Masateru Ikehata

Increase in the mitotic recombination frequency in Drosophila melanogaster by magnetic field exposure and its suppression by vitamin E supplement

In order to estimate possible mutagenic and/or carcinogenic activity of electromagnetic fields, wing spot tests were performed in Drosophila melanogaster. A DNA repair defective mutation mei-41D5 was introduced into the conventional mwh/flr test system to enhance mutant spot frequency. Third instar larvae were exposed to a 5-Tesla static magnetic field for 24 h, and after molting, wings were examined under a microscope to detect hair spots with mutant morphology. The exposure caused a statistically significant enhancement of somatic recombination compared with the unexposed control. This enhancement was suppressed to the control level by supplement of vitamin E, a non-specific antioxidant. It is inferred that the magnetic field enhanced the genotoxic effect of spontaneously produced free radicals, possibly by affecting the lifetime of the radicals. Enhancement of non-disjunction, terminal deletions and gene mutations were not detected.

Involvement of eddy currents in the mutagenicity of ELF magnetic fields.

Possible carcinogenic and/or mutagenic activity of extremely low frequency magnetic fields was examined using somatic mutation and recombination test system of Drosophila melanogaster. An X-linked semi-dominant DNA repair defective mutation mei-41(D5) was introduced into the conventional mwh/flr test system to enhance mutant spot frequency. Virgin females of w mei-41(D5)/FM6; flr/TM6 were crossed with w mei-41(D5)/Y; mwh jv; spa(pol) males. The F(1) third instar larvae were exposed to a 50Hz, 20mT sinusoidal AC magnetic field for 24h. After moulting from pupal cases, their wings were examined under a bright field microscope to detect hair spots with mwh or flr mutant morphology. The exposure caused a statistically significant enhancement in somatic recombination spot frequency. Mutant spots arising due to chromosomal non-disjunction or terminal deletion also increased but the frequency of spots resulting from point mutation was not altered. The enhancement in the recombination spot frequency was suppressed to the control level when a culture medium without electrolytes was used during exposure. When larvae were exposed to a magnetic field in an annular dish, flies from the outer ring showed more mutant spots compared to those from the inner ring. These results suggest that the detected mutagenic activity was that of the induced eddy current, rather than that of the magnetic field itself.

A Threshold Exists in the Dose–Response Relationship for Somatic Mutation Frequency Induced by X Irradiation of Drosophila

Abstract Koana, T., Takashima, Y., Okada, M. O., Ikehata, M., Miyakoshi, J. and Sakai, K. A Threshold Exists in the Dose– Response Relationship for Somatic Mutation Frequency Induced by X Irradiation of Drosophila. Radiat. Res. 161, 391– 396 (2004). The dose–response relationship of ionizing radiation and its stochastic effects has been thought to be linear without any thresholds. The basic data for this model were obtained from mutational assays in the male germ cells of the fruit fly Drosophila melanogaster. However, it is more appropriate to examine carcinogenic activity in somatic cells than in germ cells. Here the dose–response relationship of X irradiation and somatic mutation was examined in Drosophila. A threshold at approximately 1 Gy was observed in DNA repair-proficient flies. In the repair-deficient siblings, the threshold was smaller and the inclination of the dose–response curve was much steeper. These results suggest that the dose–response relationship between X irradiation and somatic mutation has a threshold and that the DNA repair function contributes to its formation.

Estimation of genetic effects of a static magnetic field by a somatic cell test using mutagen-sensitive mutants of Drosophila melanogaster

Abstract In order to estimate the genetic effects of magnetic fields, a somatic cell test was performed using mutants of the fruit fly Drosophila melanogaster which lack repair functions for damage to their cellular deoxyribonucleic acid (DNA). Young larvae of mutant and normal genotypes were exposed to a homogeneuos 0.6 T magnetic field for 24 h and were then allowed to continue development under normal culture condition until they moulted and finally emerged from their pupal cases. After eclosion, the number of surviving adults was counted. The number of adults of the mutant genotype decreased by about 8% compared with unexposed controls, while that of normal siblings remained unchanged. This suggests that exposure to a static magnetic field resulted in damage to larval cellular DNA, and that somatic cells without normal DNA repair functions failed to continue cell division which resulted in developmental lethality of mutant larvae. DNA damage occurring in normal larvae should have been repaired, so that their survival rate was not altered. The effect was compared with that of UV irradiation, and the genotoxic activity of the 0.6 T static magnetic field was estimated to be the same as that of UV light with an intensity of 0.14 mJ m−2 s−1. Possible mechanisms in which DNA damage is caused by magnetic field exposure are discussed.

Effects of intense magnetic fields on sedimentation pattern and gene expression profile in budding yeast

Effects of magnetic fields (MFs) on biological systems are usually investigated using biological indices such as gene expression profiles. However, to precisely evaluate the biological effects of MF, the effects of intense MFs on systematic material transport processes including experimental environment must be seriously taken into consideration. In this study, a culture of the budding yeast, Saccharomyces cerevisiae, was used as a model for an in vitro biological test system. After exposure to 5 T static vertical MF, we found a difference in the sedimentation pattern of cells depending on the location of the dish in the magnet bore. Sedimented cells were localized in the center of the dish when they were placed in the lower part of the magnet bore while the sedimentation of the cells was uniform in dishes placed in the upper part of the bore because of the diamagnetic force. Genome wide gene expression profile of the yeast cells after exposure to 5 T static MF for 2 h suggested that the MF did not affect the expression level of any gene in yeast cells although the sedimentation pattern was altered. In addition, exposure to 10 T for 1 h and 5 T for 24 h also did not affect the gene expression. On the other hand, a slight change in expressions of several genes which are related to respiration was observed by exposure to a 14 T static MF for 24 h. The necessity of estimating the indirect effects of MFs on a study of its biological effect of MF in vitro will be discussed.

Inhibition of UV-induced G1 arrest by exposure to 50 Hz magnetic fields in repair-proficient and -deficient yeast strains.

Purpose: To assess the possibility that extremely low frequency (ELF) magnetic fields obstruct the damage repair process, the gene conversion frequency and cell cycle kinetics in a DNA repair‐proficient and nucleotide excision repair (NER)‐deficient strain of diploid yeast Saccharomyces cerevisiae. Materials and methods: DNA repair‐ or NER‐deficient cells were irradiated with sublethal doses of ultraviolet light (UV) radiation followed by exposure to 50 Hz magnetic fields up to 30 mT for 48 h. After exposure, colony‐forming ability was scored as revertants in which gene conversion had restored the functional allele of the ARG4 gene conversion hotspot. Cell cycle analysis was performed using flow cytometry. Results: Gene conversion rate was increased by the combined exposure in DNA repair‐proficient cells, whereas it remained unchanged between UV alone and the combined exposure in NER‐deficient cells. The UV‐induced G1 arrest was inhibited by exposure to 30 mT ELF magnetic fields in both repair‐proficient and ‐deficient cells. Conclusions: The results suggest that exposure to high‐density (30 mT) ELF magnetic fields decreases the efficiency of NER by suppressing G1 arrest, which in turn led to enhancement of the UV‐induced gene conversion.

Effect of static magnetic field on the induction of micronuclei by some mutagens.

ObjectivesIt is important to assess the risk of static magnetic fields (SMFs) on human health, because epidemiological studies have indicated that SMFs play a role in the development of diseases such as leukemia and brain tumor. In our environment, we have numerous chances to be exposed to not only SMFs but also many chemicals containing mutagens. The aim of this study is to investigate the effect of SMFs on the induction of micronuclei induced by some mutagens.MethodsBALB/c mice were exposed to 4.7 tesla (T) SMF for 24 hr immediately after the injection of carboquone (alkylating agent), colcemid (spindle poison), mitomycin C (cross-linking agent), vincristine (spindle poison), sodium fluoride (a byproduct of aluminum plants under strong SMF) or 1-ethyl-1-nitrosourea (brain tumor-, gliomas- and thymic lymphoma-inducing chemical).ResultsThe frequency of micronuclei induced by six mutagens increased after co-exposure to SMF.ConclusionsAn additive/synergistic effect of SMF and chemicals was observed from the results of increased frequency of micronuclei induced by mutagens in mouse bone marrow erythrocytes.

Evaluation of biological effects of intermediate frequency magnetic field on differentiation of embryonic stem cell

The embryotoxic effect of intermediate frequency (IF) magnetic field (MF) was evaluated using murine embryonic stem (ES) cells and fibroblast cells based on the embryonic stem cell test (EST). The cells were exposed to 21 kHz IF–MF up to magnetic flux density of 3.9 mT during the cell proliferation process (7 days) or the cell differentiation process (10 days) during which an embryonic body differentiated into myocardial cells. As a result, there was no significant difference in the cell proliferation between sham- and IF–MF-exposed cells for both ES and fibroblast cells. Similarly, the ratio of the number of ES-derived cell aggregates differentiated to myocardial cells to total number of cell aggregates was not changed by IF–MF exposure. In addition, the expressions of a cardiomyocytes-specific gene, Myl2, and an early developmental gene, Hba-x, in the exposed cell aggregate were not altered. Since the magnetic flux density adopted in this study is much higher than that generated by an inverter of the electrical railway, an induction heating (IH) cooktop, etc. in our daily lives, these results suggested that IF–MF in which the public is exposed to in general living environment would not have embryotoxic effect.

Design and implementation of multi-frequency magnetic field generator producing sinusoidal current waveform for biological researches

Recently, induction heating systems are being widely used in many applications, and wireless power transmission systems are being introduced into commercially available electrical vehicles. These applications use time varying magnetic fields (under 100 kHz) which are in close proximity of the human body. Therefore, it should be discussed an influence of the time varying magnetic fields around 100 kHz to the human body. This paper presents design and implementation of a magnetic field generator for biological research. While conventional magnetic field generators only produce one frequency component, this paper discusses a multi-frequency range magnetic field generator with frequencies of 200 Hz and 20 kHz. In addition, actual systems with the inverter circuits for the purpose of biological evaluation are presented.

Evaluation of mutagenicity and co-mutagenicity of strong static magnetic fields up to 13 Tesla in Escherichia coli deficient in superoxide dismutase.

To evaluate the biological effects of static magnetic fields (SMFs) up to 13 Tesla (T), with respect to superoxide behavior, by determining the effect on mutagenicity in superoxide dismutase (SOD)‐deficient Escherichia coli strain QC774, and its parental strain GC4468.