Stereological analysis of thyroid mast cells in rats after exposure to extremely low frequency electromagnetic field and the following "off" field period

Influence of extremely low frequency electromagnetic field (ELF-EMF) on thyroid gland mast cells was investigated on male Mill Hill rats. Animals were exposed to EMF (50 Hz, 50 microT to 500 microT, 10 V/m) from 24 hours after birth, 7 hours/day, 5 days/week for three months when a part of animals (group I) was sacrificed, while the rest of them were subjected to recovery evaluation and sacrificed after one (group II), two (group II) and three (group IV) weeks following the exposure. Stereological analysis on toluidine blue-stained paraffin sections showed increased volume density of degranulated mast cells in all groups and, except in group III, and numerical density as well, implicating the sensitivity of thyroidal mast cells to power frequency EMFs. Since in our previous investigations, morphofunctional alterations of thyroid gland in rats exposed to ELF-EMF were found the contribution of released mast cell mediators to these changes could be presumed.     

The effect on rat thymocytes of the simultaneous in vivo exposure to 50-Hz electric and magnetic field and to continuous light

Thymus plays an important role in the immune system and can be modulated by numerous environmental factors, including electromagnetic fields (EMF). The present study has been undertaken with the aim to investigate the role of long-term exposure to extremely low frequency electric and magnetic fields (ELF-EMF) on thymocytes of rats housed in a regular dark/light cycle or under continuous light. Male Sprague-Dawley rats, 2 months old, were exposed or sham exposed for 8 months to 50-Hz sinusoidal EMF at two levels of field strength (1 kV/m, 5 microT and 5 kV/m, 100 microT, respectively). Thymus from adult animals exhibits signs of gradual atrophy mainly due to collagen deposition and fat substitution. This physiological involution may be accelerated by continuous light exposure that induces a massive death of thymocytes. The concurrent exposure to continuous light and to ELF-EMF did not change significantly the rate of mitoses compared to sham-exposed rats, whereas the amount of cell death was significantly increased, also in comparison with animals exposed to EMF in a 12-h dark-light cycle. In conclusion, long-term exposure to ELF-EMF, in animals housed under continuous light, may reinforce the alterations due to a photic stress, suggesting that, in vivo, stress and ELF-EMF exposure can act in synergy determining a more rapid involution of the thymus and might be responsible for an increased susceptibility to the potentially hazardous effects of ELF-EMF.     

A study on specific behavioral effects of formaldehyde in the rat

It has been reported that single exposure of rats of low-level formaldehyde vapor concentrations causes significant alteration in their motor activity in the inhalation chamber. In this study, we determined the effects of formaldehyde on the locomotor activity and behavior of adult male and female Lew. 1K rats in an open field two hours after termination of a single two hours lasting inhalative exposure to approximately 0.1, 0.5, or 5 ppm. Following behavioral parameters were quantitatively examined: numbers of crossed floor squares, occurrence frequencies of air and floor sniffing, grooming, rearing, and wall climbing, as well as the incidence of fecal boli. In the open field situation, the males of all formaldehyde groups crossed significantly lower numbers of floor squares. Furthermore, significant decrease in the occurrence frequencies of floor sniffing, rearing, and wall climbing were observed. Within the female rat groups exposed to 0.5 or 5 ppm formaldehyde, a significantly decreased numbers of crossed squares were registered, while this parameter remained unchanged in the 0.1 ppm group. Other parameters were also affected by the formaldehyde inhalation (e.g. significant increase in the occurrence frequencies of air sniffing in the 0.1 and 0.5 ppm groups and significant decrease in the numbers of floor sniffing in the 0.5 and 5 ppm groups, respectively). The incidence of fecal boli was not affected in any exposure group neither in males nor in females. It is concluded from the results obtained that formaldehyde significantly affects the locomotor behavior of adult male and female rats in the open field after a single inhalative exposure to the above mentioned concentrations.     

Effect of intermittent chronic exposure to hypoxia on feeding behaviour of rats

Healthy albino male rats were exposed to a simulated high altitude (HA) equivalent to 25000 ft (7620 m) for 6 h daily, continuously for 21 days to study the feeding behaviour. The 24-h food and water intake and body weight once in 3 days were recorded. Blood samples were drawn once a week from the retro-orbital venous plexus for blood sugar analysis. All the parameters were recorded before, during and after exposure to simulated HA. The results show a decrease in 24-h food and water intake and decreased gain in body weight during hypoxic exposure, which showed a tendency to come back to control during the post-exposure period. The blood sugar reflected a state of mild hyperglycaemia during exposure to HA.     

Evaluation of rat thyroid gland morphophysiological status after three months exposure to 50 Hz electromagnetic field

Objective of our study was to use morphophysiological criteria in order to determine the sensitivity of male rat thyroid gland to an extremely low frequency electromagnetic field (ELF-EMF) influence and the ability of the gland to repair after period of exposure. Animals were exposed to 50Hz, 50-500 microT ELF-EMF for 3 months when a part of them (group I) were sacrificed, while the rest of animals were subjected to recovery evaluation of the gland and sacrificed after 1 (group II), 2 (group III) and 3 (group IV) weeks. Histological and stereological analyses were performed on paraffin and semifine thyroid gland sections. Serum T3 and T4 were also determined. Histological and stereological analyses showed that the volume density of follicular epithelium and thyroid activation index decreased, while the volume density of colloid and capillary network increased in group I, II and III. The values of all these parameters in group IV were similar to corresponding controls. Serum T3 and T4 concentrations were significantly lower in all exposed animals, except in group I. Results of this study demonstrate that after significant morphophysiological changes caused by ELF-EMF exposure thyroid gland recovered morphologically, but not physiologically, during the investigated repair period.     

Extremely low frequency variable electromagnetic fields affect cancer and noncancerous cells in vitro differently: Preliminary study

The exposure to extremely low frequency electromagnetic field (ELF-EMF) may result in various changes at the cellular level. To identify the effect of ELF-EMF exposure on viability of cells, cancer cells (U87-MG; 143B) and noncancerous cells (BJ; HEK) in exponential growth phase were exposed or sham-exposed to different values of frequency (2, 20, 30, 50 and 60 Hz), different shapes (sinusoidal, square and triangular) and time of exposure (0.5, 1, 2, 3 h) to electromagnetic field. After exposure, viability of cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). We found a different effect of exposition of cancer and noncancerous cells to ELF-EMF on viability of cells. This preliminary study revealed that electro magentic field(EMF) might serve as a potential tool for manipulating viability of cells.     

Possible combined effects of 900 MHZ continuous-wave electromagnetic fields and gentamicin on the auditory system of rats

The aim of this study was to evaluate the cochlear functionality of Sprague-Dawley rats exposed to electromagnetic fields at 900 MHz and to gentamicin by distortion product otoacoustic emissions, which are a well-known indicator of the status of the cochlea's outer hair cells. A population of 32 rats was divided into four groups: group 1 was treated with daily intramuscular injections of 150 mg/kg body weight gentamicin for 15 days; group 2 was treated with daily intramuscular injections of 150 mg/kg body weight gentamicin for 15 days and exposed to electromagnetic fields; group 3 was exposed to electromagnetic fields; group 4 was sham-exposed. Rats were exposed 2 h/day, 5 days/week for 4 weeks at a local SAR of 4 W/kg in the ear (continuous wave at 900 MHz). Distortion product otoacoustic emissions tests were carried out before, during and after the combined exposure. The analysis of the data showed no subchronic exposure to electromagnetic fields on the inner auditory system of rats in either normal ears or ears exposed to a well-recognized pathological agent.     

Extremely Low-Frequency Electromagnetic Fields Promote In Vitro Neuronal Differentiation and Neurite Outgrowth of Embryonic Neural Stem Cells via Up-Regulating TRPC1

Exposure to extremely low-frequency electromagnetic fields (ELF-EMFs) can enhance hippocampal neurogenesis in adult mice. However, little is focused on the effects of ELF-EMFs on embryonic neurogenesis. Here, we studied the potential effects of ELF-EMFs on embryonic neural stem cells (eNSCs). We exposed eNSCs to ELF-EMF (50 Hz, 1 mT) for 1, 2, and 3 days with 4 hours per day. We found that eNSC proliferation and maintenance were significantly enhanced after ELF-EMF exposure in proliferation medium. ELF-EMF exposure increased the ratio of differentiated neurons and promoted the neurite outgrowth of eNSC-derived neurons without influencing astrocyes differentiation and the cell apoptosis. In addition, the expression of the proneural genes, NeuroD and Ngn1, which are crucial for neuronal differentiation and neurite outgrowth, was increased after ELF-EMF exposure. Moreover, the expression of transient receptor potential canonical 1 (TRPC1) was significantly up-regulated accompanied by increased the peak amplitude of intracellular calcium level induced by ELF-EMF. Furthermore, silencing TRPC1 expression eliminated the up-regulation of the proneural genes and the promotion of neuronal differentiation and neurite outgrowth induced by ELF-EMF. These results suggest that ELF-EMF exposure promotes the neuronal differentiation and neurite outgrowth of eNSCs via up-regulation the expression of TRPC1 and proneural genes (NeuroD and Ngn1). These findings also provide new insights in understanding the effects of ELF-EMF exposure on embryonic brain development.     

Effects of 60 Hz electric and magnetic fields on the development of the rat cerebellum

The effects of extremely low frequency (ELF) electromagnetic (EM) fields on the maturation of the rat cerebellum were studied. Newborn rats were exposed to 60 Hz electric and magnetic fields under three different combinations in a specially constructed apparatus. The pups were irradiated for 7–8 h daily, with a 30-min interruption for nursing. Pups were kept with their mothers for the remainder of the time. After approximately 1, 2, or 3 weeks of exposure, the pups were killed. Control pups were sham exposed. The somatic growth of the irradiated rats did not show any significant difference from shamexposed controls. At 1 kV/m and 10 gauss exposure, there was a small but statistically significant decrease in cerebellar mass. In rats exposed at 1 kV/m and 10 gauss, DNA and RNA levels were significantly higher than those in shara-exposed controls at 6 and 13 days of age, but at 20 days, these two biochemical constituents were similar in both groups of rats. The ELF-EM treatment had no effect on protein and cerebroside concentrations. In terms of age effects. DNA and RNA exhibited increases from 6 to 13 days of age, and declined from 13 to 20 days. Protein and cerebroside levels exhibited increases during the 6–20 day periods. In rats exposed at 100 kV/m and 1 gauss, the DNA levels were initially less than those of sham-exposed controls at 8 days of age, reached approximately the same levels at 14 days, and then were higher than those of controls at 22 days. There was. therefore, a significant ELF-EM effect as well as a significant interaction between age and ELF-EM exposure. In terms of age effects, DNA levels for both control and exposed animals increased from 8 to 14 days. From 14 to 22 days, DNA levels of exposed rats continued to increase while those of the controls decreased. This age effect was significant. RNA levels in both groups of animals showed increases from 8 to 14 days of age, but the increase was less for the irradiated animals than for the controls. From days 14 to 22. RNA levels for both groups showed a reduction, but the decrease was greater in the irradiated than in control rats. ELF-EM treatment significantly reduced protein levels at 8 days of age. but at 14 to 22 days, protein levels of exposed rats were higher than those of controls. The cerebroside levels were not affected by exposure treatments but increased with the age of the animals. Exposure to 100 kV/m and 10 gauss did not exert any effect on the concentrations of DNA, RNA, protein, and cerebroside at all three time points examined. Both DNA and RNA exhibited increases with age from 6 to 13 days, and leveled off from 13 to 20 days. Protein and cerebroside levels also showed corresponding increases with the age of the animals. Morphological observations revealed no detectable changes in the irradiated animals in any experimental group. Thus, only biochemical studies indicate that exposure at certain ELF-EM field combinations induces alterations in cerebellar maturation. These changes were clearly detectable in the early postnatal period but gradually diminished with time. ©1993 Wiley-Liss, Inc.     

50-Hz extremely low frequency electromagnetic fields enhance cell proliferation and DNA damage: possible involvement of a redox mechanism

HL-60 leukemia cells, Rat-1 fibroblasts and WI-38 diploid fibroblasts were exposed for 24-72 h to 0.5-1.0-mT 50-Hz extremely low frequency electromagnetic field (ELF-EMF). This treatment induced a dose-dependent increase in the proliferation rate of all cell types, namely about 30% increase of cell proliferation after 72-h exposure to 1.0 mT. This was accompanied by increased percentage of cells in the S-phase after 12- and 48-h exposure. The ability of ELF-EMF to induce DNA damage was also investigated by measuring DNA strand breaks. A dose-dependent increase in DNA damage was observed in all cell lines, with two peaks occurring at 24 and 72 h. A similar pattern of DNA damage was observed by measuring formation of 8-OHdG adducts. The effects of ELF-EMF on cell proliferation and DNA damage were prevented by pretreatment of cells with an antioxidant like alpha-tocopherol, suggesting that redox reactions were involved. Accordingly, Rat-1 fibroblasts that had been exposed to ELF-EMF for 3 or 24 h exhibited a significant increase in dichlorofluorescein-detectable reactive oxygen species, which was blunted by alpha-tocopherol pretreatment. Cells exposed to ELF-EMF and examined as early as 6 h after treatment initiation also exhibited modifications of NF kappa B-related proteins (p65-p50 and I kappa B alpha), which were suggestive of increased formation of p65-p50 or p65-p65 active forms, a process usually attributed to redox reactions. These results suggest that ELF-EMF influence proliferation and DNA damage in both normal and tumor cells through the action of free radical species. This information may be of value for appraising the pathophysiologic consequences of an exposure to ELF-EMF.     

Oxidative DNA damage in rats exposed to extremely low frequency electro magnetic fields

Extremely low frequency (ELF) electromagnetic field (EMF) is thought to prolong the life of free radicals and can act as a promoter or co-promoter of cancer. 8-hydroxy-2'-deoxyguanosine (8OHdG) is one of the predominant forms of radical-induced lesions to DNA and is a potential tool to asses the cancer risk. We examined the effects of extremely low frequency electro magnetic field (ELF-EMF) (50 Hz, 0.97 mT) on 8OHdG levels in DNA and thiobarbituric acid reactive substances (TBARS) in plasma. To examine the possible time-dependent changes resulting from magnetic field, 8OHdG and TBARS were quantitated at 50 and 100 days. Our results showed that the exposure to ELF-EMF induced oxidative DNA damage and lipid peroxidation (LPO). The 8OHdG levels of exposed group (4.39+/-0.88 and 5.29+/-1.16 8OHdG/dG.10(5), respectively) were significantly higher than sham group at 50 and 100 days (3.02+/-0.63 and 3.46+/-0.38 8OHdG/dG.10(5)) (p<0.001, p<0.001). The higher TBARS levels were also detected in the exposure group both on 50 and 100 days (p<0.001, p<0.001). In addition, the extent of DNA damage and LPO would depend on the exposure time (p<0.05 and p<0.05). Our data may have important implications for the long-term exposure to ELF-EMF which may cause oxidative DNA damage.     

Differential suppression of hyperglycemic,feeding, and neuroendocrine responses in anorexia

We have used the anorexia shown by rats given hypertonic saline to drink to investigate central mechanisms that can inhibit feeding. Rats dehydrated in this manner for 3 or 5 days showed a severe attenuation of the compensatory feeding observed after an overnight fast compared with control euhydrated rats or rats whose food was restricted to match the intake of anorexic rats. Food intake after injections of 2-deoxy-d-glucose (2-DG) was also significantly decreased in dehydrated animals compared with that after a 2-DG injection given before dehydration. However, all the dehydrated animals demonstrated a robust eating response after water was returned whether they had received injection of 2-DG or vehicle. Despite a profound reduction in 2-DG-induced feeding, other glucoregulatory responses to 2-DG remained intact in dehydrated animals. After 2-DG injection, corticosterone secretion and blood glucose were significantly elevated from preinjection values whether or not animals were dehydrated. Thus the mechanisms responsible for anorexia in dehydrated animals specifically target stimulatory feeding pathways but leave intact other counterregulatory glucometabolic motor events.     

Improvement of Spatial Memory Disorder and Hippocampal Damage by Exposure to Electromagnetic Fields in an Alzheimer’s Disease Rat Model

Although some epidemiological investigations showed a potential association between long-term exposure of extremely low frequency electromagnetic fields (ELF-EMF) and Alzheimer’s disease (AD), no reasonable mechanism can explain this association, and the related animal experiments are rare. In this study, ELF-EMF exposure (50Hz 400µT 60d) combined with D-galactose intraperitoneal (50mg/kg, q.d., 42d) and Aβ_25–35 hippocampal (5μl/unilateral, bilateral, single-dose) injection was implemented to establish a complex rat model. Then the effects of ELF-EMF exposure on AD development was studied by using the Morris water maze, pathological analysis, and comparative proteomics. The results showed that ELF-EMF exposure delayed the weight gain of rats, and partially improved cognitive and clinicopathologic symptoms of AD rats. The differential proteomic analysis results suggest that synaptic transmission, oxidative stress, protein degradation, energy metabolism, Tau aggregation, and inflammation involved in the effects mentioned above. Therefore, our findings indicate that certain conditions of ELF-EMF exposure could delay the development of AD in rats.     

Circulating antidiuretic hormone in the X-irradiated rat

Male rats exposed to 500 R of whole-body x-irradiation were allowed food and water ad libitum and housed in metabolism cages; water and food intake and urinary and fecal excretion were recorded daily. Urine output increased 200% during the first 24 hours after irradiation. No significant changes occurred in daily sodium, potassium, urea, or total solute excretion, although calcium excretion approximately doubled after irradiation. The marked increase in free water excretion implicates antidiuretic hormone (ADH) in this phenomenon. Application of a sensitive bioassay for ADH permitted measurement of plasma ADH concentrations in undisturbed, unanesthetized rats before and after irradiation. ADH levels were lower and frequently not detectable 24 hours after exposure. High ADH levels, however, could be provoked in irradiated rats by hemorrhage, indicating that the receptor cells and secretory ability of the posterior pituitary remained intact. Furthermore, irradiated rats responded normally to small intravenous injections (4 to 8 microU) of exogenous ADH. Rats with congenital diabetes insipidus given daily injections of Pitressin showed no postirradiation diuresis. Lastly, increased urinary calcium excretion may result from hypercalcemia which is known to induce diuresis through calcium-vasopressin antagonism. These results further suggest that the diuretic response is due to decreased circulating ADH.     

Age-dependent effects of chronic hypoxia on renin-angiotensin and urinary excretions

The mechanisms regulating water, electrolyte, and blood volume homeostasis continue to mature in early postnatal life, and this maturation may be altered by perturbations of volume or cardiovascular status. To evaluate the long-term effects of chronic hypoxia on water balance, urinary electrolyte excretion, heart weights, systemic arterial pressure, and components of the renin-angiotensin system, male Sprague-Dawley rats were exposed to periods of simulated altitude of 10,000 ft up to 90 days of age beginning at 2 or 30 days of age. Altitude exposure of both neonatal and adult rats was associated with increases in urine output and water intake after 30 days of exposure, and right ventricular (RV) hypertrophy at all ages was examined. However, the percent increase in urine output, water intake, and RV hypertrophy was numerically greater in neonates. Neonates also had increases in urinary sodium and potassium excretion after 30 days of exposure. Plasma renin activity and serum angiotensin-converting enzyme (ACE) activity were not affected, but plasma renin substrate was reduced in both neonatal and adult altitude-exposed rats. Lung ACE activity was also decreased in altitude-exposed neonates. These data indicate that the degree and, in some cases, the nature of these homeostatic responses varies with age during long-term hypoxia.     

Concentration of T3 and T4 in blood of non-irradiated and irradiated with different doses rats fasted for two days beforehand

There were no changes in concentration of T3 and T4 in blood of the rats that were irradiated with a dose of 0.5, 1, 2, 4 and 6 Gy in comparison with non-irradiated rats, if the animals were not fed for two days before decapitation. This suggests that the effect of ionizing radiation on thyroid function is mediated by anorexia syndrome. The decrease in concentration of T4 after exposure to 8 Gy cannot be explained by postradiative anorexia and most likely is connected with starting enterotoxemy in difficult cases of acute radiation sickness.     

A strong static-magnetic field alters operant responding by rats

Forty male rats of the Wistar ST strain were trained and observed for Sidman avoidance (SA) for 7 weeks or for discriminative avoidance (DA) for 14 weeks to determine the effects of exposure to a strong static-magnetic field. Before avoidance conditioning was completed, rats in the SA group were exposed to the static field at 0.6 T, 16 h/day for 4 days during the fifth week, and those in the DA group were exposed for 6 h/day for 4 days during the seventh week. In the SA conditioning, frequency of lever-pressing by exposed rats gradually decreased during 1 week of exposure and stayed low for at least 2 weeks after exposure. Frequencies of electric shocks received by the rats increased dramatically during the second day of exposure and consistently stayed higher than those of control rats. In the DA condition, exposed rats responded at lower rates than did control rats throughout the observation period. They received more shocks during the 2 weeks following exposure. The data indicate that performance of avoidance responses was inhibited by a comparatively long exposure to a strong magnetic field.     

Effect of extremely low frequency electromagnetic field on brain histopathology of Caspian Sea Cyprinus carpio

There is limited research on the effect of electromagnetic field on aquatic organisms, especially freshwater fish species. This study was conducted to evaluate the effect of extremely low frequency electromagnetic field (ELF-EMF) (50 Hz) exposure on brain histopathology of Cyprinus carpio, one of the important species of Caspian Sea with significant economic value. A total of 200 healthy fish were used in this study. They were classified randomly in two groups: sham-exposed group and experimental group, which were exposed to five different magnetic field intensities (0.1, 1, 3, 5, and 7 mT) at two different exposure times (0.5 and 1 h). Histologic results indicate that exposure of C. carpio to artificial ELF-EMF caused severe histopathological changes in the brain at field intensities ≥3 mT leading to brain necrosis. Field intensity and duration of exposure were key parameters in induction of lesion in the brain. Further studies are needed to elucidate exact mechanism of EMF exposure on the brain.     

Effects of Extremely Low-Frequency Electromagnetic Fields on Helicobacter pylori Biofilm

The aim of this work was to investigate the effects of exposure to extremely low-frequency electromagnetic fields (ELF-EMF) both on biofilm formation and on mature biofilm of Helicobacter pylori. Bacterial cultures and 2-day-old biofilm of H. pylori ATCC 43629 were exposed to ELF-EMF (50 Hz frequency–1 mT intensity) for 2 days to assess their effect on the cell adhesion and on the mature biofilm detachment, respectively. All the exposed cultures and the respective sham exposed controls were studied for: the cell viability status, the cell morphological analysis, the biofilm mass measurement, the genotypic profile, and the luxS and amiA gene expression. The ELF-EMF acted on the bacterial population during the biofilm formation displaying significant differences in cell viability, as well as, in morphotypes measured by the prevalence of spiral forms (58.41%) in respect to the controls (33.14%), whereas, on mature biofilm, no significant differences were found when compared to the controls. The measurement of biofilm cell mass was significantly reduced in exposed cultures in both examined experimental conditions. No changes in DNA patterns were recorded, whereas a modulation in amiA gene expression was detected. An exposure to ELF-EMF of H. pylori biofilm induces phenotypic changes on adhering bacteria and decreases the cell adhesion unbalancing the bacterial population therefore reducing the H. pylori capability to protect itself.     

A preliminary study of oscillating electromagnetic field effects on human spermatozoon motility

Some effects of extremely low frequency electromagnetic fields (ELF-EMFs) on human spermatozoa are reported. Significant increases in the values of the motility and of the other kinematic parameters have been observed when spermatozoa were exposed to an ELF-EMF with a square waveform of 5 mT amplitude and frequency of 50 Hz. By contrast, a 5 mT sine wave (50 Hz) and a 2.5 mT square wave (50 Hz) exposure did not produce any significant effect on sperm motility. The effects induced by ELF-EMF (50 Hz; 5 mT) during the first 3 h of exposure persisted for 21 h after the end of the treatment. These results indicate that ELF-EMF exposure can improve spermatozoa motility and that this effect depends on the field characteristics.