Chapter One Review

Introduction and Literature

1-1 Introduction:
Numerous sources of magnetic field exist in nature and in occupational and residential environments. Magnetic fields from permanent magnet and from electromagnet have the same effect on the surrounding objects, both fields from both sources we are called electromagnetic field (EMF). Probably non-ionizing radiation from electromagnetic field interact with biological system in many different ways depending on their intensities and frequencies, as well as on physiological properties of the biological system [1]. The low frequency region, whether the magnetic, or the electrical component of the field is responsible for primary reaction, there are experiment support both kinds of interactions. In nearly, all instances these fields pose no obvious threat to human health or safety and are generally discussed as an inevitable by product of modern technology. However public awareness of the nature of these fields and the growing controversy over their potential effect on living system and adversely effect on immune system function have stimulated the researcher to define more precisely the physical properties of these fields and to delineated the threshold for their possible effect on human health [2].

1-2 Mechanism of Interaction Between Extremely Low Frequency (ELF) Field and Living System
Research in the effects of magnetic field, starts with extremely low frequency (ELF) electromagnetic field, which has very low energy at low power densities (<1mT).However, epidemiological studies over the past years have identified exposure to environmental ELF magnetic field (either job related or residential) as a possible factor in promotion of malignancy [3]. The most 1

Chapter One Review

Introduction and Literature

frequently considered mechanisms are [2]:1-The existence of transient free radicals which interact with the magnetic field. 2-The rate of production of hormone secretion, like oxidative processes may be altered, influencing the oxidation rate of unsaturated steriods. 3- A change in the rate or mechanism of diffusion across a membrane. 4-A distortion of the band angles via paramagnetic molecules, which change the fit between enzyme and substrate. 5-A change in rotational polarization of molecules with specific reactive sites.

1-3 Literature Review

In 1964 Beischer and Miller [4] studied the behavior of nervous, cardiac function, respiration, and leukocytes for exposure to 0.5mG for 10 days and found no significant changes. Green and Halpern in1966 [5] found no effect on growth of tissue culture size exposed to 0.5mG for few days. Conley [6] studied the human enzyme reactivity exposed to <0.5mG-5kG for several minutes, there was no significant alteration of enzyme reactivity in high or very low fields. Leo D Souza et al [7] found that exposure of a sites sarcoma 37 cells to 7300G (0.7T) magnetic field for periods of 13 h produces a decrease of Deoxyribonucleic acid (DNA) synthesis of about 18% to 24% as determined by the tritiated thymidine uptake. In 1987 Babji et al [8], investigated the exposure of fresh human lymphocytes to a power frequency (60Hz) magnetic field (MF) for times ranging

from 45 min. to 68 hour, which was significantly inhibited immunogloblin (Ig) 2

Chapter One Review

Introduction and Literature

aggregation at pole on the surface of B lymphocytes. No change in H3 thymidine incorporation of phaytohaemaglutinin (PHA) stimulated lymphocyte was observed during 17 h exposure to a magnetic field. However, after 40 h, thymidine incorporation was enhanced in exposed cell. Gohary, et al in 1987[9], studied the human triceps muscle exposed by a constant magnetic field of 500G for both short duration (15-min.) and long (10 days). The smallest effect on the muscles ability observed for the short period was 2%; however, if the interaction is continued for 10 days, the effect climbed to 25%. Wolf in 1987[10], found that magnetic field has a significant effect on the dynamics of living system. This field influences cell division and substantially affects neurological processes. It has shown both clinically and experimentally that magnetic fields of high intensity (BkG) produce a variety of biological pathologies. At low magnetic field intensities, such as near the earths magnetic field living system seems to experience euphoria. Reese, et al in 1988[11], investigated Chinese hamster oval (CHO) cells were exposed for 1h to 60 Hz magnetic fields (0.1 or 2mT) , electric fields (1 or 38 V/m), or to combined magnetic and electric fields (2mT and 38V/m, respectively). Following the exposure, the cells were lysed and the DNA was analyzed for the presence of single-strand break (SSB), using alkaline elution technique. No significant differences in numbers of DNA SSB were detected between exposed and sham exposed cell. in 1991 Marino et al [12], studied the human sensitivity to weak magnetic fields 60Hz, (0.25G-0.5G) magnetic field measured the power by electroncephalogram (EEG) in v2 for 2s with field on and off. The effect of electromagnetic field (EMF) was assessed by comparing recorded the EEG during 2s,they found that half of the volunteers were responded to the electromagnetic field by significant changes in EEG, for frequencies 3

Chapter One Review

Introduction and Literature

significantly affected in three volunteers in all cases less power was observed during the stimulus epochs compared with Control epochs. Luban, in 1991[13], showed that electromagnetic field might produce metabolic responses in the body. Since many low energy electromagnetic fields have too little energy to directly traverse the membrane, it is possible that they may modify the existing signal transduction processes in cell membranes, thus producing both transduction and biochemical amplification of the effect of the field itself. Marino et al in 1992[14], investigated in the effect of static and 60 Hz magnetic field, 0.78G, applied individually and combined to each 20 human subjects during 2s epochs, spectrum obtained during field exposure with that from control epochs, all but one subject exhibited field induced alterations in the EEG; most subjects exhibited increased EEG activity following brief exposure to static and low frequency magnetic field. Also Marino in 1993[15], illustrated in his paper that environmental electromagnetic field were predicated to increase the risk for cancer in chronically exposed human subjects because of impaired immunosurveillance mediated by the neuroendocrine system .Such as electromagnetic field induced changes in melatonin, or ion resonance interactions. Feychting In 1994[16], showed with experimental evidence has implicated the cellular membrane as a primary target of biological interaction with ELF fields. Electromagnetic fields in the frequency range below 100 Hz have been demonstrated to alter the release of Ca++ from cell surface binding sites in chick brain and various other form of tissue.

Marino in 1995[17],study the average daily exposure of the workers to electromagnetic field varied from 1-8h per 8h shift, and the months of exposure varied from 2-25 months for 44 subjects and was 108 months for 1 subject. 4

Chapter One Review

Introduction and Literature

These data suggests that the amount of time of exposure to non-ionizing electromagnetic field proportionately affected white cell count, with higher counts being manifested in subjects who had a greater cumulative exposure. Holmberg in 1995[18], have been designed to reveal a possible tumor promotion obtained by applying continuous or pulsed alternating fields at flux densities varying between 0.5T and 30mT on mice or rats initiated with different initiators. Cell studies showed that magnetic fields at some frequencies, amplitudes and waveforms interact with biological system. Cellular and physiological studies suggest that this effect may be related to cell multiplication and tumor promotion. Balcavage, et al. in 1996[19], reported that the static and low frequencies (50-60Hz ) pulsed electromagnetic field (PEMFs) can modify cation flow a cross biological membranes and alter cell metabolism. Charles in 1996[20], studied the EMF effect on melatonin (MT) hormones and immunity on men and women, statistical analysis showed no evidence of exposure related suppression of MT compared to men, women showed a much greater range and variability in MT over night, women did exhibit the same pattern of change in the CD4/CD8 ratio as men, and this approached statistical significance. Henderson [21] in 1996 determine if there exists a relationship between exposure to electric and magnetic fields (EMF) and cancer at the cellular level. He found that:1-Rebionucleic acid (RNA) transcripts (steady state level) in HL-60 cells are increased in the presence of 60 Hz continues wave sinusoidal fields, increase

in transcript for c-myc and c-fos in HL-60 cells exposed to 60 Hz EMF for 20 minute. 2-The response to EMF is rapid four minutes is the earliest time point after exposure. 5

Chapter One Review

Introduction and Literature

3-in the field strength used in the experiment (8 or 80T) the increase in c-myc transcripts peak in 20min. following 20min. exposure. 4- The cell membrane is important in the response to EMF. Treatment of cells with colchicines disturbs microtubules and decreases plasma membrane fluctuations. 5- Reduction or absence of extra cellular calcium negates the effect of EMF on transcript levels. These studies suggested that EMF can enhance and down regulate c-fos gene in a manner which is analogs to other growth factors. Jeffrey in 1997[22], studied the role of ELF field induced changes in gene expression in enhanced transformation. He found that ELF (Extremely Low frequency) magnetic fields do not alter the expression of the c-myc protooncogene in HL-60 cells, a human leukemia line. Ubeda, et al in 1997[23] showed a failure to independently replicate a study showing in vivo exposure to pulsed magnetic field of 15mT caused significant changes on plasma proteins in rats. Shashikant in 1997[24] investigated how EMF exposure typical of home and work environment could promote the incidence of B-cell related cancers and to determine the role of cell surface receptors in the growth and development of normal and abnormal lymphocytes under influence of 60 Hz fields. He showed that human lymphocyte cultures stimulated by a plant derived product (PHA) exhibit 1.4 to 1.7 fold increase in proliferation when exposed to a magnetic field for 3 days, compared to unexposed cells from the same donor he showed that secretion of at least one regulatory protein (IL-2) is affected by a 1G(1.21G peak), 60 Hz field when the induced electric field varies over a 5.5 to 1 range in amplitude, he suggest that the effect is directly due to the magnetic field. Kleinman, et al. in 1998[25] illustrated that the effect of magnetic fields can be studied for the product distribution of any radical reaction in biological 6

Chapter One Review

Introduction and Literature

system as long as mobility between different solubilization sites occurs. This new model predicts that magnetic field effect can be observed by enhanced cross-terminal product (CTP) formation from radical generated in an aqueous phase. Sek wen Hui in 1998 [26] investigated the effect of exposure to environment level (EMF) on cellular signal transduction and possible effects on a known model for carcinogenesis. The objective of this study is to reproduce the 60 Hz magnetic field induced time depend transcription changes of c-fos, c-myc and c-jun oncogenes in CEM-CM3 cells. Cells were exposed to 60 Hz magnetic field at (1G) (rms) for 15,30, 60,120min.The experiment was repeated 9 times, there was no significant difference between MF and sham exposed cells at all time points. In carcinogenic promotion experiment, the objective of this study is to test the environmental strength (1-4G) 60Hz magnetic field acts as a carcinogenic promoter. Cells INIT/C3H/10T1/2 transformed mouse fibroblasts cell line were exposed to (1-4G)(rms) magnetic field for various durations and sequences. He found that MF- exposed cell show slightly, but persistently lower counts than did sham exposed cells and both cells had counts significantly higher than those of unexposed cells. In 1999 Nakasono [27]evaluated the existence of genome scale gene expression change in yeast as a model for eukaryotic cells, under exposure to 50 Hz, 10mT sinusoidal magnetic field , no significant changes was detected in 2-D gel under exposure to 50Hz,10mT magnetic field for 24h. In 1999, the specific aims of Martin [28] in his study are to determine, first the relation between changes in cytochrome oxidase function in electric and magnetic fields and rate of electron transport, second the electric and magnetic field thresholds for changes in cytochrome oxidase function, the effect of magnetic field on this enzyme reaction are similar to effects on the Na+, K+ ATPase reaction in that, 7

Chapter One Review

Introduction and Literature

60Hz magnetic field accelerates the reaction rate in magnetic field divided by the control rate (M/C>1); apparently in competition with normal electron transport by the enzyme. M/C data at low field strength suggest that the threshold is below 1T.Under this condition, the rate data yield reaction rate constants, fundamental chemical reaction properties. These experiments show similar effects of magnetic field. 60Hz magnetic field increases the oxidation rate constant at field strength below 3T and by a factor of about 2 between 6-10T. Data at low field strength suggest that the threshold is below 0.5T, in the same range as the threshold determined by the M/C extrapolation as well as earlier results for Na, K ATPsa function 0.3T and stimulation of transcription (0.8T).Magnetic field stimulation of transcription could occur by an MCI mechanism, where the field interact with electrons moving along stacked base pairs in DNA. Kazuhika and Masao in 1999[29], studied the effect of static magnetic field on cells.They are interested in the non-thermal effects of magnetic field on cells, rather than stimulation or thermal effects. The cells are exposed to the magnetic field and are observed while focusing on the mitochondria energy activity of the cell. From the data the researcher concluded that, when the magnetic field is exposed to the cells in rest phase to first growth phase (G0/G1), the respiration phase of the mitochondria is enhanced, which enhanced the energy activity of the mitochondria. This result gives a little inside into the mechanism by which the magnetic field affects the mitochondria energy. Nakasono in 2000[30], demonstrated the effect of 50Hz, 10mT sinusoidal magnetic field on mRNA synthesis was estimated by using DNA microarrary. 8

Chapter One Review

Introduction and Literature

No significant or consistent was detected between control and the magnetic field exposure for 24h. Ruiying et al. in 2000 [31] investigated the biological effect of 50Hz, 0.8mT ELF magnetic field in molecular mechanisms of Daudi cell using the differential display (DD) technologies, the results indicated that DNA sequence analysis and Genbank research demonstrated that the cDNA fragment (MF-CA) is a novel gene in human cell which is induced by ELF magnetic field (MF) . The aim of Zeni et al researcher in 2001[32] was to investigate the induction of genotoxic and cytotoxic effects in culture human lymphocyte from healthy donors following combined exposure to 50Hz sinusoidal magnetic field (1mT field intensity) and a chemical mutagen. The result obtained indicate that magnetic field exposure dose not affect spontaneous micronucleus frequency, although a slight increase in cell proliferation was observed with probability (p=0.031), mutagens induce a statistically significant increase in micronucleus frequency. The combined exposure to both magnetic and chemical treatments alone, both in terms of genotoxicity and cell proliferation with probability (p=0.05 in all cases), in the experiment condition, a adopted, no cooperative effects are induced . Jajte et al in 2001[33], in this study indicated that exposure to 7mT static magnetic field (SMF) combined with simultaneous treatment of cells with iron ion (FeCl2) significantly, enhances lymphocyte death 18h after exposure. This effect not caused by either 7mT SMF or ferrous ions alone. Marino et al in 2001[34], hypothesized a nonlinear dynamical low converse magnetic field induced changes in lymphoid phenotype, the hypothesis was tested by analyzing biological data using a noval statistical procedure that could be adjusted to detect either nonlinear or linear effects. The results obtained from sampling a known nonlinear system, in 4 independent experiments, male and 9

Chapter One Review

Introduction and Literature

female mice were exposed continuously to 0.1-0.5mT, 60Hz for 175 days and the effect on 20 immune parameters was measured by flow cytometer and functional assays. In each experiment, electromagnetic field exposure resulted in statically significant changes in lymphoid phenotype when and only when the response of the animals to the fields was analyzed as if it were governed by nonlinear law. Ivan et al. in 2001[35], in their paper examine the dose response relationship between magnetic exposure of adults in their homes and prevalence of immune related and other chronic illnesses. Marino et al. in 2001[36] found, using a noval analytical method, that exposure of mice to 5G, 60Hz, for 1-105 days in 6 independent experiments consistently affected abroad panel of immune variables. Gulsasola et al. 2002[37] checked the bioeffect of the components of magnetic resonance imaging (MRI) on cells (L-132 cells) were used as biosensors and different cellular compounds were used as biomarker. The biosensors were placed in the bore of 1.5T MRI machine; although MRI procedures do not induce any cellular stress response, it may cause an unfathomable calcium increase in vitro.

1-4 Aim of the Project

In all previous, electromagnetic field around 50-60Hz were used with high ranges which is currently practiced in the modern systems was not evaluated. Therefore, the aim of this research is to study the biological effect of high intensity static magnetic field used in some applications as in diagnosis like electron spin resonance (ESR) or magnetic resonance imaging (MRI) device, which used different high intensity magnetic field (0.3-3)T. By studying the 10

Chapter One Review

Introduction and Literature

biological effects of different intensity magnetic field (MF) (0.1-0.5)T on immune system functions such as lymphocyte blastogenic response to phytoheamagglutinin (PHA) and non PHA stimulated peripheral blood monocyte cultures, and also the phagocytosis activity of peripheral blood leukocytes to staphylococcus bacteria at different exposure intervals.

11