04/10/2012 - September 2012 - Science Update

The following is a quick summary of another twenty one papers that have come out over the last few months related to effects of electromagnetic radiation.

Adult stem cells are considered multipotent. Especially, human bone marrow-derived mesenchymal stem cells (hBM-MSCs) have the potential to differentiate into nerve type cells. Electromagnetic fields (EMFs) are widely distributed in the environment, and recently there have been many reports on the biological effects of EMFs. hBM-MSCs are weak and sensitive pluripotent stem cells, therefore extremely low frequency-electromagnetic fields (ELF-EMFs) could be affect the changes of biological functions within the cells. In our experiments, ELF-EMFs inhibited the growth of hBM-MSCs in 12 days exposure. Their gene level was changed and expression of the neural stem cell marker like nestin was decreased but the neural cell markers like MAP2, NEUROD1, NF-L, and Tau were induced. In immunofluorescence study, we confirmed the expression of each protein of neural cells. And also both oligodendrocyte and astrocyte related proteins like O4 and GFAP were expressed by ELF-EMFs. We suggest that EMFs can induce neural differentiation in BM-MSCs without any chemicals or differentiation factors.

This paper presents the development of an in vivo exposure system for exposing small rodents. The system consists of four identical cylindrical waveguide chambers, each with a plastic cage for housing the animal. The chamber is fed by circularly polarized radiofrequency power in the 1.9 GHz cellular frequency band and is vertically mounted so that the long axis of the animal is co-planar with the rotating incident electric field. Power sensors were used along with directional or hybrid couplers and a digital voltmeter for data acquisition for real-time dose rate monitoring. The system was tested to evaluate its dose rate performance when a mouse phantom or a mouse cadaver was inside the cage. The dose rate was quantified in terms of whole-body-average (WBA) specific absorption rate (SAR) per input power using both measurement and computational methods. The exposures of the mouse phantom and cadaver were evaluated for various possible postures and positions. The measurement results showed that the highest WBA-SAR was 16.9 W kg per 1 W incident power when the cadaver was lying prone against the cage wall and the lowest WBA-SAR was 10.4 W kg per 1 W incident power when the cadaver was standing upright in the cage center. These results were found to be in good agreement with those obtained from the computational method.

Consistent and independently replicated laboratory evidence to support a causative relationship between environmental exposure to extremely low-frequency electromagnetic fields (EMFs) at power line frequencies and the associated increase in risk of childhood leukemia has not been obtained. In particular, although gene expression responses have been reported in a wide variety of cells, none has emerged as robust, widely replicated effects. DNA microarrays facilitate comprehensive searches for changes in gene expression without a requirement to select candidate responsive genes. To determine if gene expression changes occur in white blood cells of volunteers exposed to an ELF-EMF, each of 17 pairs of male volunteers age 20-30 was subjected either to a 50 Hz EMF exposure of 62.0 ± 7.1 µT for 2 h or to a sham exposure (0.21 ± 0.05 µT) at the same time (11:00 a.m. to 13:00 p.m.). The alternative regime for each volunteer was repeated on the following day and the two-day sequence was repeated 6 days later, with the exception that a null exposure (0.085 ± 0.01 µT) replaced the sham exposure. Five blood samples (10 ml) were collected at 2 h intervals from 9:00 to 17:00 with five additional samples during the exposure and sham or null exposure periods on each study day. RNA samples were pooled for the same time on each study day for the group of 17 volunteers that were subjected to the ELF-EMF exposure/sham or null exposure sequence and were analyzed on Illumina microarrays. Time courses for 16 mammalian genes previously reported to be responsive to ELF-EMF exposure, including immediate early genes, stress response, cell proliferation and apoptotic genes were examined in detail. No genes or gene sets showed consistent response profiles to repeated ELF-EMF exposures. A stress response was detected as a transient increase in plasma cortisol at the onset of either exposure or sham exposure on the first study day. The cortisol response diminished progressively on subsequent exposures or sham exposures, and was attributable to mild stress associated with the experimental protocol.

This population-based case-control study in Taiwan considered incident cases aged 15years or less and admitted in 2003 to 2007 for all neoplasm (ICD-9-CM: 140-239) (n=2606), including 939 leukemia and 394 brain neoplasm cases. Controls were randomly selected, with a case/control ratio of 1:30 and matched on year of birth, from all non-neoplasm children insured in the same year when the index case was admitted. Annual summarized power (ASP, watt-year) was calculated for each of the 71,185 mobile phone base stations (MPBS) in service between 1998 and 2007. Then, the annual power density (APD, watt-year/km(2)) of each township (n=367) was computed as a ratio of the total ASP of all MPBS in a township to the area of that particular township. Exposure of each study subject to radio frequency (RF) was indicated by the averaged APD within 5years prior to the neoplasm diagnosis (cases) or July 1st of the year when the index case was admitted (controls) in the township where the subject lived. Unconditional logistic regression model with generalized estimation equation was employed to calculate the covariate-adjusted odds ratio [AOR] of childhood neoplasm in relation to RF exposure. A higher than median averaged APD (approximately 168WYs/km(2)) was significantly associated with an increased AOR for all neoplasms (1.13; 1.01 to 1.28), but not for leukemia (1.23; 0.99 to 1.52) or brain neoplasm (1.14, 0.83 to 1.55). This study noted a significantly increased risk of all neoplasms in children with higher-than-median RF exposure to MPBS. The slightly elevated risk was seen for leukemia and brain neoplasm, but was not statistically significant. These results may occur due to several methodological limitations.

Increasing use of mobile phones creates growing concern regarding harmful effects of radiofrequency non-ionizing electromagnetic radiation (NIER) on human tissues located close to the ear where phones are commonly held for long periods of time. We studied 20 subjects in the 'mobile phone group' who had a mean duration of mobile phone use of 12.5 years (range 8-15) and a mean time use of 29.6 hours per month (range 8-100). Deaf individuals served as controls. We compared salivary outcomes (secretion, oxidative damage indices, flow rate and composition) between mobile phone users and non-users. We report significant increase in all salivary oxidative stress indices studied in mobile phone users. Salivary flow, total protein, albumin and amylase activity were decreased in mobile phone users. These observations lead to the hypothesis that the use of mobile phones may cause oxidative stress and modify salivary function.

Clinically effective modalities of treatment for spinal cord injury (SCI) still remain unsatisfactory and are largely invasive in nature. There are reports of accelerated regeneration in injured peripheral nerves by extremely low-frequency pulsed electromagnetic field (ELF-EMF) in the rat. In the present study, the effect of (50 Hz), low-intensity (17.96 µT) magnetic field (MF) exposure of rats after-hemisection of T13 spinal cord (hSCI) was investigated on sensori-motor and locomotor functions. Rats were divided into hSCI (sham-exposed) and hSCI+MF (MF: 2 h/d X 6 weeks) groups. Besides their general conditions, locomotor function by Basso, Beattie, and Brenahan (BBB) score; motor responses to noxious stimuli by threshold of tail flick (TTF), simple vocalization (TSV), tail flick latency (TFL), and neuronal excitability by H-reflex were noted. It is found that, in the hSCI+MF group, a statistically significant improvement over the hSCI control group was noted in BBB score from post-SCI wk2 and TFL and TTF by post-hSCI wk1 and wk3, respectively. Correspondingly, TSV gradually restored by post-hSCI wk5.The threshold of H-reflex was reduced on ipsilateral side vs. contralateral side in hSCI and hSCI+MF group. A complete bladder control was dramatically restored on post-hSCI day4 (vs. day7 of hSCI group) and the survival rate was 100% in the hSCI+MF group (vs. 90% of hSCI group). The results of our study suggest that extremely low-frequency (50 Hz), low-intensity (17.96 µT) MF exposure for 2 h/d x 6wks promotes recovery of sensori-motor behavior including locomotion and bladder control both in terms of temporal pattern and magnitude in hemisection injury of (T13) spinal cord rats.

The relationship between radiofrequency electromagnetic fields emitted from mobile phone and infertility is a matter of continuing debate. It is postulated that these radiations may affect the reproduction pattern spell by targeting biochemistry of sperm. In an attempt to expedite the issue, 70 days old Wistar rats (n = 6) were exposed to mobile phone radiofrequency (RF) radiation for 2 h per day for 45 days and data compared with sham exposed (n = 6) group. A significant decrease (P < 0.05) in the level of testosterone and an increase in caspase-3 activity were found in the RF-exposed animals. Distortions in sperm head and mid piece of sperm mitochondrial sheath were also observed as captured by Transmission Electron Microscope (TEM). In addition, progeny from RF-exposed rats showed significant decreases in number and weight as compared with that of sham-exposed animals. A reduction in testosterone, an increase in caspase-3, and distortion in spermatozoa could be caused by overproduction of reactive oxygen species (ROS) in animals under mobile phone radiation exposure. Our findings on these biomarkers are clear indications of possible health implications of repeated exposure to mobile phone radiation.

Length and level of exposure to electromagnetic fields (EMFs) is increasing in association with the widespread use of electrical and electronic devices and technological progress. The undesirable effects of extremely low frequency magnetic fields (ELF-MFs) on health have attracted considerable interest. Sixty-four four-month-old male Wistar rats divided into eight groups of eight rats each were used. Seven groups were exposed to varying dosages of manganese (Mn) and a 50 Hz magnetic field (MF) of approximately 1 mT, while the last group was set aside as the cage control group and not subjected to any procedure. This study was intended to investigate the interactions between the application of MF and Mn and the elements Ca, Zn, Mg, and P thought to be involved in caries, in rat teeth. Levels of Ca, Mg, Zn, and P in the experimental group rats were different to those in the control group. The results demonstrate that ELF-MF and Mn can have significant effects on levels of elements in rat teeth. Further experimental and epidemiological studies of ELF-MF and Mn are needed in order to evaluate their dental effects.

Knowledge of the exposure to radiofrequency electromagnetic fields is necessary for epidemiological studies on possible health effects. The main goal of this study is to determine the exposure level and spatial and temporal variances during 39 everyday activities in 12 frequency bands used in mobile telecommunication and broadcasting. Therefore, 24h measurements were gathered from 98 volunteers living in or near Amsterdam and Purmerend, The Netherlands. They carried an activity diary to be kept to the minute, a GPS logger sampling at an interval of 1s, and an EME Spy exposimeter with a detection limit of 0.0066mW/m(2) sampling at an interval of 10s in 12 frequency bands. The mean exposure over 24h, excluding own mobile phone use, was 0.180mW/m(2). During daytime exposure was about the same, but during night it was about half, and in the evening it was about twice as high. The main contribution to environmental exposure (calling by participant not included) is from calling with mobile phones (37.5%), from cordless DECT phones and their docking stations (31.7%), and from the base stations (12.7%). The exposure to mobile phone base stations increases with the percentage of urban ground use, which is an indication for high people density. In agreement, the highest mean exposure relates to the activities with high people density, such as travelling by public transport, visiting social events, pubs or shopping malls. Exposure at home depends mainly on exposure from people calling in the neighbourhood of the participant and thus on the number of persons in a household. In addition just the possession of DECT docking stations leads to exposure as most models transmit continuously in stand-by. Also wireless internet routers continuously transmit in the WiFi band. Though the highest exposure peaks in the WiFi band, up to 0.265W/m(2), come from stray radiation of microwave ovens. The mean total exposure largely depends on phone calls of a high exposure level and short duration. These calls lead to potentially high contrasts as well in exposure levels between sessions of the same activity as between persons, thus posing a challenge for personal exposure prediction.

There are possible hazardous health effects of exposure to radiofrequency electromagnetic radiations emitted from mobile phone on the human reproductive pattern. It is more effective while keeping mobile phones in pocket or near testicular organs. Present review examines the possible concern on radio frequency radiation interaction and biological effects such as enzyme induction, and toxicological effects, including genotoxicity and carcinogenicity, testicular cancer, and reproductive outcomes. Testicular infertility or testicular cancer due to mobile phone or microwave radiations suggests an increased level of reactive oxygen species (ROS). Though generation of ROS in testis has been responsible for possible toxic effects on physiology of reproduction, the reviews of last few decades have well established that these radiations are very harmful and cause mutagenic changes in reproductive pattern and leads to infertility. The debate will be focused on bio-interaction mechanism between mobile phone and testicular cancer due to ROS formation. This causes the biological damage and leads to several changes like decreased sperm count, enzymatic and hormonal changes, DNA damage, and apoptosis formation. In the present review, physics of mobile phone including future research on various aspects has been discussed.

Extremely low frequency electro magnetic fields (EMFs) have been classified as possibly carcinogenic to humans by the International Agency for Research on Cancer. An increased number of chromosomal alterations in peripheral lymphocytes are correlated with elevated incidence of cancer. The aim of the present study was to assess occupationally induced chromosomal damage in EMF workers exposed to low levels of radiation. We used conventional metaphase chromosome aberration (CA) analysis and the micronucleus (MN) assay as biological indicators of non ionizing radiation exposure. In the present study totally 70 subjects were selected including 50 exposed and 20 controls. Informed written consent was obtained from all participants and the study was performed in accordance with the Declaration of Helsinki and the approval of the local ethical committee. A higher degree of CA and MN was observed in exposed subjects compared to controls, the frequency of CA being significantly enhanced with long years of exposure (P < 0.05). Moreover increase in CA and MN with age was noted in both exposed subjects and controls, but was significantly greater in the former. The results of this study demonstrated that a significant induction of cytogenetic damage in peripheral lymphocytes of workers occupationally exposed to EMFs in electric transformer and distribution stations. In conclusion, our findings suggest that EMFs possess genotoxic capability, as measured by CA and MN assays; CA analysis appeared more sensitive than other cytogenetic end-points. It can be concluded that chronic occupational exposure to EMFs may lead to an increased risk of genetic damage among electrical workers.

Self-reported recall data are often used in wireless phone epidemiological studies, which in turn are used to indicate relative risk of health outcomes from extended radiofrequency exposure. We sought to explain features commonly observed in wireless phone recall data and to improve analytical procedures. Each of the 16 schools selected a year 7 and/or 8 class to participate, providing a representative regional sample based on socioeconomic school ratings, school type and urban/rural balance. There was an 85% participation rate (N=373). Planned: the distribution of participants' estimated extent of SMS-texting and cordless phone calls, and the extent of rounding to a final zero or five within the full set of recall data and within each order of magnitude. Unplanned: the distribution of the leading digits of these raw data, compared with that of billed data in each order of magnitude. The nature and extent of number-rounding, and the distribution of data across each order in recall data indicated a logarithmic (ratio-based) mental process for assigning values. Responses became less specific as the leading-digit increased from 1 to 9, and 69% of responses for weekly texts sent were rounded by participants to a single non-zero digit (eg, 2, 20 and 200). Adolescents' estimation of their cellphone use indicated that it was performed on a mental logarithmic scale. This is the first time this phenomenon has been observed in the estimation of recalled, as opposed to observed, numerical quantities. Our findings provide empirical justification for log-transforming data for analysis. We recommend the use of the geometric rather than arithmetic mean when a recalled numerical range is provided. A point of calibration may improve recall.

Mobile genetic elements represent an important source of mutation and genomic instability, and their activity can be influenced by several chemical and physical agents. In this research we address the question whether exposure to extremely low-frequency pulsed magnetic fields (EMF-PMF) could affect the mobility of the human LINE-1(RP) retrotransposon. To this purpose, an in vitro retrotransposition assay was used on human neuroblastoma BE(2) cells exposed for 48h to 1mT, 50Hz PMF, or sham-exposed. Moreover, since it is well known that retrotransposition causes DNA double-strand breaks (DSB), an estimation of γ-H2AX foci, which is a marker of DNA DSB, was carried out on PMF- and sham-exposed samples. The results show that PMF-exposed cells had a lower number of both retrotransposition events and DNA DSB compared with sham-exposed samples. These results suggest that exposure to PMF can interfere with retrotransposition activity by inducing a decrease of retrotransposition events.

Purpose: The purpose of this study was to investigate the behavioral changes induced by 50 Hz, 10 mT flux density Sinusoidal Magnetic Field (MF). Material and methods: Seventy-six young adult male Wistar albino rats were used in the study. They were separated into two groups: control group (C) n=38; MF group n=38. C animals were left under the same conditions with the MF group for 21 days but with prevented or avoided exposure to MF. Anxiety and stress-related behavioral changes were investigated by elevated plus-maze and hole-board systems. Just before being tested in the maze, each animal was tested by means of the hole-board method in order to separate the directed exploration behavior and locomotion activity changes from anxiety-related behavior. Results: In the hole-board system parameters there were no statistically significant differences between the two groups. There was a statistically significant difference between MF and C groups when the ratio of time spent on open arms to the total time spent on all arms was evaluated (0.12±0.08 and 0.34±0.18 respectively and p < 0.01). Conclusion: Our results suggest that after 21 days, a continuous exposure to extremely low frequency of magnetic field (50 Hz, 10 mT) has no significant effect on activity and exploration activity but significantly induces stress and anxiety-related behavior in rats (Tab. 2, Fig. 9, Ref. 19). Keywords: magnetic field, anxiety, elevated plus-maze, hole board, stress, behavior, rat.

The present work aimed to investigate whether exposure to static magnetic field (SMF) and extremely low frequency magnetic field (ELF-MF) can induce biomolecular changes on Tuber borchii hyphal growth. Tuber borchii mycelium was exposed for 1 h for 3 consecutive days to a SMF of 300 mT or an ELF-MF of 0.1 mT 50 Hz. Gene expression and biochemical analyses were performed. In mycelia exposed to ELF-MF, some genes involved in hyphal growth, investigated using quantitative real-time polymerase chain reaction, were upregulated, and the activity of many glycolytic enzymes was increased. On the contrary, no differences were observed in gene expression after exposure to SMF treatment, and only the activities of glucose 6-phosphate dehydrogenase and hexokinase increased. The data herein presented suggest that the electromagnetic field can act as an environmental factor in promoting hyphal growth and can be used for applicative purposes, such as the set up of new in vitro cultivation techniques.

Electromagnetic fields (EMFs) originating both from both natural and manmade sources permeate our environment. As people are continuously exposed to EMFs in everyday life, it is a matter of great debate whether they can be harmful to human health. On the basis of two decades of epidemiological studies, an increased risk for childhood leukemia associated with Extremely Low Frequency fields has been consistently assessed, inducing the International Agency for Research on Cancer to insert them in the 2B section of carcinogens in 2001. EMFs interaction with biological systems may cause oxidative stress under certain circumstances. Since free radicals are essential for brain physiological processes and pathological degeneration, research focusing on the possible influence of the EMFs-driven oxidative stress is still in progress, especially in the light of recent studies suggesting that EMFs may contribute to the etiology of neurodegenerative disorders. This review synthesizes the emerging evidences about this topic, highlighting the wide data uncertainty that still characterizes the EMFs effect on oxidative stress modulation, as both pro-oxidant and neuroprotective effects have been documented. Care should be taken to avoid methodological limitations and to determine the patho-physiological relevance of any alteration found in EMFs-exposed biological system.

Based on the 'limited' evidence suggesting an association between exposure to radiofrequency fields (RF) emitted from mobile phones and two types of brain cancer, glioma and acoustic neuroma, the International Agency for Research on Cancer has classified RF as 'possibly carcinogenic to humans' in group 2B. In view of this classification and the positive correlation between increased genetic damage and carcinogenesis, a meta-analysis was conducted to determine whether a significant increase in genetic damage in human cells exposed to RF provides a potential mechanism for its carcinogenetic potential. The extent of genetic damage in human cells, assessed from various end-points, viz., single-/double-strand breaks in the DNA, incidence of chromosomal aberrations, micronuclei and sister chromatid exchanges, reported in a total of 88 peer-reviewed scientific publications during 1990-2011 was considered in the meta-analysis. Among the several variables in the experimental protocols used, the influence of five specific variables related to RF exposure characteristics was investigated: (i) frequency, (ii) specific absorption rate, (iii) exposure as continuous wave, pulsed wave and occupationally exposed/mobile phone users, (iv) duration of exposure, and (v) different cell types. The data indicated the following. (1) The magnitude of difference between RF-exposed and sham-/un-exposed controls was small with some exceptions. (2) In certain RF exposure conditions there was a statistically significant increase in genotoxicity assessed from some end-points: the effect was observed in studies with small sample size and was largely influenced by publication bias. Studies conducted within the generally recommended RF exposure guidelines showed a smaller effect. (3) The multiple regression analyses and heterogeneity goodness of fit data indicated that factors other than the above five variables as well as the quality of publications have contributed to the overall results. (4) More importantly, the mean indices for chromosomal aberrations, micronuclei and sister chromatid exchange end-points in RF-exposed and sham-/un-exposed controls were within the spontaneous levels reported in a large data-base. Thus, the classification of RF as possibly carcinogenic to humans in group 2B was not supported by genotoxicity-based mechanistic evidence.

Studies have repeatedly shown that electroencephalographic power during sleep is enhanced in the spindle frequency range following radio frequency electromagnetic field exposures pulse-modulated with fundamental frequency components of 2, 8, 14 or 217 Hz and combinations of these. However, signals used in previous studies also had significant harmonic components above 20 Hz. The current study aimed: (i) to determine if modulation components above 20 Hz, in combination with radio frequency, are necessary to alter the electroencephalogram; and (ii) to test the demodulation hypothesis, if the same effects occur after magnetic field exposure with the same pulse sequence used in the pulse-modulated radio frequency exposure. In a randomized double-blind crossover design, 25 young healthy men were exposed at weekly intervals to three different conditions for 30 min before sleep. Cognitive tasks were also performed during exposure. The conditions were a 2-Hz pulse-modulated radio frequency field, a 2-Hz pulsed magnetic field, and sham. Radio frequency exposure increased electroencephalogram power in the spindle frequency range. Furthermore, delta and theta activity (non-rapid eye movement sleep), and alpha and delta activity (rapid eye movement sleep) were affected following both exposure conditions. No effect on sleep architecture and no clear impact of exposure on cognition was observed. These results demonstrate that both pulse-modulated radio frequency and pulsed magnetic fields affect brain physiology, and the presence of significant frequency components above 20 Hz are not fundamental for these effects to occur. Because responses were not identical for all exposures, the study does not support the hypothesis that effects of radio frequency exposure are based on demodulation of the signal only.

Idiopathic environmental intolerance attributed to electromagnetic fields (IEI-EMF) remains a complex and unclear phenomenon, often characterized by the report of various, non-specific physical symptoms (NSPS) when an EMF source is present or perceived by the individual. The lack of validated criteria for defining and assessing IEI-EMF affects the quality of the relevant research, hindering not only the comparison or integration of study findings, but also the identification and management of patients by health care providers. The objective of this review was to evaluate and summarize the criteria that previous studies employed to identify IEI-EMF participants. An extensive literature search was performed for studies published up to June 2011. We searched EMBASE, Medline, Psychinfo, Scopus and Web of Science. Additionally, citation analyses were performed for key papers, reference sections of relevant papers were searched, conference proceedings were examined and a literature database held by the Mobile Phones Research Unit of King's College London was reviewed. Sixty-three studies were included. "Hypersensitivity to EMF" was the most frequently used descriptive term. Despite heterogeneity, the criteria predominantly used to identify IEI-EMF individuals were: 1. Self-report of being (hyper)sensitive to EMF. 2. Attribution of NSPS to at least one EMF source. 3. Absence of medical or psychiatric/psychological disorder capable of accounting for these symptoms 4. Symptoms should occur soon (up to 24 hours) after the individual perceives an exposure source or exposed area. (Hyper)sensitivity to EMF was either generalized (attribution to various EMF sources) or source-specific. Experimental studies used a larger number of criteria than those of observational design and performed more frequently a medical examination or interview as prerequisite for inclusion. Considerable heterogeneity exists in the criteria used by the researchers to identify IEI-EMF, due to explicit differences in their conceptual frameworks. Further work is required to produce consensus criteria not only for research purposes but also for use in clinical practice. This could be achieved by the development of an international protocol enabling a clearly defined case definition for IEI-EMF and a validated screening tool, with active involvement of medical practitioners.

Classifying evidence of causality between a risk factor and its potential health effect is challenging, in particular in an already emotional situation. Even the assessment of health risks by designated bodies may still depend on their composition of individuals with their background, bias, and, in worst case, their interests. This may explain opposing conclusions from the same pool of data which, consequently, may undermine credibility if not communicated properly. To overcome existing weakness in classifying and communicating evidence of health risks such as from electromagnetic fields, a new rule-based approach is presented. Developed by the German Commission on Radiological Protection (SSK), it discloses step-by-step the criteria for weighing scientific data and pools partial evidences of different scientific approaches to conclude on the overall evidence of causality between risk factor and effects. The validity of the approach is demonstrated by analyzing evidence of carcinogenicity of ionizing radiation, mobile phone use, and nocturnal exposure to visible light.

This study shows that a non-thermal pulse-modulated RF signal (PRF), configured to modulate calmodulin (CaM) activation via acceleration of Ca(2+) binding kinetics, produced an immediate nearly 3-fold increase in nitric oxide (NO) from dopaminergic MN9D cultures (P < 0.001). NO was measured electrochemically in real-time using a NO selective membrane electrode, which showed the PRF effect occurred within the first seconds after lipopolysaccharide (LPS) challenge. Further support that the site of action of PRF involves CaM is provided in human fibroblast cultures challenged with low serum and exposed for 15min to the identical PRF signal. In this case a CaM antagonist W-7 could be added to the culture 3h prior to PRF exposure. Those results showed the PRF signal produced nearly a two-fold increase in NO, which could be blocked by W-7 (P < 0.001). To the authors' knowledge this is the first report of a real-time effect of non-thermal electromagnetic fields (EMF) on NO release from challenged cells. The results provide mechanistic support for the many reported bioeffects of EMF in which NO plays a role. Thus, in a typical clinical application for acute post operative pain, or chronic pain from, e.g., osteoarthritis, EMF therapy could be employed to modulate the dynamics of NO via Ca/CaM-dependent constitutive nitric oxide synthase (cNOS) in the target tissue. This, in turn, would modulate the dynamics of the signaling pathways the body uses in response to the various phases of healing after physical or chemical insult or injury.