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1.  Alterations in Adenylate Kinase Activity in Human PBMCs after In Vitro Exposure to Electromagnetic Field: Comparison between Extremely Low Frequency Electromagnetic Field (ELF) and Therapeutic Application of a Musically Modulated Electromagnetic Field (TAMMEF) 
This study investigated the effects of electromagnetic fields on enzymes involved in purine metabolism in human peripheral blood mononuclear cells in vitro. Cells were obtained from 20 volunteers. We tested both low-energy, extremely low frequency (ELF; 100-Hz) electromagnetic fields and the Therapeutic Application of Musically Modulated Electromagnetic Fields (TAMMEFs); the latter is characterized by variable frequencies, intensities, and wave shapes. Adenylate kinase activity was increased after ELF field exposure but decreased slightly after TAMMEF exposure. Neither of the two electromagnetic field affected the activities of the purine metabolism enzymes ecto-5′-nucleotidase, adenosine deaminase, and adenosine kinase. We concluded that ELF fields may influence cellular electrical charge stability; stimulation of adenylate kinase activity could restore the cell to a state of equilibrium. In contrast, TAMMEF fields may be useful for maintaining and regulating the cellular electrical charge.
doi:10.1155/2009/717941
PMCID: PMC2744884  PMID: 19763276
2.  Neuronal Cellular Responses to Extremely Low Frequency Electromagnetic Field Exposure: Implications Regarding Oxidative Stress and Neurodegeneration 
PLoS ONE  2014;9(8):e104973.
Neurodegenerative diseases comprise both hereditary and sporadic conditions characterized by an identifying progressive nervous system dysfunction and distinctive neuopathophysiology. The majority are of non-familial etiology and hence environmental factors and lifestyle play key roles in their pathogenesis. The extensive use of and ever increasing worldwide demand for electricity has stimulated societal and scientific interest on the environmental exposure to low frequency electromagnetic fields (EMFs) on human health. Epidemiological studies suggest a positive association between 50/60-Hz power transmission fields and leukemia or lymphoma development. Consequent to the association between EMFs and induction of oxidative stress, concerns relating to development of neurodegenerative diseases, such as Alzheimer disease (AD), have been voiced as the brain consumes the greatest fraction of oxygen and is particularly vulnerable to oxidative stress. Exposure to extremely low frequency (ELF)-EMFs are reported to alter animal behavior and modulate biological variables, including gene expression, regulation of cell survival, promotion of cellular differentiation, and changes in cerebral blood flow in aged AD transgenic mice. Alterations in inflammatory responses have also been reported, but how these actions impact human health remains unknown. We hence evaluated the effects of an electromagnetic wave (magnetic field intensity 1mT; frequency, 50-Hz) on a well-characterized immortalized neuronal cell model, human SH-SY5Y cells. ELF-EMF exposure elevated the expession of NOS and O2−, which were countered by compensatory changes in antioxidant catylase (CAT) activity and enzymatic kinetic parameters related to CYP-450 and CAT activity. Actions of ELF-EMFs on cytokine gene expression were additionally evaluated and found rapidly modified. Confronted with co-exposure to H2O2-induced oxidative stress, ELF-EMF proved not as well counteracted and resulted in a decline in CAT activity and a rise in O2− levels. Together these studies support the further evaluation of ELF-EMF exposure in cellular and in vivo preclinical models to define mechanisms potentially impacted in humans.
doi:10.1371/journal.pone.0104973
PMCID: PMC4134243  PMID: 25127118
3.  Characterisation of weak magnetic field effects in an aqueous glutamic acid solution by nonlinear dielectric spectroscopy and voltammetry 
Background
Previous reports indicate altered metabolism and enzyme kinetics for various organisms, as well as changes of neuronal functions and behaviour of higher animals, when they were exposed to specific combinations of weak static and alternating low frequency electromagnetic fields. Field strengths and frequencies, as well as properties of involved ions were related by a linear equation, known as the formula of ion cyclotron resonance (ICR, abbreviation mentioned first by Liboff). Under certain conditions already a aqueous solution of the amino acid and neurotransmitter glutamate shows this effect.
Methods
An aqueous solution of glutamate was exposed to a combination of a static magnetic field of 40 μT and a sinusoidal electromagnetic magnetic field (EMF) with variable frequency (2–7 Hz) and an amplitude of 50 nT. The electric conductivity and dielectric properties of the solution were investigated by voltammetric techniques in combination with non linear dielectric spectroscopy (NLDS), which allow the examination of the dielectric properties of macromolecules and molecular aggregates in water. The experiments target to elucidate the biological relevance of the observed EMF effect on molecular level.
Results
An ion cyclotron resonance (ICR) effect of glutamate previously reported by the Fesenko laboratory 1998 could be confirmed. Frequency resolution of the sample currents was possible by NLDS techniques. The spectrum peaks when the conditions for ion cyclotron resonance (ICR) of glutamate are matched. Furthermore, the NLDS spectra are different under ICR- and non-ICR conditions: NLDS measurements with rising control voltages from 100–1100 mV show different courses of the intensities of the low order harmonics, which could possibly indicate "intensity windows". Furthermore, the observed magnetic field effects are pH dependent with a narrow optimum around pH 2.85.
Conclusions
Data will be discussed in the context with recent published models for the interaction of weak EMF with biological matter including ICR. A medical and health relevant aspect of such sensitive effects might be given insofar, because electromagnetic conditions for it occur at many occasions in our electromagnetic all day environment, concerning ion involvement of different biochemical pathways.
doi:10.1186/1477-044X-2-8
PMCID: PMC538269  PMID: 15571630
4.  Effects of Electromagnetic Radiation Exposure on Stress-Related Behaviors and Stress Hormones in Male Wistar Rats 
Biomolecules & Therapeutics  2014;22(6):570-576.
Studies have demonstrated that electromagnetic waves, as the one of the most important physical factors, may alter cognitive and non-cognitive behaviors, depending on the frequency and energy. Moreover, non-ionizing radiation of low energy waves e.g. very low frequency waves could alter this phenomenon via alterations in neurotransmitters and neurohormones. In this study, short, medium, and long-term exposure to the extremely low frequency electromagnetic field (ELF-EMF) (1 and 5 Hz radiation) on behavioral, hormonal, and metabolic changes in male Wistar rats (250 g) were studied. In addition, changes in plasma concentrations for two main stress hormones, noradrenaline and adrenocorticotropic hormone (ACTH) were evaluated. ELF-EMF exposure did not alter body weight, and food and water intake. Plasma glucose level was increased and decreased in the groups which exposed to the 5 and 1Hz wave, respectively. Plasma ACTH concentration increased in both using frequencies, whereas nor-adrenaline concentration showed overall reduction. At last, numbers of rearing, sniffing, locomotor activity was increased in group receiving 5 Hz wave over the time. In conclusions, these data showed that the effects of 1 and 5 Hz on the hormonal, metabolic and stress-like behaviors may be different. Moreover, the influence of waves on stress system is depending on time of exposure.
doi:10.4062/biomolther.2014.054
PMCID: PMC4256039  PMID: 25489427
Low-frequency electro-magnetic field; Corticosterone; Adrenaline; Adrenocorticotropic Hormone (ACTH); Stress
5.  The therapeutic effect of a pulsed electromagnetic field on the reproductive patterns of male Wistar rats exposed to a 2.45-GHz microwave field 
Clinics  2011;66(7):1237-1245.
INTRODUCTION:
Environmental exposure to man-made electromagnetic fields has been steadily increasing with the growing demand for electronic items that are operational at various frequencies. Testicular function is particularly susceptible to radiation emitted by electromagnetic fields.
OBJECTIVES:
This study aimed to examine the therapeutic effects of a pulsed electromagnetic field (100 Hz) on the reproductive systems of male Wistar rats (70 days old).
METHODS:
The experiments were divided into five groups: microwave sham, microwave exposure (2.45 GHz), pulsed electromagnetic field sham, pulsed electromagnetic field (100 Hz) exposure, and microwave/pulsed electromagnetic field exposure. The animals were exposed for 2 hours/day for 60 days. After exposure, the animals were sacrificed, their sperm was used for creatine and caspase assays, and their serum was used for melatonin and testosterone assays.
RESULTS:
The results showed significant increases in caspase and creatine kinase and significant decreases in testosterone and melatonin in the exposed groups. This finding emphasizes that reactive oxygen species (a potential inducer of cancer) are the primary cause of DNA damage. However, pulsed electromagnetic field exposure relieves the effect of microwave exposure by inducing Faraday currents.
CONCLUSIONS:
Electromagnetic fields are recognized as hazards that affect testicular function by generating reactive oxygen species and reduce the bioavailability of androgen to maturing spermatozoa. Thus, microwave exposure adversely affects male fertility, whereas pulsed electromagnetic field therapy is a non-invasive, simple technique that can be used as a scavenger agent to combat oxidative stress.
doi:10.1590/S1807-59322011000700020
PMCID: PMC3148471  PMID: 21876981
Microwave; Caspases; Creatine kinase; Testosterone; Infertility
6.  Influence of constant, alternating and cyclotron low-intensity electromagnetic fields on fibroblast proliferative activity in vitro 
Available data allow assuming the presence of stimulation of reparative processes under influence of low-intensity electromagnetic field, commensurable with a magnetic field of the Earth. Research of effects of low-intensity electromagnetic fields on fibroblast proliferative activity in human lungs in cell culture was performed.
The influence of a constant electromagnetic field, an alternating electromagnetic field by frequency of 50 Hz and cyclotron electromagnetic field with identical intensity for all kinds of fields – 80 mcTl – on value of cellular mass and a correlation of live and dead cells in culture is investigated in three series of experiments. We used the universal electromagnetic radiator generating all three kinds of fields and supplied by a magnetometer which allows measuring the intensity of accurate within 0.1 mcTl including taking into account the Earth’s magnetic field intensity.
The peak value for stimulation cellular proliferation in the present experiences was two-hour influence by any of the specified kinds of electromagnetic fields. The irradiation by cyclotron electromagnetic field conducts positive dynamics in growth of live cells (up to 206±22%) and decreases the number of dead cells (down to 31±6%). Application of cyclotron magnetic fields promoted creation of optimum conditions for proliferation. As a result of researches we observed the reliable 30% increase of nitro-tetrazolium index (in nitro-tetrazolium blue test) after irradiation by cyclotron electromagnetic field in experience that testifies to strengthening of the cell breathing of living cells.
In our opinion, it is necessary to pay attention not only to a pure gain of cells, but also to reduction of number dead cells that can be criterion of creation of optimum conditions for their specific development and valuable functioning.
doi:10.3205/dgkh000132
PMCID: PMC2831248  PMID: 20204088
electromagnetic field; human lung fibroblasts; cell culture; stimulation of growth; decrease of number of dead cells
7.  Exposure to GSM RF Fields Does Not Affect Calcium Homeostasis in Human Endothelial Cells, Rat Pheocromocytoma Cells or Rat Hippocampal Neurons 
PLoS ONE  2010;5(7):e11828.
In the course of modern daily life, individuals are exposed to numerous sources of electromagnetic radiation that are not present in the natural environment. The strength of the electromagnetic fields from sources such as hairdryers, computer display units and other electrical devices is modest. However, in many home and office environments, individuals can experience perpetual exposure to an “electromagnetic smog”, with occasional peaks of relatively high electromagnetic field intensity. This has led to concerns that such radiation can affect health. In particular, emissions from mobile phones or mobile phone masts have been invoked as a potential source of pathological electromagnetic radiation. Previous reports have suggested that cellular calcium (Ca2+) homeostasis is affected by the types of radiofrequency fields emitted by mobile phones. In the present study, we used a high-throughput imaging platform to monitor putative changes in cellular Ca2+ during exposure of cells to 900 MHz GSM fields of differing power (specific absorption rate 0.012–2 W/Kg), thus mimicking the type of radiation emitted by current mobile phone handsets. Data from cells experiencing the 900 Mhz GSM fields were compared with data obtained from paired experiments using continuous wave fields or no field. We employed three cell types (human endothelial cells, PC-12 neuroblastoma and primary hippocampal neurons) that have previously been suggested to be sensitive to radiofrequency fields. Experiments were designed to examine putative effects of radiofrequency fields on resting Ca2+, in addition to Ca2+ signals evoked by an InsP3-generating agonist. Furthermore, we examined putative effects of radiofrequency field exposure on Ca2+ store emptying and store-operated Ca2+ entry following application of the Ca2+ATPase inhibitor thapsigargin. Multiple parameters (e.g., peak amplitude, integrated Ca2+ signal, recovery rates) were analysed to explore potential impact of radiofrequency field exposure on Ca2+ signals. Our data indicate that 900 MHz GSM fields do not affect either basal Ca2+ homeostasis or provoked Ca2+ signals. Even at the highest field strengths applied, which exceed typical phone exposure levels, we did not observe any changes in cellular Ca2+ signals. We conclude that under the conditions employed in our experiments, and using a highly-sensitive assay, we could not detect any consequence of RF exposure.
doi:10.1371/journal.pone.0011828
PMCID: PMC2910734  PMID: 20676401
8.  Distinct Epidermal Keratinocytes Respond to Extremely Low-Frequency Electromagnetic Fields Differently 
PLoS ONE  2014;9(11):e113424.
Following an increase in the use of electric appliances that can generate 50 or 60 Hz electromagnetic fields, concerns have intensified regarding the biological effects of extremely low-frequency electromagnetic fields (ELF-EMFs) on human health. Previous epidemiological studies have suggested the carcinogenic potential of environmental exposure to ELF-EMFs, specifically at 50 or 60 Hz. However, the biological mechanism facilitating the effects of ELF-EMFs remains unclear. Cellular studies have yielded inconsistent results regarding the biological effects of ELF-EMFs. The inconsistent results might have been due to diverse cell types. In our previous study, we indicated that 1.5 mT, 60 Hz ELF-EMFs will cause G1 arrest through the activation of the ATM-Chk2-p21 pathway in human keratinocyte HaCaT cells. The aim of the current study was to investigate whether ELF-EMFs cause similar effects in a distinct epidermal keratinocyte, primary normal human epidermal keratinocytes (NHEK), by using the same ELF-EMF exposure system and experimental design. We observed that ELF-EMFs exerted no effects on cell growth, cell proliferation, cell cycle distribution, and the activation of ATM signaling pathway in NHEK cells. We demonstrated that the 2 epidermal keratinocytes responded to ELF-EMFs differently. To further validate this finding, we simultaneously exposed the NHEK and HaCaT cells to ELF-EMFs in the same incubator for 168 h and observed the cell growths. The simultaneous exposure of the two cell types results showed that the NHEK and HaCaT cells exhibited distinct responses to ELF-EMFs. Thus, we confirmed that the biological effects of ELF-EMFs in epidermal keratinocytes are cell type specific. Our findings may partially explain the inconsistent results of previous studies when comparing results across various experimental models.
doi:10.1371/journal.pone.0113424
PMCID: PMC4237442  PMID: 25409520
9.  Exposure to Extremely Low-Frequency Electromagnetic Fields Modulates Na+ Currents in Rat Cerebellar Granule Cells through Increase of AA/PGE2 and EP Receptor-Mediated cAMP/PKA Pathway 
PLoS ONE  2013;8(1):e54376.
Although the modulation of Ca2+ channel activity by extremely low-frequency electromagnetic fields (ELF-EMF) has been studied previously, few reports have addressed the effects of such fields on the activity of voltage-activated Na+ channels (Nav). Here, we investigated the effects of ELF-EMF on Nav activity in rat cerebellar granule cells (GCs). Our results reveal that exposing cerebellar GCs to ELF-EMF for 10–60 min significantly increased Nav currents (INa) by 30–125% in a time- and intensity-dependent manner. The Nav channel steady-state activation curve, but not the steady-state inactivation curve, was significantly shifted (by 5.2 mV) towards hyperpolarization by ELF-EMF stimulation. This phenomenon is similar to the effect of intracellular application of arachidonic acid (AA) and prostaglandin E2 (PGE2) on INa in cerebellar GCs. Increases in intracellular AA, PGE2 and phosphorylated PKA levels in cerebellar GCs were observed following ELF-EMF exposure. Western blottings indicated that the NaV 1.2 protein on the cerebellar GCs membrane was increased, the total expression levels of NaV 1.2 protein were not affected after exposure to ELF-EMF. Cyclooxygenase inhibitors and PGE2 receptor (EP) antagonists were able to eliminate this ELF-EMF-induced increase in phosphorylated PKA and INa. In addition, ELF-EMF exposure significantly enhanced the activity of PLA2 in cerebellar GCs but did not affect COX-1 or COX-2 activity. Together, these data demonstrate for the first time that neuronal INa is significantly increased by ELF-EMF exposure via a cPLA2 AA PGE2 EP receptors PKA signaling pathway.
doi:10.1371/journal.pone.0054376
PMCID: PMC3551899  PMID: 23349866
10.  Transient effect of weak electromagnetic fields on calcium ion concentration in Arabidopsis thaliana 
BMC Plant Biology  2009;9:47.
Background
Weak magnetic and electromagnetic fields can influence physiological processes in animals, plants and microorganisms, but the underlying way of perception is poorly understood. The ion cyclotron resonance is one of the discussed mechanisms, predicting biological effects for definite frequencies and intensities of electromagnetic fields possibly by affecting the physiological availability of small ions. Above all an influence on Calcium, which is crucial for many life processes, is in the focus of interest. We show that in Arabidopsis thaliana, changes in Ca2+-concentrations can be induced by combinations of magnetic and electromagnetic fields that match Ca2+-ion cyclotron resonance conditions.
Results
An aequorin expressing Arabidopsis thaliana mutant (Col0-1 Aeq Cy+) was subjected to a magnetic field around 65 microtesla (0.65 Gauss) and an electromagnetic field with the corresponding Ca2+ cyclotron frequency of 50 Hz. The resulting changes in free Ca2+ were monitored by aequorin bioluminescence, using a high sensitive photomultiplier unit. The experiments were referenced by the additional use of wild type plants. Transient increases of cytosolic Ca2+ were observed both after switching the electromagnetic field on and off, with the latter effect decreasing with increasing duration of the electromagnetic impact. Compared with this the uninfluenced long-term loss of bioluminescence activity without any exogenic impact was negligible. The magnetic field effect rapidly decreased if ion cyclotron resonance conditions were mismatched by varying the magnetic fieldstrength, also a dependence on the amplitude of the electromagnetic component was seen.
Conclusion
Considering the various functions of Ca2+ as a second messenger in plants, this mechanism may be relevant for perception of these combined fields. The applicability of recently hypothesized mechanisms for the ion cyclotron resonance effect in biological systems is discussed considering it's operating at magnetic field strengths weak enough, to occur occasionally in our all day environment.
doi:10.1186/1471-2229-9-47
PMCID: PMC2681476  PMID: 19405943
11.  Experimental model for ELF-EMF exposure: Concern for human health 
Low frequency (LF) electromagnetic fields (EMFs) are abundantly present in modern society and in the last 20 years the interest about the possible effect of extremely low frequency (ELF) EMFs on human health has increased progressively. Epidemiological studies, designed to verify whether EMF exposure may be a potential risk factor for health, have led to controversial results. The possible association between EMFs and an increased incidence of childhood leukemia, brain tumors or neurodegenerative diseases was not fully elucidated. On the other hand, EMFs are widely used, in neurology, psychiatry, rheumatology, orthopedics and dermatology, both in diagnosis and in therapy.
In vitro studies may help to evaluate the mechanism by which LF-EMFs affect biological systems.
Invitro model of wound healing used keratinocytes (HaCaT), neuroblastoma cell line (SH-SY5Y) as a model for analysis of differentiation, metabolism and functions related to neurodegenerative processes, and monocytic cell line (THP-1) was used as a model for inflammation and cytokines production, while leukemic cell line (K562) was used as a model for hematopoietic differentiation.
MCP-1, a chemokine that regulates the migration and infiltration of memory T cells, natural killer (NK), monocytes and epithelial cells, has been demonstrated to be induced and involved in various diseases.
Since, varying the parameters of EMFs different effects may be observed, we have studied MCP-1 expression in HaCaT, SH-SY5Y, THP-1 and K562 exposed to a sinusoidal EMF at 50 Hz frequency with a flux density of 1 mT (rms).
Our preliminary results showed that EMF-exposure differently modifies the expression of MCP-1 in different cell types. Thus, the MCP-1 expression needs to be better determined, with additional studies, with different parameters and times of exposure to ELF-EMF.
doi:10.1016/j.sjbs.2014.07.006
PMCID: PMC4281612  PMID: 25561888
AD, Alzheimer’s disease; ELF, extremely low frequency; EMFs, electromagnetic fields; HD, Huntington disease; LF, low frequency; MCP-1, monocyte chemoattractant protein-1; PMA, phorbol-12-myristate-13-acetate; PEMF, pulsed EMF; ELF-EMF; HaCaT; K562; MCP-1; SH-SY5Y; THP-1
12.  Extremely Low-Frequency Electromagnetic Fields Cause G1 Phase Arrest through the Activation of the ATM-Chk2-p21 Pathway 
PLoS ONE  2014;9(8):e104732.
In daily life, humans are exposed to the extremely low-frequency electromagnetic fields (ELF-EMFs) generated by electric appliances, and public concern is increasing regarding the biological effects of such exposure. Numerous studies have yielded inconsistent results regarding the biological effects of ELF-EMF exposure. Here we show that ELF-EMFs activate the ATM-Chk2-p21 pathway in HaCaT cells, inhibiting cell proliferation. To present well-founded results, we comprehensively evaluated the biological effects of ELF-EMFs at the transcriptional, protein, and cellular levels. Human HaCaT cells from an immortalized epidermal keratinocyte cell line were exposed to a 1.5 mT, 60 Hz ELF-EMF for 144 h. The ELF-EMF could cause G1 arrest and decrease colony formation. Protein expression experiments revealed that ELF-EMFs induced the activation of the ATM/Chk2 signaling cascades. In addition, the p21 protein, a regulator of cell cycle progression at G1 and G2/M, exhibited a higher level of expression in exposed HaCaT cells compared with the expression of sham-exposed cells. The ELF-EMF-induced G1 arrest was diminished when the CHK2 gene expression (which encodes checkpoint kinase 2; Chk2) was suppressed by specific small interfering RNA (siRNA). These findings indicate that ELF-EMFs activate the ATM-Chk2-p21 pathway in HaCaT cells, resulting in cell cycle arrest at the G1 phase. Based on the precise control of the ELF-EMF exposure and rigorous sham-exposure experiments, all transcriptional, protein, and cellular level experiments consistently supported the conclusion. This is the first study to confirm that a specific pathway is triggered by ELF-EMF exposure.
doi:10.1371/journal.pone.0104732
PMCID: PMC4128733  PMID: 25111195
13.  Effects of 60 Hz magnetic fields on teenagers and adults 
Environmental Health  2013;12:42.
Background
As use of electrical devices has increased, social concerns about the possible effects of 60 Hz electromagnetic fields on human health have increased. Accordingly, the number of people who complain of various symptoms such as headache and insomnia has risen. Many previous studies of the effects of extremely low frequency (ELF) magnetic field exposure on children have focused on the occurrence of childhood leukaemia and central nervous system cancers. However, very few provocation studies have examined the health effects of ELF magnetic fields on teenagers.
Methods
In this double-blind study, we simultaneously investigated physiological changes (heart rate, respiration rate, and heart rate variability), subjective symptoms, and magnetic field perception to determine the reliable effects of 60 Hz 12.5 μT magnetic fields on teenagers. Two volunteer groups of 30 adults and 30 teenagers were tested with exposure to sham and real magnetic fields for 32 min.
Results
ELF magnetic field exposure did not have any effects on the physiological parameters or eight subjective symptoms in either group. Neither group correctly perceived the magnetic fields.
Conclusions
Physiological data were analysed, subjective symptoms surveyed, and the percentages of those who believed they were being exposed were measured. No effects were observed in adults or teenagers resulting from 32 min of 60 Hz 12.5 μT magnetic field exposure.
doi:10.1186/1476-069X-12-42
PMCID: PMC3681577  PMID: 23705754
ELF; Physiological changes; Subjective symptoms; Perception
14.  Double tuning a single input probe for heteronuclear NMR spectroscopy at low field 
Applications of PASADENA in biomedicine are continuing to emerge due to recent demonstrations that hyperpolarized metabolic substrates and the corresponding reaction products persist sufficiently long to be detected in vivo. Biomedical applications of PASADENA typically differ from their basic science counterparts in that the polarization endowed by addition of parahydrogen is usually transferred from nascent protons to coupled storage nuclei for subsequent detection on a higher field imaging instrument. These pre-imaging preparations usually take place at low field, but commercial spectrometers capable of heteronuclear pulsed NMR at frequencies in the range of 100 kHz to 1 MHz are scarce though, in comparison to single channel consoles in that field regime. Reported here is a probe circuit that can be used in conjunction with a phase and amplitude modulation scheme we have developed called PANORAMIC (Precession And Nutation for Observing Rotations At Multiple Intervals about the Carrier), that expands a single channel console capability to double or generally multiple resonance with minimal hardware modifications. The demands of this application are geared towards uniform preparation, and since the hyperpolarized molecules are being detected externally at high field, detection sensitivity is secondary to applied field uniformity over a large reaction volume to accommodate heterogeneous chemistry of gas molecules at a liquid interface. The probe circuit was therefore configured with a large (40 mL) Helmholtz sample coil for uniformity, and double-tuned to the Larmor precession frequencies of 13C/1H (128/510 kHz) within a custom solenoidal electromagnet at a static field of 12 mT. Traditional (on-resonant) as well as PANORAMIC NMR signals with signal to noise ratios of approximately 75 have been routinely acquired with this probe and spectrometer setup from 1024 repetitions on the high frequency channel. The proton excitation pulse width was 240 μs at 6.31 W, compared to a carbon-13 pulse width of 220 μs at 2.51 W. When PANORAMIC refocusing waveforms were transmitted at a carrier frequency of 319 kHz, integrated signal intensities from a spin-echo sequence at both proton (510 kHz) and carbon-13 (128 kHz) frequencies were within experimental error to block pulse analogs transmitted on resonance. We anticipate that this probe circuit design could be extended to higher and lower frequencies, and that when used in conjunction with PANORAMIC phase and amplitude modulated arrays, will enable low field imaging consoles to serve as multinuclear consoles.
doi:10.1016/j.jmr.2012.06.008
PMCID: PMC4118687  PMID: 22975236
Single channel spectrometer; Low field NMR; Parahydrogen induced polarization; Dual resonant probe circuit; Double tuned; Single input; PANORAMIC; Phase and amplitude modulation; PASADENA; Hyperpolarization
15.  Extremely Low Frequency Magnetic Fields Induce Spermatogenic Germ Cell Apoptosis: Possible Mechanism 
BioMed Research International  2014;2014:567183.
The energy generated by an extremely low frequency electromagnetic field (ELF-EMF) is too weak to directly induce genotoxicity. However, it is reported that an extremely low frequency magnetic field (ELF-MF) is related to DNA strand breakage and apoptosis. The testes that conduct spermatogenesis through a dynamic cellular process involving meiosis and mitosis seem vulnerable to external stress such as heat, MF exposure, and chemical or physical agents. Nevertheless the results regarding adverse effects of ELF-EMF on human or animal reproductive functions are inconclusive. According to the guideline of the International Commission on Non-Ionizing Radiation Protection (ICNIRP; 2010) for limiting exposure to time-varying MF (1 Hz to 100 kHz), overall conclusion of epidemiologic studies has not consistently shown an association between human adverse reproductive outcomes and maternal or paternal exposure to low frequency fields. In animal studies there is no compelling evidence of causal relationship between prenatal development and ELF-MF exposure. However there is increasing evidence that EL-EMF exposure is involved with germ cell apoptosis in testes. Biophysical mechanism by which ELF-MF induces germ cell apoptosis has not been established. This review proposes the possible mechanism of germ cell apoptosis in testes induced by ELF-MF.
doi:10.1155/2014/567183
PMCID: PMC4082851  PMID: 25025060
16.  Growth of etiolated barley plants in weak static and 50 Hz electromagnetic fields tuned to calcium ion cyclotron resonance 
Background
The effects of weak magnetic and electromagnetic fields in biology have been intensively studied on animals, microorganisms and humans, but comparably less on plants. Perception mechanisms were attributed originally to ferrimagnetism, but later discoveries required additional explanations like the "radical pair mechanism" and the "Ion cyclotron resonance" (ICR), primarily considered by Liboff. The latter predicts effects by small ions involved in biological processes, that occur in definite frequency- and intensity ranges ("windows") of simultaneously impacting magnetic and electromagnetic fields related by a linear equation, which meanwhile is proven by a number of in vivo and in vitro experiments.
Methods
Barley seedlings (Hordeum vulgare, L. var. Steffi) were grown in the dark for 5 and 6 days under static magnetic and 50 Hz electromagnetic fields matching the ICR conditions of Ca2+. Control cultures were grown under normal geomagnetic conditions, not matching this ICR. Morphology, pigmentation and long-term development of the adult plants were subsequently investigated.
Results
The shoots of plants exposed to Ca2+-ICR exposed grew 15–20% shorter compared to the controls, the plant weight was 10–12% lower, and they had longer coleoptiles that were adhering stronger to the primary leaf tissue. The total pigment contents of protochlorophyllide (PChlide) and carotenoids were significantly decreased. The rate of PChlide regeneration after light irradiation was reduced for the Ca2+-ICR exposed plants, also the Shibata shift was slightly delayed. Even a longer subsequent natural growing phase without any additional fields could only partially eliminate these effects: the plants initially exposed to Ca2+-ICR were still significantly shorter and had a lower chlorophyll (a+b) content compared to the controls. A continued cultivation and observation of the adult plants under natural conditions without any artificial electromagnetic fields showed a retardation of the originally Ca2+-ICR exposed plants compared to control cultures lasting several weeks, with an increased tendency for dehydration.
Conclusion
A direct influence of the applied MF and EMF is discussed affecting Ca2+ levels via the ICR mechanism. It influences the available Ca2+ and thereby regulatory processes. Theoretical considerations on molecular level focus on ionic interactions with water related to models using quantum electrodynamics.
doi:10.1186/1477-044X-4-1
PMCID: PMC1403775  PMID: 16457719
17.  The Influence of Electromagnetic Radiation Generated by a Mobile Phone on the Skeletal System of Rats 
BioMed Research International  2015;2015:896019.
The study was focused on the influence of electromagnetic field generated by mobile phone on the skeletal system of rats, assessed by measuring the macrometric parameters of bones, mechanical properties of long bones, calcium and phosphorus content in bones, and the concentration of osteogenesis (osteocalcin) and bone resorption (NTX, pyridinoline) markers in blood serum. The study was carried out on male rats divided into two groups: experimental group subjected to 28-day cycle of exposures in electromagnetic field of 900 MHz frequency generated by mobile phone and a control, sham-exposed one. The mobile phone-generated electromagnetic field did not influence the macrometric parameters of long bones and L4 vertebra, it altered mechanical properties of bones (stress and energy at maximum bending force, stress at fracture), it decreased the content of calcium in long bones and L4 vertebra, and it altered the concentration of osteogenesis and bone resorption markers in rats. On the basis of obtained results, it was concluded that electromagnetic field generated by 900 MHz mobile phone does not have a direct impact on macrometric parameters of bones; however, it alters the processes of bone mineralization and the intensity of bone turnover processes and thus influences the mechanical strength of bones.
doi:10.1155/2015/896019
PMCID: PMC4331479
18.  Anti-proliferative effect of extremely low frequency electromagnetic field on preneoplastic lesions formation in the rat liver 
BMC Cancer  2010;10:159.
Background
Recently, extremely low frequency electromagnetic fields (ELF-EMF) have been studied with great interest due to their possible effects on human health. In this study, we evaluated the effect of 4.5 mT - 120 Hz ELF-EMF on the development of preneoplastic lesions in experimental hepatocarcinogenesis.
Methods
Male Fischer-344 rats were subjected to the modified resistant hepatocyte model and were exposed to 4.5 mT - 120 Hz ELF-EMF. The effects of the ELF-EMF on hepatocarcinogenesis, apoptosis, proliferation and cell cycle progression were evaluated by histochemical, TUNEL assay, caspase 3 levels, immunohistochemical and western blot analyses.
Results
The application of the ELF-EMF resulted in a decrease of more than 50% of the number and the area of γ-glutamyl transpeptidase-positive preneoplastic lesions (P = 0.01 and P = 0.03, respectively) and glutathione S-transferase placental expression (P = 0.01). The number of TUNEL-positive cells and the cleaved caspase 3 levels were unaffected; however, the proliferating cell nuclear antigen, Ki-67, and cyclin D1 expression decreased significantly (P ≤ 0.03), as compared to the sham-exposure group.
Conclusion
The application of 4.5 mT - 120 Hz ELF-EMF inhibits preneoplastic lesions chemically induced in the rat liver through the reduction of cell proliferation, without altering the apoptosis process.
doi:10.1186/1471-2407-10-159
PMCID: PMC2873390  PMID: 20416104
19.  Effects of power frequency electromagnetic fields on melatonin and sleep in the rat 
Emerging Health Threats Journal  2012;5:10.3402/ehtj.v5i0.10904.
Background
Studies investigating the effect of power frequency (50–60 Hz) electromagnetic fields (EMF) on melatonin synthesis in rats have been inconsistent with several showing suppression of melatonin synthesis, others showing no effect and a few actually demonstrating small increases. Scant research has focused on the ensuing sleep patterns of EMF exposed rats. The present study was designed to examine the effects of extremely low power frequency electromagnetic fields (EMF) on the production of melatonin and the subsequent sleep structure in rats.
Methods
Eighteen male Sprague-Dawley rats were exposed to a 1000 milligauss (mG) magnetic field for 1 month. Urine was collected for the final 3 days of the exposure period for analysis of 6-sulphatoxymelatonin, the major catabolic product of melatonin found in urine. Subsequent sleep was analyzed over a 24-hour period.
Results
Melatonin production was mildly increased in exposed animals. Although there were no statistically significant changes in sleep structure, exposed animals showed slight decreases in REM (rapid eye movement) sleep as compared to sham (non-exposed) animals.
Conclusions
Power frequency magnetic fields induced a marginally statistically significant increase in melatonin levels in exposed rats compared to control. Subsequent sleep analysis indicated little effect on the sleep architecture of rats, at least not within the first day after 1 month's continuous exposure. Varying results in the literature are discussed and future research suggested.
doi:10.3402/ehtj.v5i0.10904
PMCID: PMC3334267  PMID: 22529876
electromagnetic fields; 6-sulphatoxymelatonin; Sprague-Dawley; sleep
20.  Non-Thermal Electromagnetic Radiation Damage to Lens Epithelium 
High frequency microwave electromagnetic radiation from mobile phones and other modern devices has the potential to damage eye tissues, but its effect on the lens epithelium is unknown at present. The objective of this study was to investigate the non-thermal effects of high frequency microwave electromagnetic radiation (1.1GHz, 2.22 mW) on the eye lens epithelium in situ. Bovine lenses were incubated in organ culture at 35°C for 10-15 days. A novel computer-controlled microwave source was used to investigate the effects of microwave radiation on the lenses. 58 lenses were used in this study. The lenses were divided into four groups: (1) Control lenses incubated in organ culture for 10 to15 days. (2) Electromagnetic radiation exposure group treated with 1.1 GHz, 2.22 mW microwave radiation for 90 cycles of 50 minutes irradiation followed by 10 minutes pause and cultured up to 10 days. (3) Electromagnetic radiation exposure group treated as group 2 with 192 cycles of radiation and cultured for 15 days. (4) Lenses exposed to 39.5ºC for 2 hours 3 times with 24 hours interval after each treatment beginning on the second day of the culture and cultured for 11 days. During the culture period, lens optical quality was followed daily by a computer-operated scanning laser beam. At the end of the culture period, control and treated lenses were analyzed morphologically and by assessment of the lens epithelial ATPase activity. Exposure to 1.1 GHz, 2.22 mW microwaves caused a reversible decrease in lens optical quality accompanied by irreversible morphological and biochemical damage to the lens epithelial cell layer. The effect of the electromagnetic radiation on the lens epithelium was remarkably different from those of conductive heat. The results of this investigation showed that electromagnetic fields from microwave radiation have a negative impact on the eye lens. The lens damage by electromagnetic fields was distinctly different from that caused by conductive heat.
doi:10.2174/1874364100802010102
PMCID: PMC2694600  PMID: 19517034
21.  Idiopathic Environmental Intolerance Attributed to Electromagnetic Fields: A Content Analysis of British Newspaper Reports 
PLoS ONE  2013;8(6):e65713.
Idiopathic environmental intolerance attributed to electromagnetic fields (IEI-EMF) is a controversial condition in which people describe symptoms following exposure to electromagnetic fields from everyday electrical devices. However, double-blind experiments have found no convincing evidence that electromagnetic fields cause these symptoms. In this study, we assessed whether recent newspaper reporting in the UK reflected this scientific evidence. We searched a database of newspaper articles to identify all those that contained IEI-EMF related keywords and selected a random sample of 60 for content analysis. For our primary outcomes, we assessed how many articles mainly or wholly presented an electromagnetic cause for IEI-EMF and how many discussed unproven treatments for the condition such as strategies intended to reduce exposure to electromagnetic fields or the use of complementary and alternative therapies. We also assessed whether the type of information source used by a newspaper article (e.g. scientist, person with IEI-EMF, politician) or the type of newspaper (broadsheet, tabloid, local or regional) was associated with either outcome. Of the 60 articles, 43 (71.7%) presented a mainly electromagnetic cause, compared to 13 (21.7%) which presented mainly non-electromagnetic causes and 4 (6.7%) which did not discuss a cause. 29 (48.3%) did not mention any potential treatment, while 24 (40.0%) mentioned eletromagnetic field related strategies and 12 (20.0%) mentioned complementary or alternative therapies. Articles which quoted someone with IEI-EMF were significantly more likely to report an electromagnetic cause and to present unproven treatments. Those which used a scientist as a source were more likely to present a non-electromagnetic cause for the condition. The widespread poor reporting we identified is disappointing and has the potential for to encourage more people to misattribute their symptoms to electromagnetic fields. Scientists should remain engaged with the media to counteract this effect.
doi:10.1371/journal.pone.0065713
PMCID: PMC3683033  PMID: 23799038
22.  Terahertz Electromagnetic Fields (0.106 THz) Do Not Induce Manifest Genomic Damage In Vitro 
PLoS ONE  2012;7(9):e46397.
Terahertz electromagnetic fields are non-ionizing electromagnetic fields in the frequency range from 0.1 to 10 THz. Potential applications of these electromagnetic fields include the whole body scanners, which currently apply millimeter waves just below the terahertz range, but future scanners will use higher frequencies in the terahertz range. These and other applications will bring along human exposure to these fields. Up to now, only a limited number of investigations on biological effects of terahertz electromagnetic fields have been performed. Therefore, research is strongly needed to enable reliable risk assessment.
Cells were exposed for 2 h, 8 h, and 24 h with different power intensities ranging from 0.04 mW/cm2 to 2 mW/cm2, representing levels below, at, and above current safety limits. Genomic damage on the chromosomal level was measured as micronucleus formation. DNA strand breaks and alkali-labile sites were quantified with the comet assay. No DNA strand breaks or alkali-labile sites were observed as a consequence of exposure to terahertz electromagnetic fields in the comet assay. The fields did not cause chromosomal damage in the form of micronucleus induction.
doi:10.1371/journal.pone.0046397
PMCID: PMC3459899  PMID: 23029508
23.  Effects of whole body exposure to extremely low frequency electromagnetic fields (ELF-EMF) on serum and liver lipid levels, in the rat 
Backgound
The effects of extremely low-frequency electromagnetic fields (ELF-EMF) on the blood serum and liver lipid concentrations of male Wistar rats were assessed.
Methods
Animals were exposed to a single stimulation (2 h) of ELF-EMF (60 Hz, 2.4 mT) or sham-stimulated and thereafter sacrificed at different times (24, 48 or 96 h after beginning the exposure).
Results
Blood lipids showed, at 48 h stimulated animals, a significant increase of cholesterol associated to high density lipoproteins (HDL-C) than those observed at any other studied time. Free fatty acid serum presented at 24 h significant increases in comparison with control group. The other serum lipids, triacylglycerols and total cholesterol did not show differences between groups, at any time evaluated. No statistical differences were shown on total lipids of the liver but total cholesterol was elevated at 24 h with a significant decrease at 96 h (p = 0.026). The ELF-EMF stimulation increased the liver content of lipoperoxides at 24 h.
Conclusion
Single exposures to ELF-EMF increases the serum values of HDL-C, the liver content of lipoperoxides and decreases total cholesterol of the liver. The mechanisms for the effects of ELF-EMF on lipid metabolism are not well understand yet, but could be associated to the nitric oxide synthase EMF-stimulation.
doi:10.1186/1476-511X-6-31
PMCID: PMC2203969  PMID: 18021407
24.  Evaluation of Specific Absorption Rate as a Dosimetric Quantity for Electromagnetic Fields Bioeffects 
PLoS ONE  2013;8(6):e62663.
Purpose
To evaluate SAR as a dosimetric quantity for EMF bioeffects, and identify ways for increasing the precision in EMF dosimetry and bioactivity assessment.
Methods
We discuss the interaction of man-made electromagnetic waves with biological matter and calculate the energy transferred to a single free ion within a cell. We analyze the physics and biology of SAR and evaluate the methods of its estimation. We discuss the experimentally observed non-linearity between electromagnetic exposure and biological effect.
Results
We find that: a) The energy absorbed by living matter during exposure to environmentally accounted EMFs is normally well below the thermal level. b) All existing methods for SAR estimation, especially those based upon tissue conductivity and internal electric field, have serious deficiencies. c) The only method to estimate SAR without large error is by measuring temperature increases within biological tissue, which normally are negligible for environmental EMF intensities, and thus cannot be measured.
Conclusions
SAR actually refers to thermal effects, while the vast majority of the recorded biological effects from man-made non-ionizing environmental radiation are non-thermal. Even if SAR could be accurately estimated for a whole tissue, organ, or body, the biological/health effect is determined by tiny amounts of energy/power absorbed by specific biomolecules, which cannot be calculated. Moreover, it depends upon field parameters not taken into account in SAR calculation. Thus, SAR should not be used as the primary dosimetric quantity, but used only as a complementary measure, always reporting the estimating method and the corresponding error. Radiation/field intensity along with additional physical parameters (such as frequency, modulation etc) which can be directly and in any case more accurately measured on the surface of biological tissues, should constitute the primary measure for EMF exposures, in spite of similar uncertainty to predict the biological effect due to non-linearity.
doi:10.1371/journal.pone.0062663
PMCID: PMC3672148  PMID: 23750202
25.  Non Ionising Radiation as a Non Chemical Strategy in Regenerative Medicine: Ca2+-ICR “In Vitro” Effect on Neuronal Differentiation and Tumorigenicity Modulation in NT2 Cells 
PLoS ONE  2013;8(4):e61535.
In regenerative medicine finding a new method for cell differentiation without pharmacological treatment or gene modification and minimal cell manipulation is a challenging goal. In this work we reported a neuronal induced differentiation and consequent reduction of tumorigenicity in NT2 human pluripotent embryonal carcinoma cells exposed to an extremely low frequency electromagnetic field (ELF-EMF), matching the cyclotron frequency corresponding to the charge/mass ratio of calcium ion (Ca2+-ICR). These cells, capable of differentiating into post-mitotic neurons following treatment with Retinoic Acid (RA), were placed in a solenoid and exposed for 5 weeks to Ca2+-ICR. The solenoid was installed in a μ-metal shielded room to avoid the effect of the geomagnetic field and obtained totally controlled and reproducible conditions. Contrast microscopy analysis reveled, in the NT2 exposed cells, an important change in shape and morphology with the outgrowth of neuritic-like structures together with a lower proliferation rate and metabolic activity alike those found in the RA treated cells. A significant up-regulation of early and late neuronal differentiation markers and a significant down-regulation of the transforming growth factor-α (TGF-α) and the fibroblast growth factor-4 (FGF-4) were also observed in the exposed cells. The decreased protein expression of the transforming gene Cripto-1 and the reduced capability of the exposed NT2 cells to form colonies in soft agar supported these last results. In conclusion, our findings demonstrate that the Ca2+-ICR frequency is able to induce differentiation and reduction of tumorigenicity in NT2 exposed cells suggesting a new potential therapeutic use in regenerative medicine.
doi:10.1371/journal.pone.0061535
PMCID: PMC3621667  PMID: 23585910

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