Effects of acute low-level microwaves on pentobarbital-induced hypothermia depend on exposure orientation

Two series of experiments were performed to study the effects of acute exposure (45 min) to 2,450-MHz circularly polarized, pulsed microwaves [1 mW/cm², 2-μs pulses, 500 pps, specific absorption rate (SAR) 0.6 W/kg] on the actions of pentobarbital in the rat. In the first experiment, rats were irradiated with microwaves and then immediately injected with pentobarbital. Microwave exposure did not significantly affect the extent of the pentobarbital-induced fall in colonic temperature. However, the rate of recovery from the hypothermia was significantly slower in the microwave-irradiated rats and they also took a significantly longer time to regain their righting reflex. In a second experiment, rats were first anesthetized with pentobarbital and then exposed to microwaves with their heads either pointing toward the source of microwaves (anterior exposure) or pointing away (posterior exposure). Microwave radiation significantly retarded the pentobarbital-induced fall in colonic temperature regardless of the orientation of exposure. However, the recovery from hypothermia was significantly faster in posterior-exposed animals compared to those of the anterior-exposed and sham-irradiated animals. Furthermore, the posterior-exposed rats took a significantly shorter time to regain their righting reflex than both the anterior-exposed and sham-irradiated animals.     

Effects of 2.45-GHz microwave radiation on embryonic quail hearts

Although exposure to nonionizing electromagnetic radiation has been reported to cause a variety of systemic alterations during embryonic development, there are few reports of the induction of specific physiologic or morphologic changes in the myocardium. This study was designed to examine the effects of microwave radiation on cardiogenesis in Japanese quail embryos exposed during the first eight days of development to 2.45-GHz continuous-wave microwaves at power densities of 5 or 20 mW/cm². The specific absorption rates were 4.0 and 16.2 mW/g, respectively. The ambient temperature for each exposure was set to maintain the embryonated eggs at 37.5 °C. This did not preclude thermal gradients in the irradiated embryos since microwaves may not be uniformly absorbed. The test exposure levels did not induce changes in either the morphology of the embryonic heart or the ultrastructure of the myocardial cells. Analysis of the enzymatic activities of lactate dehydrogenase, glutamic oxaloacetic transaminase, and creatine phosphokinase failed to reveal any statistically significant differences between the nonexposed controls and those groups exposed to either 5 or 20 mW/cm². The data indicate that 2.45-GHz microwave radiation at 5 or 20 mW/cm² has no effect on the measured variables of the Japanese quail myocardium exposed during the first eight days of development.     

In vitro microwave effects on human neutrophil precursor cells (CFU-C)

Human marrow cells were irradiated with 2450-MHz CW microwaves in a fluid-filled waveguide irradiation system. Cell exposure was conducted by placing a marrow cell suspension in 20-μl glass microcapillary tubes that were positioned in the exposure chamber, and irradiated at power densities from 31 to 1,000 mW/cm² (with corresponding specific absorption rates of 62 to 2,000 mW/g) for 15 minutes. The temperature of the sample was maintained at a fixed point. Sham-irradiated (SI) and microwave-irradiated (MWI) cells were cultured in a methylcellulose culture system for neutrophil colony proliferation. There was no reduction in neutrophil colony number on days 6–7 or 12–14 in cells exposed at 31 or 62 mW/cm², but as the power density was increased to 1,000 mW/cm², there was a reduction in colony number of MWI cells compared with SI cells. The microwave interaction with the human neutrophil colony-forming cells was apparently not related to temperature rise, or to the state of cell cycle, and was irreversible.     

Ethanol-induced hypothermia and ethanol consumption in the rat are affected by low-level microwave irradiation

Microwave irradiation of rats by circularly polarized, 2,450-MHz, pulsed waves (2-μs pulses; 500 pps) was performed in waveguides to determine effects on ethanol-induced hypothermia and on ethanol consumption. Rats injected intraperitoneally with ethanol (3 g/kg in a 25% v/v water solution) immediately after 45 min of microwave irradiation exhibited attenuation of the initial rate of fall in body temperature, which was elicited by the ethanol, but exhibited no significant difference in maximal hypothermia as compared with that of sham-irradiated rats. Microwave irradiation did not affect the consumption of a 10% sucrose (w/v) solution by water-deprived rats. However, it enhanced the consumption of a solution of 10% sucrose (w/v) + 15% ethanol (v/v) by water-deprived animals. These results were obtained at a specific absorption rate (SAR) of 0.6 W/kg, which rate of energy dosing would require a power density of 3–6 mW/cm² if exposure of the animals had occurred to a 12-cm plane wave.     

Alteration of circulating antibody response of mice exposed to 9-GHz pulsed microwaves

A significant increase was observed in the circulating antibody titers of mice exposed to 9-GHz pulsed microwaves at an average power density of 10 mW/ cm², two hours per day for five days compared with sham-irradiated animals. The mice were previously immunized with type III pneumococcal polysaccharide. Following irradiation, a portion of the immunized animals were challenged with virulent ^{Streptococcus pneumoniae}, type III. Ten days after challenge, mortality was essentially the same in the two groups, but during the ten day period, there was a noticeable increase in the survival time of the irradiated animals compared with the sham-irradiated animals, suggesting that the increased circulating antibody response afforded some degree of temporary protection to the animals.     

Microwave effects on energy metabolism of rat brain

Rat brain was exposed to 591-MHz, continuous-wave (CW) microwaves at 13.8 or 5.0 mW/cm² to determine the effect on nicotinamide adenine dinucleotide, reduced (NADH), adenosine triphosphate (ATP) and creatine phosphate (CP) levels. On initiation of the in vivo microwave exposures, fluorimetrically determined NADH rapidly increased to a maximum of 4.0%–12.5% above pre-exposure control levels at one-half minute, then decreased slowly to 2% above control at three minutes, finally increasing slowly to 5% above control level at five minutes. ATP and CP assays were performed on sham- and microwave-exposed brain at each exposure time. At 13.8 mW/cm², brain CP level was decreased an average of 39.4%, 41.1%, 18.2%, 13.1%, and 36.4% of control at exposure points one-half, one, two three, and five minutes, respectively, and brain ATP concentration was decreased an average of 25.2%, 15.2%, 17.8%, 7.4%, and 11.2% of control at the corresponding exposure periods. ATP and CP levels of rat brain exposed to 591-MHz cw microwaves at 5 mW/cm² for one-half and one minute were decreased significantly below control levels at these exposure times, but were not significantly different from the 13.8 mW/cm² exposures. For all exposures, rectal temperature remained constant. Heat loss through the skull aperture caused brain temperature to decrease during the five-minute exposures. This decrease was the same in magnitude for experimental and control subjects. Changes in NADH, ATP, and CP levels during microwave exposure cannot be attributed to general tissue hyperthermia. The data support the hypothesis that microwave exposure inhibits mitochondrial electron transport chain function, which results in decreased ATP and CP levels in brain.     

Microwaves modify thermoregulatory behavior in squirrel monkey

Squirrel monkeys (Saimiri sciureus) trained to regulate environmental temperature (T_a) behaviorally were exposed in the far field of a horn antenna to ten-minute periods of 2,450 MH_z CW microwaves. Incident power density ranged from 1 to 22 mW/cm². The corresponding specific absorption rate (SAR), derived from temperature increments in saline-filled styrofoam models, ranged from 0.15 to 3.25 W/kg. Controls included exposure to infrared radiation of equivalent incident energy and no radiation exposure. Normal thermoregulatory behavior produces tight control over environmental and body temperatures; most monkeys select a T_a of 34–36°C. Ten-minute exposures to 2,450 MH_z CW microwaves at an incident power density of 6–8 mW/cm² stimulated all animals to select a lower T_a. This threshold energy represents a whole-body SAR of 1.1 W/kg, about 20% of the resting metabolic rate of the monkey. Thermoregulatory behavior was highly efficient, and skin and rectal temperatures remained stable, even at 22 mW/cm² where the preferred T_a was lowered by as much as 4°C. No comparable reduction in selected T_a below control levels occurred during exposure to infrared radiation of equal incident power density.     

Effect of nonionizing radiation on the purkinje cells of the rat cerebellum

In one experiment, Sprague Dawley rats (16–21 days of gestation) and their offspring were exposed to 100-MHz (CW) electromagnetic radiation at 46 mW/cm² (SAR 2.77 mW/g) for 4 h/day for 97 days. In another experiment, the pregnant rats were irradiated daily from 17 to 21 days of gestation with 2450-MHz (CW) microwaves at 10 mW/cm² (SAR 2 mW/g) for 21 h/day. In a third experiment, 6-day-old rat pups were irradiated 7 h/day for five days with 2450-MHz radiation at 10 mW/cm². Equal numbers of animals were sham irradiated in each group. Quantitative studies of Purkinje cells showed a significant and irreversible decrease in rats irradiated during fetal or fetal and early postnatal life. In animals exposed postnatally, and euthanized immediately after irradiation, significant decrease in the relative number of Purkinje cells was apparent. However, restoration apparently occurred after forty days of recovery.     

Tests of mutagenesis and reproduction in male rats exposed to 2,450-MHz (CW) microwaves

Tests of mutagenesis and reproduction were conducted in male rats which were irradiated by 2,450-MHz, continuous-wave (CW) microwaves, 4 hr/day from day 6 of gestation to 90 days of age at 5 mW/cm²; or 5 hr/day for five days beginning on the 90th day of age at 10 mW/cm²; or 4 hr/day, 5 days/ wk for four weeks, beginning on the 90th day of age. During selected weekly periods after treatment, the rats were bred to pairs of untreated, normal female rats that were examined in late pregnancy by means of the dominant lethal assay. The reproductive efficiency of these males, as reflected in their breeding, was also examined for changes relating to their microwave experience. No significant evidence of germ-cell mutagenesis was detected when data of microwave-exposed males were compared with those of sham-exposed males, even though there were significant increases in rectal and intra-testicular temperatures at a power density of 28 mW/cm². Temporary sterility, as indexed by fewer pregnancies, was seen at the highest power density.     

Absence of corneal endothelium injury in non‐human primates treated with and without ophthalmologic drugs and exposed to 2.8 GHz pulsed microwaves

Microwave‐induced corneal endothelial damage was reported to have a low threshold (2.6 W/kg), and vasoactive ophthalmologic medications lowered the threshold by a factor of 10–0.26 W/kg. In an attempt to confirm these observations, four adult male Rhesus monkeys (Macaca mulatta) under propofol anesthesia were exposed to pulsed microwaves in the far field of a 2.8 GHz signal (1.43 ± 0.06 µs pulse width, 34 Hz pulse repetition frequency, 13.0 mW/cm² spatial and temporal average, and 464 W/cm² spatial and temporal peak (291 W/cm² square wave equivalent) power densities). Corneal‐specific absorption rate was 5.07 W/kg (0.39 W/kg/mW/cm²). The exposure resulted in a 1.0–1.2 °C increase in eyelid temperature. In Experiment I, exposures were 4 h/day, 3 days/week for 3 weeks (nine exposures and 36 h total). In Experiment II, these subjects were pretreated with 0.5% Timolol maleate and 0.005% Xalatan® followed by 3 or 7 4‐h pulsed microwave exposures. Under ketamine–xylazine anesthesia, a non‐contact specular microscope was used to obtain corneal endothelium images, corneal endothelial cell density, and pachymetry at the center and four peripheral areas of the cornea. Ophthalmologic measurements were done before and 7, 30, 90, and 180 days after exposures. Pulsed microwave exposure did not cause alterations in corneal endothelial cell density and corneal thickness with or without ophthalmologic drugs. Therefore, previously reported changes in the cornea exposed to pulsed microwaves were not confirmed at exposure levels that are more than an order of magnitude higher. Bioelectromagnetics 31:324–333, 2010. Published 2010 Wiley‐Liss, Inc.     

Measurement of blood-brain barrier permeation in rats during exposure to 2450-MHz microwaves

Adult rats anesthesized with pentobarbital and injected intravenously with a mixture of [¹⁴C]sucrose and [³H]inulin were exposed for 30 min to an environment at an ambient temperature of 22, 30, or 40 °C, or were exposed at 22 °C to 2450-MHz CW microwave radiation at power densities of 0, 10, 20, or 30 mW/cm². Following exposure, the brain was perfused and sectioned into eight regions, and the radioactivity in each region was counted. The data were analyzed by two methods. First, the data for each of the eight regions and for each of the two radioactive tracers were analyzed by regression analysis for a total of 16 analyses and Bonferroni's Inequality was applied to prevent false positive results from numerous analyses. By this conservative test, no statistically significant increase in permeation was found for either tracer in any brain region of rats exposed to microwaves. Second, a profile analysis was used to test for a general change in tracer uptake across all brain regions. Using this statistical method, a significant increase in permeation was found for sucrose but not for inulin. A correction factor was then derived from the warm-air experiments to correct for the increase in permeation of the brain associated with change in body temperature. This correction factor was applied to the data for the irradiated animals. After correcting the data for thermal effects of the microwave radiation, no significant increase in permeation was found.     

Thermal action of 2.45 GHz microwaves on the cytoplasm of Chinese hamster cells

In order to demonstrate possible specific effects of microwaves at the cellular level V-79 Chinese hamster cells were exposed to 2.45-GHz radiation at power levels of 20–200 mW/cm² and at specific absorption rates of 10–100 mW/g. Intracellular cytoplasmic changes were observed by fluorescence polarization using a method based on the intracellular enzymatic hydrolysis of nonfluorescent fluorescein diacetate (FDA). At levels of absorbed energy below 90 J/g, modifications of microviscosity and mitochondrial state were absent, but a slight stimulation of enzymatic hydrolysis of FDA was observed which may be explained by microwave-induced alterations of cellular membranes possibly due to differences in heating pattern of microwaves compared to water-bath heating. At levels of absorbed energy above 90 J/g, the decrease of enzymatic hydrolysis of FDA, increase in degree of polarization, and increase of permeation of the fluorescent marker correlated well with the decrease in cell viability as measured by the exclusion of trypan blue. At equal absorbed energy, microwaves were found to exert effects comparable to classical heating except that permeation was slightly more affected by microwave than by classical heating. This suggests that membrane alteration produced by microwaves might differ from those induced by classical heating or that microwaves may have heated the membrane to higher temperatures than did classical heating.     

EFFECTS OF MICROWAVES AND ELF MAGNETIC FIELD ON THE PHAGOCYTIC ACTIVITY OF VARIOUSLY TREATED RAT MACROPHAGES

The purpose of this study was to investigate the effects of 9450-MHz microwaves and extremely low frequency magnetic fields (ELFMF) on the phagocytic activity of rat macrophages in control rats and those treated with vitamins C and E. In the microwave group, 24 albino Wistar rats were exposed to microwaves (2.65 mW/cm², specific absorption rate [SAR]: 1.80 W/kg) for 1 h/day for 21 days. Thirty-two albino Wistar rats were divided into four groups (one control, three experimental) (n = 8). The rats in the first exposure group were only exposed to microwaves for 1 h per day for 21 days. In addition to exposure with microwaves as in the first experimental group, vitamins E and C (150 mg/kg/day) were injected intraperitoneally into the rats in the second and third exposure groups, respectively. In the magnetic field exposure group, 26 albino Wistar rats were divided into two groups: the sham (n = 12) and exposed groups (n = 14). The rats in the experimental group were exposed to ELFMF (50 Hz, 0.75 mT) for 3 h/day for 3 weeks. After completing the exposure period, the rats were sacrificed under ketalar anesthesia. The viability of isolated alveolar macrophages of rats in the microwave and ELF groups was determined and compared to sham groups. The results were analyzed with the Mann–Whitney U test. In the microwave group, the phagocytic activity in the experimental groups was found to be higher than the sham groups. However, with phagocytic activity in rats treated with both microwaves and vitamins, only the vitamin C group was significant (p < 0.05). In the magnetic field group, the phagocytic activity of rats exposed to ELFMF was lower than that of the sham group, but the results were not significant (p > 0.05). Rectal temperatures of microwaveexposed groups were found to be significantly higher compared to the control group (p < 0.05).     

Alteration of life span of mice chronically exposed to 2.45 GHz CW microwaves

Female CD 1 mice were exposed from the thirty-fifth day of age for the remainder of their lives to 2.45 GHz, CW-microwave radiation at a power density of 3 or 10 m W/cm² (SAR = 2.0 or 6.8 W/kg). Exposures took place 1 h/day, 5 day/week in an anechoic chamber at an ambient temperature of 22 °C and a relative humidity of 50%. There were 25 animals in each exposure group, and an equal number of controls were concurrently sham exposed. The average life span of animals exposed at 10 mW/cm² was significantly shorter than that of sham-exposed controls (572 days vs. 706 days; P = .049; truncation >20%). In contrast, the average lifespan of the animals exposed at 3 mW/cm² was slightly, but not significantly, longer (738 days) than that of controls (706 days). © 1994 Wiley-Liss, Inc.

1 This article is a US Government work and, as such, is in the public domain in the United States of America.

     

Accelerated development of spontaneous and benzopyrene-induced skin cancer in mice exposed to 2450-MHz microwave radiation

C₃H/HeA mice with high incidence of spontaneous breast cancer and Balb/c mice treated with 3,4-benzopyrene (BP) (by painting of the skin resulting in the development of skin cancer) were irradiated with 2,450-MHz microwaves (MW) in an anechoic chamber at 5 or 15 mW/cm² (2 h daily, 6 sessions per week). C₃H/HeA mice were irradiated from the 6th week of life, up to the 12th month of life. Balb/c mice treated with BP were irradiated either prior to (over 1 or 3 months) or simultaneously with BP treatment (over 5 months). The appearance of palpable tumors in C₃H/HeA mice and of skin cancer in BP-treated Balb/c mice was checked every 2 weeks for 12 months. Two additional groups of mice were exposed to chronic stress caused by confinement or to sham-irradiation in an anechoic chamber; these served as controls. Irradiation with MWs at either 5 or 15 mW/cm² for 3 months resulted in a significant lowering of natural antineoplastic resistance (mean number of lung neoplastic colonies was 2.8 ± 1.6 (SD) in controls, 6.1 ± 1.8 in mice exposed at 5 mW/cm² and 10.8 ± 2.1 in those irradiated at 15 mW/cm²) and acceleration of development of BP-induced skin cancer (285 days in controls, 230 days for 5 mW/cm² and 160 days for 15 mW/cm²). Microwave-exposed C₃H/HeA mice developed breast tumors earlier than controls (322 days in controls, 261 days for 5 mW/cm² and 219 days for 15 mW/cm²). A similar acceleration was observed in the development of BP-induced skin cancer in mice exposed simultaneously to BP and MWs (285 days in controls, 220 day for 5 mW/cm² and 121 days for 15 mW/cm²). The acceleration of cancer development in all tested systems and lowering of natural antineoplastic resistance was similar in mice exposed to MW at 5 mW/cm² or to chronic stress caused by confinement but differed significantly from the data obtained on animals exposed at 15 mW/cm², where local thermal effects (“hot” spots) were possible.     

Effects of Pulsed Microwave Radiation Pre -and Post-Natally on the Developing Brain in Mice

Pregnant mice were irradiated for 5 hours daily throughout the pregnancy with pulsed microwaves at an incident average power density of 8 mW/cm², or were sham irradiated. After birth, from day 3 to day 20, half the offspring delivered by irradiated mice were irradiated (RR group) and half were sham-irradiated (RC group). The same procedure was used for offspring delivered by sham-irradiated mice (CR and CC). All offspring were sacrificed at 22 days of age. Histochemical analyses of the hypothalamus and liver were performed with a microspectrophotometer. The data suggested that succinate dehydrogenase in the hypothalamus was reduced by either pre-or post-natal microwave exposure. Similar changes occurred in the liver. The same pattern appeared with hypothalamic catechol-amine and monoamine oxidase. The data indicate that low-intensity microwave exposure can induce subtle alterations in offspring not detected with previously used techniques.     

THE EFFECT OF LOW LEVEL CONTINUOUS 2.45 GHz WAVES ON ENZYMES OF DEVELOPING RAT BRAIN

The present work describes the effect of low level continuous microwaves (2.45 GHz) on developing rat brain. Some 35-day-old Wistar rats were used for this study. The animals were exposed 2 hr/day for 35 days at a power density of 0.34 mW/cm² [specific absorption rate (SAR), 0.1 W/kg] in a specially made anechoic chamber. After the exposure, the rats were sacrificed and the brain tissue was dissected out and used for various biochemical assays. A significant increase in calcium ion efflux and ornithine decarboxylase (ODC) activity was observed in the exposed group as compared to the control. Correspondingly, a significant decrease in the calcium-dependent protein kinase activity was observed. These results indicate that this type of radiation affects the membrane bound enzymes, which are associated with cell proliferation and differentiation, thereby pointing out its possible role as a tumor promoter.     

Transbilayer movement of 24Na in sonicated phosphatidylcholine vesicles exposed to frequency-modulated microwave radiation

Sonicated egg phosphatidylcholine vesicles loaded with ²⁴Na⁺ were exposed at 20mW to a frequency-modulated (3 Hz) microwave field in the range of 2350 to 2550 MHz, or at 80 mW to a 2450-MHz CW (continuous wave) field, in a waveguide. The vesicle suspension absorbed microwaves at about 1 mW/ml and 25 mW/ml (CW experiment). The average temperature change of the irradiated suspension was < 0.1 °C from ambient. Leakage of ²⁴Na⁺ from the vesicles for up to 19 hours was measured. No difference was noted in the movement of ²⁴Na⁺ from the vesicles in the irradiated and control dispersions.     

Reduced weight in mice offspring after in utero exposure to 2450-MHz (CW) microwaves

Time-bred CD-1 mice (100) were sham-irradiated or irradiated with 2450-MHz (CW) microwaves at 28 mW/cm² for 100 minutes daily from the 6th through 17th day of gestation. The offspring were examined either as fetuses after hysterotomy on the 18th day of gestation or as naturally born neonates on the 1st and 7th day of age. Fetuses of half of the dams were examined on the 18th day of gestation. The incidence of pregnancy and the numbers of live, dead, resorbed, and total fetuses were similar in both groups. The mean weight was significantly lower (10%) in live microwave-irradiated fetuses, and ossification of sternal centers was significantly delayed. In the offspring that were born naturally, the mean weight of microwave-irradiated 7-day-old suckling mice was significantly lower (10%) than that of the sham-irradiated group. Survival rates of neonates in these two groups were not different. These data demonstrate that the decreased fetal weight seen in microwave-irradiated mice is retained at least 7 days after birth. Evidence from other published studies is presented to show that the retarded growth is persistent and might be interpreted as permanent stunting.     

Comparative effects of pulsed and continuous-wave 2.8-GHz microwaves on temporally defined behavior

The effects of pulsed-(PW) and continuous-wave (CW) 2.8-GHz microwaves were compared on the performance of rodents maintained by a temporally defined schedule of positive reinforcement. The schedule involved food-pellet reinforcement of behavior according to a differential-reinforcement-of-low-rate (DRL) contingency. The rats were independently exposed to PW and to CW fields at power densities ranging from 1 to 15 mW/cm². Alterations of normal performance were more pronounced after a 30-minute exposure to the PW field than to the CW field. The rate of emission of appropriately timed responses declined after exposure to PW at 10 and 15 mW/cm², whereas exposure at the same power levels to the CW field did not consistently affect the rate of responding. Change in performance associated with microwave exposure was not necessarily related to a general decline in responding: in some instances, increases in overall rates of responding were observed.