of this residue combined with voltage clamp experiments. By dynamic homology modeling, we further hypothesized that the conductive pore is flanked by Glu-90 and located between helices A, B, C, and G.