Pilot study of extremely low frequency magnetic fields emitted by transformers in dwellings. Social aspects

A large number of epidemiologic studies examining the potential effect of residential exposure to extremely-low frequency (ELF) magnetic fields and childhood leukemia have been published. Two pooled analyses [Ahlbom A, Day N, Feychting M, Roman E, Skinner J, Dockerty J, Linet M, et al. (2000). A pooled analysis of magnetic fields and childhood leukaemia. Br J Cancer. 83(5):692–698; Greenland S, Sheppard AR, Kaune WT, Poole C, Kelsh AM (2000). A pooled analysis of magnetic fields, wire codes, and childhood leukemia. Epidemiology. 11(6):624–634], which included the major epidemiologic studies on ELF magnetic fields and childhood leukemia showed twofold increase in childhood leukemia risk in association with residential ELF exposure above 0.3–0.4 μT. Based on “limited” epidemiologic evidence linking ELF exposure to childhood leukemia and “inadequate evidence” for carcinogenicity of ELF in rodent bioassays, the International Agency for Research on Cancer (IARC) classified ELF magnetic fields as a possible human carcinogen (2B classification) [International Agency for Research on Cancer (IARC) (2002). Non-ionizing radiation, Part 1: Static and extremely low-frequency (ELF) electric and magnetic fields. IARC monographs on the evaluation of carcinogenic risks to humans. Vol. 80. IARC Press: Lyon], confirmed by WHO on the basis of studies published after 2000 [World Health Organization. Extremely low frequency fields. In: 238 Environmental health criteria, Geneva: WHO; 2007]. The analysis of more recent studies of ELF magnetic fields and childhood leukemia had small findings and propose methodological improvements concerning the uncertainties in epidemiological approaches and exposure assessment, bias in selection of controls [Kheifets L, Oksuzyan S (2008). Exposure assessment and other challenges in non-ionizing radiation studies of childhood leukaemia. Radiat Prot Dosimetry. 132(2):139–147]. By the end of 2010, 37 countries had been identified for possible participation in the International study TRANSEXPO. The pilot work has been completed in five countries (Finland, Hungary, Israel, Switzerland and Bulgaria). In 2008, Bulgaria through the National Centre of Public Health Protection joined with pilot study in TRANSEXPO Project. At this first stage of the project our investigation was directed to performing measurements in dwellings with built-in transformer stations, collecting data of population and cancer registry and choosing the epidemiology design feasible for continuing the project. Taking into account the available sources of information in Bulgaria (different registers of the population) needed for epidemiological approach, it was found that the most appropriate epidemiology design would be the nested case-control study. Control group could be collected in accordance with the international requirements for such epidemiological studies. This approach could be modified in the course of the further study in order to ensure achievement of the purposes of the main international requirements of the study.     

Proteomic analysis of rat pheochromocytoma PC12 cells

PC12 cell line is well documented and widely applied as many kinds of models in neurobiological and neurochemical studies. Yet a thorough proteomic analysis has not been performed so far. Here we report the construction of a large-scale 2-D protein database for PC12 cells. The proteins extracted from PC12 cells were separated by 2-DE and identified by MALDI-TOF/TOF MS. A total of 1080 protein spots, excised from three different 2-D gels, were identified with high confidence. These proteins represent 474 different gene products, mainly binding proteins and enzymes. Three hundred and seven identified protein spots were located in the low-molecular weight region below 20 kDa. This database today represents one of the largest 2-D databases for higher eukaryotic cell proteomes and for low-molecular weight proteins. In addition, fragment ion spectra obtained by TOF/TOF confirmed that calcylin in PC12 cells was N-acetylated. The database of PC12 proteome is expected to be a powerful tool for neuroscientists.     

The detection threshold for extremely low frequency magnetic fields may be below 1000 nT-Hz in mice

Previous experiments with mice have shown that a repeated 1 h daily exposure to an ambient magnetic field shielded environment induces analgesia (anti-nociception). This shielding reduces ambient static and extremely low frequency magnetic fields (ELF-MF) by approximately 100 times for frequencies below 120 Hz. To determine the threshold of ELF-MF amplitude that would attenuate or abolish this effect, 30 and 120 Hz magnetic fields were introduced into the shielded environment at peak amplitudes of 25, 50, 100 and 500 nT. At 30 Hz, peak amplitudes of 50, 100, and 500 nT attenuated this effect in proportion to the amplitude magnitude. At 120 Hz, significant attenuation was observed at all amplitudes. Exposures at 10, 60, 100, and 240 Hz with peak amplitudes of 500, 300, 500, and 300 nT, respectively, also attenuated the induced analgesia. No exposure abolished this effect except perhaps at 120 Hz, 500 nT. If the peak amplitude frequency product was kept constant at 6000 nT-Hz for frequencies of 12.5, 25, 50, and 100 Hz, the extent of attenuation was constant, indicating that the detection mechanism is dependent on the nT-Hz product. A plot of effect versus the induced current metric nT-Hz suggests a threshold of ELF-MF detection in mice at or below 1000 nT-Hz.     

Effects of vibration on differentiation of cultured PC12 cells

Different types of physiological‐mechanical stress, such as shear stress in vascular endothelial cells or hydrostatic pressure in chondrocytes are well known as regulators of cell function. In this study, the effects of vibration, a type of non‐physiological mechanical stimulation, on differentiation of rat pheochromocytoma (PC12) cells are reported. A nano‐vibration system was designed to produce nanometer‐scale vibration. The frequency and amplitude of the nano‐vibrations were monitored by a capacitance displacement sensor connected to an oscilloscope. When PC12 cells exposed to nerve growth factor were subjected to vibration at 10 kHz, differentiation and elongation of their neurites were promoted earlier in the culture. Vibration promoted differentiation of PC12 cells. This approach could therefore also be promising for determining of the effects of the physical environment on cell differentiation. Biotechnol. Bioeng. 2011; 108:592–599. © 2010 Wiley Periodicals, Inc.     

Neural stimulation on human bone marrow‐derived mesenchymal stem cells by extremely low frequency electromagnetic fields

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. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

极低频正弦磁场对大鼠痛阈及氨基酸神经递质的影响

目的:探讨极低频正弦磁场对痛阈的影响以及脑内氨基酸神经递质在磁场镇痛中的作用。方法:20只SD大鼠暴露于55.6Hz、8.1mT正弦磁场中并利用辐射热甩尾法检测痛阈,同时用高效液相色谱检测暴露第11天和第14天大脑皮层和延髓谷氨酸和γ-氨基丁酸(GABA)含量。结果:与对照组相比大鼠磁场暴露第10天和第11天痛阈明显提高(P0.05)。第11天大脑皮层和延髓的γ-氨基丁酸水平有明显改变(P0.05)。结论:极低频正弦磁场有镇痛作用,调节γ-氨基丁酸可能是其作用机制之一。     

Effect of extremely low frequency magnetic fields on calpain activation

The effects of low intensity, low frequency magnetic fields (MFs) on catalytic activity of the calcium dependent protease calpain was determined following the enzyme activation both in "in vitro" and "in vivo" conditions. We have observed that a 0.3 mT MF induces a significant increase in the requirement of the protease for this metal ion. This change is detectable at low [Ca(2+)] and disappears when the level of Ca(2+) is raised to saturating amounts. The observed effects are not due to transient MF(-) induced conformational changes occurring in calpain, but to direct effects of the MF on Ca(2+) ions, which become less available for the binding sites present in calpain. Altogether, these results indicate that exposure to low intensity, low frequency MFs alters the intracellular Ca(2+) "availability," thereby modifying the related cell response.     

Binding-induced activation of overexpressed p185HER2 is essential in triggering neuronal differentiation of PC12 cells

To determine whether p185^HER2 overexpression per se triggers p185^HER2 cellular signaling or whether an extracellular signal is required, we transfected PC12 cells with the human erbB-2 proto-oncogene, and established a cell line that overexpresses p185^HER2. PC12-HER2 cells, maintained in suspension culture or plated on a collagen layer, showed the same morphology and growth rate as PC12 and PC12 mock-transfected control cells. When treated with monoclonal antibody (MAb) MGr6 or other anti-p185^HER2 MAbs, PC12–HER2 cells specifically underwent neuronal differentiation comparable to that induced by nerve growth factor (NGF), and the differentiation-inducing effect of the MAb was dramatically enhanced by the addition of a second anti-mouse IgG. MAb-induced cell differentiation correlated with p185^HER2 phosphorylation, recruitment of Shc and Grb-2 transducer molecules into complexes, and MAPK phosphorylation. These data indicate the requirement for a specific binding-induced activation of the overexpressed p185^HER2 receptor in inducing PC12 cell differentiation. PC12-HER2 cells represent a suitable system for selection of p185^HER2-activating ligands (peptides, phage-displayed peptides or proteins) or specific inhibitors of its tyrosine kinase activity. J. Cell. Biochem. 67:316–326, 1997. © 1997 Wiley-Liss, Inc.     

Experimental studies on extremely low frequency pulsed magnetic field inhibiting sarcoma and enhancing cellular immune functions

The previous observation with an electron microscope showed that extremely low frequency (ELF) pulsed magnetic field (PMF) (with the maximum intensity of 0. 6-2. 0 T, gradient of 10-100 T. M-1, pulse width of 20-200 ms and frequency of 0. 16-1. 34 Hz) inhibited the growth of S-180 sarcoma in mice and enhanced the ability of immune cell's dissolving sarcoma cells. In this study, the DNA contents of nuclei were assayed by using Faulgen Staining method. With an electron microscope and cell stereoscopy technology it was observed that magnetic field affected the sarcoma cell's metabolism, lowered its malignancy, and restrained its rapid and heteromorphic growth. The magnetic field enhanced the cellular immune ability and the reaction of lymphocytes and plasma. Since ELF pulsed magnetic fields can inhibit the growth of sarcomas and enhance the cellular immune ability, it is possible to use it as a new method to treat cancer.     

Analysis of the structure of magnetic fields that induced inhibition of stimulated neurite outgrowth

The important experiments showing nonlinear amplitude dependences of the neurite outgrowth in pheochromocytoma nerve cells due to ELF magnetic field exposure had been carried out in a nonuniform ac magnetic field. The nonuniformity entailed larger than expected variances in magnetic field magnitudes associated with specific levels of biological effects, thereby evoking a question about validity of the interpretations formulated for the case of a uniform field. In this work, we calculate the relative value of nonuniformity and deviations in ac magnetic field. It is shown that these factors do not affect the main conclusion in the original papers about the form of the amplitude dependence of the observed biological effect.     

Inability of 50 Hz magnetic fields to regulate PKC- and Ca(2+)-dependent gene expression in Jurkat cells

We have previously reported that the T cell line Jurkat registers the exposure of a sinusoidal extremely low frequency magnetic field at the level of the plasma membrane, resulting in activation of the tyrosine kinase p56(lck), increase in inositol-3-phosphate levels and increase in intracellular calcium concentration within minutes. To elucidate if these events associated with changes in intracellular calcium ion levels were biologically significant, transient transfections of Jurkat cells were performed with calcium-ion dependent reporter constructs. Three different enhancer/promoter constructs were studied coupled to the luciferase reporter gene. The luciferase activity of each construct was measured after treatment of transfected cells to EMF exposure alone, or in combination with ionomycin, phorbol ester or cross-linking anti-CD3 antibodies. There was no indication that the used EMFs could influence any of these reporter constructs.     

Analysis of individual- and school-level clustering of power frequency magnetic fields

This study reports the continuous 8-h monitoring of data on extremely low-frequency magnetic fields (ELF-MF) relating to 14 children and 35 teachers in 11 elementary schools in Northern Taiwan. It was anticipated that the subjects in two of these campuses would have elevated exposure to ELF-MF as a result of their close proximity to high-voltage (161 kilo-Volt, kV) power lines. The results of our analysis reveal that in those schools with high-voltage power lines running through the campuses, the mean ELF-MF exposure level (0.38 +/- 0.51 micro-Tesla (microT), or 0.15, 0.25 and 0.44 microT at the respective 25th, 50th and 75th percentiles) was higher than the mean ELF-MF exposure level for campuses situated far away from such high-voltage power lines (0.14 +/- 0.27 microT, or 0.04, 0.06 and 0.10 microT at the respective 25th, 50th and 75th percentiles). The multi-level analytical technique, which takes individual measurements as the analytical unit, and which also takes into consideration the inter-correlation between measurements from the same individual and/or campus, was also applied to the analysis of the data. We conclude that individual-level and school-level clustering of the measurements, both of which were discernible in this study, should be taken into consideration in any future analysis of data obtained from the continuous monitoring of exposure to ELF-MF.     

Effects of an extremely low frequency electromagnetic field on the cell division rate and plasma membrane of Paramecium tetraurelia

The eukaryotic protozoan, Paramecium, was examined as a model for effects of pulsated electromagnetic fields (PEMF) on cells. A 72-Hz PEMF similar to fields employed clinically increased cell division rates in Paramecium by 8.5%. Two calcium transport mutants of these organisms showed differential responses to the same field. Verapamil, a calcium channel blocker, abolished any effect of PEMFs on cell division rates. A fluorescent probe that is thought to sense changes in membrane potential also manifested an altered response in the PEMF-exposed cells whereas a fluorescent lipid bilayer fluidity probe produced evidence of decreased membrane fluidity in the exposed cells. An effect of PEMFs on ion transport mediated by either a direct or indirect effect on the cell membrane is suggested by these studies.     

Overexpression of miR-26b-5p regulates the cell cycle by targeting CCND2 in GC-2 cells under exposure to extremely low frequency electromagnetic fields

The increasing prevalence of extremely low frequency electromagnetic fields (ELF-EMFs) exposure has raised considerable public concern regarding the potential hazardous effects of ELF-EMFs on male reproductive function. Increasing evidence indicates that miRNAs are necessary for spermatogenesis and male fertility. However, the regulation of miRNA expression and the roles of miRNAs in response to ELF-EMFs remain unclear. In our study, mouse spermatocyte-derived GC-2 cells were intermittently exposed to a 50 Hz ELF-EMF for 72 h (5 min on/10 min off) at magnetic field intensities of 1 mT, 2 mT and 3 mT. MiR-26b-5p was differentially expressed in response to different magnetic field intensities of ELF-EMFs. The host gene CTDSP1 showed an unmethylation status in GC-2 cells at different magnetic field intensities of ELF-EMF exposure. MiR-26b-5p had no significant, obvious influence on the cell viability, apoptosis or cell cycle of GC-2 cells. However, the overexpression of miR-26b-5p significantly decreased the percentage of G0/G1 phase cells and slightly increased the percentage of S phase cells compared to the sham group that was exposed to a 50 Hz ELF-EMF. Computational algorithms identified Cyclin D2 (CCND2) as a direct target of miR-26b-5p. MiR-26b-5p and a 50 Hz ELF-EMF altered the expression of CCND2 at both the mRNA and protein levels. Overexpressed miR-26b-5p in GC-2 cells can change the mRNA expression of CCND2 following 50 Hz ELF-EMF at 3 mT. These findings demonstrate that miR-26b-5p could serve as a potential biomarker following 50 Hz ELF-EMF exposure, and miR-26b-5p-CCND2-mediated cell cycle regulation might play a pivotal role in the biological effects of ELF-EMFs.     

Extremely low frequency (ELF) pulsed-gradient magnetic fields inhibit malignant tumour growth at different biological levels

Extremely low frequency (ELF) pulsed-gradient magnetic field (with the maximum intensity of 0.6-2.0 T, gradient of 10-100 T.M(-1), pulse width of 20-200 ms and frequency of 0.16-1.34 Hz treatment of mice can inhibit murine malignant tumour growth, as seen from analyses at different hierarchical levels, from organism, organ, to tissue, and down to cell and macromolecules. Such magnetic fields induce apoptosis of cancer cells, and arrest neoangiogenesis, preventing a supply developing to the tumour. The growth of sarcomas might be amenable to such new method of treatment.     

Effects of combined DC and AC magnetic fields on germination of hornwort seeds

Seeds of hornwort (Cryptotaenia japonica Hassk) were exposed to sinusoidally time-varying extremely low frequency (ELF) magnetic fields (AC fields) in combination with the local geomagnetic field (DC field). Exposure lasted 24 h/day for 16 days. Three directions of the AC magnetic fields were considered; the vertical (magnetic flux density B ACV, the directions parallel B ACparallel), and perpendicular B ACperpendicular to the direction of total geomagnetic field (magnetic flux density BG) in the geomagnetic plane (GP). Controls consisted of seeds exposed to zero AC magnetic fields in combination with the DC magnetic field. The B ACV in combination with BG effectively promoted the germination of hornwort seeds when applied at 750 microT (RMS) at 7 Hz or 500 microT (RMS) at 14 Hz from among the cases of individual frequencies f = 3.5, 7.0, 10.5, 14.0 Hz at 500 and 750 microT. The B ACparallel promoted the germination of hornwort seeds more effectively than the B ACperpendicular in combination with BG when 500 and 750 microT at 7 Hz were applied.